WO2013176916A1 - Use of pre t alpha or functional variant thereof for expanding tcr alpha deficient t cells - Google Patents

Use of pre t alpha or functional variant thereof for expanding tcr alpha deficient t cells Download PDF

Info

Publication number
WO2013176916A1
WO2013176916A1 PCT/US2013/040766 US2013040766W WO2013176916A1 WO 2013176916 A1 WO2013176916 A1 WO 2013176916A1 US 2013040766 W US2013040766 W US 2013040766W WO 2013176916 A1 WO2013176916 A1 WO 2013176916A1
Authority
WO
WIPO (PCT)
Prior art keywords
cells
cell
ptalpha
tale
seq
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2013/040766
Other languages
English (en)
French (fr)
Other versions
WO2013176916A8 (en
Inventor
Roman Galetto
Agnes Gouble
Stephanie Grosse
Cécile MANNIOUI
Laurent Poirot
Andrew Scharenberg
Julianne Smith
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=48579464&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=WO2013176916(A1) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Application filed by Individual filed Critical Individual
Priority to JP2015514051A priority Critical patent/JP6463672B2/ja
Priority to US14/403,937 priority patent/US10426795B2/en
Priority to AU2013266734A priority patent/AU2013266734B2/en
Priority to EP13728017.8A priority patent/EP2855666B1/en
Priority to CA2874611A priority patent/CA2874611C/en
Priority to AU2013312838A priority patent/AU2013312838B2/en
Priority to KR1020157008739A priority patent/KR102141259B1/ko
Priority to CN201380057661.1A priority patent/CN104769103B/zh
Priority to SG11201501471VA priority patent/SG11201501471VA/en
Priority to RU2015112125A priority patent/RU2663725C2/ru
Priority to PT13776597T priority patent/PT2893004T/pt
Priority to CA2883502A priority patent/CA2883502C/en
Priority to DK13776597.0T priority patent/DK2893004T3/en
Priority to BR112015004522A priority patent/BR112015004522A2/pt
Priority to EP13776597.0A priority patent/EP2893004B1/en
Priority to US14/018,021 priority patent/US10342829B2/en
Priority to MX2015002760A priority patent/MX367730B/es
Priority to PCT/US2013/058005 priority patent/WO2014039523A1/en
Priority to ES13776597T priority patent/ES2714523T3/es
Priority to HK16100443.5A priority patent/HK1212728B/en
Priority to JP2015530148A priority patent/JP6352920B2/ja
Publication of WO2013176916A1 publication Critical patent/WO2013176916A1/en
Priority to ES14723436T priority patent/ES2930431T3/es
Priority to HUE14723436A priority patent/HUE060901T2/hu
Priority to PT191618610T priority patent/PT3546572T/pt
Priority to AU2014267436A priority patent/AU2014267436B2/en
Priority to PT147234363T priority patent/PT2997141T/pt
Priority to LTEP19161861.0T priority patent/LT3546572T/lt
Priority to RS20240486A priority patent/RS65484B1/sr
Priority to PCT/EP2014/059662 priority patent/WO2014184143A1/en
Priority to CN201910222072.3A priority patent/CN109897100A/zh
Priority to TW103116744A priority patent/TWI636992B/zh
Priority to PL14723436.3T priority patent/PL2997141T3/pl
Priority to HK16110847.6A priority patent/HK1222679B/en
Priority to KR1020157035319A priority patent/KR102248157B1/ko
Priority to JP2016513321A priority patent/JP6491643B2/ja
Priority to DK19161861.0T priority patent/DK3546572T3/da
Priority to SI201431999T priority patent/SI2997141T1/sl
Priority to SG10201708896WA priority patent/SG10201708896WA/en
Priority to LTEPPCT/EP2014/059662T priority patent/LT2997141T/lt
Priority to HK16110821.6A priority patent/HK1222678B/zh
Priority to EP14723436.3A priority patent/EP2997141B1/en
Priority to EP19161861.0A priority patent/EP3546572B9/en
Priority to FIEP14723436.3T priority patent/FI2997141T3/fi
Priority to RU2015153250A priority patent/RU2727447C2/ru
Priority to FIEP19161861.0T priority patent/FI3546572T3/fi
Priority to HUE19161861A priority patent/HUE067258T2/hu
Priority to CA2911292A priority patent/CA2911292C/en
Priority to ES19161861T priority patent/ES2978123T3/es
Priority to NZ714044A priority patent/NZ714044B2/en
Priority to SI201432070T priority patent/SI3546572T1/sl
Priority to PL19161861.0T priority patent/PL3546572T3/pl
Priority to HRP20221393TT priority patent/HRP20221393T1/hr
Priority to EA201501109A priority patent/EA036200B1/ru
Priority to CN201480027071.9A priority patent/CN105431532B/zh
Priority to BR112015028387-0A priority patent/BR112015028387B1/pt
Priority to EP24163057.3A priority patent/EP4364809A3/en
Priority to SG11201508805UA priority patent/SG11201508805UA/en
Priority to US14/891,296 priority patent/US10874693B2/en
Priority to RS20221084A priority patent/RS63798B1/sr
Priority to MYPI2015703966A priority patent/MY172897A/en
Priority to HRP20240576TT priority patent/HRP20240576T1/hr
Priority to MX2015015662A priority patent/MX374681B/es
Priority to CN201480039866.1A priority patent/CN105378067A/zh
Priority to ES14727626T priority patent/ES2828669T3/es
Priority to CA2912373A priority patent/CA2912373A1/en
Priority to RU2015153245A priority patent/RU2736616C2/ru
Priority to KR1020217037383A priority patent/KR20210143926A/ko
Priority to RU2015153241A priority patent/RU2725542C2/ru
Priority to PCT/IB2014/061412 priority patent/WO2014184744A1/en
Priority to CA2912375A priority patent/CA2912375C/en
Priority to EP20190570.0A priority patent/EP3936612A1/en
Priority to EP14727627.3A priority patent/EP2997133B1/en
Priority to US14/894,426 priority patent/US11311575B2/en
Priority to KR1020157035439A priority patent/KR102220382B1/ko
Priority to AU2014266833A priority patent/AU2014266833B2/en
Priority to EP14727626.5A priority patent/EP2997132B1/en
Priority to MX2015015638A priority patent/MX2015015638A/es
Priority to JP2016513481A priority patent/JP2016524464A/ja
Priority to MX2015015639A priority patent/MX380738B/es
Priority to BR112015028483-3A priority patent/BR112015028483B1/pt
Priority to KR1020237030308A priority patent/KR20230144570A/ko
Priority to KR1020157035331A priority patent/KR102329704B1/ko
Priority to BR112015028493A priority patent/BR112015028493A2/pt
Priority to AU2014266830A priority patent/AU2014266830B2/en
Priority to CN201480039917.0A priority patent/CN105378068A/zh
Priority to DK14727626.5T priority patent/DK2997132T3/da
Priority to JP2016513480A priority patent/JP6875124B2/ja
Priority to US14/889,686 priority patent/US11304975B2/en
Priority to PCT/IB2014/061409 priority patent/WO2014184741A1/en
Publication of WO2013176916A8 publication Critical patent/WO2013176916A8/en
Anticipated expiration legal-status Critical
Priority to UAA201512065A priority patent/UA118106C2/uk
Priority to IL237576A priority patent/IL237576B/en
Priority to PH12015502479A priority patent/PH12015502479B1/en
Priority to SA515370135A priority patent/SA515370135B1/ar
Priority to US15/659,792 priority patent/US10286007B2/en
Priority to JP2019037174A priority patent/JP6854307B2/ja
Priority to AU2019201818A priority patent/AU2019201818B2/en
Priority to US16/361,370 priority patent/US10517896B2/en
Priority to US16/361,438 priority patent/US11414674B2/en
Priority to US16/365,588 priority patent/US20210000869A9/en
Priority to JP2019114231A priority patent/JP6998917B2/ja
Priority to US17/099,614 priority patent/US11007224B2/en
Priority to US17/099,608 priority patent/US11077144B2/en
Priority to US17/198,505 priority patent/US11274316B2/en
Priority to JP2021182495A priority patent/JP7479339B2/ja
Priority to US17/674,436 priority patent/US20220177914A1/en
Priority to US17/715,218 priority patent/US20230050345A1/en
Priority to US17/716,102 priority patent/US20230056268A1/en
Priority to US17/848,590 priority patent/US12577581B2/en
Priority to US18/056,544 priority patent/US20230201260A1/en
Priority to US18/451,816 priority patent/US20240269178A1/en
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0636—T lymphocytes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/14—Blood; Artificial blood
    • A61K35/17—Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/10—Cellular immunotherapy characterised by the cell type used
    • A61K40/11—T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/30—Cellular immunotherapy characterised by the recombinant expression of specific molecules in the cells of the immune system
    • A61K40/31—Chimeric antigen receptors [CAR]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/40—Cellular immunotherapy characterised by antigens that are targeted or presented by cells of the immune system
    • A61K40/41—Vertebrate antigens
    • A61K40/42—Cancer antigens
    • A61K40/4202—Receptors, cell surface antigens or cell surface determinants
    • A61K40/421—Immunoglobulin superfamily
    • A61K40/4211—CD19 or B4
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/38—Drugs for disorders of the endocrine system of the suprarenal hormones
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/7051—T-cell receptor (TcR)-CD3 complex
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/70517—CD8
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/70521—CD28, CD152
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70578—NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2121/00—Preparations for use in therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
    • C07K2317/14—Specific host cells or culture conditions, e.g. components, pH or temperature
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/569—Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/03—Fusion polypeptide containing a localisation/targetting motif containing a transmembrane segment
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/74—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/30—Hormones
    • C12N2501/38—Hormones with nuclear receptors
    • C12N2501/39—Steroid hormones
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/50—Cell markers; Cell surface determinants
    • C12N2501/51—B7 molecules, e.g. CD80, CD86, CD28 (ligand), CD152 (ligand)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/50—Cell markers; Cell surface determinants
    • C12N2501/515—CD3, T-cell receptor complex
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/50—Cell markers; Cell surface determinants
    • C12N2501/599—Cell markers; Cell surface determinants with CD designations not provided for elsewhere
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2502/00—Coculture with; Conditioned medium produced by
    • C12N2502/99—Coculture with; Conditioned medium produced by genetically modified cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2510/00—Genetically modified cells

Definitions

  • the present invention concerns cell therapy and more specifically relates to a method of expanding TCRalpha deficient T-cells by expressing preTCRa ("pTalpha") or functional variants thereof into said cells, thereby restoring a functional CD3 complex.
  • This method is particularly useful to enhance the efficiency of immunotherapy using primary T-cells from donors.
  • This method involves the use of pTalpha or functional variants thereof and polynucleotides encoding such polypeptides to expand TCRalpha deficient T- cells.
  • Such engineered cells can be obtained by using specific rare-cutting endonuclease, preferably TALE-nucleases.
  • the invention further relates to the use of Chimeric Antigen Receptor (CAR), especially multi-chain CAR, in such engineered cells to target malignant or infected cells.
  • CAR Chimeric Antigen Receptor
  • Adoptive immunotherapy which involves the transfer of autologous antigen-specific T cells generated ex vivo, is a promising strategy to treat viral infections and cancer.
  • the T cells used for adoptive immunotherapy can be generated either by expansion of antigen-specific T cells or redirection of T cells through genetic engineering (Park, Rosenberg et al. 2011). Transfer of viral antigen specific T cells is a well-established procedure used for the treatment of transplant associated viral infections and rare viral-related malignancies. Similarly, isolation and transfer of tumor specific T cells has been shown to be successful in treating melanoma. Novel specificities in T cells have been successfully generated through the genetic transfer of transgenic T cell receptors or chimeric antigen receptors (CARs) (Jena, Dotti et al. 2010).
  • CARs chimeric antigen receptors
  • CARs are synthetic receptors consisting of a targeting moiety that is associated with one or more signaling domains in a single fusion molecule.
  • the binding moiety of a CAR consists of an antigen-binding domain of a single-chain antibody (scFv), comprising the light and variable fragments of a monoclonal antibody joined by a flexible linker. Binding moieties based on receptor or ligand domains have also been used successfully.
  • the signaling domains for first generation CARs are derived from the cytoplasmic region of the CD3zeta or the Fc receptor gamma chains. First generation CARs have been shown to successfully redirect T cell cytotoxicity, however, they failed to provide prolonged expansion and anti-tumor activity in vivo.
  • CARs have successfully allowed T cells to be redirected against antigens expressed at the surface of tumor cells from various malignancies including lymphomas and solid tumors (Jena, Dotti et al. 2010).
  • FceRI is a tetrameric receptor complex consisting of ligand binding alpha subunit, a beta subunit and a homodimer of two signal-transducing gamma subunits (Metzger, Alcaraz et al. 1986).
  • FcsRI alpha domain consists of an extracellular domain containing two Ig-like domains that bind IgE, a transmembrane domain and a short cytoplasmic tail.
  • Beta subunit contains four transmembrane segments separating amino and carboxy terminal cytoplasmic tails.
  • the gamma chain consists essentially of a transmembrane region and cytoplasmic tail containing one immunoreceptor tyrosine-based activation motif (IT AM) (Cambier 1995).
  • the zeta chain of the TCR complex is closely related to the gamma chain and can substitute for the gamma chain of FceRI (Howard, Rodewald et al. 1990).
  • the current protocol for treatment of patients using adoptive immunotherapy is based on autologous cell transfer.
  • T lymphocytes are recovered from patients, genetically modified or selected ex vivo, cultivated in vitro in order to amplify the number of cells if necessary and finally infused into the patient.
  • the host may be manipulated in other ways that support the engraftment of the T cells or their participation in an immune response, for example pre-conditioning (with radiation or chemotherapy) and administration of lymphocyte growth factors (such as IL-2).
  • Each patient receives an individually fabricated treatment, using the patient's own lymphocytes (i.e. an autologous therapy).
  • Autologous therapies face substantial technical and logistic hurdles to practical application, their generation requires expensive dedicated facilities and expert personnel, they must be generated in a short time following a patient's diagnosis, and in many cases, pretreatment of the patient has resulted in degraded immune function, such that the patient's lymphocytes may be poorly functional and present in very low numbers. Because of these hurdles, each patient's autologous cell preparation is effectively a new product, resulting in substantial variations in efficacy and safety. Ideally, one would like to use a standardized therapy in which allogeneic therapeutic cells could be pre-manufactured, characterized in detail, and available for immediate administration to patients.
  • allogeneic it is meant that the cells are obtained from individuals belonging to the same species but are genetically dissimilar.
  • allogeneic cells presently has many drawbacks.
  • HvG host versus graft rejection
  • allogeneic cells are able to engraft, but their endogenous TCR specificities recognize the host tissue as foreign, resulting in graft versus host disease (GvHD), which can lead to serious tissue damage and death.
  • GvHD graft versus host disease
  • glucocorticoid receptor This class of steroid hormones binds to the glucocorticoid receptor (GR) present in the cytosol of T cells resulting in the translocation into the nucleus and the binding of specific DNA motifs that regulate the expression of a number of genes involved in the immunologic process.
  • Treatment of T cells with glucocorticoid steroids results in reduced levels of cytokine production leading to T cell anergy and interfering in T cell activation.
  • Alemtuzumab also known as CAMPATH1-H, is a humanized monoclonal antibody targeting CD52, a 12 amino acid glycosylphosphatidyl-inositol- (GPI) linked glycoprotein (Waldmann and Hale 2005).
  • CD52 is expressed at high levels on T and B lymphocytes and lower levels on monocytes while being absent on granulocytes and bone marrow precursors.
  • Treatment with Alemtuzumab a humanized monoclonal antibody directed against CD52, has been shown to induce a rapid depletion of circulating lymphocytes and monocytes. It is frequently used in the treatment of T cell lymphomas and in certain cases as part of a conditioning regimen for transplantation.
  • the use of immunosuppressive drugs will also have a detrimental effect on the introduced therapeutic T cells. Therefore, to effectively use an adoptive immunotherapy approach in these conditions, the introduced cells would need to be resistant to the immunosuppressive treatment.
  • T cell receptors are cell surface receptors that participate in the activation of T cells in response to the presentation of antigen.
  • the TCR is generally made from two chains, alpha and beta, which assemble to form a heterodimer and associates with the CD3 -transducing subunits to form the T-cell receptor complex present on the cell surface.
  • Each alpha and beta chain of the TCR consists of an immunoglobulin-like N-terminal variable (V) and constant (C) region, a hydrophobic transmembrane domain, and a short cytoplasmic region.
  • variable region of the alpha and beta chains are generated by V(D)J recombination, creating a large diversity of antigen specificities within the population of T cells.
  • T cells are activated by processed peptide fragments in association with an MHC molecule, introducing an extra dimension to antigen recognition by T cells, known as MHC restriction.
  • MHC restriction Recognition of MHC disparities between the donor and recipient through the T cell receptor leads to T cell proliferation and the potential development of GVHD. It has been shown that normal surface expression of the TCR depends on the coordinated synthesis and assembly of all seven components of the complex (Ashwell and Klusner 1990).
  • TCRalpha or TCRbeta can result in the elimination of the TCR from the surface of T cells preventing recognition of alloantigen and thus GVHD.
  • TCR disruption results in the elimination of the CD3 signaling component and alters the means of further T cell expansion.
  • T cell receptors emanate from the pre-T cell receptors (pTCR) which are expressed by immature thymocytes and are crucial for T cell development from the double negative (CD4- CD8-) to the double-positive (CD4+ CD8+) stages.
  • pTCR pre-T cell receptors
  • Pre-T cells that succeed in productive rearrangements of the TCRbeta locus express a functional TCRbeta chain which pairs with an invariant preTalpha chain and CD3 signaling components to form the pre-TCR complex.
  • the expression of the preTCR at the cell surface is necessary for triggering beta- selection, a process that induces the expansion of developing T cells, enforces allelic exclusion of the TCRbeta locus and results in the induction of rearrangements at the TCRalpha locus (von Boehmer 2005).
  • thymocytes undergo a second step of selection, referred to as positive or TCRalpha/beta selection upon binding of self peptide MHC complexes expressed on thymic epithelial cells.
  • TCRalpha/beta selection upon binding of self peptide MHC complexes expressed on thymic epithelial cells.
  • TCR activation is the initiation of signaling pathways via the associated CD3 subunits that result in multiple events including clonal expansion of T cells, upregulation of activation markers on the cell surface and induction of cytotoxicity or cytokine secretion.
  • the heterologous introduction of the pTalpha transgene can result in the formation of a preTCR.
  • This pTCR can serve as a means of T cell activation or stimulation in a manner that is non-MHC dependent, thus for example allowing continued expansion of alpha/beta T-cells following TCRalpha inactivation.
  • the pTCR complex displays a similar biochemical composition as the TCR in terms of associated CD3 subunits (Carrasco, Ramiro et al. 2001).
  • pre-TCR signaling may occur in part by a ligand independent event.
  • the crystal structure of the pTCR extracellular domain has provided a structural basis for the possible ligand-independence of pTCR signaling.
  • the pTCR has been shown to form a head to tail dimer where two pTalpha-TCRbeta heterodimers associate (Pang, Berry et al. 2010).
  • the inventors have achieved the production of genetically modified T-cells, which overcome the limitations of present immunotherapy strategies, allowing them to be both non-alloreactive and resistant to immunosuppressive agents. This was made possible by gene inactivation using specific TALE-nucleases directed against TCRalpha or TCRbeta, coupled with inactivation of genes encoding targets for different immunosuppressive agents, in particular CD52 and GR.
  • TCRalpha or TCRbeta coupled with inactivation of CD52 or the glucocorticoid receptor in T lymphocytes derived from an allogeneic donor significantly reduces the risk of GVHD, by eliminating the TCR, responsible for recognition of MHC disparities, while permitting proliferation and activity of the introduced lymphocytes in the presence of immunosuppressive drugs, such as Alemtuzumab or glucocorticoid steroids, that prevent rejection of these cells.
  • immunosuppressive drugs such as Alemtuzumab or glucocorticoid steroids
  • TALE-nucleases In addition to the above conception of genetically modified T cells, which can be both non alloreactive and immunosuppressive resistant, the inventors, by the use and design of specific TALE-nucleases, have concomitantly inactivated these different genes in T-cells, thereby obtaining double mutants.
  • double gene targeting by DSB has been so far unachieved in T cells due to the difficulty of yielding and maintaining T-cells in culture over time, to their low transformation rates, and loss during selection procedures. These difficulties result in a low probability of success for obtaining such cells.
  • one significant part of the invention is to have designed specific TALE-nucleases, allowing higher rates of DSB events within the T-cells, which are well tolerated by the cells, (especially upon co-transfection), able to target the selection of genes according to the invention.
  • rare cutting endonucleases such as the TALE-nucleases described therein, the probability of obtaining double inactivation of the genes in the transfected T-cells was significantly increased, so that it now appears possible to produce engineered T cells available from donors on a regular basis, using standard procedures.
  • the present invention proposes an embodiment where T-cells are engineered to allow proliferation when TCRalpha is inactivated.
  • T-cells that have undergone TCR subunit inactivation are further transformed with CAR to redirect allogeneic cells specificity towards tumor associated antigens independent of MHC.
  • the invention relates to a multi-chain CAR, in which costimulatory domains are placed in their normal juxtamembrane positions to improve their functions and so enhance survival and increase proliferation of engineered T-cells.
  • the invention provides methods, polypeptides and polynucleotides that allow the effective transformation of allogeneic T cells for adoptive immunotherapy, and their facile expansion through the CD3 complex.
  • the present invention discloses methods to engineer T cells, in particular allogeneic T cells obtainable from donors, to make them suitable for immunotherapy purposes.
  • the methods of the present invention more particularly allow the precise modification of the genome of cells relevant for immunotherapy by inactivating or replacing genes involved in MHC recognition and or targets of immunosuppressive drugs for the treatment of cancer and/or viral infections.
  • the modified cells relevant for immunotherapy further comprise exogenous recombinant polynucleotides encoding CARs for specific cell recognition.
  • Present CARs are single fusion molecules that necessitate serial appending of signaling domains. Moving signaling domains from their natural juxtamembrane position may interfere with their function.
  • the inventors design a multi-chain CAR derived from FceRI to allow normal juxtamembrane position of all relevant signaling domains.
  • the high affinity IgE binding domain of FCERI alpha chain is replaced by an extracellular ligand-binding domain such as scFv to redirect T-cell specificity to cell targets and the N and/or C-termini tails of FcsRI beta chain is used to place costimulatory signals in normal juxtamembrane positions.
  • pTalpha or functional variant thereof are introduced into the engineered T-cells.
  • the pTalpha or functional variant thereof used can be either full-length pTalpha, a splice variant (Saint-Ruf, Lechner et al. 1998), a C-terminal truncated version that has been shown to increase preTCR cell surface expression (Carrasco, Ramiro et al. 2001). Other additional truncations either smaller or larger than that described could be used.
  • Different preTalpha versions may further comprise signaling moieties from other molecules (CD28, CD137, CD8, TCRalpha, etc.) to promote proliferation and survival or comprise mutations that affect its ability to dimerize, such as the D22A, R24A, R102A or R117A mutations previously described in mice (Yamasaki, Ishikawa et al. 2006) or the W46R mutation described in humans (Pang, Berry et al. 2010) to decrease the proliferation potential.
  • the scFv portion of the CAR may also be fused to the extracellular domain of a pTalpha or a functional variant thereof, thus coupling the specificity towards target antigens directly with the proliferative activity of the preTCR.
  • the present invention relates to the polypeptides and the polynucleotides, which encode the rare-cutting endonucleases, to precisely target the above genes of interest, in particular TCRalpha, TCRbeta, GR and CD52, thereby enabling the genetic modification of the T-cells for immunotherapy.
  • the present invention provides more particularly specific target sequences within these genes and TALE -nucleases designed to respectively target those genes.
  • the present invention also relates to the isolated cells or cell lines comprising any of the proteins, polypeptides or vectors described herein.
  • the T cells of the present invention comprise inactivated TCRalpha, TCRbeta, GR or CD52 genes for their use in immunotherapy.
  • the isolated cells of the present invention or cell lines can further comprise exogenous recombinant polynucleotides, in particular polynucleotides encoding pTalpha or functional variant thereof, CARs or multi-chain CARs.
  • the modified T cells are used as a therapeutic product, ideally as an "off the shelf product.
  • the present invention concerns the method for treating or preventing cancer or infections in the patient by administrating an engineered T-cell obtainable by the above methods.
  • Figure 1 Schematic representation of the normal relationship between T-cells and antigen presenting cell.
  • Figure 2 Schematic representation of the genetically modified therapeutic T-cells according to the invention and the patient's tumor cells.
  • FIG. 3 Schematic representation of multi-chain CAR.
  • Figure 4 Schematic of different versions of multi-chain CARs.
  • A Schematic of the FceRI receptor.
  • B-C Different versions of multi-chain CARs (csml to csmlO) comprising a scFv and a CD8 stalk region fused to the transmembrane domain of FceRI alpha chain. At least one 4 IBB, CD28 and/or CD3 zeta domains can be fused to a FcsRI alpha, beta and/or gamma chain.
  • Figure 5 Schematic representation of one example of the method of engineering human allogenic cells for immunotherapy
  • Figure 6 Concentration in cells per milliliter of live CD52-positive or CD52-negative cells after treatment with anti-CD52 antibody (CAMPATH1-H) with complement or controls.
  • Figure 7 Comparison of the forward side scatter (FSC) distribution, an indicator of cell size, between TCR-positive and TCR-negative cells, or between CD52-positive and CD52- negative cells, and non activated cells as control.
  • Figure 8 Flow cytometry analysis of CD 107a expression (marker of degranulation) on targeted CD52 and TCRalpha inactivated T cells.
  • CD 107 expression is analyzed on CD52+TCRaP+ cells (first column), CD52-TCRap- cells (second column), CD52-TCRap+ cells (third column) and CD52+TCRaP- cells (fourth column) before (A) and after incubation with Daudi cells (B); C) represents flow cytometry analysis of T cells further transfected with a CAR and incubated with Daudi cells; D) represents flow cytometry analysis of T cells transfected with a CAR but not incubated with Daudi cells and E) represents flow cytometry analysis of T cells transfected with a CAR and treated to PMA/ionomycin (positive control).
  • Figure 9 Deep sequencing analysis of CD52 and TRAC TALE-nucleases potential off-site targets.
  • Figure 10 Analysis of PDCD1 and CTLA-4 genomic locus by T7-endonuclease assay. Arrows point to digested PCR products.
  • Figure 11 Schematic representation of some examples of preTalpha constructs.
  • Figure 12 Flow cytometry analysis of transduction efficiency (% BFP+ cells) and activity of the FL, ⁇ 18, ⁇ 48 pTalpha constructs (% CD3 surface expression) in TCR alpha inactivated Jurkat cells.
  • Figure 13 Schematic representation of a lentiviral construct coding for pTalpha protein (preTCRa).
  • FIG 14 A. Representation of the experimental protocol.
  • NEP represents non electroporated cells with TRAC TALE-nucleases.
  • ⁇ - ⁇ 48 histograms correspond to the signal detected in TCR inactivated cells expressing ⁇ - ⁇ 48 (BFP+ cells) while the KO histograms correspond to TCRalpha inactivated cells which do not express ⁇ - ⁇ 48 (BFP- cells) ⁇ - ⁇ 48.
  • ⁇ histograms correspond to the signal detected in TCR inactivated cells expressing ⁇ - ⁇ 48.41 ⁇ (BFP+ cells) while the KO histograms correspond to TCRalpha inactivated cells which do not express ⁇ - ⁇ 48.41 ⁇ (BFP- cells).
  • NEP (non electroporated) histograms correspond to signal detected in non engineered cells.
  • FIG 16 Cell growth analysis of TCR alpha inactivated cells (KO) transduced with pTalpha-A48 (pTaA48) or control BFP vector (BFP) maintained in IL2 or in IL2 with anti- CD3/CD28 beads at different time points (x-axis).
  • the BFP+ cells number is estimated at different time points for each condition and the fold induction of these cells (y-axis) was estimated with respect to the value obtained at day 2 post re-activation.
  • the results are obtained from two independent donors.
  • cell growth was also determined for cells transduced with pTalpha-A48.41BB (pTa-A48.BB) and full-length pTalpha- (pTa-FL).
  • Figure 17 Flow cytometry analysis of GFP positive cells on PBMCs electroporated with the five different Cytopulse programs. The upper line corresponds to transfection of 6xl0 6 cells per cuvette, while the lower line corresponds to transfection of 3x10 6 cells per cuvette.
  • Figure 18 Flow cytometry analysis of purified T cell mortality using viability dye (eFluor- 450) and of GFP positive cells among the viable population after electroporation with GFP mRNA, GFP DNA and control pUC DNA.
  • NEP corresponds to cells that were maintained in electroporation buffer but were not electroporated and NT corresponds to non electroporated cells maintained in culture medium.
  • Figure 19 Flow cytometry analysis of TCR alpha/beta and CD3 expression on human primary T cells following TRAC TALE-nuclease mRNA electroporation (top). Deep sequencing analysis of genomic DNA extracted from human primary T cells following TRAC TALE-nuclease mRNA electroporation (bottom).
  • Figure 20 A. Flow cytometry analysis of CAR expression (anti F(ab')2 ) after electroporation of T cells with or without mRNA encoding a single chain CAR.
  • B Flow cytometry analysis of CD 107a expression (marker of degranulation) on electroporated T cells cocultured with daudi cells.
  • Figure 21 A. Representation of mRNA encoding a multi-chain CAR.
  • Table 1 Description of the GR TALE-nucleases and sequences of the TALE-nucleases target sites in the human GR gene.
  • Table 2 Cleavage activity of the GR TALE-nucleases in yeast. Values are comprised between 0 and 1. Maximal value is 1.
  • Table 3 Percentage of targeted mutagenesis at endogenous TALE-nuclease target sites in 293 cells.
  • Table 4 Percentage of targeted mutagenesis at endogenous TALE-nuclease target sites in primary T lymphocytes.
  • Table 5 Description of the CD52, TRAC and TRBC TALE-nucleases and sequences of the TALE-nucleases target sites in the human corresponding genes.
  • Table 6 Additional target sequences for TRAC and CD52 TALE-nucleases.
  • Table 7 Percentage of indels for TALE-nuclease targeting CD52 T02, TRAC_T01, TRBC_T01 and TRBC_T02 targets.
  • Table 8 Percentages of CD52- negative, TCR-negative and CD52/TCR-double negative T lymphocytes after transfection of corresponding TALE-nuclease-expressing polynucleotides.
  • Table 9 Percentages of TCR-negative T lymphocytes after transfection of TRBC TALE- nuclease-expressing polynucleotides.
  • Table 10 Description of the CTLA4 and PDCD1 TALE-nucleases and sequences of the TALE-nucleases target sites in the human corresponding genes.
  • Table 11 Description of a subset of pTalpha constructs.
  • Table 12 Activity of the different pTalpha constructs in Jurkat TCR alpha inactivated cell. Activity was measured by flow cytometry analysis of CD3 expression on jurkat TCR alpha inactivated cell transfected with the different preTalpha constructs.
  • Table 13 Different cytopulse programs used to determine the minimal voltage required for electroporation in PBMC derived T-cells.
  • Table 14 Cytopulse program used to electroporate purified T-cells. Detailed description of the invention Unless specifically defined herein, all technical and scientific terms used have the same meaning as commonly understood by a skilled artisan in the fields of gene therapy, biochemistry, genetics, and molecular biology.
  • the present invention relates to methods for new adoptive immunotherapy strategies in treating cancer and infections.
  • Non alloreactive and immunosuppressive resistant T cells relates to a method of engineering T-cells, especially for immunotherapy.
  • this method comprises:
  • TCR T-cell receptor
  • An immunosuppressive agent is an agent that suppresses immune function by one of several mechanisms of action.
  • an immunosuppressive agent is a role played by a compound which is exhibited by a capability to diminish the extent and/or voracity of an immune response.
  • an immunosuppressive agent can be a calcineurin inhibitor, a target of rapamycin, an interleukin-2 a-chain blocker, an inhibitor of inosine monophosphate dehydrogenase, an inhibitor of dihydrofolic acid reductase, a corticosteroid or an immunosuppressive antimetabolite.
  • Classical cytotoxic immunosuppressants act by inhibiting DNA synthesis.
  • targets for immunosuppressive agent can be a receptor for an immunosuppressive agent such as: CD52, glucocorticoid receptor (GR), a FKBP family gene member and a cyclophilin family gene member.
  • the genetic modification step of the method relies on the inactivation of one gene selected from the group consisting of CD52, GR, TCR alpha and TCR beta. In another embodiment, the genetic modification step of the method relies on the inactivation of two genes selected from the group consisting of CD52 and GR, CD52 and TCR alpha, CDR52 and TCR beta, GR and TCR alpha, GR and TCR beta, TCR alpha and TCR beta. In another embodiment, the genetic modification step of the method relies on the inactivation of more than two genes. The genetic modification is preferably operated ex-vivo.
  • the genetic modification of the method relies on the expression, in provided cells to engineer, of one rare-cutting endonuclease such that said rare- cutting endonuclease specifically catalyzes cleavage in one targeted gene thereby inactivating said targeted gene.
  • the nucleic acid strand breaks caused by the rare-cutting endonuclease are commonly repaired through the distinct mechanisms of homologous recombination or nonhomologous end joining (NHEJ).
  • NHEJ nonhomologous end joining
  • NHEJ non-homologous end joining
  • Said modification may be a substitution, deletion, or addition of at least one nucleotide.
  • Cells in which a cleavage- induced mutagenesis event, i.e a mutagenesis event consecutive to an NHEJ event, has occurred can be identified and/or selected by well-known method in the art.
  • said method to engineer cells comprises at least one of the following steps:
  • T-cell preferably from a cell culture or from a blood sample
  • said method comprises:
  • T-cell preferably from a cell culture or from a blood sample
  • T cell Transforming said T cell with nucleic acid encoding a rare-cutting endonuclease able to selectively inactivate by DNA cleavage, preferably by double-strand break respectively: said gene encoding a target for said immunosuppressive agent, and - at least one gene encoding a component of the T-cell receptor (TCR);
  • said rare-cutting endonuclease specifically targets one gene selected from the group consisting of CD52, GR, TCR alpha and TCR beta.
  • the genetic modification of the method relies on the expression, in provided cells to engineer, of two rare-cutting endonucleases such that said each of the two rare-cutting endonucleases specifically and respectively catalyzes cleavage in each of the pairs of genes selected from the group consisting of CD52 and GR, CD52 and TCR alpha, CDR52 and TCR beta, GR and TCR alpha, GR and TCR beta, TCR alpha and TCR beta, thereby inactivating said targeted genes.
  • more than two rare-cutting endonucleases can be expressed in cells to engineer in order to target and/or inactivate more than two genes.
  • said gene of step (b), specific for an immunosuppressive treatment is CD52 and the immunosuppressive treatment of step (d) or (e) comprises a humanized antibody targeting CD52 antigen.
  • said gene of step (b), specific for an immunosuppressive treatment is a glucocorticoid receptor (GR) and the immunosuppressive treatment of step d) or (e) comprises a corticosteroid such as dexamethasone.
  • GR glucocorticoid receptor
  • said target gene of step (b), specific for an immunosuppressive treatment is a FKBP family gene member or a variant thereof and the immunosuppressive treatment of step (d) or (e) comprises FK506 also known as Tacrolimus or fujimycin.
  • said FKBP family gene member is FKBP12 or a variant thereof.
  • said gene of step (b), specific for an immunosuppressive treatment is a cyclophilin family gene member or a variant thereof and the immunosuppressive treatment of step (d) or (e) comprises cyclosporine.
  • said rare-cutting endonuclease can be a meganuclease, a Zinc finger nuclease or a TALE-nuclease.
  • said rare-cutting endonuclease is a TALE-nuclease.
  • TALE-nuclease is intended a fusion protein consisting of a DNA-binding domain derived from a Transcription Activator Like Effector (TALE) and one nuclease catalytic domain to cleave a nucleic acid target sequence. (Boch, Scholze et al. 2009; Moscou and Bogdanove 2009)(Deng, Yan et al.
  • Preferred TALE-nucleases according to the invention are those recognizing and cleaving the target sequence selected from the group consisting of: - SEQ ID NO: 1 to 6 (GR),
  • Said TALE-nucleases preferably comprise a polypeptide sequence selected from SEQ ID NO: 7 to SEQ ID NO: 18 and SEQ ID NO: 41 to SEQ ID NO: 48, in order to cleave the respective target sequences SEQ ID NO: 1 to 6 and SEQ ID NO: 37 to 40.
  • additional catalytic domain can be further introduced into the cell with said rare-cutting endonuclease to increase mutagenesis in order to enhance their capacity to inactivate targeted genes.
  • said additional catalytic domain is a DNA end processing enzyme.
  • DNA end-processing enzymes include 5-3' exonucleases, 3-5' exonucleases, 5-3' alkaline exonucleases, 5' flap endonucleases, helicases, hosphatase, hydrolases and template-independent DNA polymerases.
  • Non limiting examples of such catalytic domain comprise of a protein domain or catalytically active derivate of the protein domain seleced from the group consisting of hExoI (EX01_HUMAN), Yeast Exol (EX01_YEAST), E.coli Exol, Human TREX2, Mouse TREX1, Human TREX1, Bovine TREX1, Rat TREX1, TdT (terminal deoxynucleotidyl transferase) Human DNA2, Yeast DNA2 (DNA2 YEAST).
  • said additional catalytic domain has a 3'-5'-exonuclease activity, and in a more preferred embodiment, said additional catalytic domain is TREX, more preferably TREX2 catalytic domain (WO2012/058458). In another preferred embodiment, said catalytic domain is encoded by a single chain TREX polypeptide. Said additional catalytic domain may be fused to a nuclease fusion protein or chimeric protein according to the invention optionally by a peptide linker.
  • the genetic modification step of the method further comprises a step of introduction into cells an exogeneous nucleic acid comprising at least a sequence homologous to a portion of the target nucleic acid sequence, such that homologous recombination occurs between the target nucleic acid sequence and the exogeneous nucleic acid.
  • said exogenous nucleic acid comprises first and second portions which are homologous to region 5' and 3' of the target nucleic acid sequence, respectively.
  • Said exogenous nucleic acid in these embodiments also comprises a third portion positioned between the first and the second portion which comprises no homology with the regions 5' and 3' of the target nucleic acid sequence.
  • a homologous recombination event is stimulated between the target nucleic acid sequence and the exogenous nucleic acid.
  • homologous sequences of at least 50 bp, preferably more than 100 bp and more preferably more than 200 bp are used within said donor matrix. Therefore, the exogenous nucleic acid is preferably from 200 bp to 6000 bp, more preferably from 1000 bp to 2000 bp. Indeed, shared nucleic acid homologies are located in regions flanking upstream and downstream the site of the break and the nucleic acid sequence to be introduced should be located between the two arms.
  • said exogenous nucleic acid successively comprises a first region of homology to sequences upstream of said cleavage, a sequence to inactivate one targeted gene selected from the group consisting of CD52, GR, TCR alpha and TCR beta and a second region of homology to sequences downstream of the cleavage.
  • Said polynucleotide introduction step can be simultaneous, before or after the introduction or expression of said rare-cutting endonuclease.
  • exogenous nucleic acid can be used to knock-out a gene, e.g.
  • sequence insertions by using such exogenous nucleic acid can be used to modify a targeted existing gene, by correction or replacement of said gene (allele swap as a non-limiting example), or to up- or down-regulate the expression of the targeted gene (promoter swap as non-limiting example), said targeted gene correction or replacement.
  • inactivation of genes from the group consisting of CD52, GR, TCR alpha and TCR beta can be done at a precise genomic location targeted by a specific TALE-nuclease, wherein said specific TALE-nuclease catalyzes a cleavage and wherein said exogenous nucleic acid successively comprising at least a region of homology and a sequence to inactivate one targeted gene selected from the group consisting of CD52, GR, TCR alpha and TCR beta which is integrated by homologous recombination.
  • several genes can be, successively or at the same time, inactivated by using several TALE-nucleases respectively and specifically targeting one defined gene and several specific polynucleotides for specific gene inactivation.
  • additional genomic modification step can be intended also the inactivation of another gene selected from the group consisting of CD52, GR, TCR alpha and TCR beta.
  • additional genomic modification step can be an inactivation step comprising:
  • exogenous nucleic acid successively comprising a first region of homology to sequences upstream of said cleavage, a sequence to be inserted in the genome of said cell and a second region of homology to sequences downstream of said cleavage, wherein said introduced exogenous nucleic acid inactivates a gene and integrates at least one exogenous polynucleotide sequence encoding at least one recombinant protein of interest.
  • said exogenous polynucleotide sequence is integrated within a gene selected from the group consisting of CD52, GR, TCR alpha and TCR beta.
  • said method to engineer cell further comprises an additional genomic modification step.
  • Said protein of interest can be, as non limiting examples, pTalpha or functional variant thereof, a Chimeric Antigen Receptor (CAR), a multi-chain CAR, a bispecific antibody or rare-cutting endonuclease targeting PDCD1 or CTLA-4 as described in the present disclosure.
  • CAR Chimeric Antigen Receptor
  • the invention also relates to TALE-nucleases.
  • the invention relates to TALE- nuclease comprising:
  • TALE Transcription Activator-Like Effector
  • Preferred TALE-nucleases according to the invention are those recognizing and cleaving the target sequence selected from the group consisting of: - SEQ ID NO: 1 to 6 (GR),
  • Said TALE-nucleases preferably comprise a polypeptide sequence selected from SEQ ID NO: 7 to SEQ ID NO: 18 and SEQ ID NO: 41 to SEQ ID NO: 48, in order to cleave the respective target sequences SEQ ID NO: 1 to 6 and SEQ ID NO: 37 to 40.
  • the invention encompasses polypeptides variants of the above polypeptides that share at least 70%, preferably at least 80 %, more preferably at least 90 % and even more preferably at least 95 % identity with the sequences provided in this patent application.
  • polypeptides comprising a polypeptide sequence that has at least 70%, preferably at least 80%, more preferably at least 90 %, 95 % 97 % or 99 % sequence identity with amino acid sequence selected from the group consisting of SEQ ID NO:7 to SEQ ID NO: 18 and SEQ ID NO: 41 to SEQ ID NO: 48.
  • polynucleotides, vectors encoding the above described rare-cutting endonucleases according to the invention are also encompassed isolated cells or cell lines susceptible to be obtained by said method to engineer cells, in particular T cells, in which at least one gene selected from the group consisting of CD52, GR, TCR alpha and TCR beta has been inactivated.
  • T cells in which at least one gene selected from the group consisting of CD52, GR, TCR alpha and TCR beta has been inactivated.
  • two genes selected from the group consisting of CD52 and GR, CD52 and TCR alpha, CDR52 and TCR beta, GR and TCR alpha, GR and TCR beta, TCR alpha and TCR beta have been inactivated.
  • those genes are preferably inactivated by at least one rare-cutting endonuclease. It has been shown by the inventors that the use of TALE-nucleases was particularly advantageous to achieve double inactivation in T-cells.
  • the invention encompasses an isolated T-cell comprising at least two polynucleotides, said polynucleotides encoding at least a first and second TALE-nucleases, preferably the first TALE-nuclease being directed against a gene encoding TCR and the second being directed against a gene encoding a receptor for an immunosuppressive agent, such as CD52 or GR.
  • said isolated cell further comprises one additional genomic modification.
  • said additional genomic modification is the integration of at least one exogenous polynucleotide sequence.
  • said exogenous sequence is integrated into one gene selected from the group consisting of CD52, GR, TCR alpha and TCR beta.
  • the invention in another aspect, relates to a method of expanding TCR alpha deficient T-cell comprising introducing into said T-cell pTalpha (also named preTCRa) or a functional variant thereof and expanding said cells, optionally through stimulation of the CD3 complex.
  • the method comprises: a) Transforming said cells with nucleic acid encoding at least a fragment of pTalpha to support CD3 surface expression b) Expressing said pTalpha into said cells c) Expanding said cells optionally, optionally through stimulation of the CD3 complex.
  • the invention also relates to a method of preparing T-cells for immunotherapy comprising steps of the method for expansion for T-cell.
  • the pTalpha polynucleotide sequence can be introduced randomly or else through homologous recombination, in particular the insertion could be associated with the inactivation of the TCRalpha gene.
  • a “functional variant” of the peptide refers to a molecule substantially similar to either the entire peptide or a fragment thereof.
  • a “fragment” of the pTalpha or functional variant thereof of the present Invention refers to any subset of the molecule, that is, a shorter peptide.
  • Preferred pTalpha or functional variants can be full length pTalpha or a C-terminal truncated pTalpha version. C- terminal truncated pTalpha lacks in C-terminal end one or more residues.
  • C-terminal truncated pTalpha version lacks 18 , 48, 62, 78, 92, 110 or 114 residues from the C-terminus of the protein (SEQ ID NO: 107 to SEQ ID NO: 114).
  • amino acid sequence variants of the peptide can be prepared by mutations in the DNA which encodes the peptide. Such functional variants include, for example, deletions from, or insertions or substitutions of, residues within the amino acid sequence. Any combination of deletion, insertion, and substitution may also be made to arrive at the final construct, provided that the final construct possesses the desired activity, in particular the restoration of a functional CD3 complex.
  • mutated residue can be at least W46R, D22A, K24A, R102A or R117A of the human pTalpha protein or aligned positions using CLUSTALW method on pTalpha family or homologue member.
  • pTalpha or variant thereof as described above comprise the mutated residue W46R (SEQ ID NO: 123) or the mutated residues D22A, K24A, R102A and Rl 17A (SEQ ID NO: 124 ).
  • said pTalpha or variants are also fused to a signal-transducing domain such as CD28, OX40, ICOS, CD27, CD137 (4-1BB) and CD8 as non limiting examples (SEQ ID NO: 1 15 to SEQ ID NO: 120).
  • the extracellular domain of pTalpha or variants as described above can be fused to a fragment of the TCRalpha protein, particularly the transmembrane and intracellular domain of TCRalpha (SEQ ID NO: 122).
  • pTalpha variants can also be fused to the intracellular domain of TCRalpha (SEQ ID NO: 121 ).
  • said pTalpha versions are fused to an extracellular ligand-binding domain and more preferably pTalpha or functional variant thereof is fused to a single chain antibody fragment (scFV) comprising the light (VL) and the heavy (VH) variable fragment of a target antigen specific monoclonal antibody joined by a flexible linker.
  • scFV single chain antibody fragment
  • amino acid sequence of pTalpha or functional variant thereof is selected from the group consisting of SEQ ID NO: 107 to SEQ ID NO: 124.
  • the invention encompasses polypeptides variants of the above polypeptides that share at least 70%, preferably at least 80 %, more preferably at least 90 % and even more preferably at least 95 % identity with the sequences provided in this patent application.
  • polypeptides comprising a polypeptide sequence that has at least 70%, preferably at least 80%, more preferably at least 90 %, 95 % 97 % or 99 % sequence identity with amino acid sequence selected from the group consisting of SEQ ID NO: 107 to SEQ ID NO: 124.
  • TCR alpha deficient T cell an isolated T cell that lacks expression of a functional TCR alpha chain. This may be accomplished by different means, as non limiting examples, by engineering a T cell such that it does not express any functional TCR alpha on its cell surface or by engineering a T cell such that it produces very little functional TCR alpha chain on its surface or by engineering a T cell to express mutated or truncated form of TCR alpha chain. TCR alpha deficient cells can no longer be expanded through CD3 complex. Thus, to overcome this problem and to allow proliferation of TCR alpha deficient cells, pTalpha or functional variant thereof is introduced into said cells, thus restoring a functional CD3 complex.
  • the method further comprises introducing into said T cells rare-cutting endonucleases able to selectively inactivate by DNA cleavage one gene encoding one component of the T-cell receptor (TCR).
  • said rare- cutting endonuclease is a TALE-nucleases.
  • TALE-nuclease is directed against one of the gene target sequences of TCRalpha selected from the group consisting of SEQ ID NO: 37 and SEQ ID NO: 57 to 60.
  • TALE-nucleases are selected from the group consisting of SEQ ID NO: 41 and SEQ ID NO: 42.
  • said method for expansion of TCR alpha deficient T-cells comprises an additional genomic modification step.
  • additional genomic modification step can be intended the introduction into cells to engineer of one protein of interest.
  • Said protein of interest can be, as non limiting examples, a Chimeric Antigen Receptor (CAR), particularly CAR comprising amino acid sequence SEQ ID NO: 73, a multi-chain CAR, particularly multi-chain CAR comprising amino acid sequence SEQ ID NO: 125 a bispecific antibody, rare-cutting endonucleases targeting PDCD1 or CTLA-4, particularly targeting nucleic acid sequence SEQ ID NO: 74 to SEQ ID NO: 78 or a rare-cutting endonuclease targeting a target for immunosuppressive agent as described in the present disclosure.
  • CAR Chimeric Antigen Receptor
  • polypeptides encoding pTalpha particularly functional variants described above.
  • the invention relates to a pTalpha or functional variant thereof fused to a signal transducing domain such as CD28, OX40, ICOS, CD137 and CD8. More particularly, the invention relates to pTalpha functional variant comprising amino acid sequence selected form the group consisting of SEQ ID NO: 107 to SEQ ID NO: 124.
  • isolated cells or cell lines susceptible to be obtained by said method.
  • said isolated cells or cell lines are obtained by introducing into said cells a pTalpha or a functional variant thereof to support CD3 surface expression.
  • said isolated cell or cell line are further genetically modified by inactivating TCRalpha gene. This gene is preferably inactivating by at least one rare-cutting endonuclease.
  • said rare-cutting endonuclease is TALE-nuclease.
  • the invention in another embodiment, relates to a multi-chain chimeric antigen receptor (CAR) particularly adapted to the production and expansion of engineered T-cells of the present invention.
  • the multi-chain CAR comprising at least two of the following components: a) one polypeptide comprising the transmusinembrane domain of FcsRI alpha chain and an extracellular ligand-binding domain, b) one polypeptide comprising a part of N- and C- terminal cytoplasmic tail and the transmembrane domain of FcsRI beta chain and/or c) two polypeptides comprising each a part of intracytoplasmic tail and the transmembrane domain of FceRI gamma chain, whereby different polypeptides multimerize together spontaneously to form dimeric, trimeric or tetrameric CAR.
  • multi-chain CAR comprises amino acid sequence SEQ ID NO: 125.
  • the term "a part of used herein refers to any subset of the molecule, that is a shorter peptide.
  • amino acid sequence functional variants of the polypeptide can be prepared by mutations in the DNA which encodes the polypeptide. Such functional variants include, for example, deletions from, or insertions or substitutions of, residues within the amino acid sequence. Any combination of deletion, insertion, and substitution may also be made to arrive at the final construct, provided that the final construct possesses the desired activity, especially to exhibit a specific anti-target cellular immune activity.
  • said extracellular ligand-binding domain is a scFv.
  • Other binding domain than scFv can also be used for predefined targeting of lymphocytes, such as camelid single-domain antibody fragments or receptor ligands like a vascular endothelial growth factor polypeptide, an integrin-binding peptide, heregulin or an IL-13 mutein, antibody binding domains, antibody hypervariable loops or CDRs as non limiting examples.
  • said polypeptide of a) further comprises a stalk region between said extracellular ligand-binding domain and said transmembrane domain.
  • stalk region generally means any oligo- or polypeptide that functions to link the transmembrane domain to the extracellular ligand-binding domain. In particular, stalk region are used to provide more flexibility and accessibility for the extracellular ligand-binding domain.
  • a stalk region may comprise up to 300 amino acids, preferably 10 to 100 amino acids and most preferably 25 to 50 amino acids.
  • Stalk region may be derived from all or part of naturally occurring molecules, such as from all or part of the extracellular region of CD8, CD4 or CD28, or from all or part of an antibody constant region. Alternatively the stalk region may be a synthetic sequence that corresponds to a naturally occurring stalk sequence, or may be an entirely synthetic stalk sequence.
  • said polypeptide of a), b) and/or c) further comprises at least one signal-transducing domain.
  • said signal-transducing domain is selected from the group consisting of CD28, OX40, ICOS, CD137 and CD8.
  • said C-terminal cytoplasmic tail of FCERI alpha, beta and/or gamma chain fragment further comprises TNFR-associated Factor 2 (TRAF2) binding motifs.
  • said C- terminal cytoplasmic tail of FceRI alpha, beta and/or gamma chain is replaced by intracytoplasmic tail of costimulatory TNFR member family.
  • Cytoplasmic tail of costimulatory TNFR family member contains TRAF2 binding motifs consisting of the major conserved motif (P/S/A)X(Q/E)E) or the minor motif (PXQXXD), wherein X is any amino acid.
  • TRAF proteins are recruited to the intracellular tails of many TNFRs in response to receptor trimerization.
  • said intracytoplasmic domain of FceRI alpha, beta and/or gamma chain is replaced by intracytoplasmic domain of TCR zeta chain (also named CD3 zeta).
  • said intracytoplasmic domain of FCERI alpha, beta and/or gamma chain comprises at least one additional immunoreceptor tyrosine-based activation motif (ITAM).
  • ITAMs are well defined signaling motifs found in the intracytoplasmic tail of a variety of receptors that serve as binding sites for syk/zap70 class tyrosine kinases.
  • ITAM used in the invention examples include those derived from TCRzeta, FCRgamma, FCRbeta, CD3gamma, CD3delta, CD3epsilon, CD5, CD22, CD79a, CD79b, and CD66d.
  • TCRzeta FCRgamma
  • FCRbeta FCRgamma
  • CD3gamma CD3delta
  • CD3epsilon CD5, CD22, CD79a, CD79b, and CD66d.
  • Figure 4 different versions of multi-chain CAR are illustrated in Figure 4.
  • the multi-chain CAR comprise the amino acid sequence SEQ ID NO: 125.
  • the present invention relates to polypeptides comprising a polypeptide sequence that has at least 70%, preferably at least 80%, more preferably at least 90 %, 95 % 97 % or 99 % sequence identity with amino acid sequence selected from the group consisting of SEQ ID NO: 125.
  • polynucleotides are also comprised in the scope of the present invention, polynucleotides, vectors encoding the above described multi-chain CAR according to the invention.
  • the invention relates to a method of preparing T-cells for immunotherapy comprising introducing into said T-cells the different polypeptides composing said multi-chain CAR and expanding said cells.
  • said method further comprises a step of genetically modifying said cells by inactivating at least one gene expressing one component of the TCR and/or a target for an immunosuppressive agent.
  • said gene is selected from the group consisting of TCRalpha, TCRbeta, CD52 and GR.
  • said method further comprises introducing into said T cells a rare-cutting endonuclease able to selectively inactivate by DNA cleavage said genes.
  • said rare- cutting endonuclease is TALE-nuclease.
  • Preferred TALE-nucleases according to the invention are those recognizing and cleaving the target sequence selected from the group consisting of: SEQ ID NO: 1 to 6 (GR), SEQ ID NO: 37, 57 to 60 (TCRalpha), SEQ ID NO: 38 or 39 (TCRbeta), and SEQ ID NO: 40, SEQ ID NO: 61 to SEQ ID NO: 65 (CD52).
  • said method further comprises an additional genomic modification step.
  • additional genomic modification step can be intended the introduction into cells to engineer of one protein of interest.
  • Said protein of interest can be, as non limiting examples a bispecific antibody, rare-cutting endonuclease targeting PDCD1 or CTLA-4, a pTalpha or a functional variant thereof as described in the present disclosure.
  • the present invention also relates isolated cells or cell lines susceptible to be obtained by said method to engineer cells.
  • said isolated cell comprises exogenous polynucleotide sequences encoding polypeptides composing said multi-chain CAR.
  • T-cell activation is regulated by the counterbalancing of stimulatory and inhibitory signal.
  • the expression of immune-checkpoint proteins can be dysregulated by tumours and can be an important immune resistance mechanism.
  • Negative regulators of T- cell function include molecules such as CTLA-4, a key negative regulatory molecule that down-regulates pathways of T-cell activation and programmed death- 1 (PD1) also known as PDCD1, a transmembrane receptor up-regulated on activated T cells that when bound to its ligand (programmed death ligand-1, PD-L1) leads to decreased cytokine production and proliferation of T cells (Pardoll 2012).
  • PD1 programmed death- 1
  • PD-L1 transmembrane receptor up-regulated on activated T cells that when bound to its ligand
  • the present invention relates to a method of engineering T-cells, especially for immunotherapy, comprising genetically modifying T-cells by inactivating at least one protein involved in the immune check-point, in particular PDCD1 and/or CTLA-4.
  • the method comprises one of the following steps:
  • said rare-cutting endonuclease is a TALE-nuclease.
  • new TALE-nucleases have been designed for precisely targeting relevant genes for adoptive immunotherapy strategies.
  • Preferred TALE-nucleases according to the invention are those recognizing and cleaving the target sequence selected from the group consisting of SEQ ID NO: 77 and SEQ ID NO: 78 (PDCD-1), SEQ ID NO: 74 to SEQ ID NO: 76 (CTLA-4).
  • the present invention also relates to TALE-nucleases polypeptides which comprise an amino acid sequence selected from the group consisting of SEQ ID NO: 79 to SEQ ID NO: 88.
  • the present invention also relates to polypeptides comprising an amino acid sequence that has at least 70%, preferably at least 80%, more preferably at least 90 %, 95 % 97 % or 99 % sequence identity with amino acid sequence selected from the group consisting of SEQ ID NO: 79 to SEQ ID NO: 88.
  • polypeptides comprising an amino acid sequence that has at least 70%, preferably at least 80%, more preferably at least 90 %, 95 % 97 % or 99 % sequence identity with amino acid sequence selected from the group consisting of SEQ ID NO: 79 to SEQ ID NO: 88.
  • polynucleotides, vectors encoding the above described rare-cutting endonucleases according to the invention This method can be associated with any one of the different methods described in the present disclosure.
  • bispecific antibodies According to a further embodiment, engineered T cells obtained by the different methods as previously described can be further exposed with bispecific antibodies. Said T-cells could be exposed to bispecific antibodies ex vivo prior to administration to a patient or in vivo following administration to a patient. Said bispecific antibodies comprise two variable regions with distinct antigen properties that allow bringing the engineered cells into proximity to a target antigen. As a non limiting example, said bispecific antibody is directed against a tumor marker and lymphocyte antigen such as CD3 and has the potential to redirect and activate any circulating T cells against tumors.
  • a tumor marker and lymphocyte antigen such as CD3
  • the different methods described above involve introducing pTalpha or functional variants thereof, rare cutting endonuclease, TALE-nuclease, CAR or multi-chain CAR optionally with DNA-end processing enzyme or exogenous nucleic acid into a cell.
  • said pTalpha or functional variant thereof, rare cutting endonucleases, TALE-nucleases, CAR or multi-chain CAR optionally with DNA-end processing enzyme or exogenous nucleic acid can be introduced as transgenes encoded by one or as different plasmidic vectors.
  • Different transgenes can be included in one vector which comprises a nucleic acid sequence encoding ribosomal skip sequence such as a sequence encoding a 2A peptide.
  • 2A peptides which were identified in the Aphthovirus subgroup of picomaviruses, causes a ribosomal "skip" from one codon to the next without the formation of a peptide bond between the two amino acids encoded by the codons (see Donnelly et al., J. of General Virology 82: 1013-1025 (2001); Donnelly et al., J. of Gen. Virology 78: 13-21 (1997); Doronina et al., Mol. And. Cell. Biology 28(13): 4227-4239 (2008); Atkins et al., RNA 13: 803-810 (2007)).
  • cognate is meant three nucleotides on an mRNA (or on the sense strand of a DNA molecule) that are translated by a ribosome into one amino acid residue.
  • two polypeptides can be synthesized from a single, contiguous open reading frame within an mRNA when the polypeptides are separated by a 2A oligopeptide sequence that is in frame.
  • Such ribosomal skip mechanisms are well known in the art and are known to be used by several vectors for the expression of several proteins encoded by a single messenger RNA.
  • 2A peptides have been used to express into the cell the rare-cutting endonuclease and a DNA end-processing enzyme or the different polypeptides of the multi-chain CAR.
  • Said plasmid vector can contain a selection marker which provides for identification and/or selection of cells which received said vector.
  • Polypeptides may be synthesized in situ in the cell as a result of the introduction of polynucleotides encoding said polypeptides into the cell. Alternatively, said polypeptides could be produced outside the cell and then introduced thereto.
  • Methods for introducing a polynucleotide construct into animal cells are known in the art and including as non limiting examples stable transformation methods wherein the polynucleotide construct is integrated into the genome of the cell, transient transformation methods wherein the polynucleotide construct is not integrated into the genome of the cell and virus mediated methods.
  • Said polynucleotides may be introduced into a cell by for example, recombinant viral vectors (e.g.
  • retroviruses adenoviruses
  • liposome adenoviruses
  • transient transformation methods include for example microinjection, electroporation or particle bombardment.
  • Said polynucleotides may be included in vectors, more particularly plasmids or virus, in view of being expressed in cells.
  • polynucleotides encoding polypeptides according to the present invention can be mRNA which is introduced directly into the cells, for example by electroporation.
  • the inventors determined the optimal condition for mRNA electroporation in T-cell.
  • the inventor used the cytoPulse technology which allows, by the use of pulsed electric fields, to transiently permeabilize living cells for delivery of material into the cells.
  • the technology based on the use of PulseAgile (Cellectis property) electroporation waveforms grants the precise control of pulse duration, intensity as well as the interval between pulses (U.S. patent 6,010,613 and International PCT application WO2004083379). All these parameters can be modified in order to reach the best conditions for high transfection efficiency with minimal mortality.
  • the first high electric field pulses allow pore formation, while subsequent lower electric field pulses allow to move the polynucleotide into the cell.
  • the inventor describe the steps that led to achievement of >95% transfection efficiency of mRNA in T cells, and the use of the electroporation protocol to transiently express different kind of proteins in T cells.
  • the invention relates to a method of transforming T cell comprising contacting said T cell with RNA and applying to T cell an agile pulse sequence consisting of: (a) one electrical pulse with a voltage range from 2250 to 3000 V per centimeter, a pulse width of 0.1 ms and a pulse interval of 0.2 to 10 ms between the electrical pulses of step (a) and (b);
  • step (b) one electrical pulse with a voltage range from 2250 to 3000 V with a pulse width of 100 ms and a pulse interval of 100 ms between the electrical pulse of step (b) and the first electrical pulse of step (c) ;
  • the method of transforming T cell comprising contacting said T cell with RNA and applying to T cell an agile pulse sequence consisting of: (a) one electrical pulse with a voltage of 2250, 2300, 2350, 2400, 2450, 2500, 2550,
  • step (b) one electrical pulse with a voltage range from 2250, of 2250, 2300, 2350, 2400, 2450, 2500, 2550, 2400, 2450, 2500, 2600, 2700, 2800, 2900 or 3000V with a pulse width of 100 ms and a pulse interval of 100 ms between the electrical pulse of step (b) and the first electrical pulse of step (c); and
  • Electroporation medium can be any suitable medium known in the art.
  • the electroporation medium has conductivity in a range spanning 0.01 to 1.0 milliSiemens.
  • said RNA encodes a rare-cutting endonuclase, one monomer of the rare-cutting endonuclease such as Half-TALE-nuclease, a Chimeric Antigen Receptor, at least one component of the multi-chain chimeric antigen receptor, a pTalpha or functional variant thereof, an exogenous nucleic acid, one additional catalytic domain.
  • a rare-cutting endonuclase one monomer of the rare-cutting endonuclease such as Half-TALE-nuclease, a Chimeric Antigen Receptor, at least one component of the multi-chain chimeric antigen receptor, a pTalpha or functional variant thereof, an exogenous nucleic acid, one additional catalytic domain.
  • the T cells can be activated and expanded generally using methods as described, for example, in U.S. Patents 6,352,694; 6,534,055; 6,905,680; 6,692,964; 5,858,358; 6,887,466; 6,905,681; 7,144,575; 7,067,318; 7,172,869; 7,232,566; 7,175,843; 5,883,223; 6,905,874; 6,797,514; 6,867,041 ; and U.S. Patent Application Publication No. 20060121005.
  • T cells can be expanded in vitro or in vivo.
  • the T cells of the invention are expanded by contact with a surface having attached thereto an agent that stimulates a CD3 TCR complex associated signal and a ligand that stimulates a co-stimulatory molecule on the surface of the T cells.
  • T cell populations may be stimulated in vitro such as by contact with an anti- CD3 antibody, or antigen-binding fragment thereof, or an anti-CD2 antibody immobilized on a surface, or by contact with a protein kinase C activator (e.g., bryostatin) in conjunction with a calcium ionophore.
  • a protein kinase C activator e.g., bryostatin
  • a ligand that binds the accessory molecule is used for co-stimulation of an accessory molecule on the surface of the T cells.
  • a population of T cells can be contacted with an anti-CD3 antibody and an anti-CD28 antibody, under conditions appropriate for stimulating proliferation of the T cells.
  • an anti-CD3 antibody and an anti-CD28 antibody may be in solution or coupled to a surface.
  • the ratio of particles to cells may depend on particle size relative to the target cell.
  • the cells such as T cells
  • the agents providing each signal may be in solution or coupled to a surface.
  • the ratio of particles to cells may depend on particle size relative to the target cell.
  • the cells such as T cells
  • the agents-coated beads and cells are subsequently separated, and then the cells are cultured.
  • the agent-coated beads and cells prior to culture, are not separated but are cultured together.
  • Cell surface proteins may be ligated by allowing paramagnetic beads to which anti-CD3 and anti- CD28 are attached (3x28 beads) to contact the T cells.
  • the cells for example, 4 to 10 T cells
  • beads for example, DYNABEADS® M-450 CD3/CD28 T paramagnetic beads at a ratio of 1 : 1
  • a buffer preferably PBS (without divalent cations such as, calcium and magnesium).
  • PBS without divalent cations such as, calcium and magnesium
  • any cell concentration may be used.
  • the mixture may be cultured for several hours (about 3 hours) to about 14 days or any hourly integer value in between. In another embodiment, the mixture may be cultured for 21 days.
  • Conditions appropriate for T cell culture include an appropriate media (e.g., Minimal Essential Media or RPMI Media 1640 or, X-vivo 5, (Lonza)) that may contain factors necessary for proliferation and viability, including serum (e.g., fetal bovine or human serum), interleukin-2 (IL-2), insulin, IFN-g , 1L- 4, 1L-7, GM-CSF, -10, - 2, 1L-15, TGFp, and TNF- or any other additives for the growth of cells known to the skilled artisan.
  • Other additives for the growth of cells include, but are not limited to, surfactant, plasmanate, and reducing agents such as N-acetyl-cysteine and 2- mercaptoethanoi.
  • Media can include RPMI 1640, A1M-V, DMEM, MEM, a-MEM, F-12, X- Vivo 1 , and X-Vivo 20, Optimizer, with added amino acids, sodium pyruvate, and vitamins, either serum-free or supplemented with an appropriate amount of serum (or plasma) or a defined set of hormones, and/or an amount of cytokine(s) sufficient for the growth and expansion of T cells.
  • Antibiotics e.g., penicillin and streptomycin, are included only in experimental cultures, not in cultures of cells that are to be infused into a subject.
  • the target cells are maintained under conditions necessary to support growth, for example, an appropriate temperature (e.g., 37° C) and atmosphere (e.g., air plus 5% C02) .
  • T cells that have been exposed to varied stimulation times may exhibit different characteristics
  • said cells can be expanded by co-culturing with tissue or cells. Said cells can also be expanded in vivo, for example in the subject's blood after administrating said cell into the subject.
  • T-cell obtained according to any one of the methods previously described.
  • Said T-cell according to the present invention can be derived from a stem cell.
  • the stem cells can be adult stem cells, embryonic stem cells, more particularly non-human stem cells, cord blood stem cells, progenitor cells, bone marrow stem cells, induced pluripotent stem cells, totipotent stem cells or hematopoietic stem cells.
  • Representative human cells are CD34+ cells.
  • Said isolated cell can also be a dendritic cell, a NK-cell, a B-cell or a T-cell selected from the group consisting of inflammatory T-lymphocytes, cytotoxic T-lymphocytes, regulatory T-lymphocytes or helper T-lymphocytes.
  • said cell can be derived from the group consisting of CD4+ T-lymphocytes and CD8+ T-lymphocytes.
  • T cells can be obtained from a number of non- limiting sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, cord blood, thymus tissue, tissue from a site of infection, ascites, pleural effusion, spleen tissue, and tumors.
  • any number of T cell lines available and known to those skilled in the art may be used.
  • said cell can be derived from a healthy donor, from a patient diagnosed with cancer or from a patient diagnosed with an infection.
  • said cell is part of a mixed population of cells which present different phenotypic characteristics.
  • said isolated cell according to the present invention comprises one inactivated gene selected from the group consisting of CD52, GR, TCR alpha and TCR beta and/or expresses a CAR, a multi-chain CAR and/or a pTalpha transgene.
  • said isolated cell according to the present invention comprises two inactivated genes selected from the group consisting of CD52 and GR, CD52 and TCR alpha, CDR52 and TCR beta, GR and TCR alpha, GR and TCR beta, TCR alpha and TCR beta and/or expresses a CAR, a multi-chain CAR and/or a pTalpha transgene.
  • TCR is rendered not functional in the cells according to the invention by inactivating TCR alpha gene and/or TCR beta gene(s).
  • the above strategies are used more particularly to avoid GvHD.
  • a particular aspect of the present invention is a method to obtain modified cells derived from an individual, wherein said cells can proliferate independently of the Major Histocompatibility Complex signaling pathway. Said method comprises the following steps: (a) Recovering cells from said individual;
  • Modified cells which can proliferate independently of the Major Histocompatibility Complex signaling pathway, susceptible to be obtained by this method are encompassed in the scope of the present invention.
  • Said modified cells can be used in a particular aspect of the invention for treating patients in need thereof against Host versus Graft (HvG) rejection and Graft versus Host Disease (GvHD); therefore in the scope of the present invention is a method of treating patients in need thereof against Host versus Graft (HvG) rejection and Graft versus Host Disease (GvHD) comprising treating said patient by administering to said patient an effective amount of modified cells comprising inactivated TCR alpha and/or TCR beta genes.
  • isolated cell obtained by the different methods or cell line derived from said isolated cell as previously described can be used as a medicament.
  • said medicament can be used for treating cancer or infections in a patient in need thereof.
  • said isolated cell according to the invention or cell line derived from said isolated cell can be used in the manufacture of a medicament for treatment of a cancer or a viral infection in a patient in need thereof.
  • the present invention relies on methods for treating patients in need thereof, said method comprising at least one of the following steps:
  • said T cells of the invention can undergo robust in vivo T cell expansion and can persist for an extended amount of time.
  • Said treatment can be ameliorating, curative or prophylactic. It may be either part of an autologous immunotherapy or part of an allogenic immunotherapy treatment.
  • autologous it is meant that cells, cell line or population of cells used for treating patients are originating from said patient or from a Human Leucocyte Antigen (HLA) compatible donor.
  • HLA Human Leucocyte Antigen
  • allogeneic is meant that the cells or population of cells used for treating patients are not originating from said patient but from a donor.
  • the invention is particularly suited for allogenic immunotherapy, insofar as it enables the transformation of T-cells, typically obtained from donors, into non-alloreactive cells. This may be done under standard protocols and reproduced as many times as needed.
  • the resulted modified T cells may be pooled and administrated to one or several patients, being made available as an "off the shelf therapeutic product.
  • Cancers that can be used with the disclosed methods are described in the previous section. Said treatment can be used to treat patients diagnosed with cancer, viral infection, autoimmune disorders or Graft versus Host Disease (GvHD). Cancers that may be treated include tumors that are not vascularized, or not yet substantially vascularized, as well as vascularized tumors.
  • the cancers may comprise nonsolid tumors (such as hematological tumors, for example, leukemias and lymphomas) or may comprise solid tumors.
  • Types of cancers to be treated with the CARs of the invention include, but are not limited to, carcinoma, blastoma, and sarcoma, and certain leukemia or lymphoid malignancies, benign and malignant tumors, and malignancies e.g., sarcomas, carcinomas, and melanomas.
  • carcinoma a malignant neoplasm originating from a malignant neoplasm originating from tumors.
  • sarcomas e.g., sarcomas, carcinomas, and melanomas.
  • adult tumors/cancers and pediatric tumors/cancers are also included.
  • the present invention can be a treatment in combination with one or more therapies against cancer selected from the group of antibodies therapy, chemotherapy, cytokines therapy, dendritic cell therapy, gene therapy, hormone therapy, laser light therapy and radiation therapy.
  • said treatment can be administrated into patients undergoing an immunosuppressive treatment.
  • the present invention preferably relies on cells or population of cells, which have been made resistant to at least one immunosuppressive agent due to the inactivation of a gene encoding a receptor for such immunosuppressive agent.
  • the immunosuppressive treatment should help the selection and expansion of the T-cells according to the invention within the patient.
  • the administration of the cells or population of cells according to the present invention may be carried out in any convenient manner, including by aerosol inhalation, injection, ingestion, transfusion, implantation or transplantation.
  • the compositions described herein may be administered to a patient subcutaneously, intradermaliy, intratumorally, intranodally, intramedullary, intramuscularly, by intravenous or intralymphatic injection, or intraperitoneally.
  • the cell compositions of the present invention are preferably administered by intravenous injection.
  • the administration of the cells or population of cells can consist of the administration of 10 4 - 10 9 cells per kg body weight, preferably 10 5 to 10 6 cells/kg body weight including all integer values of cell numbers within those ranges.
  • the cells or population of cells can be administrated in one or more doses.
  • said effective amount of cells are administrated as a single dose.
  • said effective amount of cells are administrated as more than one dose over a period time. Timing of administration is within the judgment of managing physician and depends on the clinical condition of the patient.
  • the cells or population of cells may be obtained from any source, such as a blood bank or a donor. While individual needs vary, determination of optimal ranges of effective amounts of a given cell type for a particular disease or conditions within the skill of the art.
  • An effective amount means an amount which provides a therapeutic or prophylactic benefit.
  • the dosage administrated will be dependent upon the age, health and weight of the recipient, kind of concurrent treatment, if any, frequency of treatment and the nature of the effect desired.
  • said effective amount of cells or composition comprising those cells are administrated parenterally.
  • Said administration can be an intravenous administration.
  • Said administration can be directly done by injection within a tumor.
  • cells are administered to a patient in conjunction with (e.g., before, simultaneously or following) any number of relevant treatment modalities, including but not limited to treatment with agents such as antiviral therapy, cidofovir and interleukin-2, Cytarabine (also known as ARA-C) or nataliziimab treatment for MS patients or efaliztimab treatment for psoriasis patients or other treatments for PML patients.
  • agents such as antiviral therapy, cidofovir and interleukin-2, Cytarabine (also known as ARA-C) or nataliziimab treatment for MS patients or efaliztimab treatment for psoriasis patients or other treatments for PML patients.
  • the T cells of the invention may be used in combination with chemotherapy, radiation, immunosuppressive agents, such as cyclosporin, azathioprine, methotrexate, mycophenolate, and FK506, antibodies, or other immunoablative agents such as CAM PATH, anti-CD3 antibodies or other antibody therapies, cytoxin, fludaribine, cyclosporin, FK506, rapamycin, mycoplienolic acid, steroids, FR901228, cytokines, and irradiation.
  • immunosuppressive agents such as cyclosporin, azathioprine, methotrexate, mycophenolate, and FK506, antibodies
  • other immunoablative agents such as CAM PATH, anti-CD3 antibodies or other antibody therapies, cytoxin, fludaribine, cyclosporin, FK506, rapamycin, mycoplienolic acid, steroids, FR901228, cytokines, and irradiation.
  • the cell compositions of the present invention are administered to a patient in conjunction with (e.g., before, simultaneously or following) bone marrow transplantation, T cell ablative therapy using either chemotherapy agents such as, fludarabine, external-beam radiation therapy (XRT), cyclophosphamide, or antibodies such as OKT3 or CAMPATH,
  • chemotherapy agents such as, fludarabine, external-beam radiation therapy (XRT), cyclophosphamide, or antibodies such as OKT3 or CAMPATH
  • the cell compositions of the present invention are administered following B-cell ablative therapy such as agents that react with CD20, e.g., Rituxan.
  • subjects may undergo standard treatment with high dose chemotherapy followed by peripheral blood stem cell transplantation.
  • subjects receive an infusion of the expanded immune cells of the present invention.
  • expanded cells are administered before or following surgeiy.
  • Said modified cells obtained by any one of the methods described here can be used in a particular aspect of the invention for treating patients in need thereof against Host versus Graft (HvG) rejection and Graft versus Host Disease (GvHD); therefore in the scope of the present invention is a method of treating patients in need thereof against Host versus Graft (HvG) rejection and Graft versus Host Disease (GvHD) comprising treating said patient by administering to said patient an effective amount of modified cells comprising inactivated TCR alpha and/or TCR beta genes.
  • T-cells from a cell culture or from a blood sample from one individual patient or from blood bank and activating said T cells using anti-CD3/C28 activator beads.
  • the beads provide both the primary and co-stimulatory signals that are required for activation and expansion of T cells.
  • TCR disruption is expected to the elimination of the TCR complex and removes alloreactivity (GvHD) but may alter allogenic cells expansion due to the loss of CD3 signaling component.
  • Transduced cells are expected to express pTalpha chain or functional variant thereof. This pTalpha chain pairs with TCRbeta chain and CD3 signaling components to form the preTCR complex and, thus restore a functional CD3 complex and support activation or stimulation of inactivated TCRalpha cells. Transduction of T-cells with pTalpha lentiviral vector can be realized before or after TCRalpha inactivation.
  • Transducing said cells with multi-chain CARs allow redirecting T cells against antigens expressed at the surface of target cells from various malignancies including lymphomas and solid tumors.
  • the inventors have designed a multi-chain CAR derived from FcsRI as previously described. Transduction can be realized before or after the inactivation of TCRalpha and CD52 genes.
  • TCR alpha in said cells to eliminate the TCR from the surface of the cell and prevent recognition of host tissue as foreign by TCR of allogenic and thus to avoid GvHD.
  • target of immunosuppressive agents is CD52 and immunosuppressive agent is a humanized monoclonal anti-CD52 antibody.
  • TALE-nuclease by allowing higher rates of DSB events within T-cells was particularly advantageous to achieve the above double inactivation in T-cells.
  • TCRalpha and CD52 genes are inactivated by electoporating T cells with mRNA coding for TALE-nuclease targeting said genes. It has been found by the inventors that using mRNA resulted into high transformation rate was less harmful to T-cells and so, was critical in the process of engineering T-cells. Then, inactivated T cells are sorted using magnetic beads. For example, T cells expressing CD52 are removed by fixation on a solid surface, and inactivated cells are not exposed of the stress of being passed through a column. This gentle method increases the concentration of properly engineered T-cells.
  • - Amino acid residues in a polypeptide sequence are designated herein according to the one- letter code, in which, for example, Q means Gin or Glutamine residue, R means Arg or Arginine residue and D means Asp or Aspartic acid residue.
  • - Amino acid substitution means the replacement of one amino acid residue with another, for instance the replacement of an Arginine residue with a Glutamine residue in a peptide sequence is an amino acid substitution.
  • nucleosides are designated as follows: one-letter code is used for designating the base of a nucleoside: a is adenine, t is thymine, c is cytosine, and g is guanine.
  • r represents g or a (purine nucleotides)
  • k represents g or t
  • s represents g or c
  • w represents a or t
  • m represents a or c
  • y represents t or c (pyrimidine nucleotides)
  • d represents g, a or t
  • v represents g, a or c
  • b represents g, t or c
  • h represents a, t or c
  • n represents g, a, t or c.
  • nucleic acid or “polynucleotides” refers to nucleotides and/or polynucleotides, such as deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), oligonucleotides, fragments generated by the polymerase chain reaction (PCR), and fragments generated by any of ligation, scission, endonuclease action, and exonuclease action.
  • DNA deoxyribonucleic acid
  • RNA ribonucleic acid
  • PCR polymerase chain reaction
  • Nucleic acid molecules can be composed of monomers that are naturally-occurring nucleotides (such as DNA and RNA), or analogs of naturally-occurring nucleotides (e.g., enantiomeric forms of naturally-occurring nucleotides), or a combination of both.
  • Modified nucleotides can have alterations in sugar moieties and/or in pyrimidine or purine base moieties.
  • Sugar modifications include, for example, replacement of one or more hydroxyl groups with halogens, alkyl groups, amines, and azido groups, or sugars can be functionalized as ethers or esters.
  • sugar moiety can be replaced with sterically and electronically similar structures, such as aza-sugars and carbocyclic sugar analogs.
  • modifications in a base moiety include alkylated purines and pyrimidines, acylated purines or pyrimidines, or other well-known heterocyclic substitutes.
  • Nucleic acid monomers can be linked by phosphodiester bonds or analogs of such linkages. Nucleic acids can be either single stranded or double stranded.
  • polynucleotide successively comprising a first region of homology to sequences upstream of said double-stranded break, a sequence to be inserted in the genome of said cell and a second region of homology to sequences downstream of said double-stranded break it is intended to mean a DNA construct or a matrix comprising a first and second portion that are homologous to regions 5' and 3' of a DNA target in situ.
  • the DNA construct also comprises a third portion positioned between the first and second portion which comprise some homology with the corresponding DNA sequence in situ or alternatively comprise no homology with the regions 5' and 3' of the DNA target in situ.
  • a homologous recombination event is stimulated between the genome containing the targeted gene comprised in the locus of interest and this matrix, wherein the genomic sequence containing the DNA target is replaced by the third portion of the matrix and a variable part of the first and second portions of said matrix.
  • DNA target a DNA target sequence
  • target DNA sequence a DNA sequence that can be targeted and processed by a rare-cutting endonuclease according to the present invention.
  • processing site a polynucleotide sequence that can be targeted and processed by a rare-cutting endonuclease according to the present invention.
  • These terms refer to a specific DNA location, preferably a genomic location in a cell, but also a portion of genetic material that can exist independently to the main body of genetic material such as plasmids, episomes, virus, transposons or in organelles such as mitochondria as non-limiting example.
  • targeted genomic sequences generally consist of two 17-bp long sequences (called half targets) separated by a 15-bp spacer. Each half-target is recognized by repeats of TALE- nucleases listed in tables 1, 5, 6 and 10 as non-limiting examples, encoded in plasmids, under the control of EF1 -alpha promoter or T7 promoter.
  • the nucleic acid target sequence is defined by the 5' to 3' sequence of one strand of said target, as indicated in tables 1, 5, 6 and 10.
  • CAR chimeric antigen receptor
  • a component present on the target cell for example an antibody-based specificity for a desired antigen (e.g., tumor antigen) with a T cell receptor-activating intracellular domain to generate a chimeric protein that exhibits a specific anti-target cellular immune activity.
  • CAR consists of an extracellular single chain antibody (scFvFc) fused to the intracellular signaling domain of the T cell antigen receptor complex zeta chain (scFvFc ⁇ ) and have the ability, when expressed in T cells, to redirect antigen recognition based on the monoclonal antibody's specificity.
  • scFvFc ⁇ extracellular single chain antibody
  • scFvFc ⁇ extracellular single chain antibody
  • scFvFc ⁇ extracellular signaling domain of the T cell antigen receptor complex zeta chain
  • One example of CAR used in the present invention is a CAR directing against CD 19 antigen and can comprise as non limiting example the amino acid sequence :
  • delivery vector or “ delivery vectors” is intended any delivery vector which can be used in the present invention to put into cell contact ( i.e “contacting") or deliver inside cells or subcellular compartments (i.e “introducing") agents/chemicals and molecules (proteins or nucleic acids) needed in the present invention. It includes, but is not limited to liposomal delivery vectors, viral delivery vectors, drug delivery vectors, chemical carriers, polymeric carriers, lipoplexes, polyplexes, dendrimers, microbubbles (ultrasound contrast agents), nanoparticles, emulsions or other appropriate transfer vectors.
  • delivery vectors allow delivery of molecules, chemicals, macromolecules (genes, proteins), or other vectors such as plasmids, peptides developed by Diatos. In these cases, delivery vectors are molecule carriers. By “delivery vector” or “delivery vectors” is also intended delivery methods to perform transfection. - The terms “vector” or “vectors” refer to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked.
  • a "vector” in the present invention includes, but is not limited to, a viral vector, a plasmid, a RNA vector or a linear or circular DNA or RNA molecule which may consists of a chromosomal, non chromosomal, semi-synthetic or synthetic nucleic acids.
  • Preferred vectors are those capable of autonomous replication (episomal vector) and/or expression of nucleic acids to which they are linked (expression vectors). Large numbers of suitable vectors are known to those of skill in the art and commercially available.
  • Viral vectors include retrovirus, adenovirus, parvovirus (e. g. adenoassociated viruses), coronavirus, negative strand RNA viruses such as orthomyxovirus (e. g., influenza virus), rhabdovirus (e. g., rabies and vesicular stomatitis virus), paramyxovirus (e. g. measles and Sendai), positive strand RNA viruses such as picornavirus and alphavirus, and double- stranded DNA viruses including adenovirus, herpesvirus (e. g., Herpes Simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus), and poxvirus (e.
  • orthomyxovirus e. g., influenza virus
  • rhabdovirus e. g., rabies and vesicular stomatitis virus
  • paramyxovirus e. g. measles and Sendai
  • viruses include Norwalk virus, togavirus, flavivirus, reoviruses, papovavirus, hepadnavirus, and hepatitis virus, for example.
  • retroviruses include: avian leukosis-sarcoma, mammalian C-type, B-type viruses, D type viruses, HTLV- BLV group, lentivirus, spumavirus (Coffin, J. M., Retroviridae: The viruses and their replication, In Fundamental Virology, Third Edition, B. N. Fields, et al., Eds., Lippincott- Raven Publishers, Philadelphia, 1996).
  • lentiviral vector HIV-Based lentiviral vectors that are very promising for gene delivery because of their relatively large packaging capacity, reduced immunogenicity and their ability to stably transduce with high efficiency a large range of different cell types.
  • Lentiviral vectors are usually generated following transient transfection of three (packaging, envelope and transfer) or more plasmids into producer cells.
  • lentiviral vectors enter the target cell through the interaction of viral surface glycoproteins with receptors on the cell surface.
  • the viral RNA undergoes reverse transcription, which is mediated by the viral reverse transcriptase complex.
  • the product of reverse transcription is a double-stranded linear viral DNA, which is the substrate for viral integration in the DNA of infected cells.
  • integrative lentiviral vectors or LV
  • NILV non integrative lentiviral vectors
  • efficient gene delivery vectors that do not integrate the genome of a target cell through the action of the virus integrase.
  • - Delivery vectors and vectors can be associated or combined with any cellular permeabilization techniques such as sonoporation or electroporation or derivatives of these techniques.
  • cell or cells any eukaryotic living cells, primary cells and cell lines derived from these organisms for in vitro cultures.
  • primary cell or “primary cells” are intended cells taken directly from living tissue (i.e. biopsy material) and established for growth in vitro, that have undergone very few population doublings and are therefore more representative of the main functional components and characteristics of tissues from which they are derived from, in comparison to continuous tumorigenic or artificially immortalized cell lines.
  • cell lines can be selected from the group consisting of CHO-K1 cells; HEK293 cells; Caco2 cells; U2-OS cells; NIH 3T3 cells; NSO cells; SP2 cells; CHO-S cells; DG44 cells; K-562 cells, U-937 cells; MRC5 cells; IMR90 cells; Jurkat cells; HepG2 cells; HeLa cells; HT-1080 cells; HCT-116 cells; Hu-h7 cells; Huvec cells; Molt 4 cells.
  • All these cell lines can be modified by the method of the present invention to provide cell line models to produce, express, quantify, detect, study a gene or a protein of interest; these models can also be used to screen biologically active molecules of interest in research and production and various fields such as chemical, biofuels, therapeutics and agronomy as non- limiting examples.
  • - by “mutation” is intended the substitution, deletion, insertion of up to one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, twenty, twenty five, thirty, fourty, fifty, or more nucleotides/amino acids in a polynucleotide (cDNA, gene) or a polypeptide sequence.
  • the mutation can affect the coding sequence of a gene or its regulatory sequence. It may also affect the structure of the genomic sequence or the structure/stability of the encoded mRNA.
  • variant(s) it is intended a repeat variant, a variant, a DNA binding variant, a TALE- nuclease variant, a polypeptide variant obtained by mutation or replacement of at least one residue in the amino acid sequence of the parent molecule.
  • - by "functional variant” is intended a catalytically active mutant of a protein or a protein domain; such mutant may have the same activity compared to its parent protein or protein domain or additional properties, or higher or lower activity.
  • gene is meant the basic unit of heredity, consisting of a segment of DNA arranged in a linear manner along a chromosome, which codes for a specific protein or segment of protein.
  • a gene typically includes a promoter, a 5' untranslated region, one or more coding sequences (exons), optionally introns, a 3' untranslated region.
  • the gene may further comprise a terminator, enhancers and/or silencers.
  • locus is the specific physical location of a DNA sequence (e.g. of a gene) on a chromosome.
  • locus can refer to the specific physical location of a rare-cutting endonuclease target sequence on a chromosome.
  • Such a locus can comprise a target sequence that is recognized and/or cleaved by a rare-cutting endonuclease according to the invention. It is understood that the locus of interest of the present invention can not only qualify a nucleic acid sequence that exists in the main body of genetic material (i.e.
  • endonuclease refers to any wild-type or variant enzyme capable of catalyzing the hydrolysis (cleavage) of bonds between nucleic acids within a DNA or RNA molecule, preferably a DNA molecule. Endonucleases do not cleave the DNA or RNA molecule irrespective of its sequence, but recognize and cleave the DNA or RNA molecule at specific polynucleotide sequences, further referred to as "target sequences" or "target sites”.
  • Endonucleases can be classified as rare-cutting endonucleases when having typically a polynucleotide recognition site greater than 12 base pairs (bp) in length, more preferably of 14-55 bp.
  • Rare-cutting endonucleases significantly increase HR by inducing DNA double- strand breaks (DSBs) at a defined locus (Rouet, Smih et al. 1994; Choulika, Perrin et al. 1995; Pingoud and Silva 2007).
  • Rare-cutting endonucleases can for example be a homing endonuclease (Paques and Duchateau 2007), a chimeric Zinc-Finger nuclease (ZFN) resulting from the fusion of engineered zinc-finger domains with the catalytic domain of a restriction enzyme such as Fokl (Porteus and Carroll 2005) or a chemical endonuclease (Eisenschmidt, Lanio et al. 2005; Arimondo, Thomas et al. 2006).
  • a chemical or peptidic cleaver is conjugated either to a polymer of nucleic acids or to another DNA recognizing a specific target sequence, thereby targeting the cleavage activity to a specific sequence.
  • Chemical endonucleases also encompass synthetic nucleases like conjugates of orthophenanthroline, a DNA cleaving molecule, and triplex-forming oligonucleotides (TFOs), known to bind specific DNA sequences (Kalish and Glazer 2005). Such chemical endonucleases are comprised in the term "endonuclease" according to the present invention.
  • Rare-cutting endonucleases can also be for example TALE-nucleases, a new class of chimeric nucleases using a Fokl catalytic domain and a DNA binding domain derived from Transcription Activator Like Effector (TALE), a family of proteins used in the infection process by plant pathogens of the Xanthomonas genus (Boch, Scholze et al. 2009; Moscou and Bogdanove 2009; Christian, Cermak et al. 2010; Li, Huang et al.).
  • TALE Transcription Activator Like Effector
  • TALE-nuclease Fokl-based TALE-nuclease
  • ZFN Zinc- finger DNA binding domain
  • TALE-nuclease DNA cleavage by a TALE-nuclease requires two DNA recognition regions flanking an unspecific central region.
  • Rare-cutting endonucleases encompassed in the present invention can also be derived from TALE-nucleases.
  • Rare-cutting endonuclease can be a homing endonuclease, also known under the name of meganuclease. Such homing endonucleases are well-known to the art (Stoddard 2005). Homing endonucleases recognize a DNA target sequence and generate a single- or double- strand break. Homing endonucleases are highly specific, recognizing DNA target sites ranging from 12 to 45 base pairs (bp) in length, usually ranging from 14 to 40 bp in length.
  • the homing endonuclease according to the invention may for example correspond to a LAGLIDADG endonuclease, to a HNH endonuclease, or to a GIY-YIG endonuclease.
  • Preferred homing endonuclease according to the present invention can be an l-Crel variant.
  • TALE-nuclease TALEN
  • TALEN Transcription Activator Like Effector
  • the catalytic domain is preferably a nuclease domain and more preferably a domain having endonuclease activity, like for instance I-Tevl, ColE7, NucA and Fok-I.
  • the TALE domain can be fused to a meganuclease like for instance I-Crel and I-Onul or functional variant thereof.
  • said nuclease is a monomeric TALE- Nuclease.
  • a monomeric TALE-Nuclease is a TALE-Nuclease that does not require dimerization for specific recognition and cleavage, such as the fusions of engineered TAL repeats with the catalytic domain of I-Tevl described in WO2012138927.
  • Transcription Activator like Effector are proteins from the bacterial species Xanthomonas comprise a plurality of repeated sequences, each repeat comprising di -residues in position 12 and 13 (RVD) that are specific to each nucleotide base of the nucleic acid targeted sequence.
  • Binding domains with similar modular base-per-base nucleic acid binding properties can also be derived from new modular proteins recently discovered by the applicant in a different bacterial species.
  • the new modular proteins have the advantage of displaying more sequence variability than TAL repeats.
  • RVDs associated with recognition of the different nucleotides are HD for recognizing C, NG for recognizing T, NI for recognizing A, for recognizing G or A, NS for recognizing A, C, G or T, HG for recognizing T, IG for recognizing T, NK for recognizing G, HA for recognizing C, ND for recognizing C, HI for recognizing C, HN for recognizing G, NA for recognizing G, SN for recognizing G or A and YG for recognizing T, TL for recognizing A, VT for recognizing A or G and SW for recognizing A.
  • critical amino acids 12 and 13 can be mutated towards other amino acid residues in order to modulate their specificity towards nucleotides A, T, C and G and in particular to enhance this specificity.
  • TALE-nuclease have been already described and used to stimulate gene targeting and gene modifications (Boch, Scholze et al. 2009; Moscou and Bogdanove 2009; Christian, Cermak et al. 2010; Li, Huang et al.).
  • Engineered TAL-nucleases are commercially available under the trade name TALENTM (Cellectis, 8 rue de la Croix Jarry, 75013 Paris, France).
  • cleavage refers to the breakage of the covalent backbone of a polynucleotide. Cleavage can be initiated by a variety of methods including, but not limited to, enzymatic or chemical hydrolysis of a phosphodiester bond. Both single-stranded cleavage and double- stranded cleavage are possible, and double-stranded cleavage can occur as a result of two distinct single-stranded cleavage events. Double stranded DNA, RNA, or DNA/RNA hybrid cleavage can result in the production of either blunt ends or staggered ends.
  • fusion protein is intended the result of a well-known process in the art consisting in the joining of two or more genes which originally encode for separate proteins or part of them, the translation of said "fusion gene” resulting in a single polypeptide with functional properties derived from each of the original proteins.
  • identity refers to sequence identity between two nucleic acid molecules or polypeptides. Identity can be determined by comparing a position in each sequence which may be aligned for purposes of comparison. When a position in the compared sequence is occupied by the same base, then the molecules are identical at that position. A degree of similarity or identity between nucleic acid or amino acid sequences is a function of the number of identical or matching nucleotides at positions shared by the nucleic acid sequences.
  • Various alignment algorithms and/or programs may be used to calculate the identity between two sequences, including FASTA, or BLAST which are available as a part of the GCG sequence analysis package (University of Wisconsin, Madison, Wis.), and can be used with, e.g., default setting.
  • polypeptides having at least 70%, 85%, 90%, 95%, 98% or 99% identity to specific polypeptides described herein and preferably exhibiting substantially the same functions, as well as polynucleotide encoding such polypeptides, are contemplated.
  • Similarity describes the relationship between the amino acid sequences of two or more polypeptides.
  • BLASTP may also be used to identify an amino acid sequence having at least 70%, 75%, 80%, 85%, 87.5%, 90%, 92.5%, 95%, 97.5%, 98%, 99% sequence similarity to a reference amino acid sequence using a similarity matrix such as BLOSUM45, BLOSUM62 or BLOSUM80. Unless otherwise indicated a similarity score will be based on use of BLOSUM62.
  • BLOSUM45 BLOSUM45
  • BLOSUM62 BLOSUM80
  • BLASTP "Identities” shows the number and fraction of total residues in the high scoring sequence pairs which are identical; and BLASTP “Positives” shows the number and fraction of residues for which the alignment scores have positive values and which are similar to each other.
  • Amino acid sequences having these degrees of identity or similarity or any intermediate degree of identity of similarity to the amino acid sequences disclosed herein are contemplated and encompassed by this disclosure.
  • the polynucleotide sequences of similar polypeptides are deduced using the genetic code and may be obtained by conventional means.
  • a functional variant of pTalpha can have 70%, 75%, 80%, 85%, 87.5%, 90%, 92.5%, 95%, 97.5%, 98%, 99% sequence similarity to the amino acid sequence of SEQ ID NO : 107.
  • a polynucleotide encoding such a functional variant would be produced by reverse translating its amino acid sequence using the genetic code.
  • signal-transducing domain or "co-stimulatory ligand” refers to a molecule on an antigen presenting cell that specifically binds a cognate co-stimulatory molecule on a T-cell, thereby providing a signal which, in addition to the primary signal provided by, for instance, binding of a TCR/CD3 complex with an MHC molecule loaded with peptide, mediates a T cell response, including, but not limited to, proliferation activation, differentiation and the like.
  • a co-stimulatory ligand can include but is not limited to CD7, B7-1 (CD80), B7-2 (CD86), PD- LI, PD-L2, 4-1BBL, OX40L, inducible costimulatory igand (ICOS-L), intercellular adhesion molecule (ICAM, CD30L, CD40, CD70, CD83, HLA-G, MICA, M1CB, HVEM, lymphotoxin beta receptor, 3/TR6, ILT3, ILT4, an agonist or antibody that binds Toll ligand receptor and a ligand that specifically binds with B7-H3.
  • a co-stimulatory ligand also encompasses, inter alia, an antibody that specifically binds with a co-stimulatory molecule present on a T cell, such as but not limited to, CD27, CD28, 4-IBB, OX40, CD30, CD40, PD- 1, ICOS, lymphocyte function-associated antigen- 1 (LFA-1), CD2, CD7, LTGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83.
  • an antibody that specifically binds with a co-stimulatory molecule present on a T cell such as but not limited to, CD27, CD28, 4-IBB, OX40, CD30, CD40, PD- 1, ICOS, lymphocyte function-associated antigen- 1 (LFA-1), CD2, CD7, LTGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83.
  • a "co-stimulatory molecule” refers to the cognate binding partner on a Tcell that specifically binds with a co-stimulatory ligand, thereby mediating a co-stimulatory response by the cell, such as, but not limited to proliferation.
  • Co-stimulatory molecules include, but are not limited to an MHC class I molecule, BTLA and Toll ligand receptor.
  • a "co-stimulatory signal” as used herein refers to a signal, which in combination with primary signal, such as TCR/CD3 ligation, leads to T cell proliferation and/or upregulation or downregulation of key molecules.
  • bispecific antibody refers to an antibody that has binding sites for two different antigens within a single antibody molecule. It will be appreciated by those skilled in the art that other molecules in addition to the canonical antibody structure may be constructed with two binding specificities. It will further be appreciated that antigen binding by bispecific antibodies may be simultaneous or sequential. Bispecific antibodies can be produced by chemical techniques (see e.g., Kranz et al. (1981) Proc. Natl. Acad. Sci. USA 78, 5807), by "polydoma” techniques (See U.S. Pat. No. 4,474,893) or by recombinant DNA techniques, which all are known per se.
  • each binding domain comprises at least one variable region from an antibody heavy chain ("VH or H region"), wherein the VH region of the first binding domain specifically binds to the lymphocyte marker such as CD3, and the VH region of the second binding domain specifically binds to tumor antigen.
  • VH or H region an antibody heavy chain
  • extracellular ligand-binding domain is defined as an oligo- or polypeptide that is capable of binding a ligand.
  • the domain will be capable of interacting with a cell surface molecule.
  • the extracellular ligand-binding domain may be chosen to recognize a ligand that acts as a cell surface marker on target cells associated with a particular disease state.
  • cell surface markers that may act as ligands include those associated with viral, bacterial and parasitic infections, autoimmune disease and cancer cells.
  • subject or "patient” as used herein includes all members of the animal kingdom including non-human primates and humans.
  • the above written description of the invention provides a manner and process of making and using it such that any person skilled in this art is enabled to make and use the same, this enablement being provided in particular for the subject matter of the appended claims, which make up a part of the original description.
  • Example 1 TALE-nucleases cleaving the human GR gene
  • TALE-nucleases targeting exons of the human GR gene were designed and produced.
  • Table 1 indicates the target sequences cleaved by each TALE-nuclease.
  • GR TALE-nuclease was composed of two independent entities (called half TALE-nucleases) each containing a repeat sequence engineered to bind and cleave GR target sequences consisting of two 17-bp long sequences (called half targets) separated by a 15-bp spacer.
  • Table 1 Description of the GR TALE-nucleases and sequences of the TALE-nucleases target sites in the human GR gene.
  • the amino acid sequences of the N-terminal, C-terminal domains and repeat are based on the AvrBs3 TALE (ref: GenBank: X16130.1).
  • the C-terminal and the N-terminal domains are separated by two BsmBI restriction sites.
  • the repeat arrays (SEQ ID NO: 7 to 18), targeting the desired sequences (SEQ ID NO: 1 to 6) were synthesized using a solid support method composed of consecutive restriction/ligation/washing steps (International PCT application WO2013/017950).
  • the first block (coding for a di-repeat) was immobilized on a solid support through biotin/streptavidin interaction
  • the second block (tri-repeat) was then ligated to the first and after SfaNI digestion a third bloc (tri-repeat) was coupled.
  • the process was repeated using tri- or di-repeat blocks upon obtaining the desired repeat array.
  • the product was then cloned in a classical pAPGlO cloning plasmid for amplification in E. coli and sequenced.
  • the repeat array sequences thus obtained were subcloned in a yeast expression TALE vector using type IIS restriction enzymes BsmBI for the receiving plasmid and Bbvl and SfaNI for the inserted repeat sequence.
  • GR TALE-nucleases Activity of GR TALE-nucleases in yeast: Nuclease activity of the six GR-TALE-nucleases were tested at 37°C and 30°C in our yeast SSA assay previously described (International PCT Applications WO 2004/067736 and in (Epinat, Arnould et al. 2003; Chames, Epinat et al. 2005; Arnould, Chames et al. 2006; Smith, Grizot et al. 2006) on targets containing the two TALE target sequences facing each other on the DNA strand separated by a spacer of 15 bps resulting in SEQ ID NO: 1 to 6.
  • yeast target reporter plasmids containing the TALE-nuclease DNA target sequences were constructed as previously described (International PCT Applications WO 2004/067736 and in (Epinat, Arnould et al. 2003; Chames, Epinat et al. 2005; Arnould, Chames et al. 2006; Smith, Grizot et al. 2006). TALE-nuclease cleavage activity levels, in yeast, of individual clones on the targets are presented in table 2.
  • Table 2 Cleavage activity of the GR TALE-nucleases in yeast.
  • Each TALE-nuclease construct was subcloned using restriction enzyme digestion in a mammalian expression vector under the control of a pEFl alpha long promoter.
  • HEK293 cells were seeded one day prior to transfection.
  • Cells were co- transfected with 2.5 ⁇ g of each of two plasmids encoding left and right half of GRex2, GRex3T2, GRex3T4, GRex5Tl, GRex5T2 or GRex5T3 TALE-nuclease recognizing the two half targets genomic sequences of interest in the GR gene under the control of EF1 alpha promoter using 25 ⁇ of lipofectamine (Invitrogen) according to the manufacturer's instructions.
  • TALE-nucleases targeting the T-cell receptor alpha constant chain region (TRAC_T01) target site ((TRAC_T01-L and -R TALE-nuclease (SEQ ID NO: 41 and SEQ ID NO: 42, TRAC T01 target site (SEQ ID NO: 37)) under the control of EF1 alpha promoter.
  • TALE-T01 target site (TRAC_T01-L and -R TALE-nuclease (SEQ ID NO: 41 and SEQ ID NO: 42, TRAC T01 target site (SEQ ID NO: 37)) under the control of EF1 alpha promoter.
  • the double strand break generated by TALE-nucleases in GR coding sequence induces non homologous end joining (NHEJ), which is an error-prone mechanism.
  • NHEJ non homologous end joining
  • TALE-nucleases Activity of TALE-nucleases is measured by the frequency of insertions or deletions at the genomic locus targeted. 2 or 7 days post transfection cells were harvested and locus specific PCRs were performed on genomic DNA extracted using the following primers: 5'- CCATCTCATCCCTGCGTGTCTCCGACTCAG-3' (forward adaptator sequence)- 10N (TAG)- locus specific forward sequence for GR exon 2: 5 ' -GGTTC ATTTAACAAGCTGCC- 3' (SEQ ID NO: 31), for GR exon 3: 5 ' -GC ATTCTGACTATGAAGTGA-3 ' (SEQ ID NO: 32) and for GR exon 5: 5 ' -TC AGC AGGCC ACTAC AGGAGTCTCAC AAG-3 ' (SEQ ID NO: 33) and the reverse primer 5 ' -CCTATCCCCTGTGTGCCTTGGC AGTCTC AG-3 ' (reverse adaptor sequence)- locus specific reverse sequence for
  • PCR products were sequenced by a 454 sequencing system (454 Life Sciences). Approximately 10,000 sequences were obtained per PCR product and then analyzed for the presence of site-specific insertion or deletion events. Table 3 indicates the percentage of the sequences showing insertions or deletions at the TALE-nuclease target site among the total number of sequences in the sample. In table 3 are listed for GRex2, GRex3T2 and GRex3T4 the results of a representative experiment.
  • TALE-nuclease construct was subcloned using restriction enzyme digestion in an expression vector under the control of a T7 promoter.
  • mRNA encoding TALE-nucleases cleaving GR genomic sequences were synthesized from each plasmid carrying the coding sequences downstream from the T7 promoter.
  • T lymphocytes isolated from peripheral blood were activated for 5 days using anti-CD3/CD28 activator beads (Life technologies) and 5 million cells were transfected by electroporation with 10 ⁇ g of each of 2 mRNAs encoding both half TALE-nucleases using a CytoLVT-P instrument (BTX-Harvard apparatus).
  • T cells transfected with 10 ⁇ g of each of the 2 mRNAs encoding both half TALE-nucleases targeting the CD52 gene (CD52 T02-L and -R TALEN (SEQ ID NO: 55 and 56), target sequence CD52 T02 SEQ ID NO: 40) are used as a control.
  • Table 4 Percentage of targeted mutagenesis at endogenous TALE-nuclease target sites in primary T lymphocytes.
  • Example 2 TALE-nucleases cleaving the human CD52 gene, the human T-cell receptor alpha constant chain (TRAC) and the human T-cell receptor beta constant chains 1 and 2 (TRBC)
  • heterodimeric TALE-nucleases targeting respectively CD52, TRAC and TRBC genes were designed and produced.
  • the targeted genomic sequences consist of two 17-bp long sequences (called half targets) separated by an 11 or 15-bp spacer. Each half-target is recognized by repeats of half TALE-nucleases listed in table 5.
  • the human genome contains two functional T-cell receptor beta chains (TRBC1 and TRBC2). During the development of alpha/beta T lymphocytes, one of these two constant chains is selected in each cell to be spliced to the variable region of TCR-beta and form a functional full length beta chain.
  • TRBC1 and TRBC2 T-cell receptor beta chains
  • the 2 TRBC targets were chosen in sequences conserved between TRBCl and TRBC2 so that the corresponding TALE-nuclease would cleave both TRBCl and TRBC2 at the same time.
  • Table 6 Additional target sequences for TRAC and CD52 TALE-nucleases.
  • HEK293 cells Each TALE-nuclease construct was subcloned using restriction enzyme digestion in a mammalian expression vector under the control of pEFl alpha long promoter. One million HEK293 cells were seeded one day prior to transfection.
  • TALE-nucleases recognizing the two half targets in the genomic sequence of interest in the CD52 gene, T-cell receptor alpha constant chain region (TRAC) or T-cell receptor beta constant chain region (TRBC) under the control of the EFl -alpha promoter or 5 ⁇ g of a control pUC vector (pCLS0003) using 25 ⁇ of lipofectamine (Invitrogen) according to the manufacturer's instructions.
  • TALE-nucleases in CD52 or TRAC coding sequences is repaired in live cells by non homologous end joining (NHEJ), which is an error-prone mechanism.
  • NHEJ non homologous end joining
  • TALE-nucleases Activity of TALE-nucleases in live cells is measured by the frequency of insertions or deletions at the genomic locus targeted. 48 hours after transfection, genomic DNA was isolated from transfected cells and locus specific PCRs were performed using the following primers: 5'-CCATCTCATCCCTGCGTGTCTCCGACTCAG (forward adaptor sequence)- ION (TAG)- locus specific forward sequence for CD52: 5'- CAGATCTGCAGAAAGGAAGC-3 ' (SEQ ID NO: 66), for TRAC: 5'- ATC ACTGGC ATCTGGACTCCA-3 ' (SEQ ID NO: 67), for TRBC1 : 5'- AGAGCCCCTACCAGAACCAGAC-3 ' (SEQ ID NO: 68), or for TRBC2: 5'- GGACCTAGTAACATAATTGTGC-3 ' (SEQ ID NO: 69), and the reverse primer 5'- CCTATCCCCTGTGTGCCTTGGCAGTCTCAG
  • Table 7 Percentages of indels for TALE-nuclease targeting CD52_T02, TRAC T01,
  • TRBC_T01 and TRBC_T02 targets.
  • CD52-TALE-nuclease Activity of CD52-TALE-nuclease, TRBC-TALE-nuclease and TRAC-TALE-nuclease in primary T lymphocytes
  • TALE-nuclease construct was subcloned using restriction enzyme digestion in a mammalian expression vector under the control of the T7 promoter.
  • mRNA encoding TALE-nuclease cleaving CD52 TRAC and TRBC genomic sequence were synthesized from plasmid carrying the coding sequences downstream from the T7 promoter.
  • T lymphocytes isolated from peripheral blood were activated for 5 days using anti-CD3/CD28 activator beads (Life technologies) and 5 million cells were then transfected by electroporation with 10 ⁇ g of each of 2 mRNAs encoding both half TALE-nuclease (or non coding RNA as controls) using a CytoLVT-P instrument.
  • the coding sequence for CD52 and/or TRAC will be out of frame in a fraction of the cells resulting in non-functional genes.
  • 5 days after electroporation cells were labeled with fluorochrome-conjugated anti-CD52 or anti-TCR antibody by flow cytometry for the presence of CD52 or TCR at their cell surface. Since all T lymphocytes expanded from peripheral blood normally express CD52 and TCR, the proportion of CD52- negative or TCR-negative cells is a direct measure of TALE-nuclease activity.
  • table 8 are listed the results of a representative experiment.
  • the table 9 shows the results of a representative experiment testing the efficiency of TRBC TALE-nucleases. % % %
  • Table 8 Percentages of CD52- negative, TCR-negative and CD52/TCR-double negative T lymphocytes after transfection of corresponding TALE-nuclease-expressing polynucleotides.
  • Table 9 Percentages of TCR-negative T lymphocytes after transfection of TRBC TALE- nuclease-expressing polynucleotides.
  • T lymphocytes were transfected with mRNA encoding TALE-nuclease cleaving CD52. 7 days after transfection, cells were treated with 50 ⁇ g/ml anti-CD52 monoclonal antibody (or rat IgG as control) with or without 30% rabbit complement (Cedarlane).
  • TRAC gene inactivation is to render T lymphocytes unresponsive to T-cell receptor stimulation.
  • T lymphocytes were transfected with mRNA encoding TALE-nuclease cleaving TRAC or CD52. 16 days after transfection, cells were treated with up to 5 ⁇ g/ml of phytohemagglutinin (PHA, Sigma- Aldrich), a T-cell mitogen acting through the T cell receptor. Cells with a functional T-cell receptor should increase in size following PHA treatment.
  • PHA phytohemagglutinin
  • TCR-positive and TCR-negative cells were labeled with a fluorochrome-conjugated anti-CD52 or anti-TCR antibody and analyzed by flow cytometry to compare the cell size distribution between TCR-positive and TCR-negative cells, or between CD52-positive and CD52-negative cells.
  • Figure 7 shows that TCR-positive cells significantly increase in size after PHA treatment whereas TCR-negative cells have the same size as untreated cells indicating that TRAC inactivation rendered them unresponsive to TCR-signaling.
  • CD52-positive and CD52-negative increase in size to same extent. Functional analysis of T cells with targeted CD52 and TRAC genes
  • T cells that had been targeted with CD52-TALE-nuclease and TRAC-TALE-nuclease with l( ⁇ g of RNA encoding an anti-CD19 CAR (SEQ ID NO: 73). 24 hours later, T cells were incubated for 4 hours with CD 19 expressing Daudi cells. The cell surface upregulation of CD 107a, a marker of cytotoxic granule release by T lymphocytes (called degranulation) was measured by flow cytometry analysis (Betts, Brenchley et al. 2003).
  • Example 3 TALE-nucleases cleaving the human CTLA4 gene and the human PDCDl gene.
  • heterodimeric TALE-nucleases targeting respectively PDCDl and CTLA4 genes were designed and produced.
  • the targeted genomic sequences consist of two 17-bp long sequences (called half targets) separated by an 1 1 or 15-bp spacer. Each half-target is recognized by repeats of half TALE-nucleases listed in table 10.
  • CTLA4 T01 TGGCCCTGCACTCTCCT Repeat CTLA4 T01-L gttttttcttctcttt CTLA4 T01-L TALEN CATCCCTGTCTTCTGCA (SEQ ID NO: 79 ) (SEQ ID NO: 89) (SEQ ID NO: 74) Repeat CTLA4 T01-R
  • CTLA4 T01-R TALEN SEQ ID NO: 80
  • SEQ ID NO: 90 SEQ ID NO: 90
  • CTLA4 T03 TTTTCCATGCTAGCAAT Repeat CTLA4 T03-L gcacgtggcccagcc CTLA4 T03-L TALEN
  • CTLA4 T03-R TALEN (SEQ ID NO: 82) (SEQ ID NO: 92)
  • CTLA4_T04 TCCATGCTAGCAATGCA Repeat CTLA4 T04-L cgtggcccagcctgc CTLA4 T04-L TALEN TGTGGTACTGGCCAGCA (SEQ ID NO: 84) (SEQ ID NO: 93) (SEQ ID NO: 76) Repeat CTLA4 T04-R
  • CTLA4 T04-R TALEN (SEQ ID NO: 85) (SEQ ID NO: 94)
  • PDCD1_T01 TTCTCCCCAGCCCTGCT Repeat PDCD1 T01-L cgtggtgaccgaagg PDCD1 T01-L TALEN GGACAACGCCACCTTCA (SEQ ID NO: 86) (SEQ ID NO: 95) (SEQ ID NO : 77) Repeat PDCD1 T01-R
  • PDCD1 T03 TACCTCTGTGGGGCCAT Repeat PDCD1 T03-L ctccctggcccccaa PDCD1 T03-L TALEN GGCGCAGATCAAAGAGA (SEQ ID NO: 88) (SEQ ID NO: 97) (SEQ ID NO : 78) Repeat PDCD1 T03-R
  • Table 10 Description of the CTLA4 and PDCD1 TALE-nucleases and sequences of the
  • TALE-nucleases target sites in the human corresponding genes.
  • TALE-nuclease construct Activity of CTLA4-TALE-nuclease and PDCD 1 -TALE-nuclease in HEK293 cells
  • Each TALE-nuclease construct was subcloned using restriction enzyme digestion in a mammalian expression vector under the control of the pEFl alpha long promoter.
  • One million HEK293 cells were seeded one day prior to transfection.
  • Cells were co-transfected with 2.5 ⁇ g of each of two plasmids encoding the TALE-nucleases recognizing the two half targets in the genomic sequence of interest in the PDCD1 and CTLA-4 gene under the control of the EFl -alpha promoter or 5 ⁇ g of a control pUC vector (pCLS0003) using 25 ⁇ of lipofectamine (Invitrogen) according to the manufacturer's instructions.
  • the double stranded cleavage generated by TALE-nucleases in PDCD1 or CTLA-4 coding sequences is repaired in live cells by non homologous end joining (NHEJ), which is an error-prone mechanism.
  • NHEJ non homologous end joining
  • TALE-nucleases Activity of TALE-nucleases in live cells is measured by the frequency of insertions or deletions at the genomic locus targeted. 48 hours after transfection, genomic DNA was isolated from transfected cells and locus specific PCRs were performed using the following primers: 5 ' -CC ATCTC ATCCCTGCGTGTCTCCG ACTC AG (forward adaptor sequence)- 10N (TAG)- locus specific forward sequence for CTLA4 T01 : 5'- CTCTACTTCCTGAAGACCTG-3 ' (SEQ ID NO: 99) , for CTLA4 T03/T04: 5'- ACAGTTGAGAGATGGAGGGG-3 ' (SEQ ID NO: 100), for PDCD1_T01 : 5'- CCACAGAGGTAGGTGCCGC-3 ' (SEQ ID NO: 101) or for PDCD1_T03: 5'- GAC AGAGATGCCGGTC ACC A-3 ' (SEQ ID NO: 102) and the reverse primer 5'
  • PCR products were analyzed by T7-endonuclease assay: briefly, after denaturation and reannealing of the PCR product, T7 endonuclease will specifically digest mismatched DNA composed of wild type and mutated strands. The digestion product is then resolved by polyacrylamide gel electrophoresis. The presence of a digested product is indicative of mutated sequences induced by TALE-nuclease activity. Results are displayed in Figure 10 where arrows point to the digested PCR products. They demonstrate that PDCD1 T1, PDCD1 T3, CTLA4 T1 , CTLA4 T3 and CTLA4_T4 TALE-nucleases all exhibit mutagenic nuclease activity at their target sites.
  • Example 4 pTalpha permits CD3 surface expression in inactivated TCR alpha T lymphocytes:
  • the human pTalpha gene encodes a transmembrane glycoprotein comprising an extracellular Ig-like domain, a hydrophobic transmembrane domain and a large C-terminal intracytoplasmic tail.
  • Different versions derived from human pTalpha glycoprotein have been designed and are described in Table 1 1 and represented in figure 11.
  • the different preTalpha constructs tested include:
  • pTalpha deletion mutants Different deletions were generated in the intracellular cytoplasmic tail of the human pTalpha protein (which comprises 114 amino acids) (SEQ ID NO: 107). The constructs tested include the full length version of the protein
  • pTalpha mutants containing intracellular activation domains The FL and ⁇ 48 variants where fused to the CD8, CD28 or 4 IBB intracellular activation domains at their C-terminus (SEQ ID NO: 115 to SEQ ID NO: 120).
  • pTalpha/TCRa chimeric mutants In one of the constructs, the TCRa intracellular domain (IC) was fused to a tail-less version ( ⁇ 114) of pTalpha (SEQ ID NO: 121). A second construct was also generated in which the pTalpha extracellular domain was fused to the transmembrane (TM) and the IC domains from TCRa (SEQ ID NO: 122). 4) pTalpha dimerization mutants: Some mutations have been described in the literature as being capable to alter the oligomerisation/dimerisation ability of the preTCR complex. These mutants are proposed to allow preTCR expression at the cell surface, without inducing the constitutive signaling (supposed to be induced upon preTCR oligomerization). The mutations have been introduced in the pTalphaA48 variant and are:
  • a cell line was generated in which the TCRalpha gene was disrupted using TALEN targeting TRAC.
  • Jurkat cells a T-cell leukemia cell line
  • plasmids coding for the TALEN cleaving TRAC using CytoPulse electroporation and the KO cells (TCR a p NEG ; CD3 NEG ) where then purified by negative selection using CD3 magnetic beads.
  • the KO population JKT_KOx3 cells was amplified and used for screening of the different pTalpha variants.
  • FIG. 12 is a representative example of the transfection efficiencies (% of BFP+ cells) and activity of the FL, ⁇ 18 and ⁇ 48 pTalpha constructs in JKT_KOx3 cells, based on the % of CD3+ cells, determined by flow cytometry. The results from the different constructs are grouped in Table 12.
  • Table 12 Activity of the different pTalpha constructs in Jurkat TCR alpha inactivated cells. Activity was measured by flow cytometry analysis of CD3 expression in jurkat TCR alpha inactivated cells transfected with the different preTalpha constructs.
  • pTalpha-FL and pTalpha-A48 versions were cloned into a self-inactivating pLV-SFFV-BFP-2A-PCTRA lentiviral vector that codes for Blue Fluorescent protein (BFP) under the SFFV promoter followed by the self- cleaving T2A peptide (figure 13).
  • BFP Blue Fluorescent protein
  • T lymphocytes isolated from peripheral blood were activated for 72 hours using anti- CD3/CD28 activator beads (Life technologies) and 4.5 million cells were transfected by electroporation with 10 ⁇ g mRNA encoding the TALE-nuclease targeting TCR alpha constant chain region (TRAC) using a CytoLVT-S instrument (BTX-Harvard Harbour). Two days after electroporation, T cells were transduced with either the LV-SFFV-BFP-2A- pTalpha- ⁇ 48 or LV-SFFV-BFP-2A-control lentiviral vectors. CD3 negative and CD31ow T cells were then purified using anti-CD3 magnetic beads (Miltenyi Biotech). This experimental protocol is represented in Figure 14A.
  • Figure 14B represents flow cytometry analysis of TCRalpha/beta, CD3 cell surface expression, and BFP expression on TCRalpha inactivated T cells (KO) transduced with either BFP-2A-pTalphaA48 ( ⁇ / ⁇ 48) or control BFP lentiviral vector (KO/BFP) before and after purification with CD3 beads.
  • TCRalpha inactivated cells transduced with the BFP-T2A- pTalpha-A48 vector show higher levels of CD3 compared to non transduced cells (BFP- cells). No differences are observed among cells transduced with the control BFP vector.
  • TCRalpha/beta staining remains, as expected, unchanged in cells transduced or not with the pTalpha-A48 expressing vector.
  • pTalpha-mediated CD3 expression supports activation of TCR-deficient T-cells:
  • TCR alpha inactivated T cells transduced with pTalpha-A48 and pTalpha-A48.41BB were generated from primary human T-cells as described in previous section and in Figure 14 A.
  • TCR alpha inactivated cells expressing pTalpha-A48 ( ⁇ / ⁇ - ⁇ 48) or pTalpha-A48.41BB ( ⁇ / ⁇ - ⁇ 48. ⁇ ) show upregulation of the activation markers, to levels similar to those observed in TCRalpha/beta expressing cells (NEP: non electroporated cells).
  • Another indicator of T cell activation is an increase in cell size which is sometimes referred to as "blasting".
  • the capacity of the preTCR complexes to induce “blasting” was measured by flow cytometry analysis of the cell size 72 hours after re-activation using anti-CD3/CD28- beads ( Figure 15C). Stimulation with anti-CD3/CD28 beads induced comparable increases in cell size in cells expressing TCRalpha/beta complexes vs. cells expressing pTalpha-A48 or pTalpha-A48.41BB.
  • pTalpha mediated CD3 expression supports expansion of TCR-deficient primary T-cells using stimulatory anti-CD3/CD28 antibodies
  • TCRalpha inactivated cells expressing pTalpha-A48 displayed greater expansion than TCR alpha inactivated cells expressing only the BFP control vector.
  • TCRalpha inactivated cells expressing pTalpha-A48.41BB or full-length pTalpha were also included, displaying also greater expansion than TCRalpha inactivated cells expressing only the BFP control vector.
  • Example 5 optimization of mR A transfection in T cells using Cytopulse Technology.
  • Table 13 Different cytopulse programs used to determine the minimal voltage required for electroporation in PBMC derived T-cells.
  • Electroporation of mRNA of purified Tcells activated After determining the best cytopulse program that allows an efficient DNA electroporation of T cells, we tested whether this method was applicable to the mRNA electroporation.
  • T cells preactivated 6 days with PHA/IL2 were resupended in cytoporation buffer T (BTX-Harvard apparatus) and electroporated in 0.4 cm cuvettes with 10 g of mRNA encoding GFP or 20 ⁇ g of plasmids encoding GFP or pUC using the preferred cytopulse program as determined in the previous section (table 14).
  • RNA transfection has no impact on cellular viability and allows uniform expression levels of the transfected gene of interest in the cellular population.
  • Efficient transfection can be achieved early after cellular activation, independently of the activation method used (PHA/IL-2 or CD3/CD28-coated-beads).
  • the inventors have succeeded in transfecting cells from 72h after activation with efficiencies of >95%.
  • efficient transfection of T cells after thawing and activation can also be obtained using the same electroporation protocol.
  • mRNA electroporation in primary human T cells for TALE-nuclease functional expression After demonstrating that mRNA electroporation allow efficient expression of GFP in primary human T cells, we tested whether this method was applicable to the expression of other proteins of interest.
  • Transcription activator-like effector nucleases are site- specific nucleases generated by the fusion of a TAL DNA binding domain to a DNA cleavage domain. They are powerful genome editing tools as they induce double-strand breaks at practically any desired DNA sequence. These double-strand breaks activate Non-homologous end-joining (NHEJ), an error-prone DNA repair mechanism, potentially leading to inactivation of any desired gene of interest. Alternatively, if an adequate repair template is introduced into the cells at the same time, TALE-nuclease-induced DNA breaks can be repaired by homologous recombination, therefore offering the possibility of modifying at will the gene sequence.
  • NHEJ Non-homologous end-joining
  • TALE-nuclease designed to specifically cleave a sequence in the human gene coding for the alpha chain of the T cell antigen receptor (TRAC). Mutations induced in this sequence are expected to result in gene inactivation and loss of TCRaP complex from the cell surface.
  • TRAC TALE-nuclease RNA or non coding RNA as control are transfected into activated primary human T lymphocytes using Cytopulse technology.
  • the electroporation sequence consisted in 2 pulses of 1200 V followed by four pulses of 130 V as described in Table 14.
  • T cells with a monocistronic mRNA encoding for an anti-CD 19 single chain chimeric antigen receptor (CAR): 5X10 6 T cells preactivated several days (3-5) with anti-CD3/CD28 coated beads and IL2 were resuspended in cytoporation buffer T, and electroporated in 0.4cm cuvettes without mRNA or with l( ⁇ g of mRNA encoding a single chain CAR (SEQ ID NO: 73) using the program described in Table 14.
  • CAR single chain chimeric antigen receptor
  • T cells preactivated several days (3-5) with anti CD3/CD28 coated beads and IL2 were electroporated in cytoporation buffer T, and electroporated in 0.4cm cuvettes without mRNA or with 45 ⁇ g of mRNA encoding a multi-chain CAR (SEQ ID NO: 125, encoded by SEQ ID NO: 126, Figure 21 A and figure 4B (csm4)) using the program as described in Table 14.
  • 24 hours post electroporation cells were stained with a fixable viability dye eFluor-780 and a PE-conjugated goat anti mouse IgG F(ab')2 fragment specific to assess the cell surface expression of the CAR on the live cells.
  • the data shown in Figure 21 indicates that the vast majority of the live T cells electroporated with the polycistronic mRNA described previously express the CAR at their surface. 24 hours post electroporation, T cells were cocultured with Daudi (CD19 + ) for 6 hours and analyzed by flow cytometry to detect the expression of the degranulation marker CD 107a at their surface.
  • the data shown in Figure 21 indicates that the majority of the cells electroporated with the polycistronic mRNA described previously degranulate in the presence of target cells expressing CD 19.
  • CD3 zeta subunit can substitute for the gamma subunit of Fc epsilon receptor type I in assembly and functional expression of the high- affinity IgE receptor: evidence for interreceptor complementation. Proc Natl Acad Sci U S A 87(18): 7015-9.
  • TAL nucleases hybrid proteins composed of TAL effectors and Fokl DNA-cleavage domain. Nucleic Acids Res 39(1): 359-72.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Cell Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biomedical Technology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • Epidemiology (AREA)
  • Hematology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Oncology (AREA)
  • Virology (AREA)
  • Communicable Diseases (AREA)
  • Developmental Biology & Embryology (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Transplantation (AREA)
  • Physical Education & Sports Medicine (AREA)
PCT/US2013/040766 2012-05-25 2013-05-13 Use of pre t alpha or functional variant thereof for expanding tcr alpha deficient t cells Ceased WO2013176916A1 (en)

Priority Applications (110)

Application Number Priority Date Filing Date Title
JP2015514051A JP6463672B2 (ja) 2012-05-25 2013-05-13 Ｔｃｒアルファ欠損ｔ細胞を増殖させるためのプレｔアルファまたはその機能性変種の使用
US14/403,937 US10426795B2 (en) 2012-05-25 2013-05-13 Use of preTalpha or functional variant thereof for expanding TCRalpha deficient T cells
AU2013266734A AU2013266734B2 (en) 2012-05-25 2013-05-13 Use of pre T alpha or functional variant thereof for expanding TCR alpha deficient T cells
EP13728017.8A EP2855666B1 (en) 2012-05-25 2013-05-13 Use of pre t alpha or functional variant thereof for expanding tcr alpha deficient t cells
CA2874611A CA2874611C (en) 2012-05-25 2013-05-13 Use of pre t alpha or functional variant thereof for expanding tcr alpha deficient t cells
AU2013312838A AU2013312838B2 (en) 2012-09-04 2013-09-04 Multi-chain chimeric antigen receptor and uses thereof
KR1020157008739A KR102141259B1 (ko) 2012-09-04 2013-09-04 멀티―체인 키메라 항원 수용체 및 그것의 용도들
CN201380057661.1A CN104769103B (zh) 2012-09-04 2013-09-04 多链嵌合抗原受体和其用途
SG11201501471VA SG11201501471VA (en) 2012-09-04 2013-09-04 Multi-chain chimeric antigen receptor and uses thereof
RU2015112125A RU2663725C2 (ru) 2012-09-04 2013-09-04 Многоцепочечный химерный антигенный рецептор и его применения
PT13776597T PT2893004T (pt) 2012-09-04 2013-09-04 Recetor de antígeno quimérico de cadeia múltipla e utilizações do mesmo
CA2883502A CA2883502C (en) 2012-09-04 2013-09-04 Multi-chain chimeric antigen receptor and uses thereof
DK13776597.0T DK2893004T3 (en) 2012-09-04 2013-09-04 MULTI-CHAIN CHEMICAL ANTIGEN RECEPTOR AND APPLICATIONS THEREOF
BR112015004522A BR112015004522A2 (pt) 2012-09-04 2013-09-04 receptor de antígeno quimérico multicadeia e usos destes
EP13776597.0A EP2893004B1 (en) 2012-09-04 2013-09-04 Multi-chain chimeric antigen receptor and uses thereof
US14/018,021 US10342829B2 (en) 2012-05-25 2013-09-04 Multi-chain chimeric antigen receptor and uses thereof
MX2015002760A MX367730B (es) 2012-09-04 2013-09-04 Receptor de antigeno quimerico multi-cadena y usos del mismo.
PCT/US2013/058005 WO2014039523A1 (en) 2012-09-04 2013-09-04 Multi-chain chimeric antigen receptor and uses thereof
ES13776597T ES2714523T3 (es) 2012-09-04 2013-09-04 Receptor quimérico de antígenos multicatenario y usos del mismo
HK16100443.5A HK1212728B (en) 2012-09-04 2013-09-04 Multi-chain chimeric antigen receptor and uses thereof
JP2015530148A JP6352920B2 (ja) 2012-09-04 2013-09-04 多重鎖キメラ抗原受容体およびその使用
ES14723436T ES2930431T3 (es) 2013-05-13 2014-05-12 Receptor quimérico de antígeno específico para CD19 y sus usos
HUE14723436A HUE060901T2 (hu) 2013-05-13 2014-05-12 CD-19-re specifikus kiméra antigénreceptor és alkalmazásai
PT191618610T PT3546572T (pt) 2013-05-13 2014-05-12 Recetor de antigénio quimérico específico para cd19 e utilizações do mesmo
AU2014267436A AU2014267436B2 (en) 2013-05-13 2014-05-12 CD19 specific chimeric antigen receptor and uses thereof
PT147234363T PT2997141T (pt) 2013-05-13 2014-05-12 Recetor de antigénio quimérico específico para cd19 e utilizações do mesmo
LTEP19161861.0T LT3546572T (lt) 2013-05-13 2014-05-12 Cd19 specifinis chimerinis antigeno receptorius ir jo panaudojimas
RS20240486A RS65484B1 (sr) 2013-05-13 2014-05-12 Cd19 specifični himerni antigenski receptor i njegove primene
PCT/EP2014/059662 WO2014184143A1 (en) 2013-05-13 2014-05-12 Cd19 specific chimeric antigen receptor and uses thereof
CN201910222072.3A CN109897100A (zh) 2013-05-13 2014-05-12 Cd19特异性嵌合抗原受体及其用途
TW103116744A TWI636992B (zh) 2013-05-13 2014-05-12 Ｃｄ１９特異性嵌合抗原受體及其用途
PL14723436.3T PL2997141T3 (pl) 2013-05-13 2014-05-12 Chimeryczny receptor antygenowy swoisty względem cd19 i jego zastosowania
HK16110847.6A HK1222679B (en) 2013-05-13 2014-05-12 Cd19 specific chimeric antigen receptor and uses thereof
KR1020157035319A KR102248157B1 (ko) 2013-05-13 2014-05-12 Ｃｄ19 특이적 키메라 항원 수용체 및 이의 용도
JP2016513321A JP6491643B2 (ja) 2013-05-13 2014-05-12 Ｃｄ１９特異的キメラ抗原受容体およびその使用
DK19161861.0T DK3546572T3 (da) 2013-05-13 2014-05-12 CD19-specifik kimær antigenreceptor og anvendelser deraf
SI201431999T SI2997141T1 (sl) 2013-05-13 2014-05-12 CD19 specifični himerni antigenski receptor in njegova uporaba
SG10201708896WA SG10201708896WA (en) 2013-05-13 2014-05-12 Cd19 specific chimeric antigen receptor and uses thereof
LTEPPCT/EP2014/059662T LT2997141T (lt) 2013-05-13 2014-05-12 Cd19 specifinis chimerinis antigeno receptorius ir jo panaudojimai
HK16110821.6A HK1222678B (zh) 2013-05-13 2014-05-12 Cd19特异性嵌合抗原受体及其用途
EP14723436.3A EP2997141B1 (en) 2013-05-13 2014-05-12 Cd19 specific chimeric antigen receptor and uses thereof
EP19161861.0A EP3546572B9 (en) 2013-05-13 2014-05-12 Cd19 specific chimeric antigen receptor and uses thereof
FIEP14723436.3T FI2997141T3 (fi) 2013-05-13 2014-05-12 CD19-spesifinen kimeerinen antigeenireseptori ja sen käyttöjä
RU2015153250A RU2727447C2 (ru) 2013-05-13 2014-05-12 Cd19-специфический химерный антигенный рецептор и его применения
FIEP19161861.0T FI3546572T3 (fi) 2013-05-13 2014-05-12 Cd19-spesifinen kimeerinen antigeenireseptori ja sen käyttötapoja
HUE19161861A HUE067258T2 (hu) 2013-05-13 2014-05-12 CD19-re specifikus kiméra antigénreceptor és alkalmazásai
CA2911292A CA2911292C (en) 2013-05-13 2014-05-12 Cd19 specific chimeric antigen receptor and uses thereof
ES19161861T ES2978123T3 (es) 2013-05-13 2014-05-12 Receptor quimérico de antígeno específico para CD19 y sus usos
NZ714044A NZ714044B2 (en) 2013-05-13 2014-05-12 Cd19 specific chimeric antigen receptor and uses thereof
SI201432070T SI3546572T1 (sl) 2013-05-13 2014-05-12 CD19 specifični himerni antigenski receptor in njegova uporaba
PL19161861.0T PL3546572T3 (pl) 2013-05-13 2014-05-12 Chimeryczny receptor antygenowy swoisty względem cd19 i jego zastosowania
HRP20221393TT HRP20221393T1 (hr) 2013-05-13 2014-05-12 Cd19 specifični kimerni antigenski receptor i njegova uporaba
EA201501109A EA036200B1 (ru) 2013-05-13 2014-05-12 Cd19-специфический химерный антигенный рецептор и его применения
CN201480027071.9A CN105431532B (zh) 2013-05-13 2014-05-12 Cd19特异性嵌合抗原受体及其用途
BR112015028387-0A BR112015028387B1 (pt) 2013-05-13 2014-05-12 Receptor antigênico quimérico específico para cd19, polinucleotídeo, vetor de expressão, método in vitro para manipulação e uso de uma célula imune
EP24163057.3A EP4364809A3 (en) 2013-05-13 2014-05-12 Cd19 specific chimeric antigen receptor and uses thereof
SG11201508805UA SG11201508805UA (en) 2013-05-13 2014-05-12 Cd19 specific chimeric antigen receptor and uses thereof
US14/891,296 US10874693B2 (en) 2012-05-25 2014-05-12 CD19 specific chimeric antigen receptor and uses thereof
RS20221084A RS63798B1 (sr) 2013-05-13 2014-05-12 Cd19 specifični himerni antigenski receptor i njegove primene
MYPI2015703966A MY172897A (en) 2013-05-13 2014-05-12 Cd19 specific chimeric antigen receptor and uses thereof
HRP20240576TT HRP20240576T1 (hr) 2013-05-13 2014-05-12 Cd19 specifični kimerni antigenski receptor i njegova upotreba
MX2015015662A MX374681B (es) 2013-05-13 2014-05-12 Receptor quimérico de antígeno específico para cd19 y sus usos.
CN201480039866.1A CN105378067A (zh) 2013-05-13 2014-05-13 工程化用于免疫治疗的高活性t细胞的方法
ES14727626T ES2828669T3 (es) 2013-05-13 2014-05-13 Métodos para genomodificación de linfocitos T alogénicos y altamente activos para inmunoterapia
CA2912373A CA2912373A1 (en) 2013-05-13 2014-05-13 Methods for engineering allogeneic and highly active t cell for immunotherapy
RU2015153245A RU2736616C2 (ru) 2013-05-13 2014-05-13 Способы конструирования аллогенных и высокоактивных т-клеток для иммунотерапии
KR1020217037383A KR20210143926A (ko) 2013-05-13 2014-05-13 면역 요법을 위한 동종이형 및 고활성 ｔ 세포의 조작 방법들
RU2015153241A RU2725542C2 (ru) 2013-05-13 2014-05-13 Способы конструирования высокоактивных т-клеток для иммунотерапии
PCT/IB2014/061412 WO2014184744A1 (en) 2013-05-13 2014-05-13 Methods for engineering highly active t cell for immunotherapy
CA2912375A CA2912375C (en) 2013-05-13 2014-05-13 Methods for engineering highly active t cell for immunotherapy
EP20190570.0A EP3936612A1 (en) 2013-05-13 2014-05-13 Methods for engineering allogeneic and highly active t cell for immunotheraphy
EP14727627.3A EP2997133B1 (en) 2013-05-13 2014-05-13 Methods for engineering highly active t cell for immunotherapy
US14/894,426 US11311575B2 (en) 2013-05-13 2014-05-13 Methods for engineering highly active T cell for immunotherapy
KR1020157035439A KR102220382B1 (ko) 2013-05-13 2014-05-13 면역요법을 위한 매우 활성인 ｔ 세포를 조작하는 방법
AU2014266833A AU2014266833B2 (en) 2013-05-13 2014-05-13 Methods for engineering highly active T cell for immunotherapy
EP14727626.5A EP2997132B1 (en) 2013-05-13 2014-05-13 Methods for engineering allogeneic and highly active t cell for immunotherapy
MX2015015638A MX2015015638A (es) 2013-05-13 2014-05-13 Metodos para diseñar celulas t altamente activas para inmunoterapia.
JP2016513481A JP2016524464A (ja) 2013-05-13 2014-05-13 免疫療法のために高活性ｔ細胞を操作するための方法
MX2015015639A MX380738B (es) 2013-05-13 2014-05-13 Métodos para manipular células t alogénicas, altamente activas para inmunoterapia.
BR112015028483-3A BR112015028483B1 (pt) 2013-05-13 2014-05-13 Método para preparar células t para imunoterapia, e uso de células t modificadas
KR1020237030308A KR20230144570A (ko) 2013-05-13 2014-05-13 면역 요법을 위한 동종이형 및 고활성 ｔ 세포의 조작 방법들
KR1020157035331A KR102329704B1 (ko) 2013-05-13 2014-05-13 면역 요법을 위한 동종이계 및 고활성 ｔ 세포의 조작 방법들
BR112015028493A BR112015028493A2 (pt) 2013-05-13 2014-05-13 Métodos para preparar células t para imunoterapia, célula t isolada, seu uso e composição farmacêutica
AU2014266830A AU2014266830B2 (en) 2013-05-13 2014-05-13 Methods for engineering allogeneic and highly active T cell for immunotherapy
CN201480039917.0A CN105378068A (zh) 2013-05-13 2014-05-13 用于免疫治疗的同种异体且高活性的t细胞的工程化方法
DK14727626.5T DK2997132T3 (da) 2013-05-13 2014-05-13 Fremgangsmåder til konstruering af allogenisk og høj-aktiv T-celle til immunoterapi
JP2016513480A JP6875124B2 (ja) 2013-05-13 2014-05-13 免疫療法のために同種異系かつ高活性のｔ細胞を操作するための方法
US14/889,686 US11304975B2 (en) 2013-05-13 2014-05-13 Methods for engineering allogeneic and highly active t cell for immunotherapy
PCT/IB2014/061409 WO2014184741A1 (en) 2013-05-13 2014-05-13 Methods for engineering allogeneic and highly active t cell for immunotheraphy
UAA201512065A UA118106C2 (uk) 2013-05-13 2014-12-05 Химерний антигенний рецептор, специфічний до cd19, і його застосування
IL237576A IL237576B (en) 2012-09-04 2015-03-04 Multichain chimeric antigen receptor and its uses
PH12015502479A PH12015502479B1 (en) 2013-05-13 2015-10-27 Cd19 specific chimeric antigen receptor and uses thereof
SA515370135A SA515370135B1 (ar) 2013-05-13 2015-11-12 مستقبل مولد المضاد الكيميري المتخصص cd19 وإستخداماته
US15/659,792 US10286007B2 (en) 2012-05-25 2017-07-26 Use of pre T alpha or functional variant thereof for expanding TCR alpha deficient T cells
JP2019037174A JP6854307B2 (ja) 2013-05-13 2019-03-01 Ｃｄ１９特異的キメラ抗原受容体およびその使用
AU2019201818A AU2019201818B2 (en) 2013-05-13 2019-03-15 CD19 specific chimeric antigen receptor and uses thereof
US16/361,370 US10517896B2 (en) 2012-05-25 2019-03-22 Use of pre T alpha or functional variant thereof for expanding TCR alpha deficient T cells
US16/361,438 US11414674B2 (en) 2012-05-25 2019-03-22 Use of pre T alpha or functional variant thereof for expanding TCR alpha deficient T cells
US16/365,588 US20210000869A9 (en) 2013-05-13 2019-03-26 Cd19 specific chimeric antigen receptor and uses thereof
JP2019114231A JP6998917B2 (ja) 2013-05-13 2019-06-20 免疫療法のために同種異系かつ高活性のｔ細胞を操作するための方法
US17/099,614 US11007224B2 (en) 2012-05-25 2020-11-16 CD19 specific chimeric antigen receptor and uses thereof
US17/099,608 US11077144B2 (en) 2013-05-13 2020-11-16 CD19 specific chimeric antigen receptor and uses thereof
US17/198,505 US11274316B2 (en) 2012-05-25 2021-03-11 Use of pre T alpha or functional variant thereof for expanding TCR alpha deficient T cells
JP2021182495A JP7479339B2 (ja) 2013-05-13 2021-11-09 免疫療法のために同種異系かつ高活性のｔ細胞を操作するための方法
US17/674,436 US20220177914A1 (en) 2012-05-25 2022-02-17 Use of pre t alpha or functional variant thereof for expanding tcr alpha deficient t cells
US17/715,218 US20230050345A1 (en) 2013-05-13 2022-04-07 Methods for engineering allogeneic and highly active t cell for immunotheraphy
US17/716,102 US20230056268A1 (en) 2013-05-13 2022-04-08 Methods for engineering highly active t cell for immunotheraphy
US17/848,590 US12577581B2 (en) 2012-05-25 2022-06-24 Use of pre T alpha or functional variant thereof for expanding TCR alpha deficient T cells
US18/056,544 US20230201260A1 (en) 2013-05-13 2022-11-17 Methods for engineering allogeneic and highly active t cell for immunotheraphy
US18/451,816 US20240269178A1 (en) 2013-05-13 2023-08-17 Cd19 specific chimeric antigen receptor and uses thereof

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US201261651933P 2012-05-25 2012-05-25
US61/651,933 2012-05-25
US201261696612P 2012-09-04 2012-09-04
US61/696,612 2012-09-04

Related Parent Applications (3)

Application Number Title Priority Date Filing Date
PCT/US2013/040755 Continuation-In-Part WO2013176915A1 (en) 2012-05-25 2013-05-13 Methods for engineering allogeneic and immunosuppressive resistant t cell for immunotherapy
US13/892,805 Continuation-In-Part US11603539B2 (en) 2012-05-25 2013-05-13 Methods for engineering allogeneic and immunosuppressive resistant T cell for immunotherapy
US13/942,191 Continuation-In-Part US20150017136A1 (en) 2012-05-25 2013-07-15 Methods for engineering allogeneic and highly active t cell for immunotherapy

Related Child Applications (5)

Application Number Title Priority Date Filing Date
US14/403,937 A-371-Of-International US10426795B2 (en) 2012-05-25 2013-05-13 Use of preTalpha or functional variant thereof for expanding TCRalpha deficient T cells
PCT/US2013/040755 Continuation-In-Part WO2013176915A1 (en) 2012-05-25 2013-05-13 Methods for engineering allogeneic and immunosuppressive resistant t cell for immunotherapy
PCT/US2013/040755 Division WO2013176915A1 (en) 2012-05-25 2013-05-13 Methods for engineering allogeneic and immunosuppressive resistant t cell for immunotherapy
US13/892,805 Continuation-In-Part US11603539B2 (en) 2012-05-25 2013-05-13 Methods for engineering allogeneic and immunosuppressive resistant T cell for immunotherapy
US15/659,792 Division US10286007B2 (en) 2012-05-25 2017-07-26 Use of pre T alpha or functional variant thereof for expanding TCR alpha deficient T cells

Publications (2)

Publication Number Publication Date
WO2013176916A1 true WO2013176916A1 (en) 2013-11-28
WO2013176916A8 WO2013176916A8 (en) 2014-10-16

Family

ID=48579464

Family Applications (2)

Application Number Title Priority Date Filing Date
PCT/US2013/040766 Ceased WO2013176916A1 (en) 2012-05-25 2013-05-13 Use of pre t alpha or functional variant thereof for expanding tcr alpha deficient t cells
PCT/US2013/040755 Ceased WO2013176915A1 (en) 2012-05-25 2013-05-13 Methods for engineering allogeneic and immunosuppressive resistant t cell for immunotherapy

Family Applications After (1)

Application Number Title Priority Date Filing Date
PCT/US2013/040755 Ceased WO2013176915A1 (en) 2012-05-25 2013-05-13 Methods for engineering allogeneic and immunosuppressive resistant t cell for immunotherapy

Country Status (25)

Country Link
US (15) US10426795B2 (https=)
EP (7) EP3276000A3 (https=)
JP (5) JP6463672B2 (https=)
KR (2) KR102437522B1 (https=)
CN (3) CN108998418A (https=)
AR (1) AR096274A1 (https=)
AU (5) AU2013266733B2 (https=)
BR (1) BR112014029417B1 (https=)
CA (3) CA2874609C (https=)
DK (2) DK2855667T5 (https=)
EA (1) EA201492222A1 (https=)
ES (2) ES2962571T3 (https=)
HK (1) HK1245321A1 (https=)
HU (2) HUE070380T2 (https=)
IN (1) IN2014DN10991A (https=)
MA (1) MA37681B2 (https=)
MX (1) MX370265B (https=)
MY (1) MY180127A (https=)
NZ (1) NZ703281A (https=)
PL (2) PL2855667T3 (https=)
PT (2) PT3473707T (https=)
SG (2) SG11201407802WA (https=)
UA (1) UA118652C2 (https=)
WO (2) WO2013176916A1 (https=)
ZA (1) ZA201409536B (https=)

Cited By (68)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014165707A3 (en) * 2013-04-03 2015-03-12 Memorial Sloan-Kettering Cancer Center Effective generation of tumor-targeted t-cells derived from pluripotent stem cells
US9068179B1 (en) 2013-12-12 2015-06-30 President And Fellows Of Harvard College Methods for correcting presenilin point mutations
WO2015155341A1 (en) * 2014-04-11 2015-10-15 Cellectis Method for generating immune cells resistant to arginine and/or tryptophan depleted microenvironment
US9163284B2 (en) 2013-08-09 2015-10-20 President And Fellows Of Harvard College Methods for identifying a target site of a Cas9 nuclease
US9228207B2 (en) 2013-09-06 2016-01-05 President And Fellows Of Harvard College Switchable gRNAs comprising aptamers
US9322006B2 (en) 2011-07-22 2016-04-26 President And Fellows Of Harvard College Evaluation and improvement of nuclease cleavage specificity
US9322037B2 (en) 2013-09-06 2016-04-26 President And Fellows Of Harvard College Cas9-FokI fusion proteins and uses thereof
US9359599B2 (en) 2013-08-22 2016-06-07 President And Fellows Of Harvard College Engineered transcription activator-like effector (TALE) domains and uses thereof
US9526784B2 (en) 2013-09-06 2016-12-27 President And Fellows Of Harvard College Delivery system for functional nucleases
JP2017504601A (ja) * 2013-12-20 2017-02-09 セレクティスＣｅｌｌｅｃｔｉｓ 免疫療法のためにマルチインプットシグナル感受性ｔ細胞を操作する方法
US9834791B2 (en) 2013-11-07 2017-12-05 Editas Medicine, Inc. CRISPR-related methods and compositions with governing gRNAS
WO2018052828A1 (en) 2016-09-14 2018-03-22 Janssen Biotech, Inc. Chimeric antigen receptors comprising bcma-specific fibronectin type iii domains and uses thereof
CN108024544A (zh) * 2015-07-13 2018-05-11 桑格摩生物治疗股份有限公司 用于核酸酶介导的基因组工程化的递送方法及组合物
WO2018115189A1 (en) 2016-12-21 2018-06-28 Cellectis Stably enginereed proteasome inhibitor resistant immune cells for immunotherapy
US10077453B2 (en) 2014-07-30 2018-09-18 President And Fellows Of Harvard College CAS9 proteins including ligand-dependent inteins
US10113163B2 (en) 2016-08-03 2018-10-30 President And Fellows Of Harvard College Adenosine nucleobase editors and uses thereof
US10167457B2 (en) 2015-10-23 2019-01-01 President And Fellows Of Harvard College Nucleobase editors and uses thereof
WO2019002633A1 (en) 2017-06-30 2019-01-03 Cellectis CELLULAR IMMUNOTHERAPY FOR REPETITIVE ADMINISTRATION
WO2019018402A2 (en) 2017-07-17 2019-01-24 Janssen Biotech, Inc. ANTIGEN-BINDING REGIONS DIRECTED AGAINST FIBRONECTIN TYPE III DOMAINS AND METHODS OF USING THE SAME
EP3466967A1 (en) * 2015-05-18 2019-04-10 TCR2 Therapeutics Inc. Compositions and methods for tcr reprogramming using fusion proteins
WO2019072824A1 (en) 2017-10-09 2019-04-18 Cellectis IMPROVED ANTI-CD123 CAR IN UNIVERSAL MODIFIED IMMUNE T LYMPHOCYTES
US10287606B2 (en) 2015-11-04 2019-05-14 Fate Therapeutics, Inc. Genomic engineering of pluripotent cells
WO2019155191A1 (en) * 2018-02-06 2019-08-15 Autolus Limited Polypeptides and methods
EP3569619A1 (en) 2014-03-19 2019-11-20 Cellectis Cd123 specific chimeric antigen receptors for cancer immunotherapy
WO2020007593A1 (en) 2018-07-02 2020-01-09 Cellectis Chimeric antigen receptors (car)-expressing cells and combination treatment for immunotherapy of patients with relapse refractory adverse genetic risk aml
WO2020043152A1 (en) 2018-08-29 2020-03-05 Nanjing Legend Biotech Co., Ltd. Anti-mesothelin chimeric antigen receptor (car) constructs and uses thereof
US10626372B1 (en) 2015-01-26 2020-04-21 Fate Therapeutics, Inc. Methods and compositions for inducing hematopoietic cell differentiation
WO2020109953A1 (en) 2018-11-30 2020-06-04 Janssen Biotech, Inc. Gamma delta t cells and uses thereof
EP3693384A1 (en) 2014-03-11 2020-08-12 Cellectis Method for generating t-cells compatible for allogenic transplantation
US10745677B2 (en) 2016-12-23 2020-08-18 President And Fellows Of Harvard College Editing of CCR5 receptor gene to protect against HIV infection
US10858628B2 (en) 2015-11-04 2020-12-08 Fate Therapeutics, Inc. Methods and compositions for inducing hematopoietic cell differentiation
WO2021041486A1 (en) 2019-08-27 2021-03-04 Janssen Biotech, Inc. Chimeric antigen receptor system and uses thereof
US11014989B2 (en) 2015-01-26 2021-05-25 Cellectis Anti-CLL1 specific single-chain chimeric antigen receptors (scCARs) for cancer immunotherapy
US11085021B2 (en) 2016-10-07 2021-08-10 TCR2 Therapeutics Inc. Compositions and methods for TCR reprogramming using fusion proteins
WO2021176373A1 (en) 2020-03-03 2021-09-10 Janssen Biotech, Inc. ꝩδ T CELLS AND USES THEREOF
EP3765039A4 (en) * 2018-03-09 2021-12-08 TCR2 Therapeutics Inc. COMPOSITIONS AND METHODS FOR REPROGRAMMING TCR USING FUSION PROTEINS
US11203758B2 (en) 2014-10-31 2021-12-21 The Trustees Of The University Of Pennsylvania Altering gene expression in modified T cells and uses thereof
WO2022016119A1 (en) 2020-07-17 2022-01-20 Simurx, Inc. Chimeric myd88 receptors for redirecting immunosuppressive signaling and related compositions and methods
US11242376B2 (en) 2016-08-02 2022-02-08 TCR2 Therapeutics Inc. Compositions and methods for TCR reprogramming using fusion proteins
US11268082B2 (en) 2017-03-23 2022-03-08 President And Fellows Of Harvard College Nucleobase editors comprising nucleic acid programmable DNA binding proteins
US11306324B2 (en) 2016-10-14 2022-04-19 President And Fellows Of Harvard College AAV delivery of nucleobase editors
US11319532B2 (en) 2017-08-30 2022-05-03 President And Fellows Of Harvard College High efficiency base editors comprising Gam
US11447770B1 (en) 2019-03-19 2022-09-20 The Broad Institute, Inc. Methods and compositions for prime editing nucleotide sequences
US11466271B2 (en) 2017-02-06 2022-10-11 Novartis Ag Compositions and methods for the treatment of hemoglobinopathies
US11542496B2 (en) 2017-03-10 2023-01-03 President And Fellows Of Harvard College Cytosine to guanine base editor
US11542509B2 (en) 2016-08-24 2023-01-03 President And Fellows Of Harvard College Incorporation of unnatural amino acids into proteins using base editing
US11560566B2 (en) 2017-05-12 2023-01-24 President And Fellows Of Harvard College Aptazyme-embedded guide RNAs for use with CRISPR-Cas9 in genome editing and transcriptional activation
US11661590B2 (en) 2016-08-09 2023-05-30 President And Fellows Of Harvard College Programmable CAS9-recombinase fusion proteins and uses thereof
US11732274B2 (en) 2017-07-28 2023-08-22 President And Fellows Of Harvard College Methods and compositions for evolving base editors using phage-assisted continuous evolution (PACE)
JP2023129663A (ja) * 2015-03-27 2023-09-14 プレジデント アンド フェローズ オブ ハーバード カレッジ 改変ｔ細胞ならびにそれを作製および使用する方法
WO2023179766A1 (zh) 2022-03-24 2023-09-28 南京传奇生物科技有限公司 制备dna文库和检测逆转录病毒整合位点的方法
US11795443B2 (en) 2017-10-16 2023-10-24 The Broad Institute, Inc. Uses of adenosine base editors
US11851491B2 (en) 2016-11-22 2023-12-26 TCR2 Therapeutics Inc. Compositions and methods for TCR reprogramming using fusion proteins
US11851659B2 (en) 2017-03-22 2023-12-26 Novartis Ag Compositions and methods for immunooncology
US11883432B2 (en) 2020-12-18 2024-01-30 Century Therapeutics, Inc. Chimeric antigen receptor system with adaptable receptor specificity
US11898179B2 (en) 2017-03-09 2024-02-13 President And Fellows Of Harvard College Suppression of pain by gene editing
US11912985B2 (en) 2020-05-08 2024-02-27 The Broad Institute, Inc. Methods and compositions for simultaneous editing of both strands of a target double-stranded nucleotide sequence
US12031154B2 (en) 2015-05-08 2024-07-09 President And Fellows Of Harvard College Universal donor stem cells and related methods
US12037583B2 (en) 2015-12-04 2024-07-16 Novartis Ag Compositions and methods for immunooncology
EP3592380B1 (en) * 2017-03-31 2024-10-09 Cellectis SA New universal chimeric antigen receptor t cells specific for cd22
US12157760B2 (en) 2018-05-23 2024-12-03 The Broad Institute, Inc. Base editors and uses thereof
US12281338B2 (en) 2018-10-29 2025-04-22 The Broad Institute, Inc. Nucleobase editors comprising GeoCas9 and uses thereof
US12351837B2 (en) 2019-01-23 2025-07-08 The Broad Institute, Inc. Supernegatively charged proteins and uses thereof
US12390514B2 (en) 2017-03-09 2025-08-19 President And Fellows Of Harvard College Cancer vaccine
US12406749B2 (en) 2017-12-15 2025-09-02 The Broad Institute, Inc. Systems and methods for predicting repair outcomes in genetic engineering
US12435330B2 (en) 2019-10-10 2025-10-07 The Broad Institute, Inc. Methods and compositions for prime editing RNA
US12473543B2 (en) 2019-04-17 2025-11-18 The Broad Institute, Inc. Adenine base editors with reduced off-target effects
US12522807B2 (en) 2018-07-09 2026-01-13 The Broad Institute, Inc. RNA programmable epigenetic RNA modifiers and uses thereof

Families Citing this family (383)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9273283B2 (en) * 2009-10-29 2016-03-01 The Trustees Of Dartmouth College Method of producing T cell receptor-deficient T cells expressing a chimeric receptor
US10704021B2 (en) 2012-03-15 2020-07-07 Flodesign Sonics, Inc. Acoustic perfusion devices
US9458450B2 (en) 2012-03-15 2016-10-04 Flodesign Sonics, Inc. Acoustophoretic separation technology using multi-dimensional standing waves
US10967298B2 (en) 2012-03-15 2021-04-06 Flodesign Sonics, Inc. Driver and control for variable impedence load
US9950282B2 (en) 2012-03-15 2018-04-24 Flodesign Sonics, Inc. Electronic configuration and control for acoustic standing wave generation
BR112014029417B1 (pt) 2012-05-25 2023-03-07 Cellectis Método ex vivo para a preparação de células t para imunoterapia
US20150017136A1 (en) * 2013-07-15 2015-01-15 Cellectis Methods for engineering allogeneic and highly active t cell for immunotherapy
AU2013329186B2 (en) * 2012-10-10 2019-02-14 Sangamo Therapeutics, Inc. T cell modifying compounds and uses thereof
EP4282419A1 (en) 2012-12-20 2023-11-29 Purdue Research Foundation Chimeric antigen receptor-expressing t cells as anti-cancer therapeutics
DK2956175T3 (da) 2013-02-15 2017-11-27 Univ California Kimærisk antigenreceptor og fremgangsmåder til anvendelse deraf
US9587237B2 (en) 2013-03-14 2017-03-07 Elwha Llc Compositions, methods, and computer systems related to making and administering modified T cells
CA2905352A1 (en) * 2013-03-14 2014-09-25 Bellicum Pharmaceuticals, Inc. Methods for controlling t cell proliferation
US9499855B2 (en) 2013-03-14 2016-11-22 Elwha Llc Compositions, methods, and computer systems related to making and administering modified T cells
JP6005572B2 (ja) * 2013-03-28 2016-10-12 Kddi株式会社 動画像符号化装置、動画像復号装置、動画像符号化方法、動画像復号方法、およびプログラム
US11311575B2 (en) * 2013-05-13 2022-04-26 Cellectis Methods for engineering highly active T cell for immunotherapy
US11077144B2 (en) 2013-05-13 2021-08-03 Cellectis CD19 specific chimeric antigen receptor and uses thereof
US20230056268A1 (en) * 2013-05-13 2023-02-23 Cellectis Methods for engineering highly active t cell for immunotheraphy
US20230050345A1 (en) * 2013-05-13 2023-02-16 Cellectis Methods for engineering allogeneic and highly active t cell for immunotheraphy
EP3783098A1 (en) 2013-05-14 2021-02-24 Board Of Regents, The University Of Texas System Human application of engineered chimeric antigen receptor (car) t-cells
ES2883131T3 (es) 2013-05-29 2021-12-07 Cellectis Métodos para la modificación de células T para inmunoterapia utilizando el sistema de nucleasa CAS guiado por ARN
WO2014191128A1 (en) 2013-05-29 2014-12-04 Cellectis Methods for engineering t cells for immunotherapy by using rna-guided cas nuclease system
US10208125B2 (en) 2013-07-15 2019-02-19 University of Pittsburgh—of the Commonwealth System of Higher Education Anti-mucin 1 binding agents and uses thereof
EP3071686B1 (en) * 2013-11-22 2020-07-22 Cellectis SA Method for generating batches of allogeneic t-cells with averaged potency
WO2015075195A1 (en) * 2013-11-22 2015-05-28 Cellectis Method of engineering chemotherapy drug resistant t-cells for immunotherapy
RU2689558C1 (ru) * 2013-11-22 2019-05-28 Селлектис Способ конструирования аллогенных и устойчивых к лекарственным препаратам т-клеток для иммунотерапии
WO2015077717A1 (en) 2013-11-25 2015-05-28 The Broad Institute Inc. Compositions and methods for diagnosing, evaluating and treating cancer by means of the dna methylation status
WO2015085147A1 (en) 2013-12-05 2015-06-11 The Broad Institute Inc. Polymorphic gene typing and somatic change detection using sequencing data
KR20230076867A (ko) 2013-12-20 2023-05-31 더 브로드 인스티튜트, 인코퍼레이티드 신생항원 백신과의 병용 요법
WO2015105955A1 (en) 2014-01-08 2015-07-16 Flodesign Sonics, Inc. Acoustophoresis device with dual acoustophoretic chamber
ES2701846T3 (es) * 2014-01-14 2019-02-26 Cellectis Receptor de antígeno quimérico que usa dominios de reconocimiento de antígenos derivados de pez cartilaginoso
EP3690044B1 (en) 2014-02-11 2024-01-10 The Regents of the University of Colorado, a body corporate Crispr enabled multiplexed genome engineering
KR102375998B1 (ko) * 2014-02-14 2022-03-21 더 보드 오브 리젠츠 오브 더 유니버시티 오브 텍사스 시스템 키메라 항원 수용체 및 제조방법
WO2015121454A1 (en) * 2014-02-14 2015-08-20 Cellectis Cells for immunotherapy engineered for targeting antigen present both on immune cells and pathological cells
US10934346B2 (en) 2014-02-14 2021-03-02 Bellicum Pharmaceuticals, Inc. Modified T cell comprising a polynucleotide encoding an inducible stimulating molecule comprising MyD88, CD40 and FKBP12
CA2939711C (en) * 2014-02-21 2020-09-29 Cellectis Method for in situ inhibition of regulatory t cells
DK3241561T3 (da) * 2014-03-05 2025-06-30 Autolus Ltd Konjugeret antistof eller bispecifik t-celle-engager, som selektivt binder enten tcr-beta konstant region 1 (trbc1) eller trbc2
US11982673B2 (en) 2014-03-05 2024-05-14 Autolus Limited Methods
US11385233B2 (en) 2015-03-05 2022-07-12 Autolus Limited Methods of depleting malignant T-cells
CN106795221B (zh) 2014-04-03 2022-06-07 塞勒克提斯公司 用于癌症免疫治疗的cd33特异性嵌合抗原受体
EP3131927B8 (en) 2014-04-14 2020-12-23 Cellectis Bcma (cd269) specific chimeric antigen receptors for cancer immunotherapy
WO2015161276A2 (en) * 2014-04-18 2015-10-22 Editas Medicine, Inc. Crispr-cas-related methods, compositions and components for cancer immunotherapy
US20170051037A1 (en) 2014-05-02 2017-02-23 Cellectis Cs1 specific multi-chain chimeric antigen receptor
KR20170032406A (ko) 2014-07-15 2017-03-22 주노 쎄러퓨티크스 인코퍼레이티드 입양 세포 치료를 위한 조작된 세포
WO2016016344A1 (en) 2014-07-29 2016-02-04 Cellectis Ror1(ntrkr1)specific chimeric antigen receptors for cancer immunotherapy
US10544201B2 (en) 2014-07-31 2020-01-28 Cellectis ROR1 specific multi-chain chimeric antigen receptor
PT3177640T (pt) 2014-08-08 2020-08-31 Univ Leland Stanford Junior Agentes pd-1 de alta afinidade e métodos de utilização
AU2015312117A1 (en) 2014-09-02 2017-03-02 Bellicum Pharmaceuticals, Inc. Costimulation of chimeric antigen receptors by Myd88 and CD40 polypeptides
EP3189073B2 (en) 2014-09-04 2025-06-11 Cellectis Trophoblast glycoprotein (5t4, tpbg) specific chimeric antigen receptors for cancer immunotherapy
EP3209679A1 (en) * 2014-10-24 2017-08-30 BCRT Holding BV T cell-based immunotherapeutics
ES2987570T3 (es) * 2014-11-05 2024-11-15 Juno Therapeutics Inc Métodos para la transducción y el procesamiento de células
GB201421096D0 (en) * 2014-11-27 2015-01-14 Imp Innovations Ltd Genome editing methods
EP3234130B1 (en) 2014-12-19 2020-11-25 The Broad Institute, Inc. Methods for profiling the t-cell- receptor repertoire
US10975442B2 (en) 2014-12-19 2021-04-13 Massachusetts Institute Of Technology Molecular biomarkers for cancer immunotherapy
US10557140B2 (en) * 2015-02-02 2020-02-11 Industry-Academic Cooperation Foundation, Dankook University CTLA-4-targeting trans-splicing ribozyme for delivery of chimeric antigen receptor, and use thereof
WO2016124765A1 (en) * 2015-02-06 2016-08-11 Cellectis Primary hematopoietic cells genetically engineered by slow release of nucleic acids using nanoparticles
US20170151281A1 (en) 2015-02-19 2017-06-01 Batu Biologics, Inc. Chimeric antigen receptor dendritic cell (car-dc) for treatment of cancer
KR102624023B1 (ko) 2015-02-24 2024-01-11 더 리젠츠 오브 더 유니버시티 오브 캘리포니아 결합-촉발된 전사 스위치 및 이들의 이용 방법
WO2016138846A1 (en) * 2015-03-02 2016-09-09 Shanghai Sidansai Biotechnology Co., Ltd Reducing immune tolerance induced by pd‐l1
JP2018509148A (ja) * 2015-03-11 2018-04-05 セレクティスＣｅｌｌｅｃｔｉｓ 患者における持続性および／または生着を増加させるために同種ｔ細胞を改変する方法
US20180072990A1 (en) * 2015-03-20 2018-03-15 Children's National Medical Center Generating virus or other antigen-specific t cells from a naïve t cell population
GB201504840D0 (en) * 2015-03-23 2015-05-06 Ucl Business Plc Chimeric antigen receptor
WO2016166268A1 (en) 2015-04-17 2016-10-20 Cellectis Engineering animal or plant genome using dna-guided argonaute interference systems (dais) from mesophilic prokaryotes
US11021699B2 (en) 2015-04-29 2021-06-01 FioDesign Sonics, Inc. Separation using angled acoustic waves
US11708572B2 (en) 2015-04-29 2023-07-25 Flodesign Sonics, Inc. Acoustic cell separation techniques and processes
US11377651B2 (en) 2016-10-19 2022-07-05 Flodesign Sonics, Inc. Cell therapy processes utilizing acoustophoresis
WO2016179319A1 (en) * 2015-05-04 2016-11-10 Cellerant Therapeutics, Inc. Chimeric antigen receptors with ctla4 signal transduction domains
AU2016261927B2 (en) * 2015-05-12 2022-04-07 Sangamo Therapeutics, Inc. Nuclease-mediated regulation of gene expression
US10752670B2 (en) 2015-05-20 2020-08-25 Cellectis Anti-GD3 specific chimeric antigen receptors for cancer immunotherapy
IL294183B2 (en) 2015-05-20 2023-10-01 Dana Farber Cancer Inst Inc shared neoantigens
WO2016196388A1 (en) 2015-05-29 2016-12-08 Juno Therapeutics, Inc. Composition and methods for regulating inhibitory interactions in genetically engineered cells
WO2016205749A1 (en) 2015-06-18 2016-12-22 The Broad Institute Inc. Novel crispr enzymes and systems
JP7033453B2 (ja) 2015-06-30 2022-03-10 セレクティス 特異的エンドヌクレアーゼを使用した遺伝子不活化によってｎｋ細胞の機能性を改善する方法
US20170014449A1 (en) * 2015-07-13 2017-01-19 Elwha LLC, a limited liability company of the State of Delaware Site-specific epigenetic editing
HK1255161A1 (zh) * 2015-07-15 2019-08-09 Juno Therapeutics, Inc. 用於过继性细胞治疗的工程细胞
US11474085B2 (en) 2015-07-28 2022-10-18 Flodesign Sonics, Inc. Expanded bed affinity selection
US11459540B2 (en) 2015-07-28 2022-10-04 Flodesign Sonics, Inc. Expanded bed affinity selection
GB2592821B (en) 2015-07-31 2022-01-12 Univ Minnesota Modified cells and methods of therapy
WO2017025323A1 (en) 2015-08-11 2017-02-16 Cellectis Cells for immunotherapy engineered for targeting cd38 antigen and for cd38 gene inactivation
CN108348551A (zh) * 2015-08-28 2018-07-31 宾夕法尼亚大学董事会 表达嵌合细胞内信号传导分子的细胞的方法和组合物
EP3340995A4 (en) 2015-08-28 2019-04-03 The Trustees Of The University Of Pennsylvania METHOD AND COMPOSITIONS FOR CELLS FOR EXPRESSING A CHIMERIC INTRA-CELLULAR SIGNAL MOLECULE
EP3359660B1 (en) 2015-10-05 2019-12-04 Precision Biosciences, Inc. Engineered meganucleases with recognition sequences found in the human t cell receptor alpha constant region gene
JP6816133B2 (ja) * 2015-10-05 2021-01-20 プレシジョン バイオサイエンシズ，インク． 改変ヒトｔ細胞受容体アルファ定常領域遺伝子を含む遺伝子改変細胞
US12241053B2 (en) 2015-10-09 2025-03-04 The Brigham And Women's Hospital, Inc. Modulation of novel immune checkpoint targets
WO2017070429A1 (en) * 2015-10-22 2017-04-27 Regents Of The University Of Minnesota Methods involving editing polynucleotides that encode t cell receptor
WO2017075451A1 (en) 2015-10-28 2017-05-04 The Broad Institute Inc. Compositions and methods for evaluating and modulating immune responses by detecting and targeting pou2af1
WO2017075478A2 (en) 2015-10-28 2017-05-04 The Broad Institute Inc. Compositions and methods for evaluating and modulating immune responses by use of immune cell gene signatures
WO2017075465A1 (en) 2015-10-28 2017-05-04 The Broad Institute Inc. Compositions and methods for evaluating and modulating immune responses by detecting and targeting gata3
CN108289913A (zh) 2015-10-30 2018-07-17 儿童国家医疗中心 从未致敏t细胞群体产生hpv抗原特异性t细胞
EP3370514A4 (en) * 2015-11-05 2019-03-27 Baylor College of Medicine EFFICIENT SCALABLE XENOTRANAPLANT SYSTEM FROM ONE PATIENT BASED ON A CHICKEN CHORIOAL ANTI-TEMEMBRBRANE (CAM) IN VIVO MODEL
AU2016355178B9 (en) 2015-11-19 2019-05-30 Massachusetts Institute Of Technology Lymphocyte antigen CD5-like (CD5L)-interleukin 12B (p40) heterodimers in immunity
EP3389677B1 (en) * 2015-12-18 2024-06-26 Sangamo Therapeutics, Inc. Targeted disruption of the t cell receptor
CN105505869A (zh) * 2015-12-21 2016-04-20 河南大学淮河医院 一种针对肿瘤干细胞的嵌合抗原受体t细胞
EP3184548A1 (en) * 2015-12-23 2017-06-28 Miltenyi Biotec GmbH Chimeric antigen receptor with cytokine receptor activating or blocking domain
IL299616A (en) 2016-01-08 2023-03-01 Univ California Conditionally active heterodimeric polypeptides and methods of using them
GB201604213D0 (en) * 2016-03-11 2016-04-27 Proximagen Ltd Drug combination and its use in therapy
JP2019509738A (ja) * 2016-03-11 2019-04-11 ブルーバード バイオ， インコーポレイテッド ゲノム編集された免疫エフェクター細胞
US20190161530A1 (en) * 2016-04-07 2019-05-30 Bluebird Bio, Inc. Chimeric antigen receptor t cell compositions
EP3439675A4 (en) 2016-04-08 2019-12-18 Purdue Research Foundation METHOD AND COMPOSITIONS FOR CAR-T CELL THERAPY
US11446398B2 (en) 2016-04-11 2022-09-20 Obsidian Therapeutics, Inc. Regulated biocircuit systems
CA3021634A1 (en) * 2016-04-15 2017-10-19 Zymeworks Inc. Multi-specific antigen-binding constructs targeting immunotherapeutics
EP4628587A3 (en) 2016-04-15 2026-02-25 Memorial Sloan Kettering Cancer Center Transgenic t cell and chimeric antigen receptor t cell compositions and related methods
WO2017178586A1 (en) 2016-04-15 2017-10-19 Cellectis A method of engineering prodrug-specific hypersensitive t-cells for immunotherapy by gene expression
WO2017178585A1 (en) 2016-04-15 2017-10-19 Cellectis A method of engineering drug-specific hypersensitive t-cells for immunotherapy by gene inactivation
AU2017254477A1 (en) 2016-04-18 2018-11-01 Jennifer G. ABELIN Improved HLA epitope prediction
WO2017192536A1 (en) 2016-05-02 2017-11-09 University Of Kansas Eliminating mhc restriction from the t cell receptor as a strategy for immunotherapy
US11214789B2 (en) 2016-05-03 2022-01-04 Flodesign Sonics, Inc. Concentration and washing of particles with acoustics
US11085035B2 (en) 2016-05-03 2021-08-10 Flodesign Sonics, Inc. Therapeutic cell washing, concentration, and separation utilizing acoustophoresis
AU2017280353B2 (en) 2016-06-24 2021-11-11 Inscripta, Inc. Methods for generating barcoded combinatorial libraries
AU2017291851B2 (en) * 2016-07-06 2022-10-13 Cellectis Sequential gene editing in primary immune cells
US11384156B2 (en) 2016-07-25 2022-07-12 The Nemours Foundation Adoptive T-cell therapy using EMPD-specific chimeric antigen receptors for treating IgE-mediated allergic diseases
KR102546839B1 (ko) 2016-08-03 2023-06-23 워싱턴 유니버시티 키메라 항원 수용체를 이용한 t 세포 악성종양의 치료를 위한 car-t 세포의 유전자 편집
WO2018035364A1 (en) 2016-08-17 2018-02-22 The Broad Institute Inc. Product and methods useful for modulating and evaluating immune responses
EP3995574A1 (en) 2016-08-24 2022-05-11 Sangamo Therapeutics, Inc. Regulation of gene expression using engineered nucleases
KR102455249B1 (ko) * 2016-08-24 2022-10-17 상가모 테라퓨틱스, 인코포레이티드 가공된 표적 특이적 뉴클레아제
ES2981703T3 (es) 2016-09-02 2024-10-10 Lentigen Tech Inc Composiciones y métodos para tratar cáncer con DuoCars
WO2018049025A2 (en) 2016-09-07 2018-03-15 The Broad Institute Inc. Compositions and methods for evaluating and modulating immune responses
KR102451510B1 (ko) 2016-09-08 2022-10-07 2세븐티 바이오, 인코포레이티드 Pd-1 호밍 엔도뉴클레아제 변이체, 조성물 및 사용 방법
US12499971B2 (en) 2016-09-28 2025-12-16 The Broad Institute, Inc. Systematic screening and mapping of regulatory elements in non-coding genomic regions, methods, compositions, and applications thereof
MX2019003768A (es) 2016-10-03 2019-06-24 Juno Therapeutics Inc Moleculas de enlace especificas de hpv.
JP7248571B2 (ja) 2016-10-05 2023-03-29 フジフィルム セルラー ダイナミクス，インコーポレイテッド ＭｅＣＰ２が破壊された誘導多能性幹細胞からの成熟系列の生成
US12447213B2 (en) 2016-10-07 2025-10-21 The Broad Institute, Inc. Modulation of novel immune checkpoint targets
US11591582B2 (en) 2016-10-11 2023-02-28 2Seventy Bio, Inc. TCRα homing endonuclease variants
CN110520530A (zh) 2016-10-18 2019-11-29 明尼苏达大学董事会 肿瘤浸润性淋巴细胞和治疗方法
WO2018073394A1 (en) 2016-10-19 2018-04-26 Cellectis Cell death inducing chimeric antigen receptors
WO2018073391A1 (en) 2016-10-19 2018-04-26 Cellectis Targeted gene insertion for improved immune cells therapy
WO2018073393A2 (en) 2016-10-19 2018-04-26 Cellectis Tal-effector nuclease (talen) -modified allogenic cells suitable for therapy
US11873511B2 (en) * 2016-10-19 2024-01-16 Cellectis Targeted gene insertion for improved immune cells therapy
EP3529265A2 (en) * 2016-10-19 2019-08-28 Cellectis Tal-effector nuclease (talen) -modified allogenic cells suitable for therapy
WO2018075830A1 (en) 2016-10-19 2018-04-26 Flodesign Sonics, Inc. Affinity cell extraction by acoustics
EP3538550A1 (en) 2016-11-10 2019-09-18 Deutsches Krebsforschungszentrum Stiftung des Öffentlichen Rechts Or10h1 modulators and uses thereof
EP3321280B8 (en) 2016-11-10 2021-03-10 Deutsches Krebsforschungszentrum Stiftung des Öffentlichen Rechts Immune modulators for reducing immune-resistance in melanoma and other proliferative diseases
WO2018089386A1 (en) 2016-11-11 2018-05-17 The Broad Institute, Inc. Modulation of intestinal epithelial cell differentiation, maintenance and/or function through t cell action
US11793833B2 (en) 2016-12-02 2023-10-24 Juno Therapeutics, Inc. Engineered B cells and related compositions and methods
EP3548055A4 (en) 2016-12-02 2020-08-19 University of Southern California SYNTHETIC IMMUNE RECEPTORS AND THEIR PROCESSES FOR USE
CA3048211A1 (en) 2016-12-23 2018-06-28 Macrogenics, Inc. Adam9-binding molecules, and methods of use thereof
WO2018132479A1 (en) * 2017-01-10 2018-07-19 The General Hospital Corporation Modified t cells and methods of their use
US11549149B2 (en) 2017-01-24 2023-01-10 The Broad Institute, Inc. Compositions and methods for detecting a mutant variant of a polynucleotide
US11649288B2 (en) 2017-02-07 2023-05-16 Seattle Children's Hospital Phospholipid ether (PLE) CAR T cell tumor targeting (CTCT) agents
CN108395482B (zh) 2017-02-08 2021-02-05 西比曼生物科技（香港）有限公司 一种靶向cd20抗原嵌合抗原受体的构建及其工程化t细胞的活性鉴定
EP3583203B1 (en) * 2017-02-15 2023-11-01 2seventy bio, Inc. Donor repair templates multiplex genome editing
WO2018160622A1 (en) 2017-02-28 2018-09-07 Endocyte, Inc. Compositions and methods for car t cell therapy
KR20230166145A (ko) 2017-03-15 2023-12-06 옥스포드 바이오메디카(유케이) 리미티드 방법
US11963966B2 (en) 2017-03-31 2024-04-23 Dana-Farber Cancer Institute, Inc. Compositions and methods for treating ovarian tumors
WO2018183921A1 (en) 2017-04-01 2018-10-04 The Broad Institute, Inc. Methods and compositions for detecting and modulating an immunotherapy resistance gene signature in cancer
CA3058807A1 (en) 2017-04-03 2018-10-11 Biontech Us Inc. Protein antigens and uses thereof
US12590288B2 (en) 2017-04-12 2026-03-31 The Broad Institute, Inc. Method of treating an inflammatory disease by administering an agent which binds a surface receptor on a tuft cell that induces an ILC class 2 inflammatory response
EP3610266A4 (en) 2017-04-12 2021-04-21 Massachusetts Eye and Ear Infirmary TUMOR SIGNATURE OF METASTASIS, COMPOSITIONS OF SUBSTANCES AND USES THEREOF
WO2018189360A1 (en) 2017-04-13 2018-10-18 Cellectis New sequence specific reagents targeting ccr5 in primary hematopoietic cells
EP3612210A4 (en) * 2017-04-19 2021-01-27 Board Of Regents, The University Of Texas System MANIPULATED ANTIGEN RECEPTORS EXPRESSING IMMUNE CELLS
EP3391907B8 (en) 2017-04-20 2020-03-04 iOmx Therapeutics AG Intracellular kinase sik3 associated with resistance against anti-tumour immune responses, and uses thereof
EP3622092A4 (en) 2017-05-11 2021-06-23 The Broad Institute, Inc. METHODS AND COMPOSITIONS OF USE OF CD8 + TUMOR-INFILTRATING LYMPHOCYTE SUBTYPES AND GENE SIGNATURES THEREOF
MX2019013514A (es) 2017-05-12 2020-01-20 Crispr Therapeutics Ag Materiales y metodos para modificar celulas por ingenieria genetica y usos de los mismos en inmunooncologia.
US11166985B2 (en) 2017-05-12 2021-11-09 Crispr Therapeutics Ag Materials and methods for engineering cells and uses thereof in immuno-oncology
WO2018213726A1 (en) 2017-05-18 2018-11-22 The Broad Institute, Inc. Systems, methods, and compositions for targeted nucleic acid editing
WO2018231759A1 (en) * 2017-06-12 2018-12-20 Obsidian Therapeutics, Inc. Pde5 compositions and methods for immunotherapy
WO2018232195A1 (en) 2017-06-14 2018-12-20 The Broad Institute, Inc. Compositions and methods targeting complement component 3 for inhibiting tumor growth
US20210139870A1 (en) 2017-06-19 2021-05-13 Cellectis Anti-hbv combination therapies involving specific endonucleases
BR112019027133B8 (pt) 2017-06-20 2022-08-09 Inst Curie Uso de uma célula imune modificada deficiente para suv39h1
CA3068286A1 (en) * 2017-06-22 2018-12-27 Board Of Regents, The University Of Texas System Methods for producing regulatory immune cells and uses thereof
US10011849B1 (en) 2017-06-23 2018-07-03 Inscripta, Inc. Nucleic acid-guided nucleases
US9982279B1 (en) 2017-06-23 2018-05-29 Inscripta, Inc. Nucleic acid-guided nucleases
EP3645038B1 (en) 2017-06-30 2026-02-18 Precision Biosciences, Inc. Genetically-modified t cells comprising a modified intron in the t cell receptor alpha gene
EP3645021A4 (en) 2017-06-30 2021-04-21 Intima Bioscience, Inc. ADENO-ASSOCIATED VIRAL VECTORS FOR GENE THERAPY
GB201710973D0 (en) 2017-07-07 2017-08-23 Avacta Life Sciences Ltd Scaffold proteins
US12049643B2 (en) 2017-07-14 2024-07-30 The Broad Institute, Inc. Methods and compositions for modulating cytotoxic lymphocyte activity
US20210130817A1 (en) 2017-07-14 2021-05-06 Cure Genetics Co., Ltd. Gene Editing System and Gene Editing Method
EP3638260A1 (en) 2017-07-21 2020-04-22 Cellectis Engineered immune cells resistant to tumor microoenvironment
WO2019016360A1 (en) 2017-07-21 2019-01-24 Cellectis MODIFIED IMMUNE CELLS RESISTANT TO TUMOR MICRO-ENVIRONMENT
CA3070998A1 (en) * 2017-07-25 2019-01-31 Board Of Regents, The University Of Texas System Enhanced chimeric antigen receptors and use thereof
WO2019020733A1 (en) 2017-07-26 2019-01-31 Cellectis METHODS OF IMMUNE CELL SELECTION OF THE ANTIGEN-DEPENDENT CHIMERIC ANTIGENIC RECEPTOR (CAR)
KR20260026092A (ko) 2017-09-19 2026-02-25 메사추세츠 인스티튜트 오브 테크놀로지 키메라 항원 수용체 t 세포 요법을 위한 조성물 및 그의 용도
AU2018338318B2 (en) 2017-09-21 2022-12-22 Massachusetts Institute Of Technology Systems, methods, and compositions for targeted nucleic acid editing
WO2019070755A1 (en) 2017-10-02 2019-04-11 The Broad Institute, Inc. METHODS AND COMPOSITIONS FOR DETECTING AND MODULATING A GENETIC SIGNATURE OF IMMUNOTHERAPY RESISTANCE IN CANCER
JP2020537515A (ja) 2017-10-03 2020-12-24 ジュノー セラピューティクス インコーポレイテッド Ｈｐｖ特異的結合分子
KR20200075851A (ko) 2017-10-19 2020-06-26 셀렉티스 개선된 면역 세포들 치료를 위한 nk 억제제들의 타겟인 유전자 통합
MY204419A (en) 2017-10-31 2024-08-28 Servier Lab Methods and compositions for dosing of allogeneic chimeric antigen receptor t cells
WO2019094955A1 (en) 2017-11-13 2019-05-16 The Broad Institute, Inc. Methods and compositions for targeting developmental and oncogenic programs in h3k27m gliomas
EP3710039A4 (en) 2017-11-13 2021-08-04 The Broad Institute, Inc. METHODS AND COMPOSITIONS FOR TREATMENT OF CANCER BY TARGETING THE CLEC2D-KLRB1 PATH
CR20200251A (es) 2017-11-17 2020-07-17 Iovance Biotherapeutics Inc Expansión de til de aspirados con aguja fina y biopsias por punción
EP3714041A1 (en) 2017-11-22 2020-09-30 Iovance Biotherapeutics, Inc. Expansion of peripheral blood lymphocytes (pbls) from peripheral blood
WO2019106163A1 (en) 2017-12-01 2019-06-06 Cellectis Reprogramming of genetically engineered primary immune cells
KR102439221B1 (ko) 2017-12-14 2022-09-01 프로디자인 소닉스, 인크. 음향 트랜스듀서 구동기 및 제어기
TW201930340A (zh) 2017-12-18 2019-08-01 美商尼恩醫療公司 新抗原及其用途
IL316257A (en) 2017-12-22 2024-12-01 Fate Therapeutics Inc Enhanced immune effector cells and use thereof
US12570958B2 (en) * 2017-12-22 2026-03-10 Cell Design Labs, Inc. Single- and multi-chain chimeric antigen receptors
WO2019119822A1 (en) * 2017-12-23 2019-06-27 Uwell Biopharma Inc. Pharmaceutical chimeric receptor composition and method thereof
EP3684399A1 (en) 2017-12-29 2020-07-29 Cellectis Method for improving production of car t cells
WO2019129850A1 (en) 2017-12-29 2019-07-04 Cellectis Off-the-shelf engineered cells for therapy
US11994512B2 (en) 2018-01-04 2024-05-28 Massachusetts Institute Of Technology Single-cell genomic methods to generate ex vivo cell systems that recapitulate in vivo biology with improved fidelity
EP3737743A1 (en) 2018-01-08 2020-11-18 Iovance Biotherapeutics, Inc. Processes for generating til products enriched for tumor antigen-specific t-cells
WO2019136459A1 (en) 2018-01-08 2019-07-11 Iovance Biotherapeutics, Inc. Processes for generating til products enriched for tumor antigen-specific t-cells
EP3508499A1 (en) 2018-01-08 2019-07-10 iOmx Therapeutics AG Antibodies targeting, and other modulators of, an immunoglobulin gene associated with resistance against anti-tumour immune responses, and uses thereof
US11713446B2 (en) 2018-01-08 2023-08-01 Iovance Biotherapeutics, Inc. Processes for generating TIL products enriched for tumor antigen-specific T-cells
US11311576B2 (en) 2018-01-22 2022-04-26 Seattle Children's Hospital Methods of use for CAR T cells
US20230158070A1 (en) 2018-01-30 2023-05-25 Cellectis Combination comprising allogeneic immune cells deficient for an antigen present on both t-cells and pathological cells and therapeutic antibody against said antigen
US12312416B2 (en) 2018-02-06 2025-05-27 Seattle Children's Hospital Fluorescein-specific cars exhibiting optimal t cell function against FL-PLE labelled tumors
WO2019157454A1 (en) 2018-02-11 2019-08-15 Memorial Sloan-Kettering Cancer Center Non-hla restricted t cell receptors and uses thereof
US12473336B2 (en) 2018-02-21 2025-11-18 Board Of Regents, The University Of Texas System Methods for activation and expansion of natural killer cells and uses thereof
AU2019225174B2 (en) 2018-02-23 2025-11-20 Endocyte, Inc. Sequencing method for CAR T cell therapy
US20210017248A1 (en) * 2018-03-16 2021-01-21 The Regents Of The University Of California Cellular signaling domain engineering in chimeric antigen receptor-modified regulatory t cells
KR102817092B1 (ko) 2018-03-29 2025-06-09 페이트 세러퓨틱스, 인코포레이티드 조작된 면역 효과기 세포 및 이의 용도
MA52207A (fr) 2018-04-05 2021-02-17 Editas Medicine Inc Lymphocytes t exprimant un récepteur recombinant, polynucléotides et procédés associés
EP3773908A1 (en) 2018-04-05 2021-02-17 Juno Therapeutics, Inc. T cell receptors and engineered cells expressing same
CA3094468A1 (en) 2018-04-05 2019-10-10 Juno Therapeutics, Inc. Methods of producing cells expressing a recombinant receptor and related compositions
US11786554B2 (en) 2018-04-12 2023-10-17 Precision Biosciences, Inc. Optimized engineered nucleases having specificity for the human T cell receptor alpha constant region gene
US11957695B2 (en) 2018-04-26 2024-04-16 The Broad Institute, Inc. Methods and compositions targeting glucocorticoid signaling for modulating immune responses
WO2019210131A1 (en) 2018-04-27 2019-10-31 Iovance Biotherapeutics, Inc. Closed process for expansion and gene editing of tumor infiltrating lymphocytes and uses of same in immunotherapy
MX2020012028A (es) 2018-05-11 2021-03-29 Crispr Therapeutics Ag Metodos y composiciones para tratar el cancer.
WO2019232542A2 (en) 2018-06-01 2019-12-05 Massachusetts Institute Of Technology Methods and compositions for detecting and modulating microenvironment gene signatures from the csf of metastasis patients
US12036240B2 (en) 2018-06-14 2024-07-16 The Broad Institute, Inc. Compositions and methods targeting complement component 3 for inhibiting tumor growth
MX2020014243A (es) 2018-06-19 2021-05-12 Biontech Us Inc Neoantigenos y usos de los mismos.
MD3823665T2 (ro) 2018-07-19 2024-05-31 Regeneron Pharma Receptori antigenci chimerici cu specificitate BCMA și utilizările acestora
US20220177524A1 (en) * 2018-07-26 2022-06-09 Nanjing Legend Biotech Co., Ltd. Nef-containing t cells and methods of producing thereof
WO2020068304A2 (en) 2018-08-20 2020-04-02 The Broad Institute, Inc. Inhibitors of rna-guided nuclease target binding and uses thereof
WO2020041384A1 (en) 2018-08-20 2020-02-27 The Broad Institute, Inc. 3-phenyl-2-cyano-azetidine derivatives, inhibitors of rna-guided nuclease activity
US20210324357A1 (en) 2018-08-20 2021-10-21 The Brigham And Women's Hospital, Inc. Degradation domain modifications for spatio-temporal control of rna-guided nucleases
AU2019346335B2 (en) 2018-09-28 2024-07-25 Massachusetts Institute Of Technology Collagen-localized immunomodulatory molecules and methods thereof
WO2020072700A1 (en) 2018-10-02 2020-04-09 Dana-Farber Cancer Institute, Inc. Hla single allele lines
US20210379057A1 (en) 2018-10-16 2021-12-09 Massachusetts Institute Of Technology Nutlin-3a for use in treating a mycobacterium tuberculosis infection
MY210603A (en) 2018-11-05 2025-10-01 Iovance Biotherapeutics Inc Processes for production of tumor infiltrating lymphocytes and uses of the same in immunotherapy
WO2020096927A1 (en) 2018-11-05 2020-05-14 Iovance Biotherapeutics, Inc. Expansion of tils utilizing akt pathway inhibitors
JP7854297B2 (ja) 2018-11-05 2026-05-01 アイオバンス バイオセラピューティクス，インコーポレイテッド 改良された腫瘍反応性ｔ細胞の選択
US12611427B2 (en) 2018-11-05 2026-04-28 Iovance Biotherapeutics, Inc. Treatment of NSCLC patients refractory for anti-PD-1 antibody
WO2020097193A1 (en) 2018-11-06 2020-05-14 The Regents Of The University Of California Chimeric antigen receptors for phagocytosis
WO2020099572A1 (en) * 2018-11-14 2020-05-22 Fundación Pública Andaluza Progreso Y Salud Polynucleotide for safer and more effective immunotherapies
MX2021006208A (es) 2018-11-28 2021-10-01 Univ Texas Edición por multiplexación del genoma de células inmunitarias para mejorar la funcionalidad y resistencia al entorno supresor.
MX2021006393A (es) 2018-11-29 2021-10-13 Univ Texas Metodos para expansion ex vivo de celulas exterminadoras naturales y uso de las mismas.
MX2021006194A (es) 2018-12-02 2021-06-30 Fate Therapeutics Inc Inmunoterapias mediante el uso de celulas efectoras potenciadas derivadas de ipsc.
IL283724B1 (en) 2018-12-10 2026-02-01 Genetix Biotherapeutics Inc pdcd-1 homing endonuclease variants
JP2022513750A (ja) 2018-12-10 2022-02-09 ２セブンティ バイオ インコーポレイテッド ホーミングエンドヌクレアーゼバリアント
US20220062394A1 (en) 2018-12-17 2022-03-03 The Broad Institute, Inc. Methods for identifying neoantigens
CA3123392A1 (en) 2018-12-19 2020-06-25 Iovance Biotherapeutics, Inc. Methods of expanding tumor infiltrating lymphocytes using engineered cytokine receptor pairs and uses thereof
US11739156B2 (en) 2019-01-06 2023-08-29 The Broad Institute, Inc. Massachusetts Institute of Technology Methods and compositions for overcoming immunosuppression
WO2020180733A1 (en) 2019-03-01 2020-09-10 Iovance Biotherapeutics, Inc. Expansion of tumor infiltrating lymphocytes from liquid tumors and therapeutic uses thereof
US20220154282A1 (en) 2019-03-12 2022-05-19 The Broad Institute, Inc. Detection means, compositions and methods for modulating synovial sarcoma cells
EP3942023A1 (en) 2019-03-18 2022-01-26 The Broad Institute, Inc. Compositions and methods for modulating metabolic regulators of t cell pathogenicity
WO2020191378A1 (en) 2019-03-21 2020-09-24 Allogene Therapeutics, Inc. METHODS FOR ENHANCING TCRαβ+ CELL DEPLETION EFFICIENCY
KR102691932B1 (ko) 2019-04-03 2024-08-06 프리시젼 바이오사이언시스 인코포레이티드 마이크로RNA-적응 shRNA(shRNAmiR)를 포함하는 유전자-변형 면역 세포
KR20210152536A (ko) 2019-04-12 2021-12-15 더 보드 오브 리젠츠 오브 더 유니버시티 오브 텍사스 시스템 조절 b 세포의 생산 방법 및 이의 용도
SG11202111031RA (en) * 2019-04-26 2021-11-29 Allogene Therapeutics Inc Rituximab-resistant chimeric antigen receptors and uses thereof
MX2021013359A (es) 2019-04-30 2022-01-31 Crispr Therapeutics Ag Terapia de celulas alogénicas de neoplasias malignas de células b usando células t modificadas genéticamente dirigidas a cd19.
US11026973B2 (en) 2019-04-30 2021-06-08 Myeloid Therapeutics, Inc. Engineered phagocytic receptor compositions and methods of use thereof
EP3736330A1 (en) 2019-05-08 2020-11-11 European Molecular Biology Laboratory Modified adeno-associated virus (aav) particles for gene therapy
WO2020232029A1 (en) 2019-05-13 2020-11-19 Iovance Biotherapeutics, Inc. Methods and compositions for selecting tumor infiltrating lymphocytes and uses of the same in immunotherapy
US20220235340A1 (en) 2019-05-20 2022-07-28 The Broad Institute, Inc. Novel crispr-cas systems and uses thereof
US20220226464A1 (en) 2019-05-28 2022-07-21 Massachusetts Institute Of Technology Methods and compositions for modulating immune responses
JP7644722B2 (ja) * 2019-06-01 2025-03-12 シベック バイオテクノロジーズ，インコーポレイティド 真核細胞への遺伝子編集システムの送達のための細菌プラットフォーム
WO2020263399A1 (en) 2019-06-26 2020-12-30 Massachusetts Institute Of Technology Immunomodulatory fusion protein-metal hydroxide complexes and methods thereof
KR20220051164A (ko) 2019-07-05 2022-04-26 아이오엠엑스 테라퓨틱스 아게 Igsf11 (vsig3)의 항체 결합 igc2 및 이의 용도
CN114258429A (zh) 2019-07-17 2022-03-29 菲特治疗公司 免疫效应细胞工程化和其用途
BR112022001148A2 (pt) 2019-07-23 2022-03-15 Inst Nat Sante Rech Med Células imunes modificadas, composição farmacêutica, kit, uso de uma célula imune modificada e invenção de produto
EP4004049A1 (en) 2019-07-24 2022-06-01 Regeneron Pharmaceuticals, Inc. Chimeric antigen receptors with mage-a4 specificity and uses thereof
WO2021030627A1 (en) 2019-08-13 2021-02-18 The General Hospital Corporation Methods for predicting outcomes of checkpoint inhibition and treatment thereof
US12421557B2 (en) 2019-08-16 2025-09-23 The Broad Institute, Inc. Methods for predicting outcomes and treating colorectal cancer using a cell atlas
CA3148179A1 (en) * 2019-08-20 2021-02-25 Bruce J. Mccreedy Jr. Lymphodepletion dosing regimens for cellular immunotherapies
WO2021041922A1 (en) 2019-08-30 2021-03-04 The Broad Institute, Inc. Crispr-associated mu transposase systems
CN114981409A (zh) 2019-09-03 2022-08-30 美洛德生物医药公司 用于基因组整合的方法和组合物
WO2021050789A1 (en) 2019-09-10 2021-03-18 Obsidian Therapeutics, Inc. Ca2-il15 fusion proteins for tunable regulation
WO2021061648A1 (en) 2019-09-23 2021-04-01 Massachusetts Institute Of Technology Methods and compositions for stimulation of endogenous t cell responses
US12297426B2 (en) 2019-10-01 2025-05-13 The Broad Institute, Inc. DNA damage response signature guided rational design of CRISPR-based systems and therapies
US12394502B2 (en) 2019-10-02 2025-08-19 The General Hospital Corporation Method for predicting HLA-binding peptides using protein structural features
US12195725B2 (en) 2019-10-03 2025-01-14 Dana-Farber Cancer Institute, Inc. Compositions and methods for modulating and detecting tissue specific TH17 cell pathogenicity
US11981922B2 (en) 2019-10-03 2024-05-14 Dana-Farber Cancer Institute, Inc. Methods and compositions for the modulation of cell interactions and signaling in the tumor microenvironment
US11793787B2 (en) 2019-10-07 2023-10-24 The Broad Institute, Inc. Methods and compositions for enhancing anti-tumor immunity by targeting steroidogenesis
JP2023500799A (ja) 2019-10-18 2023-01-11 トラスティーズ オブ ボストン ユニバーシティ Ｃａｌ－ｔコンストラクトおよびその使用
EP4048295A1 (en) 2019-10-25 2022-08-31 Iovance Biotherapeutics, Inc. Gene editing of tumor infiltrating lymphocytes and uses of same in immunotherapy
US11844800B2 (en) 2019-10-30 2023-12-19 Massachusetts Institute Of Technology Methods and compositions for predicting and preventing relapse of acute lymphoblastic leukemia
CA3162755A1 (en) * 2019-11-25 2021-06-03 Kyoto University T-cell master cell bank
JP2023504081A (ja) * 2019-11-27 2023-02-01 ボード オブ リージェンツ，ザ ユニバーシティ オブ テキサス システム 神経膠芽細胞腫及び他の癌を処置するためのナチュラルキラー細胞免疫療法
WO2021119538A1 (en) * 2019-12-11 2021-06-17 Myeloid Therapeutics, Inc. Therapeutic cell compositions and methods for manufacture and uses thereof
US12516291B2 (en) 2019-12-11 2026-01-06 Iovance Biotherapeutics, Inc. Processes for the production of tumor infiltrating lymphocytes (TILs) and methods of using the same
US10980836B1 (en) 2019-12-11 2021-04-20 Myeloid Therapeutics, Inc. Therapeutic cell compositions and methods of manufacturing and use thereof
EP4081537A1 (en) 2019-12-23 2022-11-02 Cellectis New mesothelin specific chimeric antigen receptors (car) for solid tumors cancer immunotherapy
US11865168B2 (en) 2019-12-30 2024-01-09 Massachusetts Institute Of Technology Compositions and methods for treating bacterial infections
KR20220141299A (ko) 2020-01-14 2022-10-19 신테카인, 인크. Il2 오르토로그 및 사용 방법
WO2021146641A1 (en) 2020-01-17 2021-07-22 The Broad Institute, Inc. Small type ii-d cas proteins and methods of use thereof
US12165747B2 (en) 2020-01-23 2024-12-10 The Broad Institute, Inc. Molecular spatial mapping of metastatic tumor microenvironment
CN115210252A (zh) 2020-02-04 2022-10-18 西雅图儿童医院（Dba西雅图儿童研究所） 抗二硝基苯酚的嵌合抗原受体
MX2022010340A (es) 2020-02-24 2022-09-19 Allogene Therapeutics Inc Celulas t con car de bcma con actividades mejoradas.
CN115552015A (zh) * 2020-02-28 2022-12-30 香港中文大学 经由同时敲入和基因破坏来改造免疫细胞
JP2023517063A (ja) 2020-03-10 2023-04-21 マサチューセッツ インスティテュート オブ テクノロジー 操作されたメモリー様ｎｋ細胞を作製するための方法およびその組成物
BR112022017924A2 (pt) 2020-03-10 2022-12-20 Massachusetts Inst Technology Composições e métodos para imunoterapia de câncer npm1c-positivo
CN113402612A (zh) 2020-03-17 2021-09-17 西比曼生物科技（香港）有限公司 靶向cd19和cd20的联合嵌合抗原受体及其应用
US20210340524A1 (en) 2020-05-01 2021-11-04 Massachusetts Institute Of Technology Methods for identifying chimeric antigen receptor-targeting ligands and uses thereof
US12433954B2 (en) 2020-05-01 2025-10-07 Massachusetts Institute Of Technology Methods of activating anti-CD19 chimeric antigen receptor (CAR) T cells using amphiphilic ligand conjugates comprising CAR-targeting protein sequence motifs
CA3176826A1 (en) 2020-05-04 2021-11-11 Iovance Biotherapeutics, Inc. Processes for production of tumor infiltrating lymphocytes and uses of the same in immunotherapy
CN115803435A (zh) 2020-05-06 2023-03-14 塞勒克提斯公司 用于在细胞基因组中靶向插入外源序列的方法
EP3910331A1 (en) 2020-05-15 2021-11-17 iOmx Therapeutics AG Intracellular kinase associated with resistance against t-cell mediated cytotoxicity, and uses thereof
BR112022025806A2 (pt) 2020-06-19 2023-03-07 Fate Therapeutics Inc Combinação de tipos de células efetoras derivadas de ipsc para uso em imunoterapia
WO2022003158A1 (en) 2020-07-03 2022-01-06 Cellectis S.A. Method for determining potency of chimeric antigen receptor expressing immune cells
EP4175668A1 (en) 2020-07-06 2023-05-10 iOmx Therapeutics AG Antibodies binding igv of igsf11 (vsig3) and uses thereof
EP4185616A1 (en) 2020-07-24 2023-05-31 Cellectis S.A. T-cells expressing immune cell engagers in allogenic settings
EP4188395A1 (en) 2020-07-30 2023-06-07 Institut Curie Immune cells defective for socs1
CA3186325A1 (en) 2020-07-31 2022-02-03 Andre Choulika Dual car-t cells
US20220031751A1 (en) 2020-08-03 2022-02-03 Kyverna Therapeutics, Inc. Methods of producing t regulatory cells, methods of transducing t cells, and uses of the same
US20230321239A1 (en) 2020-08-14 2023-10-12 H. Lee Moffitt Cancer Center And Research Institute Inc. Chimeric antigen receptor t cells for treating autoimmunity
WO2022076606A1 (en) 2020-10-06 2022-04-14 Iovance Biotherapeutics, Inc. Treatment of nsclc patients with tumor infiltrating lymphocyte therapies
JP2023546359A (ja) 2020-10-06 2023-11-02 アイオバンス バイオセラピューティクス，インコーポレイテッド 腫瘍浸潤リンパ球療法によるｎｓｃｌｃ患者の治療
EP4240367A4 (en) 2020-11-04 2024-10-16 Myeloid Therapeutics, Inc. MODIFIED CHIMERIC FUSION PROTEIN COMPOSITIONS AND METHODS OF USE THEREOF
AU2021378075B2 (en) 2020-11-11 2025-03-27 Borea Therapeutics S.R.L. Modified viral particles for gene therapy
WO2022109277A1 (en) * 2020-11-20 2022-05-27 Pact Pharma, Inc. COMPOSITIONS AND METHODS FOR THE TREATMENT OF CANCER USING A TGFβRII ENGINEERED T CELL THERAPY
EP4251645A1 (en) 2020-11-25 2023-10-04 Catamaran Bio, Inc. Cellular therapeutics engineered with signal modulators and methods of use thereof
WO2022112596A1 (en) 2020-11-30 2022-06-02 Cellectis Sa Use of aminoquinoline compounds for higher gene integration
US11661459B2 (en) 2020-12-03 2023-05-30 Century Therapeutics, Inc. Artificial cell death polypeptide for chimeric antigen receptor and uses thereof
KR20230118887A (ko) 2020-12-03 2023-08-14 센츄리 쎄라퓨틱스 인코포레이티드 유전자 조작 세포 및 이의 용도
WO2022132720A1 (en) * 2020-12-14 2022-06-23 Allogene Therapeutics, Inc. Methods and reagents for characterizing car t cells for therapies
EP4262827A1 (en) 2020-12-17 2023-10-25 Iovance Biotherapeutics, Inc. Treatment of cancers with tumor infiltrating lymphocytes
AU2021401302A1 (en) 2020-12-17 2023-07-06 Iovance Biotherapeutics, Inc. Treatment with tumor infiltrating lymphocyte therapies in combination with ctla-4 and pd-1 inhibitors
TW202342542A (zh) * 2020-12-23 2023-11-01 大陸商廣東菲鵬製藥股份有限公司 抗pd-l1抗體、編碼其的核酸、包含其的載體、細胞、藥物組合物及其用途
EP4271817A2 (en) 2020-12-30 2023-11-08 Alaunos Therapeutics, Inc. Recombinant vectors comprising polycistronic expression cassettes and methods of use thereof
AU2022205377A1 (en) 2021-01-08 2023-07-20 Cellanome, Inc. Devices and methods for analyzing biological samples
US12576400B2 (en) 2021-01-08 2026-03-17 Cellanome, Inc. Methods for incubating and analyzing a cell in a compartment of a fluidic device
WO2022165260A1 (en) 2021-01-29 2022-08-04 Iovance Biotherapeutics, Inc. Methods of making modified tumor infiltrating lymphocytes and their use in adoptive cell therapy
WO2022165419A1 (en) 2021-02-01 2022-08-04 Kyverna Therapeutics, Inc. Methods for increasing t-cell function
KR20230153529A (ko) * 2021-02-19 2023-11-06 프리트 엠. 쇼드하리 다양한 면역세포를 위한 단일사슬 및 다중사슬 합성 항원 수용체
WO2022197949A2 (en) 2021-03-17 2022-09-22 Myeloid Therapeutics, Inc. Engineered chimeric fusion protein compositions and methods of use thereof
US20240191191A1 (en) 2021-03-19 2024-06-13 Iovance Biotherapeutics, Inc. Methods for infiltrating lymphocyte (til) expansion related to cd39/cd69 selection and gene knockout in tils
CA3213080A1 (en) 2021-03-23 2022-09-29 Krit RITTHIPICHAI Cish gene editing of tumor infiltrating lymphocytes and uses of same in immunotherapy
JP2024512029A (ja) 2021-03-25 2024-03-18 アイオバンス バイオセラピューティクス，インコーポレイテッド Ｔ細胞共培養効力アッセイのための方法及び組成物、ならびに細胞療法製品との使用
CA3215830A1 (en) 2021-04-19 2022-10-27 Rafael CUBAS Chimeric costimulatory receptors, chemokine receptors, and the use of same in cellular immunotherapies
US20240197880A1 (en) 2021-04-30 2024-06-20 Cellectis S.A. New anti-muc1 cars and gene edited immune cells for solid tumors cancer immunotherapy
AU2022271212A1 (en) 2021-05-04 2023-11-30 Regeneron Pharmaceuticals, Inc. Chimeric antigen receptors with mage-a4 specificity and uses thereof
BR112023023642A2 (pt) 2021-05-11 2024-01-30 Myeloid Therapeutics Inc Métodos e composições para integração genômica
JP2024519029A (ja) 2021-05-17 2024-05-08 アイオバンス バイオセラピューティクス，インコーポレイテッド Ｐｄ－１遺伝子編集された腫瘍浸潤リンパ球及び免疫療法におけるその使用
AU2022277649A1 (en) 2021-05-21 2023-11-30 Cellectis S.A. Enhancing efficacy of t-cell-mediated immunotherapy by modulating cancer-associated fibroblasts in solid tumors
US20220409665A1 (en) 2021-06-15 2022-12-29 Allogene Therapeutics, Inc. Selective targeting of host cd70+ alloreactive cells to prolong allogeneic car t cell persistence
US20250288666A1 (en) 2021-07-14 2025-09-18 Synthekine, Inc. Methods and compositions for use in cell therapy of neoplastic disease
EP4373270A2 (en) 2021-07-22 2024-05-29 Iovance Biotherapeutics, Inc. Method for cryopreservation of solid tumor fragments
JP2024527961A (ja) 2021-07-28 2024-07-26 アイオバンス バイオセラピューティクス，インコーポレイテッド Ｋｒａｓ阻害剤と併用した腫瘍浸潤リンパ球療法によるがん患者の治療
AU2022318416A1 (en) 2021-07-29 2024-01-25 Takeda Pharmaceutical Company Limited Engineered immune cell that specifically targets mesothelin and uses thereof
IL310550A (en) 2021-08-04 2024-03-01 Univ Colorado Regents LAT-activating chimeric antigen receptor T cells and methods of using them
WO2023039488A1 (en) 2021-09-09 2023-03-16 Iovance Biotherapeutics, Inc. Processes for generating til products using pd-1 talen knockdown
WO2023077015A2 (en) 2021-10-27 2023-05-04 Iovance Biotherapeutics, Inc. Systems and methods for coordinating manufacturing of cells for patient-specific immunotherapy
US20260083848A1 (en) 2021-11-03 2026-03-26 Viracta Therapeutics, Inc. Combination of car t-cell therapy with btk inhibitors and methods of use thereof
WO2023081900A1 (en) 2021-11-08 2023-05-11 Juno Therapeutics, Inc. Engineered t cells expressing a recombinant t cell receptor (tcr) and related systems and methods
EP4437091A1 (en) 2021-11-23 2024-10-02 Cellectis SA New tale protein scaffolds with improved on-target/off-target activity ratios
JP2025501272A (ja) 2021-12-28 2025-01-17 ムネモ・セラピューティクス 不活性化されたｓｕｖ３９ｈ１及び改変ｔｃｒを有する免疫細胞
EP4460564A4 (en) * 2022-01-08 2026-02-11 Carogen Corp MULTI-ANTIGEN THERAPEUTIC VACCINES TO TREAT OR PREVENT CHRONIC HEPATITIS B VIRUS INFECTION
EP4469065A1 (en) 2022-01-28 2024-12-04 Iovance Biotherapeutics, Inc. Cytokine associated tumor infiltrating lymphocytes compositions and methods
WO2023196877A1 (en) 2022-04-06 2023-10-12 Iovance Biotherapeutics, Inc. Treatment of nsclc patients with tumor infiltrating lymphocyte therapies
US20250228943A1 (en) 2022-04-08 2025-07-17 Fate Therapeutics, Inc. Cells having solid tumor targeting backbone and use thereof
US20250222029A1 (en) 2022-04-08 2025-07-10 Fate Therapeutics, Inc. Chimeric antigen receptor for tumor targeting
JP2025512401A (ja) 2022-04-15 2025-04-17 アイオバンス バイオセラピューティクス，インコーポレイテッド 特定のサイトカインの組み合わせ及び／またはＡＫＴｉ処理を使用したＴＩＬ拡張プロセス
CA3251533A1 (en) 2022-05-10 2023-11-16 Iovance Biotherapeutics, Inc. TREATMENT OF CANCER PATIENTS WITH TUMOR-INFILTRATING LYMPHOCYTE THERAPIES IN COMBINATION WITH AN IL-15R AGONIST
EP4279085A1 (en) 2022-05-20 2023-11-22 Mnemo Therapeutics Compositions and methods for treating a refractory or relapsed cancer or a chronic infectious disease
WO2023233003A1 (en) 2022-06-03 2023-12-07 Cellectis Sa Tale base editors for gene and cell therapy
AU2023298102A1 (en) 2022-06-30 2024-11-14 Cellectis S.A. Enhancing safety of t-cell-mediated immunotherapy
WO2024022509A1 (en) * 2022-07-29 2024-02-01 Nanjing Legend Biotech Co., Ltd. Methods for promoting persistence of cell therapy
EP4583890A1 (en) 2022-09-09 2025-07-16 Iovance Biotherapeutics, Inc. Processes for generating til products using pd-1/tigit talen double knockdown
EP4583889A1 (en) 2022-09-09 2025-07-16 Iovance Biotherapeutics, Inc. Processes for generating til products using pd-1/tigit talen double knockdown
US20260091113A1 (en) * 2022-09-19 2026-04-02 Emendobio Inc. Biallelic Knockout of CTLA4
WO2024062138A1 (en) 2022-09-23 2024-03-28 Mnemo Therapeutics Immune cells comprising a modified suv39h1 gene
WO2024077256A1 (en) 2022-10-07 2024-04-11 The General Hospital Corporation Methods and compositions for high-throughput discovery ofpeptide-mhc targeting binding proteins
JP2025536578A (ja) 2022-11-03 2025-11-07 セレクティス ソシエテ アノニム Ｔ細胞媒介免疫療法の有効性及び安全性の増進
WO2024098024A1 (en) 2022-11-04 2024-05-10 Iovance Biotherapeutics, Inc. Expansion of tumor infiltrating lymphocytes from liquid tumors and therapeutic uses thereof
EP4612277A1 (en) 2022-11-04 2025-09-10 Iovance Biotherapeutics, Inc. Methods for tumor infiltrating lymphocyte (til) expansion related to cd39/cd103 selection
JP2025539712A (ja) 2022-11-21 2025-12-09 アイオバンス バイオセラピューティクス，インコーポレイテッド 遺伝子編集されたｔ細胞の増殖能を評価するための方法
JP2025539816A (ja) 2022-11-21 2025-12-09 アイオバンス バイオセラピューティクス，インコーポレイテッド 腫瘍浸潤リンパ球の増幅のための２次元プロセス及びそれからの治療法
WO2024118836A1 (en) 2022-11-30 2024-06-06 Iovance Biotherapeutics, Inc. Processes for production of tumor infiltrating lymphocytes with shortened rep step
WO2024124044A1 (en) 2022-12-07 2024-06-13 The Brigham And Women’S Hospital, Inc. Compositions and methods targeting sat1 for enhancing anti¬ tumor immunity during tumor progression
WO2024121385A1 (en) 2022-12-09 2024-06-13 Cellectis S.A. Two-dose regimen in immunotherapy
CN116284370B (zh) * 2023-02-27 2024-05-28 南京立顶医疗科技有限公司 一种多聚体糖化血红蛋白单克隆抗体及制备方法
WO2024246301A1 (en) 2023-06-02 2024-12-05 Cellectis Sa Methods for identifying tale base editors off-sites
CN121532650A (zh) 2023-06-20 2026-02-13 艾洛基治疗公司 工程化的细胞的检测方法
CN121889172A (zh) 2023-07-21 2026-04-17 意大利商博雷亚医疗有限责任公司 制备表面修饰病毒衣壳的方法
WO2025021772A1 (en) 2023-07-21 2025-01-30 Borea Therapeutics S.R.L. Chemogenetic surface modified viral particles
LU504827B1 (en) 2023-07-28 2025-01-28 Univ Saarland Compounds and compositions for use in the treatment of lung diseases
CN116987699B (zh) * 2023-09-05 2024-08-27 深圳市艾迪贝克生物医药有限公司 用于制备通用型car-t细胞的基因片段、其工具系统及应用
EP4520334A1 (en) 2023-09-07 2025-03-12 Mnemo Therapeutics Methods and compositions for improving immune response
WO2025054540A1 (en) 2023-09-08 2025-03-13 Iovance Biotherapeutics, Inc. Methods of gene-editing using programmable nucleases
WO2025059533A1 (en) 2023-09-13 2025-03-20 The Broad Institute, Inc. Crispr enzymes and systems
WO2025072571A1 (en) 2023-09-27 2025-04-03 Cellanome, Inc. Cell culture within microfluidic structures
CN117164714B (zh) * 2023-10-08 2024-04-23 北京奇迈永华生物科技有限公司 一种靶向bcma的抗体及其应用
WO2025097055A2 (en) 2023-11-02 2025-05-08 The Broad Institute, Inc. Compositions and methods of use of t cells in immunotherapy
WO2025101484A1 (en) 2023-11-06 2025-05-15 Iovance Biotherapeutics, Inc. Treatment of endometrial cancers with tumor infiltrating lymphocyte therapies
WO2025117544A1 (en) 2023-11-29 2025-06-05 The Broad Institute, Inc. Engineered omega guide molecule and iscb compositions, systems, and methods of use thereof
WO2025163107A1 (en) 2024-02-01 2025-08-07 Institut Gustave Roussy Immune cells defective for znf217 and uses thereof
WO2025171182A1 (en) 2024-02-08 2025-08-14 Iovance Biotherapeutics, Inc. Treatment of cancer patients with tumor infiltrating lymphocyte therapies in combination with cancer vaccine
WO2025196087A1 (en) 2024-03-19 2025-09-25 Borea Therapeutics S.R.L. Targeting tissues in the cns
CN118027211A (zh) * 2024-04-11 2024-05-14 上海惠盾因泰生物科技有限公司 一种识别重组egf-crm197疫苗的单克隆抗体及其应用
US20250345431A1 (en) 2024-05-10 2025-11-13 Juno Therapeutics, Inc. Genetically engineered t cells expressing a cd19 chimeric antigen receptor (car) and uses thereof for allogeneic cell therapy
WO2026046724A1 (en) 2024-08-30 2026-03-05 Cellectis Sa Tale protein scaffolds involving fusions of monopartite and bipartite nls
CN120058920B (zh) * 2025-04-29 2025-09-09 山东省农业科学院家禽研究所（山东省无特定病原鸡研究中心） 抗s11型鸭疫里默氏杆菌单克隆抗体及其阻断elisa抗体检测试剂盒和应用

Family Cites Families (104)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR901228A (fr) 1943-01-16 1945-07-20 Deutsche Edelstahlwerke Ag Système d'aimant à entrefer annulaire
US4474893A (en) 1981-07-01 1984-10-02 The University of Texas System Cancer Center Recombinant monoclonal antibodies
US4683195A (en) 1986-01-30 1987-07-28 Cetus Corporation Process for amplifying, detecting, and/or-cloning nucleic acid sequences
US6905680B2 (en) 1988-11-23 2005-06-14 Genetics Institute, Inc. Methods of treating HIV infected subjects
US5858358A (en) 1992-04-07 1999-01-12 The United States Of America As Represented By The Secretary Of The Navy Methods for selectively stimulating proliferation of T cells
US6534055B1 (en) 1988-11-23 2003-03-18 Genetics Institute, Inc. Methods for selectively stimulating proliferation of T cells
US6352694B1 (en) 1994-06-03 2002-03-05 Genetics Institute, Inc. Methods for inducing a population of T cells to proliferate using agents which recognize TCR/CD3 and ligands which stimulate an accessory molecule on the surface of the T cells
WO1992010591A1 (en) 1990-12-14 1992-06-25 Cell Genesys, Inc. Chimeric chains for receptor-associated signal transduction pathways
IL103418A (en) * 1991-10-15 1997-11-20 Wellcome Found Therapeutic agent for the t-cell mediated inflammation of the joints and its production
WO1993019163A1 (en) 1992-03-18 1993-09-30 Yeda Research And Development Co, Ltd. Chimeric receptor genes and cells transformed therewith
US6129914A (en) 1992-03-27 2000-10-10 Protein Design Labs, Inc. Bispecific antibody effective to treat B-cell lymphoma and cell line
US5556763A (en) * 1992-04-06 1996-09-17 United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Evaluation and treatment of patients with progressive immunosuppression
US5770396A (en) * 1992-04-16 1998-06-23 The United States Of America As Represented By The Department Of Health And Human Services Isolation characterization, and use of the human beta subunit of the high affinity receptor for immunoglobulin E
US7175843B2 (en) 1994-06-03 2007-02-13 Genetics Institute, Llc Methods for selectively stimulating proliferation of T cells
US5668263A (en) * 1994-12-16 1997-09-16 Smithkline Beecham Corporation Conserved yeast nucleic acid sequences
US7067318B2 (en) 1995-06-07 2006-06-27 The Regents Of The University Of Michigan Methods for transfecting T cells
US6692964B1 (en) 1995-05-04 2004-02-17 The United States Of America As Represented By The Secretary Of The Navy Methods for transfecting T cells
US6010613A (en) 1995-12-08 2000-01-04 Cyto Pulse Sciences, Inc. Method of treating materials with pulsed electrical fields
US5766944A (en) * 1996-12-31 1998-06-16 Ruiz; Margaret Eileen T cell differentiation of CD34+ stem cells in cultured thymic epithelial fragments
US6004779A (en) * 1997-08-22 1999-12-21 Scriptgen Pharmaceuticals, Inc. Regulated gene expression in yeast
HUP0203035A3 (en) * 1998-07-14 2007-12-28 Corixa Corp Compositions and methods for therapy and diagnosis of prostate cancer
US6673594B1 (en) * 1998-09-29 2004-01-06 Organ Recovery Systems Apparatus and method for maintaining and/or restoring viability of organs
JP2004500029A (ja) * 1999-06-30 2004-01-08 コリクサ　コーポレイション 肺癌の治療および診断のための組成物および方法
CA2397741A1 (en) * 2000-01-14 2001-07-19 Corixa Corporation Compositions and methods for the therapy and diagnosis of prostate cancer
US6867041B2 (en) 2000-02-24 2005-03-15 Xcyte Therapies, Inc. Simultaneous stimulation and concentration of cells
KR20030032922A (ko) 2000-02-24 2003-04-26 싸이트 테라피스 인코포레이티드 세포의 동시 자극 및 농축
US7572631B2 (en) 2000-02-24 2009-08-11 Invitrogen Corporation Activation and expansion of T cells
US6797514B2 (en) 2000-02-24 2004-09-28 Xcyte Therapies, Inc. Simultaneous stimulation and concentration of cells
US7176243B2 (en) * 2000-06-02 2007-02-13 The General Hospital Corporation CaR receptor as a mediator of migratory cell chemotaxis and/or chemokinesis
EP1156062A1 (en) 2000-05-12 2001-11-21 GPC Biotech AG Immunomodulatory human MHC class II antigen-binding peptides/proteins
CA2459136A1 (en) 2001-09-04 2003-03-13 Yeda Research And Development Co. Ltd. A caspase-8 binding protein, its preparation and use
US20050130125A1 (en) * 2002-03-04 2005-06-16 Yuly Zagyansky End of aids for general virology, based on profound science as protein foldings: safe vaccines, universal antimicrobial means, mad cow end
MXPA04010927A (es) * 2002-05-02 2006-01-30 Univ Washington Metodos y composiciones para el tratamiento de enfermedades y trastornos inflamatorios/autoinmunes producidos por linfocitos t en sujetos que padecen una deficiencia en la regulacion de glucocorticoides.
AU2003281200A1 (en) * 2002-07-03 2004-01-23 Tasuku Honjo Immunopotentiating compositions
US7496848B2 (en) 2002-07-22 2009-02-24 Konica Corporation Image forming apparatus and image forming system
US7575925B2 (en) * 2002-12-10 2009-08-18 Sunnybrook Health Sciences Centre Cell preparations comprising cells of the T cell lineage and methods of making and using them
EP3202899B1 (en) * 2003-01-28 2020-10-21 Cellectis Custom-made meganuclease and use thereof
US9982251B2 (en) 2003-03-14 2018-05-29 Cellectis S.A. Large volume ex vivo electroporation method
EP1618891A1 (en) * 2003-03-31 2006-01-25 Kirin Beer Kabushiki Kaisha Method of inducing differentiation and proliferating regulatory t cell by anti-cd52 antibody and medicinal composition therefor
BRPI0417993A (pt) * 2003-12-22 2007-04-27 Genzyme Corp anticorpo anti-cd52 para tratamento da diabetes
US20060194725A1 (en) * 2004-05-07 2006-08-31 James Rasmussen Methods of treating disease with random copolymers
JP2008514702A (ja) * 2004-09-29 2008-05-08 エーエムアール テクノロジー インコーポレイテッド 新規シクロスポリン類似体およびそれらの薬学的使用
CN103710371B (zh) * 2005-08-03 2017-03-01 人类多克隆治疗股份有限公司 表达人源化免疫球蛋白的转基因动物中b细胞凋亡的抑制
KR101527334B1 (ko) * 2005-08-11 2015-06-09 알피 마토씨안-로저스 자가면역 질병의 치료 및 진단용 ｔｃｒ-ｖ-베타 관련 펩티드
WO2007043200A1 (ja) 2005-09-30 2007-04-19 Riken Ｔ細胞分化調節剤
ES2405552T3 (es) * 2005-11-29 2013-05-31 Actogenix N.V. Inducción de tolerancia mucosa a antiantígenos de células beta de islotes pancreáticos
GB0609121D0 (en) 2006-05-09 2006-06-21 Univ Birmingham Peptide Therapy
PL2383297T3 (pl) 2006-08-14 2013-06-28 Xencor Inc Zoptymalizowane przeciwciała ukierunkowane na CD19
WO2008060510A2 (en) * 2006-11-13 2008-05-22 Sangamo Biosciences, Inc. Zinc finger nuclease for targeting the human glucocorticoid receptor locus
US20080131415A1 (en) 2006-11-30 2008-06-05 Riddell Stanley R Adoptive transfer of cd8 + t cell clones derived from central memory cells
EP2123754B1 (en) * 2007-01-30 2011-04-06 Forerunner Pharma Research Co., Ltd. CHIMERIC Fc GAMMA RECEPTOR AND METHOD FOR DETERMINATION OF ADCC ACTIVITY BY USING THE RECEPTOR
US8563314B2 (en) * 2007-09-27 2013-10-22 Sangamo Biosciences, Inc. Methods and compositions for modulating PD1
WO2009091826A2 (en) 2008-01-14 2009-07-23 The Board Of Regents Of The University Of Texas System Compositions and methods related to a human cd19-specific chimeric antigen receptor (h-car)
AU2009288730B2 (en) * 2008-08-25 2013-06-20 Amplimmune, Inc. Compositions of PD-1 antagonists and methods of use
EP2331566B1 (en) 2008-08-26 2015-10-07 City of Hope Method and compositions for enhanced anti-tumor effector functioning of t cells
CA2740077A1 (en) * 2008-10-08 2010-04-15 Cambridge Enterprise Limited Methods and compositions for diagnosis and treatment
US8591905B2 (en) * 2008-10-12 2013-11-26 The Brigham And Women's Hospital, Inc. Nicotine immunonanotherapeutics
US20110030072A1 (en) * 2008-12-04 2011-02-03 Sigma-Aldrich Co. Genome editing of immunodeficiency genes in animals
JPWO2010101249A1 (ja) * 2009-03-06 2012-09-10 国立大学法人三重大学 Ｔ細胞の機能増強方法
EP2411520A2 (en) * 2009-03-27 2012-02-01 Merck Sharp&Dohme Corp. RNA INTERFERENCE MEDIATED INHIBITION OF THE THYMIC STROMAL LYMPHOPOIETIN (TSLP) GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
WO2010132697A2 (en) * 2009-05-13 2010-11-18 Genzyme Corporation Methods and compositions for treatment
US20110027881A1 (en) * 2009-07-31 2011-02-03 St. Marianna University School Of Medicine Production method of immune cells
US8586526B2 (en) * 2010-05-17 2013-11-19 Sangamo Biosciences, Inc. DNA-binding proteins and uses thereof
WO2011059836A2 (en) 2009-10-29 2011-05-19 Trustees Of Dartmouth College T cell receptor-deficient t cell compositions
US8956828B2 (en) * 2009-11-10 2015-02-17 Sangamo Biosciences, Inc. Targeted disruption of T cell receptor genes using engineered zinc finger protein nucleases
WO2011072246A2 (en) * 2009-12-10 2011-06-16 Regents Of The University Of Minnesota Tal effector-mediated dna modification
EP2392208B1 (en) * 2010-06-07 2016-05-04 Helmholtz Zentrum München Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH) Fusion proteins comprising a DNA-binding domain of a Tal effector protein and a non-specific cleavage domain of a restriction nuclease and their use
US20130189759A1 (en) * 2010-07-07 2013-07-25 Cellectis Meganucleases variants cleaving a dna target sequence in the nanog gene and uses thereof
AU2011281062B2 (en) * 2010-07-21 2015-01-22 Board Of Regents, The University Of Texas System Methods and compositions for modification of a HLA locus
US20130315933A1 (en) 2010-10-06 2013-11-28 Christoph Renner Antibodies Directed Against HLA-B27 Homodimers and Methods and Uses Thereof in Diagnosis and Therapy
WO2012050374A2 (en) 2010-10-13 2012-04-19 Innocell, Inc. Immunotherapy for solid tumors
US20130337454A1 (en) 2010-10-27 2013-12-19 Philippe Duchateau Method for increasing the efficiency of double-strand break-induced mutagenesis
PH12013501201A1 (en) 2010-12-09 2013-07-29 Univ Pennsylvania Use of chimeric antigen receptor-modified t cells to treat cancer
KR20140004174A (ko) 2011-01-18 2014-01-10 더 트러스티스 오브 더 유니버시티 오브 펜실바니아 암 치료를 위한 조성물 및 방법
MX363013B (es) * 2011-02-25 2019-03-04 Recombinetics Inc Animales genéticamente modificados y métodos para su obtención.
US9987308B2 (en) 2011-03-23 2018-06-05 Fred Hutchinson Cancer Research Center Method and compositions for cellular immunotherapy
CA3111953C (en) 2011-04-05 2023-10-24 Cellectis Method for the generation of compact tale-nucleases and uses thereof
US20130071414A1 (en) 2011-04-27 2013-03-21 Gianpietro Dotti Engineered cd19-specific t lymphocytes that coexpress il-15 and an inducible caspase-9 based suicide gene for the treatment of b-cell malignancies
CN102836441B (zh) * 2011-06-24 2019-06-11 台北荣民总医院 于感染性与恶性疾病的治疗中提升免疫反应的方法
EP2737066B1 (en) 2011-07-29 2017-11-08 Cellectis High throughput method for assembly and cloning polynucleotides comprising highly similar polynucleotidic modules
MX2014001222A (es) 2011-07-29 2014-09-15 Univ Pennsylvania Receptores coestimuladores de cambio.
CN103946952A (zh) 2011-09-16 2014-07-23 宾夕法尼亚大学董事会 用于治疗癌症的rna改造的t细胞
ES2654060T3 (es) * 2011-10-20 2018-02-12 The U.S.A. As Represented By The Secretary, Department Of Health And Human Services Receptores de antígenos quiméricos anti-CD22
HK1200871A1 (en) * 2011-11-16 2015-08-14 Sangamo Therapeutics, Inc. Modified dna-binding proteins and uses thereof
US10391126B2 (en) * 2011-11-18 2019-08-27 Board Of Regents, The University Of Texas System CAR+ T cells genetically modified to eliminate expression of T-cell receptor and/or HLA
AU2012347919B2 (en) 2011-12-05 2017-02-02 Factor Bioscience Inc. Methods and products for transfecting cells
US20130280220A1 (en) * 2012-04-20 2013-10-24 Nabil Ahmed Chimeric antigen receptor for bispecific activation and targeting of t lymphocytes
BR112014029417B1 (pt) 2012-05-25 2023-03-07 Cellectis Método ex vivo para a preparação de células t para imunoterapia
DK3187586T3 (da) * 2012-08-05 2019-10-14 Absci Llc Inducerbar co-ekspressionssystem
MX367730B (es) 2012-09-04 2019-09-04 Cellectis Receptor de antigeno quimerico multi-cadena y usos del mismo.
AU2013329186B2 (en) * 2012-10-10 2019-02-14 Sangamo Therapeutics, Inc. T cell modifying compounds and uses thereof
US9778269B2 (en) * 2012-11-15 2017-10-03 The Walter And Eliza Hall Institute Of Medical Research Method of treating sepsis by administering a soluble CD52 glycoprotein
DK2956175T3 (da) * 2013-02-15 2017-11-27 Univ California Kimærisk antigenreceptor og fremgangsmåder til anvendelse deraf
US11311575B2 (en) * 2013-05-13 2022-04-26 Cellectis Methods for engineering highly active T cell for immunotherapy
FI2997141T3 (fi) 2013-05-13 2022-12-15 CD19-spesifinen kimeerinen antigeenireseptori ja sen käyttöjä
US11077144B2 (en) 2013-05-13 2021-08-03 Cellectis CD19 specific chimeric antigen receptor and uses thereof
WO2016094880A1 (en) * 2014-12-12 2016-06-16 The Broad Institute Inc. Delivery, use and therapeutic applications of crispr systems and compositions for genome editing as to hematopoietic stem cells (hscs)
US9790490B2 (en) 2015-06-18 2017-10-17 The Broad Institute Inc. CRISPR enzymes and systems
JP6816133B2 (ja) 2015-10-05 2021-01-20 プレシジョン バイオサイエンシズ，インク． 改変ヒトｔ細胞受容体アルファ定常領域遺伝子を含む遺伝子改変細胞
EP3371300A1 (en) * 2015-11-05 2018-09-12 Centro en Investigación Biomédica en Red Process of gene-editing of cells isolated from a subject suffering from a metabolic disease affecting the erythroid lineage, cells obtained by said process and uses thereof
WO2017105110A1 (ko) 2015-12-15 2017-06-22 (주)옵토마인드 광 케이블용 송수신 장치 및 그 정렬 방법
US20190032088A1 (en) * 2016-02-26 2019-01-31 Cellectis Micelle based system nuclease encapsulation for in-vivo gene editing
WO2019016360A1 (en) * 2017-07-21 2019-01-24 Cellectis MODIFIED IMMUNE CELLS RESISTANT TO TUMOR MICRO-ENVIRONMENT
US20230158070A1 (en) * 2018-01-30 2023-05-25 Cellectis Combination comprising allogeneic immune cells deficient for an antigen present on both t-cells and pathological cells and therapeutic antibody against said antigen

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
H. TORIKAI ET AL: "A foundation for universal T-cell based immunotherapy: T cells engineered to express a CD19-specific chimeric-antigen-receptor and eliminate expression of endogenous TCR", BLOOD, PUBLISHED ONLINE APRIL 24, 2012, vol. 119, no. 24, 24 April 2012 (2012-04-24), pages 5697 - 5705, XP055071623, ISSN: 0006-4971, DOI: 10.1182/blood-2012-01-405365 *
M. M. MAHFOUZ ET AL: "De novo-engineered transcription activator-like effector (TALE) hybrid nuclease with novel DNA binding specificity creates double-strand breaks", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES, vol. 108, no. 6, 8 February 2011 (2011-02-08), pages 2623 - 2628, XP055007615, ISSN: 0027-8424, DOI: 10.1073/pnas.1019533108 *
MIZOGUCHI E ET AL: "Pathogenic role of IL-4, but not IFN-gamma in colitis of TCRalpha knockout mice", GASTROENTEROLOGY, ELSEVIER, PHILADELPHIA, PA, , ABSTRACT G4261, vol. 114, 15 April 1998 (1998-04-15), pages A1041, XP027469136, ISSN: 0016-5085, [retrieved on 19980415], DOI: 10.1016/S0016-5085(98)84235-2 *
TROP ET AL: "Competitive displacement of pT alpha by TCR-alpha during TCR assembly prevents surface coexpression of pre-TCR and alpha beta TCR.", THE JOURNAL OF IMMUNOLOGY, vol. 165, no. 10, 1 November 2000 (2000-11-01), pages 5566 - 5572, XP055079320, ISSN: 0022-1767 *

Cited By (150)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9322006B2 (en) 2011-07-22 2016-04-26 President And Fellows Of Harvard College Evaluation and improvement of nuclease cleavage specificity
US12006520B2 (en) 2011-07-22 2024-06-11 President And Fellows Of Harvard College Evaluation and improvement of nuclease cleavage specificity
US10323236B2 (en) 2011-07-22 2019-06-18 President And Fellows Of Harvard College Evaluation and improvement of nuclease cleavage specificity
WO2014165707A3 (en) * 2013-04-03 2015-03-12 Memorial Sloan-Kettering Cancer Center Effective generation of tumor-targeted t-cells derived from pluripotent stem cells
US10370452B2 (en) 2013-04-03 2019-08-06 Memorial Sloan-Kettering Cancer Center Effective generation of tumor-targeted T cells derived from pluripotent stem cells
US10954548B2 (en) 2013-08-09 2021-03-23 President And Fellows Of Harvard College Nuclease profiling system
US9163284B2 (en) 2013-08-09 2015-10-20 President And Fellows Of Harvard College Methods for identifying a target site of a Cas9 nuclease
US11920181B2 (en) 2013-08-09 2024-03-05 President And Fellows Of Harvard College Nuclease profiling system
US10508298B2 (en) 2013-08-09 2019-12-17 President And Fellows Of Harvard College Methods for identifying a target site of a CAS9 nuclease
US11046948B2 (en) 2013-08-22 2021-06-29 President And Fellows Of Harvard College Engineered transcription activator-like effector (TALE) domains and uses thereof
US10227581B2 (en) 2013-08-22 2019-03-12 President And Fellows Of Harvard College Engineered transcription activator-like effector (TALE) domains and uses thereof
US9359599B2 (en) 2013-08-22 2016-06-07 President And Fellows Of Harvard College Engineered transcription activator-like effector (TALE) domains and uses thereof
US9340799B2 (en) 2013-09-06 2016-05-17 President And Fellows Of Harvard College MRNA-sensing switchable gRNAs
US9999671B2 (en) 2013-09-06 2018-06-19 President And Fellows Of Harvard College Delivery of negatively charged proteins using cationic lipids
US9737604B2 (en) 2013-09-06 2017-08-22 President And Fellows Of Harvard College Use of cationic lipids to deliver CAS9
US10682410B2 (en) 2013-09-06 2020-06-16 President And Fellows Of Harvard College Delivery system for functional nucleases
US10858639B2 (en) 2013-09-06 2020-12-08 President And Fellows Of Harvard College CAS9 variants and uses thereof
US12584118B2 (en) 2013-09-06 2026-03-24 President And Fellows Of Harvard College Cas9 variants and uses thereof
US10597679B2 (en) 2013-09-06 2020-03-24 President And Fellows Of Harvard College Switchable Cas9 nucleases and uses thereof
US11299755B2 (en) 2013-09-06 2022-04-12 President And Fellows Of Harvard College Switchable CAS9 nucleases and uses thereof
US9526784B2 (en) 2013-09-06 2016-12-27 President And Fellows Of Harvard College Delivery system for functional nucleases
US9388430B2 (en) 2013-09-06 2016-07-12 President And Fellows Of Harvard College Cas9-recombinase fusion proteins and uses thereof
US12559737B2 (en) 2013-09-06 2026-02-24 President And Fellows Of Harvard College Cas9 variants and uses thereof
US12473573B2 (en) 2013-09-06 2025-11-18 President And Fellows Of Harvard College Switchable Cas9 nucleases and uses thereof
US9340800B2 (en) 2013-09-06 2016-05-17 President And Fellows Of Harvard College Extended DNA-sensing GRNAS
US9322037B2 (en) 2013-09-06 2016-04-26 President And Fellows Of Harvard College Cas9-FokI fusion proteins and uses thereof
US9228207B2 (en) 2013-09-06 2016-01-05 President And Fellows Of Harvard College Switchable gRNAs comprising aptamers
US10912833B2 (en) 2013-09-06 2021-02-09 President And Fellows Of Harvard College Delivery of negatively charged proteins using cationic lipids
US11390887B2 (en) 2013-11-07 2022-07-19 Editas Medicine, Inc. CRISPR-related methods and compositions with governing gRNAS
US10190137B2 (en) 2013-11-07 2019-01-29 Editas Medicine, Inc. CRISPR-related methods and compositions with governing gRNAS
US9834791B2 (en) 2013-11-07 2017-12-05 Editas Medicine, Inc. CRISPR-related methods and compositions with governing gRNAS
US10640788B2 (en) 2013-11-07 2020-05-05 Editas Medicine, Inc. CRISPR-related methods and compositions with governing gRNAs
US12215365B2 (en) 2013-12-12 2025-02-04 President And Fellows Of Harvard College Cas variants for gene editing
US9068179B1 (en) 2013-12-12 2015-06-30 President And Fellows Of Harvard College Methods for correcting presenilin point mutations
US11053481B2 (en) 2013-12-12 2021-07-06 President And Fellows Of Harvard College Fusions of Cas9 domains and nucleic acid-editing domains
US9840699B2 (en) 2013-12-12 2017-12-12 President And Fellows Of Harvard College Methods for nucleic acid editing
US11124782B2 (en) 2013-12-12 2021-09-21 President And Fellows Of Harvard College Cas variants for gene editing
US10465176B2 (en) 2013-12-12 2019-11-05 President And Fellows Of Harvard College Cas variants for gene editing
JP2017504601A (ja) * 2013-12-20 2017-02-09 セレクティスＣｅｌｌｅｃｔｉｓ 免疫療法のためにマルチインプットシグナル感受性ｔ細胞を操作する方法
EP3693384A1 (en) 2014-03-11 2020-08-12 Cellectis Method for generating t-cells compatible for allogenic transplantation
EP4368705A2 (en) 2014-03-11 2024-05-15 Cellectis Method for generating t-cells compatible for allogenic transplantation
EP3569619A1 (en) 2014-03-19 2019-11-20 Cellectis Cd123 specific chimeric antigen receptors for cancer immunotherapy
US20200384094A1 (en) * 2014-04-11 2020-12-10 Cellectis Method for generating immune cells resistant to arginine and/or tryptophan depleted microenvironment
WO2015155341A1 (en) * 2014-04-11 2015-10-15 Cellectis Method for generating immune cells resistant to arginine and/or tryptophan depleted microenvironment
US10765728B2 (en) 2014-04-11 2020-09-08 Cellectis Method for generating immune cells resistant to arginine and/or tryptophan depleted microenvironment
US12171820B2 (en) 2014-04-11 2024-12-24 Cellectis Method for generating immune cells resistant to arginine and/or tryptophan depleted microenvironment
JP2017513472A (ja) * 2014-04-11 2017-06-01 セレクティスＣｅｌｌｅｃｔｉｓ アルギニンおよび／またはトリプトファン枯渇微小環境に対して抵抗性を有する免疫細胞を作製するための方法
US11578343B2 (en) 2014-07-30 2023-02-14 President And Fellows Of Harvard College CAS9 proteins including ligand-dependent inteins
US10077453B2 (en) 2014-07-30 2018-09-18 President And Fellows Of Harvard College CAS9 proteins including ligand-dependent inteins
US12398406B2 (en) 2014-07-30 2025-08-26 President And Fellows Of Harvard College CAS9 proteins including ligand-dependent inteins
US10704062B2 (en) 2014-07-30 2020-07-07 President And Fellows Of Harvard College CAS9 proteins including ligand-dependent inteins
US11208661B2 (en) 2014-10-31 2021-12-28 The Trustees Of The University Of Pennsylvania Altering gene expression in modified T cells and uses thereof
US12577567B2 (en) 2014-10-31 2026-03-17 The Trustees Of The University Of Pennsylvania Altering gene expression in modified T cells and uses thereof
US12570982B2 (en) 2014-10-31 2026-03-10 The Trustees Of The University Of Pennsylvania Altering gene expression in modified T cells and uses thereof
US11203758B2 (en) 2014-10-31 2021-12-21 The Trustees Of The University Of Pennsylvania Altering gene expression in modified T cells and uses thereof
US12344843B2 (en) 2014-10-31 2025-07-01 The Trustees Of The University Of Pennsylvania Altering gene expression in cart cells and uses thereof
US10626372B1 (en) 2015-01-26 2020-04-21 Fate Therapeutics, Inc. Methods and compositions for inducing hematopoietic cell differentiation
US11634688B2 (en) 2015-01-26 2023-04-25 Fate Therapeutics, Inc. Methods and compositions for inducing hematopoietic cell differentiation
US12385012B2 (en) 2015-01-26 2025-08-12 Fate Therapeutics, Inc. Methods and compositions for inducing hematopoietic cell differentiation
US11014989B2 (en) 2015-01-26 2021-05-25 Cellectis Anti-CLL1 specific single-chain chimeric antigen receptors (scCARs) for cancer immunotherapy
JP2023129663A (ja) * 2015-03-27 2023-09-14 プレジデント アンド フェローズ オブ ハーバード カレッジ 改変ｔ細胞ならびにそれを作製および使用する方法
US12031154B2 (en) 2015-05-08 2024-07-09 President And Fellows Of Harvard College Universal donor stem cells and related methods
US12110500B2 (en) 2015-05-08 2024-10-08 President And Fellows Of Harvard College Universal donor stem cells and related methods
US12497590B2 (en) 2015-05-08 2025-12-16 President And Fellows Of Harvard College Universal donor stem cells and related methods
US12031155B2 (en) 2015-05-08 2024-07-09 President And Fellows Of Harvard College Universal donor stem cells and related methods
US12421493B2 (en) 2015-05-08 2025-09-23 President And Fellows Of Harvard College Universal donor stem cells and related methods
US11028142B2 (en) 2015-05-18 2021-06-08 TCR2 Therapeutics Inc. Compositions and methods for TCR reprogramming using fusion proteins
EP3466967A1 (en) * 2015-05-18 2019-04-10 TCR2 Therapeutics Inc. Compositions and methods for tcr reprogramming using fusion proteins
US10358473B2 (en) 2015-05-18 2019-07-23 TCR2 Therapeutics Inc. Compositions and methods for TCR reprogramming using fusion proteins
US10358474B2 (en) 2015-05-18 2019-07-23 TCR2 Therapeutics Inc. Compositions and methods for TCR reprogramming using fusion proteins
US11965012B2 (en) 2015-05-18 2024-04-23 TCR2 Therapeutics Inc. Compositions and methods for TCR reprogramming using fusion proteins
US10442849B2 (en) 2015-05-18 2019-10-15 Tcr2 Therabeutics Inc. Compositions and methods for TCR reprogramming using fusion proteins
CN108024544A (zh) * 2015-07-13 2018-05-11 桑格摩生物治疗股份有限公司 用于核酸酶介导的基因组工程化的递送方法及组合物
US10450585B2 (en) * 2015-07-13 2019-10-22 Sangamo Therapeutics, Inc. Delivery methods and compositions for nuclease-mediated genome engineering
US10167457B2 (en) 2015-10-23 2019-01-01 President And Fellows Of Harvard College Nucleobase editors and uses thereof
US12344869B2 (en) 2015-10-23 2025-07-01 President And Fellows Of Harvard College Nucleobase editors and uses thereof
US12043852B2 (en) 2015-10-23 2024-07-23 President And Fellows Of Harvard College Evolved Cas9 proteins for gene editing
US11214780B2 (en) 2015-10-23 2022-01-04 President And Fellows Of Harvard College Nucleobase editors and uses thereof
US10858628B2 (en) 2015-11-04 2020-12-08 Fate Therapeutics, Inc. Methods and compositions for inducing hematopoietic cell differentiation
US11162075B2 (en) 2015-11-04 2021-11-02 Fate Therapeutics, Inc. Methods and compositions for inducing hematopoietic cell differentiation
US10287606B2 (en) 2015-11-04 2019-05-14 Fate Therapeutics, Inc. Genomic engineering of pluripotent cells
US11162076B2 (en) 2015-11-04 2021-11-02 Fate Therapeutics, Inc. Methods and compositions for inducing hematopoietic cell differentiation
US11352607B2 (en) 2015-11-04 2022-06-07 Fate Therapeutics, Inc. Genomic engineering of pluripotent cells
US12410403B2 (en) 2015-11-04 2025-09-09 Fate Therapeutics, Inc. Methods and compositions for inducing hematopoietic cell differentiation
US11072781B2 (en) 2015-11-04 2021-07-27 Fate Therapeutics, Inc. Genomic engineering of pluripotent cells
US10947505B2 (en) 2015-11-04 2021-03-16 Fate Therapeutics, Inc. Methods and compositions for inducing hematopoietic cell differentiation
US12037583B2 (en) 2015-12-04 2024-07-16 Novartis Ag Compositions and methods for immunooncology
US11242376B2 (en) 2016-08-02 2022-02-08 TCR2 Therapeutics Inc. Compositions and methods for TCR reprogramming using fusion proteins
US11999947B2 (en) 2016-08-03 2024-06-04 President And Fellows Of Harvard College Adenosine nucleobase editors and uses thereof
US10947530B2 (en) 2016-08-03 2021-03-16 President And Fellows Of Harvard College Adenosine nucleobase editors and uses thereof
US10113163B2 (en) 2016-08-03 2018-10-30 President And Fellows Of Harvard College Adenosine nucleobase editors and uses thereof
US11702651B2 (en) 2016-08-03 2023-07-18 President And Fellows Of Harvard College Adenosine nucleobase editors and uses thereof
US11661590B2 (en) 2016-08-09 2023-05-30 President And Fellows Of Harvard College Programmable CAS9-recombinase fusion proteins and uses thereof
US12084663B2 (en) 2016-08-24 2024-09-10 President And Fellows Of Harvard College Incorporation of unnatural amino acids into proteins using base editing
US11542509B2 (en) 2016-08-24 2023-01-03 President And Fellows Of Harvard College Incorporation of unnatural amino acids into proteins using base editing
WO2018052828A1 (en) 2016-09-14 2018-03-22 Janssen Biotech, Inc. Chimeric antigen receptors comprising bcma-specific fibronectin type iii domains and uses thereof
US11096998B2 (en) 2016-09-14 2021-08-24 Janssen Biotech, Inc. Chimeric antigen receptors comprising BCMA-specific fibronectin type III domains and uses thereof
US11377638B2 (en) 2016-10-07 2022-07-05 TCR2 Therapeutics Inc. Compositions and methods for TCR reprogramming using fusion proteins
US11085021B2 (en) 2016-10-07 2021-08-10 TCR2 Therapeutics Inc. Compositions and methods for TCR reprogramming using fusion proteins
US11306324B2 (en) 2016-10-14 2022-04-19 President And Fellows Of Harvard College AAV delivery of nucleobase editors
US11851491B2 (en) 2016-11-22 2023-12-26 TCR2 Therapeutics Inc. Compositions and methods for TCR reprogramming using fusion proteins
WO2018115189A1 (en) 2016-12-21 2018-06-28 Cellectis Stably enginereed proteasome inhibitor resistant immune cells for immunotherapy
US11820969B2 (en) 2016-12-23 2023-11-21 President And Fellows Of Harvard College Editing of CCR2 receptor gene to protect against HIV infection
US10745677B2 (en) 2016-12-23 2020-08-18 President And Fellows Of Harvard College Editing of CCR5 receptor gene to protect against HIV infection
US11466271B2 (en) 2017-02-06 2022-10-11 Novartis Ag Compositions and methods for the treatment of hemoglobinopathies
US12559748B2 (en) 2017-02-06 2026-02-24 Novartis Ag Compositions and methods for the treatment of hemoglobinopathies
US12390514B2 (en) 2017-03-09 2025-08-19 President And Fellows Of Harvard College Cancer vaccine
US11898179B2 (en) 2017-03-09 2024-02-13 President And Fellows Of Harvard College Suppression of pain by gene editing
US12516308B2 (en) 2017-03-09 2026-01-06 President And Fellows Of Harvard College Suppression of pain by gene editing
US12435331B2 (en) 2017-03-10 2025-10-07 President And Fellows Of Harvard College Cytosine to guanine base editor
US11542496B2 (en) 2017-03-10 2023-01-03 President And Fellows Of Harvard College Cytosine to guanine base editor
US11851659B2 (en) 2017-03-22 2023-12-26 Novartis Ag Compositions and methods for immunooncology
US11268082B2 (en) 2017-03-23 2022-03-08 President And Fellows Of Harvard College Nucleobase editors comprising nucleic acid programmable DNA binding proteins
EP3592380B1 (en) * 2017-03-31 2024-10-09 Cellectis SA New universal chimeric antigen receptor t cells specific for cd22
US11560566B2 (en) 2017-05-12 2023-01-24 President And Fellows Of Harvard College Aptazyme-embedded guide RNAs for use with CRISPR-Cas9 in genome editing and transcriptional activation
WO2019002633A1 (en) 2017-06-30 2019-01-03 Cellectis CELLULAR IMMUNOTHERAPY FOR REPETITIVE ADMINISTRATION
WO2019018402A2 (en) 2017-07-17 2019-01-24 Janssen Biotech, Inc. ANTIGEN-BINDING REGIONS DIRECTED AGAINST FIBRONECTIN TYPE III DOMAINS AND METHODS OF USING THE SAME
US11161897B2 (en) 2017-07-17 2021-11-02 Janssen Biotech, Inc. Antigen binding regions against fibronectin type III domains and methods of using the same
US12269870B2 (en) 2017-07-17 2025-04-08 Janssen Biotech, Inc. Antigen binding regions against fibronectin type III domains and methods of using the same
US12359218B2 (en) 2017-07-28 2025-07-15 President And Fellows Of Harvard College Methods and compositions for evolving base editors using phage-assisted continuous evolution (PACE)
US11732274B2 (en) 2017-07-28 2023-08-22 President And Fellows Of Harvard College Methods and compositions for evolving base editors using phage-assisted continuous evolution (PACE)
US11932884B2 (en) 2017-08-30 2024-03-19 President And Fellows Of Harvard College High efficiency base editors comprising Gam
US11319532B2 (en) 2017-08-30 2022-05-03 President And Fellows Of Harvard College High efficiency base editors comprising Gam
WO2019072824A1 (en) 2017-10-09 2019-04-18 Cellectis IMPROVED ANTI-CD123 CAR IN UNIVERSAL MODIFIED IMMUNE T LYMPHOCYTES
US11795443B2 (en) 2017-10-16 2023-10-24 The Broad Institute, Inc. Uses of adenosine base editors
US12406749B2 (en) 2017-12-15 2025-09-02 The Broad Institute, Inc. Systems and methods for predicting repair outcomes in genetic engineering
WO2019155191A1 (en) * 2018-02-06 2019-08-15 Autolus Limited Polypeptides and methods
EP3765039A4 (en) * 2018-03-09 2021-12-08 TCR2 Therapeutics Inc. COMPOSITIONS AND METHODS FOR REPROGRAMMING TCR USING FUSION PROTEINS
US12157760B2 (en) 2018-05-23 2024-12-03 The Broad Institute, Inc. Base editors and uses thereof
WO2020007593A1 (en) 2018-07-02 2020-01-09 Cellectis Chimeric antigen receptors (car)-expressing cells and combination treatment for immunotherapy of patients with relapse refractory adverse genetic risk aml
US12522807B2 (en) 2018-07-09 2026-01-13 The Broad Institute, Inc. RNA programmable epigenetic RNA modifiers and uses thereof
WO2020043152A1 (en) 2018-08-29 2020-03-05 Nanjing Legend Biotech Co., Ltd. Anti-mesothelin chimeric antigen receptor (car) constructs and uses thereof
US12281338B2 (en) 2018-10-29 2025-04-22 The Broad Institute, Inc. Nucleobase editors comprising GeoCas9 and uses thereof
WO2020109953A1 (en) 2018-11-30 2020-06-04 Janssen Biotech, Inc. Gamma delta t cells and uses thereof
US12351837B2 (en) 2019-01-23 2025-07-08 The Broad Institute, Inc. Supernegatively charged proteins and uses thereof
US11795452B2 (en) 2019-03-19 2023-10-24 The Broad Institute, Inc. Methods and compositions for prime editing nucleotide sequences
US11643652B2 (en) 2019-03-19 2023-05-09 The Broad Institute, Inc. Methods and compositions for prime editing nucleotide sequences
US12281303B2 (en) 2019-03-19 2025-04-22 The Broad Institute, Inc. Methods and compositions for prime editing nucleotide sequences
US12570972B2 (en) 2019-03-19 2026-03-10 The Broad Institute, Inc. Methods and compositions for prime editing nucleotide sequences
US11447770B1 (en) 2019-03-19 2022-09-20 The Broad Institute, Inc. Methods and compositions for prime editing nucleotide sequences
US12509680B2 (en) 2019-03-19 2025-12-30 The Broad Institute, Inc. Methods and compositions for prime editing nucleotide sequences
US12473543B2 (en) 2019-04-17 2025-11-18 The Broad Institute, Inc. Adenine base editors with reduced off-target effects
WO2021041486A1 (en) 2019-08-27 2021-03-04 Janssen Biotech, Inc. Chimeric antigen receptor system and uses thereof
US12435330B2 (en) 2019-10-10 2025-10-07 The Broad Institute, Inc. Methods and compositions for prime editing RNA
WO2021176373A1 (en) 2020-03-03 2021-09-10 Janssen Biotech, Inc. ꝩδ T CELLS AND USES THEREOF
US11912985B2 (en) 2020-05-08 2024-02-27 The Broad Institute, Inc. Methods and compositions for simultaneous editing of both strands of a target double-stranded nucleotide sequence
US12031126B2 (en) 2020-05-08 2024-07-09 The Broad Institute, Inc. Methods and compositions for simultaneous editing of both strands of a target double-stranded nucleotide sequence
WO2022016119A1 (en) 2020-07-17 2022-01-20 Simurx, Inc. Chimeric myd88 receptors for redirecting immunosuppressive signaling and related compositions and methods
US11883432B2 (en) 2020-12-18 2024-01-30 Century Therapeutics, Inc. Chimeric antigen receptor system with adaptable receptor specificity
WO2023179766A1 (zh) 2022-03-24 2023-09-28 南京传奇生物科技有限公司 制备dna文库和检测逆转录病毒整合位点的方法

Also Published As

Publication number Publication date
JP6991167B2 (ja) 2022-02-03
MA37681A3 (fr) 2018-06-29
US11274316B2 (en) 2022-03-15
AU2013266734A1 (en) 2015-01-15
US20210220405A1 (en) 2021-07-22
EP2855667A1 (en) 2015-04-08
US10342829B2 (en) 2019-07-09
NZ703281A (en) 2017-01-27
US20220348955A1 (en) 2022-11-03
DK2855667T5 (da) 2024-10-14
JP2015525065A (ja) 2015-09-03
HK1208879A1 (en) 2016-03-18
US20130315884A1 (en) 2013-11-28
CN104718284A (zh) 2015-06-17
WO2013176916A8 (en) 2014-10-16
JP2019058189A (ja) 2019-04-18
US20210060080A1 (en) 2021-03-04
ES3015120T3 (en) 2025-04-29
SG11201407802WA (en) 2015-01-29
US11603539B2 (en) 2023-03-14
KR20150029651A (ko) 2015-03-18
JP2015523064A (ja) 2015-08-13
WO2013176915A1 (en) 2013-11-28
CN108998418A (zh) 2018-12-14
ZA201409536B (en) 2022-11-30
PL2855667T3 (pl) 2024-03-25
JP6463672B2 (ja) 2019-02-06
US20190216853A1 (en) 2019-07-18
US11007224B2 (en) 2021-05-18
EP4541818A2 (en) 2025-04-23
DK3473707T3 (da) 2025-03-17
US20160145337A1 (en) 2016-05-26
EP2855666A1 (en) 2015-04-08
US20190216854A1 (en) 2019-07-18
AU2018236867A1 (en) 2018-10-18
KR102247979B1 (ko) 2021-05-04
US20220177914A1 (en) 2022-06-09
US20180360883A1 (en) 2018-12-20
ES2962571T3 (es) 2024-03-19
US20180021379A1 (en) 2018-01-25
EP3279315A2 (en) 2018-02-07
KR102437522B1 (ko) 2022-08-26
US11414674B2 (en) 2022-08-16
JP2018011603A (ja) 2018-01-25
JP7257749B2 (ja) 2023-04-14
US12577581B2 (en) 2026-03-17
PT3473707T (pt) 2025-04-16
UA118652C2 (uk) 2019-02-25
AU2021201200A1 (en) 2021-03-11
US10363270B2 (en) 2019-07-30
US20140134142A1 (en) 2014-05-15
PL3473707T3 (pl) 2025-04-14
KR20210050590A (ko) 2021-05-07
AU2013266733B2 (en) 2018-07-05
US10517896B2 (en) 2019-12-31
AU2021201967B2 (en) 2024-07-18
BR112014029417B1 (pt) 2023-03-07
US10426795B2 (en) 2019-10-01
CA3133545A1 (en) 2013-11-28
US11891614B2 (en) 2024-02-06
JP6463671B2 (ja) 2019-02-06
CA2874609A1 (en) 2013-11-28
CA2874611A1 (en) 2013-11-28
US20170360835A1 (en) 2017-12-21
EP2855666B1 (en) 2019-12-04
EP2855667B1 (en) 2023-08-09
US20240309397A1 (en) 2024-09-19
CA2874611C (en) 2023-01-24
EP3473707A1 (en) 2019-04-24
AU2021201967A1 (en) 2021-04-29
MY180127A (en) 2020-11-23
EP3276000A3 (en) 2018-02-21
JP6655049B2 (ja) 2020-02-26
MX370265B (es) 2019-12-09
EA201492222A1 (ru) 2015-05-29
BR112014029417A2 (https=) 2017-08-15
US20200281979A1 (en) 2020-09-10
EP3473707B1 (en) 2024-12-18
US10286007B2 (en) 2019-05-14
MA37681B2 (fr) 2020-07-29
EP3276000A2 (en) 2018-01-31
JP2018183167A (ja) 2018-11-22
MX2014014364A (es) 2015-09-22
MA37681A2 (fr) 2016-06-30
SG10201806573TA (en) 2018-09-27
HK1245321A1 (en) 2018-08-24
EP3964567A1 (en) 2022-03-09
US10874693B2 (en) 2020-12-29
HUE064187T2 (hu) 2024-02-28
IN2014DN10991A (https=) 2015-09-25
AU2018236867B2 (en) 2021-01-21
AR096274A1 (es) 2015-12-16
US20150203817A1 (en) 2015-07-23
CA2874609C (en) 2021-12-07
EP3279315A3 (en) 2018-02-28
HUE070380T2 (hu) 2025-06-28
DK2855667T3 (da) 2023-10-30
AU2013266733A1 (en) 2015-01-15
CA3133545C (en) 2023-08-08
CN111676196A (zh) 2020-09-18
PT2855667T (pt) 2023-11-22
EP4541818A3 (en) 2025-07-23
AU2013266734B2 (en) 2018-11-22
AU2021201200B2 (en) 2023-10-19

Similar Documents

Publication Publication Date Title
US12577581B2 (en) Use of pre T alpha or functional variant thereof for expanding TCR alpha deficient T cells
HK40071469A (en) Methods for engineering allogeneic and immunosuppressive resistant t cell for immunotherapy
HK40001542A (en) Methods for engineering allogeneic and immunosuppressive resistant t cell for immunotherapy
HK40001542B (en) Methods for engineering allogeneic and immunosuppressive resistant t cell for immunotherapy
HK1208879B (en) Methods for engineering allogeneic and immunosuppressive resistant t cell for immunotherapy

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 13728017

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2874611

Country of ref document: CA

ENP Entry into the national phase

Ref document number: 2015514051

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 14403937

Country of ref document: US

WWE Wipo information: entry into national phase

Ref document number: 2013728017

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2013266734

Country of ref document: AU

Date of ref document: 20130513

Kind code of ref document: A