WO2020042647A1 - 改进的治疗性t细胞 - Google Patents
改进的治疗性t细胞 Download PDFInfo
- Publication number
- WO2020042647A1 WO2020042647A1 PCT/CN2019/084805 CN2019084805W WO2020042647A1 WO 2020042647 A1 WO2020042647 A1 WO 2020042647A1 CN 2019084805 W CN2019084805 W CN 2019084805W WO 2020042647 A1 WO2020042647 A1 WO 2020042647A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- cancer
- seq
- nucleotide sequence
- cell
- receptor
- Prior art date
Links
- 230000001225 therapeutic effect Effects 0.000 title claims abstract description 36
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 105
- 239000000427 antigen Substances 0.000 claims abstract description 91
- 108091007433 antigens Proteins 0.000 claims abstract description 91
- 102000036639 antigens Human genes 0.000 claims abstract description 91
- 210000001744 T-lymphocyte Anatomy 0.000 claims abstract description 83
- 239000013598 vector Substances 0.000 claims abstract description 83
- 201000011510 cancer Diseases 0.000 claims abstract description 71
- 102000005962 receptors Human genes 0.000 claims abstract description 28
- 108020003175 receptors Proteins 0.000 claims abstract description 28
- 102000004060 Transforming Growth Factor-beta Type II Receptor Human genes 0.000 claims abstract description 26
- 108010082684 Transforming Growth Factor-beta Type II Receptor Proteins 0.000 claims abstract description 26
- 239000002773 nucleotide Substances 0.000 claims description 74
- 125000003729 nucleotide group Chemical group 0.000 claims description 74
- 108010019670 Chimeric Antigen Receptors Proteins 0.000 claims description 55
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 53
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 34
- 108090000623 proteins and genes Proteins 0.000 claims description 34
- 229920001184 polypeptide Polymers 0.000 claims description 33
- 238000000034 method Methods 0.000 claims description 28
- 102000004169 proteins and genes Human genes 0.000 claims description 27
- 102000016266 T-Cell Antigen Receptors Human genes 0.000 claims description 22
- 108091008874 T cell receptors Proteins 0.000 claims description 21
- 230000004927 fusion Effects 0.000 claims description 19
- 108091093126 WHP Posttrascriptional Response Element Proteins 0.000 claims description 17
- -1 CD78 Proteins 0.000 claims description 16
- 102100033726 Tumor necrosis factor receptor superfamily member 17 Human genes 0.000 claims description 16
- 108090001012 Transforming Growth Factor beta Proteins 0.000 claims description 15
- 102000004887 Transforming Growth Factor beta Human genes 0.000 claims description 15
- 102100029290 Transthyretin Human genes 0.000 claims description 15
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 claims description 15
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 claims description 14
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 claims description 14
- 102000001301 EGF receptor Human genes 0.000 claims description 12
- 108060006698 EGF receptor Proteins 0.000 claims description 12
- 101000801255 Homo sapiens Tumor necrosis factor receptor superfamily member 17 Proteins 0.000 claims description 12
- 230000019491 signal transduction Effects 0.000 claims description 11
- 230000008685 targeting Effects 0.000 claims description 11
- 150000001413 amino acids Chemical group 0.000 claims description 10
- 239000008194 pharmaceutical composition Substances 0.000 claims description 10
- 108010022366 Carcinoembryonic Antigen Proteins 0.000 claims description 8
- 102100025475 Carcinoembryonic antigen-related cell adhesion molecule 5 Human genes 0.000 claims description 8
- 101000874179 Homo sapiens Syndecan-1 Proteins 0.000 claims description 8
- 101000835093 Homo sapiens Transferrin receptor protein 1 Proteins 0.000 claims description 8
- 102100035721 Syndecan-1 Human genes 0.000 claims description 8
- 102100034922 T-cell surface glycoprotein CD8 alpha chain Human genes 0.000 claims description 8
- 102100026144 Transferrin receptor protein 1 Human genes 0.000 claims description 8
- 230000000139 costimulatory effect Effects 0.000 claims description 8
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 claims description 8
- 102000018651 Epithelial Cell Adhesion Molecule Human genes 0.000 claims description 7
- 108010066687 Epithelial Cell Adhesion Molecule Proteins 0.000 claims description 7
- 108700018351 Major Histocompatibility Complex Proteins 0.000 claims description 7
- 102000017420 CD3 protein, epsilon/gamma/delta subunit Human genes 0.000 claims description 6
- 108050005493 CD3 protein, epsilon/gamma/delta subunit Proteins 0.000 claims description 6
- 208000032612 Glial tumor Diseases 0.000 claims description 6
- 206010018338 Glioma Diseases 0.000 claims description 6
- 206010060862 Prostate cancer Diseases 0.000 claims description 6
- 208000000236 Prostatic Neoplasms Diseases 0.000 claims description 6
- 230000004068 intracellular signaling Effects 0.000 claims description 6
- 230000020382 suppression by virus of host antigen processing and presentation of peptide antigen via MHC class I Effects 0.000 claims description 6
- 102100038080 B-cell receptor CD22 Human genes 0.000 claims description 5
- 101000884305 Homo sapiens B-cell receptor CD22 Proteins 0.000 claims description 5
- 102100036011 T-cell surface glycoprotein CD4 Human genes 0.000 claims description 5
- 239000003814 drug Substances 0.000 claims description 5
- 239000002245 particle Substances 0.000 claims description 5
- BGFTWECWAICPDG-UHFFFAOYSA-N 2-[bis(4-chlorophenyl)methyl]-4-n-[3-[bis(4-chlorophenyl)methyl]-4-(dimethylamino)phenyl]-1-n,1-n-dimethylbenzene-1,4-diamine Chemical compound C1=C(C(C=2C=CC(Cl)=CC=2)C=2C=CC(Cl)=CC=2)C(N(C)C)=CC=C1NC(C=1)=CC=C(N(C)C)C=1C(C=1C=CC(Cl)=CC=1)C1=CC=C(Cl)C=C1 BGFTWECWAICPDG-UHFFFAOYSA-N 0.000 claims description 4
- 101710145634 Antigen 1 Proteins 0.000 claims description 4
- 101100279855 Arabidopsis thaliana EPFL5 gene Proteins 0.000 claims description 4
- 108010008014 B-Cell Maturation Antigen Proteins 0.000 claims description 4
- 208000025324 B-cell acute lymphoblastic leukemia Diseases 0.000 claims description 4
- 102100022005 B-lymphocyte antigen CD20 Human genes 0.000 claims description 4
- 206010005003 Bladder cancer Diseases 0.000 claims description 4
- 102100026094 C-type lectin domain family 12 member A Human genes 0.000 claims description 4
- 101150013553 CD40 gene Proteins 0.000 claims description 4
- 101150031358 COLEC10 gene Proteins 0.000 claims description 4
- 102000013392 Carboxylesterase Human genes 0.000 claims description 4
- 108010051152 Carboxylesterase Proteins 0.000 claims description 4
- 102100025064 Cellular tumor antigen p53 Human genes 0.000 claims description 4
- 102000016359 Fibronectins Human genes 0.000 claims description 4
- 108010067306 Fibronectins Proteins 0.000 claims description 4
- 102100021197 G-protein coupled receptor family C group 5 member D Human genes 0.000 claims description 4
- 102000006395 Globulins Human genes 0.000 claims description 4
- 108010044091 Globulins Proteins 0.000 claims description 4
- 102100041003 Glutamate carboxypeptidase 2 Human genes 0.000 claims description 4
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 claims description 4
- 102000004457 Granulocyte-Macrophage Colony-Stimulating Factor Human genes 0.000 claims description 4
- 102100028113 Granulocyte-macrophage colony-stimulating factor receptor subunit alpha Human genes 0.000 claims description 4
- 102100031573 Hematopoietic progenitor cell antigen CD34 Human genes 0.000 claims description 4
- 102100026122 High affinity immunoglobulin gamma Fc receptor I Human genes 0.000 claims description 4
- 208000017604 Hodgkin disease Diseases 0.000 claims description 4
- 208000010747 Hodgkins lymphoma Diseases 0.000 claims description 4
- 101000897405 Homo sapiens B-lymphocyte antigen CD20 Proteins 0.000 claims description 4
- 101100496086 Homo sapiens CLEC12A gene Proteins 0.000 claims description 4
- 101000721661 Homo sapiens Cellular tumor antigen p53 Proteins 0.000 claims description 4
- 101001040713 Homo sapiens G-protein coupled receptor family C group 5 member D Proteins 0.000 claims description 4
- 101000892862 Homo sapiens Glutamate carboxypeptidase 2 Proteins 0.000 claims description 4
- 101000916625 Homo sapiens Granulocyte-macrophage colony-stimulating factor receptor subunit alpha Proteins 0.000 claims description 4
- 101000777663 Homo sapiens Hematopoietic progenitor cell antigen CD34 Proteins 0.000 claims description 4
- 101000913074 Homo sapiens High affinity immunoglobulin gamma Fc receptor I Proteins 0.000 claims description 4
- 101001103039 Homo sapiens Inactive tyrosine-protein kinase transmembrane receptor ROR1 Proteins 0.000 claims description 4
- 101000599951 Homo sapiens Insulin-like growth factor I Proteins 0.000 claims description 4
- 101001057504 Homo sapiens Interferon-stimulated gene 20 kDa protein Proteins 0.000 claims description 4
- 101001055144 Homo sapiens Interleukin-2 receptor subunit alpha Proteins 0.000 claims description 4
- 101000998120 Homo sapiens Interleukin-3 receptor subunit alpha Proteins 0.000 claims description 4
- 101000917858 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-A Proteins 0.000 claims description 4
- 101000917839 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-B Proteins 0.000 claims description 4
- 101001014223 Homo sapiens MAPK/MAK/MRK overlapping kinase Proteins 0.000 claims description 4
- 101001008874 Homo sapiens Mast/stem cell growth factor receptor Kit Proteins 0.000 claims description 4
- 101000934338 Homo sapiens Myeloid cell surface antigen CD33 Proteins 0.000 claims description 4
- 101001109501 Homo sapiens NKG2-D type II integral membrane protein Proteins 0.000 claims description 4
- 101001103036 Homo sapiens Nuclear receptor ROR-alpha Proteins 0.000 claims description 4
- 101000891028 Homo sapiens Peptidyl-prolyl cis-trans isomerase FKBP11 Proteins 0.000 claims description 4
- 101000610551 Homo sapiens Prominin-1 Proteins 0.000 claims description 4
- 101001012157 Homo sapiens Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 claims description 4
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 claims description 4
- 101000633784 Homo sapiens SLAM family member 7 Proteins 0.000 claims description 4
- 101000884271 Homo sapiens Signal transducer CD24 Proteins 0.000 claims description 4
- 101000716102 Homo sapiens T-cell surface glycoprotein CD4 Proteins 0.000 claims description 4
- 101000946843 Homo sapiens T-cell surface glycoprotein CD8 alpha chain Proteins 0.000 claims description 4
- 101000596234 Homo sapiens T-cell surface protein tactile Proteins 0.000 claims description 4
- 101000655352 Homo sapiens Telomerase reverse transcriptase Proteins 0.000 claims description 4
- 101000851376 Homo sapiens Tumor necrosis factor receptor superfamily member 8 Proteins 0.000 claims description 4
- 102100039615 Inactive tyrosine-protein kinase transmembrane receptor ROR1 Human genes 0.000 claims description 4
- 102000004877 Insulin Human genes 0.000 claims description 4
- 108090001061 Insulin Proteins 0.000 claims description 4
- 108090001117 Insulin-Like Growth Factor II Proteins 0.000 claims description 4
- 102000048143 Insulin-Like Growth Factor II Human genes 0.000 claims description 4
- 102100037852 Insulin-like growth factor I Human genes 0.000 claims description 4
- 102100027268 Interferon-stimulated gene 20 kDa protein Human genes 0.000 claims description 4
- 102100033493 Interleukin-3 receptor subunit alpha Human genes 0.000 claims description 4
- 102100034872 Kallikrein-4 Human genes 0.000 claims description 4
- 208000008839 Kidney Neoplasms Diseases 0.000 claims description 4
- 102100029185 Low affinity immunoglobulin gamma Fc region receptor III-B Human genes 0.000 claims description 4
- 102100031520 MAPK/MAK/MRK overlapping kinase Human genes 0.000 claims description 4
- 108010046938 Macrophage Colony-Stimulating Factor Proteins 0.000 claims description 4
- 102100028123 Macrophage colony-stimulating factor 1 Human genes 0.000 claims description 4
- 102100027754 Mast/stem cell growth factor receptor Kit Human genes 0.000 claims description 4
- 108010063954 Mucins Proteins 0.000 claims description 4
- 102100025243 Myeloid cell surface antigen CD33 Human genes 0.000 claims description 4
- 102100022680 NKG2-D type II integral membrane protein Human genes 0.000 claims description 4
- 102100040348 Peptidyl-prolyl cis-trans isomerase FKBP11 Human genes 0.000 claims description 4
- 102100040120 Prominin-1 Human genes 0.000 claims description 4
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 claims description 4
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 claims description 4
- 206010038389 Renal cancer Diseases 0.000 claims description 4
- 102100029198 SLAM family member 7 Human genes 0.000 claims description 4
- 102100038081 Signal transducer CD24 Human genes 0.000 claims description 4
- 102100037253 Solute carrier family 45 member 3 Human genes 0.000 claims description 4
- 102100035268 T-cell surface protein tactile Human genes 0.000 claims description 4
- 108010017842 Telomerase Proteins 0.000 claims description 4
- 102000007000 Tenascin Human genes 0.000 claims description 4
- 108010008125 Tenascin Proteins 0.000 claims description 4
- 108010034949 Thyroglobulin Proteins 0.000 claims description 4
- 102000009843 Thyroglobulin Human genes 0.000 claims description 4
- 101800000385 Transmembrane protein Proteins 0.000 claims description 4
- 102100040245 Tumor necrosis factor receptor superfamily member 5 Human genes 0.000 claims description 4
- 102100036857 Tumor necrosis factor receptor superfamily member 8 Human genes 0.000 claims description 4
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 claims description 4
- 102000002287 alpha Subunit Glycoprotein Hormones Human genes 0.000 claims description 4
- 108010000732 alpha Subunit Glycoprotein Hormones Proteins 0.000 claims description 4
- 210000003169 central nervous system Anatomy 0.000 claims description 4
- 102000003675 cytokine receptors Human genes 0.000 claims description 4
- 108010057085 cytokine receptors Proteins 0.000 claims description 4
- 230000003511 endothelial effect Effects 0.000 claims description 4
- 230000001605 fetal effect Effects 0.000 claims description 4
- 210000002950 fibroblast Anatomy 0.000 claims description 4
- 150000002339 glycosphingolipids Chemical class 0.000 claims description 4
- 239000003102 growth factor Substances 0.000 claims description 4
- 229940084986 human chorionic gonadotropin Drugs 0.000 claims description 4
- 229940125396 insulin Drugs 0.000 claims description 4
- 230000000968 intestinal effect Effects 0.000 claims description 4
- 108010024383 kallikrein 4 Proteins 0.000 claims description 4
- 201000010982 kidney cancer Diseases 0.000 claims description 4
- 239000011159 matrix material Substances 0.000 claims description 4
- 238000012737 microarray-based gene expression Methods 0.000 claims description 4
- 238000012243 multiplex automated genomic engineering Methods 0.000 claims description 4
- 229920001481 poly(stearyl methacrylate) Polymers 0.000 claims description 4
- 108010079891 prostein Proteins 0.000 claims description 4
- 230000004083 survival effect Effects 0.000 claims description 4
- 101150047061 tag-72 gene Proteins 0.000 claims description 4
- 201000005112 urinary bladder cancer Diseases 0.000 claims description 4
- 102100023635 Alpha-fetoprotein Human genes 0.000 claims description 3
- 102100025570 Cancer/testis antigen 1 Human genes 0.000 claims description 3
- 102000004190 Enzymes Human genes 0.000 claims description 3
- 108090000790 Enzymes Proteins 0.000 claims description 3
- 101000856237 Homo sapiens Cancer/testis antigen 1 Proteins 0.000 claims description 3
- 101000994322 Homo sapiens Integrin alpha-8 Proteins 0.000 claims description 3
- 101710123134 Ice-binding protein Proteins 0.000 claims description 3
- 101710082837 Ice-structuring protein Proteins 0.000 claims description 3
- 102100032825 Integrin alpha-8 Human genes 0.000 claims description 3
- 108010028275 Leukocyte Elastase Proteins 0.000 claims description 3
- 208000031422 Lymphocytic Chronic B-Cell Leukemia Diseases 0.000 claims description 3
- 206010025323 Lymphomas Diseases 0.000 claims description 3
- 108090000015 Mesothelin Proteins 0.000 claims description 3
- 102000003735 Mesothelin Human genes 0.000 claims description 3
- 102100033174 Neutrophil elastase Human genes 0.000 claims description 3
- 208000015914 Non-Hodgkin lymphomas Diseases 0.000 claims description 3
- 101710107540 Type-2 ice-structuring protein Proteins 0.000 claims description 3
- 239000003937 drug carrier Substances 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 239000002777 nucleoside Substances 0.000 claims description 3
- 150000003833 nucleoside derivatives Chemical class 0.000 claims description 3
- 229960002175 thyroglobulin Drugs 0.000 claims description 3
- 206010000830 Acute leukaemia Diseases 0.000 claims description 2
- 208000024893 Acute lymphoblastic leukemia Diseases 0.000 claims description 2
- 208000014697 Acute lymphocytic leukaemia Diseases 0.000 claims description 2
- 208000031261 Acute myeloid leukaemia Diseases 0.000 claims description 2
- 206010061424 Anal cancer Diseases 0.000 claims description 2
- 208000007860 Anus Neoplasms Diseases 0.000 claims description 2
- 208000010839 B-cell chronic lymphocytic leukemia Diseases 0.000 claims description 2
- 208000032791 BCR-ABL1 positive chronic myelogenous leukemia Diseases 0.000 claims description 2
- 206010005949 Bone cancer Diseases 0.000 claims description 2
- 208000018084 Bone neoplasm Diseases 0.000 claims description 2
- 206010006143 Brain stem glioma Diseases 0.000 claims description 2
- 206010006187 Breast cancer Diseases 0.000 claims description 2
- 208000026310 Breast neoplasm Diseases 0.000 claims description 2
- 206010007953 Central nervous system lymphoma Diseases 0.000 claims description 2
- 206010008342 Cervix carcinoma Diseases 0.000 claims description 2
- 208000010833 Chronic myeloid leukaemia Diseases 0.000 claims description 2
- 206010009944 Colon cancer Diseases 0.000 claims description 2
- 208000000461 Esophageal Neoplasms Diseases 0.000 claims description 2
- 201000001342 Fallopian tube cancer Diseases 0.000 claims description 2
- 208000013452 Fallopian tube neoplasm Diseases 0.000 claims description 2
- 201000003741 Gastrointestinal carcinoma Diseases 0.000 claims description 2
- 208000007766 Kaposi sarcoma Diseases 0.000 claims description 2
- 206010023825 Laryngeal cancer Diseases 0.000 claims description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 2
- 208000032271 Malignant tumor of penis Diseases 0.000 claims description 2
- 208000033761 Myelogenous Chronic BCR-ABL Positive Leukemia Diseases 0.000 claims description 2
- 208000033776 Myeloid Acute Leukemia Diseases 0.000 claims description 2
- 208000001894 Nasopharyngeal Neoplasms Diseases 0.000 claims description 2
- 206010061306 Nasopharyngeal cancer Diseases 0.000 claims description 2
- 206010030155 Oesophageal carcinoma Diseases 0.000 claims description 2
- 206010033128 Ovarian cancer Diseases 0.000 claims description 2
- 206010061535 Ovarian neoplasm Diseases 0.000 claims description 2
- 206010061902 Pancreatic neoplasm Diseases 0.000 claims description 2
- 208000000821 Parathyroid Neoplasms Diseases 0.000 claims description 2
- 208000002471 Penile Neoplasms Diseases 0.000 claims description 2
- 206010034299 Penile cancer Diseases 0.000 claims description 2
- 208000007913 Pituitary Neoplasms Diseases 0.000 claims description 2
- 201000005746 Pituitary adenoma Diseases 0.000 claims description 2
- 206010061538 Pituitary tumour benign Diseases 0.000 claims description 2
- 208000006664 Precursor Cell Lymphoblastic Leukemia-Lymphoma Diseases 0.000 claims description 2
- 208000015634 Rectal Neoplasms Diseases 0.000 claims description 2
- 206010039491 Sarcoma Diseases 0.000 claims description 2
- 208000000453 Skin Neoplasms Diseases 0.000 claims description 2
- 208000021712 Soft tissue sarcoma Diseases 0.000 claims description 2
- 208000005718 Stomach Neoplasms Diseases 0.000 claims description 2
- 206010042971 T-cell lymphoma Diseases 0.000 claims description 2
- 208000027585 T-cell non-Hodgkin lymphoma Diseases 0.000 claims description 2
- 208000024313 Testicular Neoplasms Diseases 0.000 claims description 2
- 206010057644 Testis cancer Diseases 0.000 claims description 2
- 208000024770 Thyroid neoplasm Diseases 0.000 claims description 2
- 208000023915 Ureteral Neoplasms Diseases 0.000 claims description 2
- 206010046392 Ureteric cancer Diseases 0.000 claims description 2
- 206010046431 Urethral cancer Diseases 0.000 claims description 2
- 206010046458 Urethral neoplasms Diseases 0.000 claims description 2
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 claims description 2
- 208000002495 Uterine Neoplasms Diseases 0.000 claims description 2
- 206010047741 Vulval cancer Diseases 0.000 claims description 2
- 208000004354 Vulvar Neoplasms Diseases 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims description 2
- 201000005188 adrenal gland cancer Diseases 0.000 claims description 2
- 208000024447 adrenal gland neoplasm Diseases 0.000 claims description 2
- 201000011165 anus cancer Diseases 0.000 claims description 2
- 239000010425 asbestos Substances 0.000 claims description 2
- 201000010881 cervical cancer Diseases 0.000 claims description 2
- 230000001684 chronic effect Effects 0.000 claims description 2
- 208000024207 chronic leukemia Diseases 0.000 claims description 2
- 208000032852 chronic lymphocytic leukemia Diseases 0.000 claims description 2
- 208000029742 colonic neoplasm Diseases 0.000 claims description 2
- 210000000750 endocrine system Anatomy 0.000 claims description 2
- 201000004101 esophageal cancer Diseases 0.000 claims description 2
- 206010017758 gastric cancer Diseases 0.000 claims description 2
- 201000010536 head and neck cancer Diseases 0.000 claims description 2
- 208000014829 head and neck neoplasm Diseases 0.000 claims description 2
- 201000002313 intestinal cancer Diseases 0.000 claims description 2
- 210000003734 kidney Anatomy 0.000 claims description 2
- 206010023841 laryngeal neoplasm Diseases 0.000 claims description 2
- 201000007270 liver cancer Diseases 0.000 claims description 2
- 208000014018 liver neoplasm Diseases 0.000 claims description 2
- 201000005202 lung cancer Diseases 0.000 claims description 2
- 208000020816 lung neoplasm Diseases 0.000 claims description 2
- 210000004698 lymphocyte Anatomy 0.000 claims description 2
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 claims description 2
- 208000020984 malignant renal pelvis neoplasm Diseases 0.000 claims description 2
- 208000026045 malignant tumor of parathyroid gland Diseases 0.000 claims description 2
- 201000001441 melanoma Diseases 0.000 claims description 2
- 201000008968 osteosarcoma Diseases 0.000 claims description 2
- 201000002528 pancreatic cancer Diseases 0.000 claims description 2
- 208000008443 pancreatic carcinoma Diseases 0.000 claims description 2
- 208000021310 pituitary gland adenoma Diseases 0.000 claims description 2
- 208000016800 primary central nervous system lymphoma Diseases 0.000 claims description 2
- 206010038038 rectal cancer Diseases 0.000 claims description 2
- 201000001275 rectum cancer Diseases 0.000 claims description 2
- 201000007444 renal pelvis carcinoma Diseases 0.000 claims description 2
- 229910052895 riebeckite Inorganic materials 0.000 claims description 2
- 201000000849 skin cancer Diseases 0.000 claims description 2
- 208000037959 spinal tumor Diseases 0.000 claims description 2
- 206010041823 squamous cell carcinoma Diseases 0.000 claims description 2
- 201000011549 stomach cancer Diseases 0.000 claims description 2
- 201000003120 testicular cancer Diseases 0.000 claims description 2
- 201000002510 thyroid cancer Diseases 0.000 claims description 2
- 230000002463 transducing effect Effects 0.000 claims description 2
- 230000005747 tumor angiogenesis Effects 0.000 claims description 2
- 201000011294 ureter cancer Diseases 0.000 claims description 2
- 206010046766 uterine cancer Diseases 0.000 claims description 2
- 206010046885 vaginal cancer Diseases 0.000 claims description 2
- 208000013139 vaginal neoplasm Diseases 0.000 claims description 2
- 201000005102 vulva cancer Diseases 0.000 claims description 2
- 102100031507 Fc receptor-like protein 5 Human genes 0.000 claims 2
- 101150032879 Fcrl5 gene Proteins 0.000 claims 2
- CXQCLLQQYTUUKJ-ALWAHNIESA-N beta-D-GalpNAc-(1->4)-[alpha-Neup5Ac-(2->8)-alpha-Neup5Ac-(2->3)]-beta-D-Galp-(1->4)-beta-D-Glcp-(1<->1')-Cer(d18:1/18:0) Chemical compound O[C@@H]1[C@@H](O)[C@H](OC[C@H](NC(=O)CCCCCCCCCCCCCCCCC)[C@H](O)\C=C\CCCCCCCCCCCCC)O[C@H](CO)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@]2(O[C@H]([C@H](NC(C)=O)[C@@H](O)C2)[C@H](O)[C@@H](CO)O[C@]2(O[C@H]([C@H](NC(C)=O)[C@@H](O)C2)[C@H](O)[C@H](O)CO)C(O)=O)C(O)=O)[C@@H](O[C@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](CO)O1 CXQCLLQQYTUUKJ-ALWAHNIESA-N 0.000 claims 2
- 201000009030 Carcinoma Diseases 0.000 claims 1
- QCDFBFJGMNKBDO-UHFFFAOYSA-N Clioquinol Chemical compound C1=CN=C2C(O)=C(I)C=C(Cl)C2=C1 QCDFBFJGMNKBDO-UHFFFAOYSA-N 0.000 claims 1
- 206010014733 Endometrial cancer Diseases 0.000 claims 1
- 206010014759 Endometrial neoplasm Diseases 0.000 claims 1
- 206010066476 Haematological malignancy Diseases 0.000 claims 1
- 208000002250 Hematologic Neoplasms Diseases 0.000 claims 1
- 102000016387 Pancreatic elastase Human genes 0.000 claims 1
- 108010067372 Pancreatic elastase Proteins 0.000 claims 1
- 210000000440 neutrophil Anatomy 0.000 claims 1
- 102000014187 peptide receptors Human genes 0.000 claims 1
- 108010011903 peptide receptors Proteins 0.000 claims 1
- 208000037965 uterine sarcoma Diseases 0.000 claims 1
- 230000026683 transduction Effects 0.000 abstract description 25
- 238000010361 transduction Methods 0.000 abstract description 25
- 230000004186 co-expression Effects 0.000 abstract description 6
- 238000002360 preparation method Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 130
- 230000014509 gene expression Effects 0.000 description 36
- 241000700605 Viruses Species 0.000 description 11
- 230000002147 killing effect Effects 0.000 description 11
- 108020004414 DNA Proteins 0.000 description 10
- 230000000694 effects Effects 0.000 description 10
- 239000012634 fragment Substances 0.000 description 10
- 230000031146 intracellular signal transduction Effects 0.000 description 9
- 241000713666 Lentivirus Species 0.000 description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 7
- 239000012528 membrane Substances 0.000 description 7
- 230000002829 reductive effect Effects 0.000 description 7
- 210000004881 tumor cell Anatomy 0.000 description 7
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 6
- 101500025614 Homo sapiens Transforming growth factor beta-1 Proteins 0.000 description 6
- 102100025751 Mothers against decapentaplegic homolog 2 Human genes 0.000 description 6
- 101710143123 Mothers against decapentaplegic homolog 2 Proteins 0.000 description 6
- 108700019146 Transgenes Proteins 0.000 description 6
- 201000010099 disease Diseases 0.000 description 6
- 230000002401 inhibitory effect Effects 0.000 description 6
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 6
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 5
- 241000283707 Capra Species 0.000 description 5
- 241000713772 Human immunodeficiency virus 1 Species 0.000 description 5
- 102100037850 Interferon gamma Human genes 0.000 description 5
- 108010074328 Interferon-gamma Proteins 0.000 description 5
- 239000005089 Luciferase Substances 0.000 description 5
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 5
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 5
- 230000009089 cytolysis Effects 0.000 description 5
- 239000012636 effector Substances 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 238000000684 flow cytometry Methods 0.000 description 5
- 210000002865 immune cell Anatomy 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 108060001084 Luciferase Proteins 0.000 description 4
- 108700026244 Open Reading Frames Proteins 0.000 description 4
- 102000046299 Transforming Growth Factor beta1 Human genes 0.000 description 4
- 101800002279 Transforming growth factor beta-1 Proteins 0.000 description 4
- 230000022534 cell killing Effects 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 230000001086 cytosolic effect Effects 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 108020004999 messenger RNA Proteins 0.000 description 4
- 230000026731 phosphorylation Effects 0.000 description 4
- 238000006366 phosphorylation reaction Methods 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 230000003612 virological effect Effects 0.000 description 4
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 3
- 102000004127 Cytokines Human genes 0.000 description 3
- 108090000695 Cytokines Proteins 0.000 description 3
- 101000914514 Homo sapiens T-cell-specific surface glycoprotein CD28 Proteins 0.000 description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 description 3
- 108010072866 Prostate-Specific Antigen Proteins 0.000 description 3
- 102100038358 Prostate-specific antigen Human genes 0.000 description 3
- 102100027213 T-cell-specific surface glycoprotein CD28 Human genes 0.000 description 3
- 108700005077 Viral Genes Proteins 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 230000002950 deficient Effects 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 230000010354 integration Effects 0.000 description 3
- 108020001756 ligand binding domains Proteins 0.000 description 3
- 230000000670 limiting effect Effects 0.000 description 3
- 238000004806 packaging method and process Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000010839 reverse transcription Methods 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 230000004614 tumor growth Effects 0.000 description 3
- 239000013603 viral vector Substances 0.000 description 3
- FFILOTSTFMXQJC-QCFYAKGBSA-N (2r,4r,5s,6s)-2-[3-[(2s,3s,4r,6s)-6-[(2s,3r,4r,5s,6r)-5-[(2s,3r,4r,5r,6r)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-2-[(2r,3s,4r,5r,6r)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(e)-3-hydroxy-2-(octadecanoylamino)octadec-4-enoxy]oxan-3-yl]oxy-3-hy Chemical compound O[C@@H]1[C@@H](O)[C@H](OCC(NC(=O)CCCCCCCCCCCCCCCCC)C(O)\C=C\CCCCCCCCCCCCC)O[C@H](CO)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@]2(O[C@@H]([C@@H](N)[C@H](O)C2)C(O)C(O)CO[C@]2(O[C@@H]([C@@H](N)[C@H](O)C2)C(O)C(O)CO)C(O)=O)C(O)=O)[C@@H](O[C@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](CO)O1 FFILOTSTFMXQJC-QCFYAKGBSA-N 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- 241000713704 Bovine immunodeficiency virus Species 0.000 description 2
- 108091007741 Chimeric antigen receptor T cells Proteins 0.000 description 2
- 108091026890 Coding region Proteins 0.000 description 2
- 102000053602 DNA Human genes 0.000 description 2
- 241000283073 Equus caballus Species 0.000 description 2
- 241000713800 Feline immunodeficiency virus Species 0.000 description 2
- 208000021519 Hodgkin lymphoma Diseases 0.000 description 2
- 239000012124 Opti-MEM Substances 0.000 description 2
- 239000002033 PVDF binder Substances 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 241000713311 Simian immunodeficiency virus Species 0.000 description 2
- 239000006180 TBST buffer Substances 0.000 description 2
- 102100033455 TGF-beta receptor type-2 Human genes 0.000 description 2
- 101710084188 TGF-beta receptor type-2 Proteins 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 230000006037 cell lysis Effects 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 230000005754 cellular signaling Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 102000027596 immune receptors Human genes 0.000 description 2
- 108091008915 immune receptors Proteins 0.000 description 2
- 238000009169 immunotherapy Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 239000013600 plasmid vector Substances 0.000 description 2
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 2
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 238000004448 titration Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- 230000006490 viral transcription Effects 0.000 description 2
- 108020004463 18S ribosomal RNA Proteins 0.000 description 1
- 239000012103 Alexa Fluor 488 Substances 0.000 description 1
- 102000006306 Antigen Receptors Human genes 0.000 description 1
- 108010083359 Antigen Receptors Proteins 0.000 description 1
- 108010039627 Aprotinin Proteins 0.000 description 1
- 206010003445 Ascites Diseases 0.000 description 1
- 238000009010 Bradford assay Methods 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 241000713756 Caprine arthritis encephalitis virus Species 0.000 description 1
- 102100025466 Carcinoembryonic antigen-related cell adhesion molecule 3 Human genes 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 241000282994 Cervidae Species 0.000 description 1
- 101800001148 Delta-peptide Proteins 0.000 description 1
- 102000016911 Deoxyribonucleases Human genes 0.000 description 1
- 108010053770 Deoxyribonucleases Proteins 0.000 description 1
- 206010015108 Epstein-Barr virus infection Diseases 0.000 description 1
- 102100031381 Fc receptor-like A Human genes 0.000 description 1
- 108010087819 Fc receptors Proteins 0.000 description 1
- 102000009109 Fc receptors Human genes 0.000 description 1
- 241000710198 Foot-and-mouth disease virus Species 0.000 description 1
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 1
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- 208000031886 HIV Infections Diseases 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000914337 Homo sapiens Carcinoembryonic antigen-related cell adhesion molecule 3 Proteins 0.000 description 1
- 101000846860 Homo sapiens Fc receptor-like A Proteins 0.000 description 1
- 101001059454 Homo sapiens Serine/threonine-protein kinase MARK2 Proteins 0.000 description 1
- 101000946860 Homo sapiens T-cell surface glycoprotein CD3 epsilon chain Proteins 0.000 description 1
- 101000934341 Homo sapiens T-cell surface glycoprotein CD5 Proteins 0.000 description 1
- 241000725303 Human immunodeficiency virus Species 0.000 description 1
- 241000713340 Human immunodeficiency virus 2 Species 0.000 description 1
- 108010073807 IgG Receptors Proteins 0.000 description 1
- 102000009490 IgG Receptors Human genes 0.000 description 1
- 208000006142 Infectious Encephalitis Diseases 0.000 description 1
- 102100034353 Integrase Human genes 0.000 description 1
- 102100026878 Interleukin-2 receptor subunit alpha Human genes 0.000 description 1
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 206010052178 Lymphocytic lymphoma Diseases 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 102000010292 Peptide Elongation Factor 1 Human genes 0.000 description 1
- 108010077524 Peptide Elongation Factor 1 Proteins 0.000 description 1
- 208000002151 Pleural effusion Diseases 0.000 description 1
- 238000012193 PureLink RNA Mini Kit Methods 0.000 description 1
- 239000013614 RNA sample Substances 0.000 description 1
- 238000010240 RT-PCR analysis Methods 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 241000555745 Sciuridae Species 0.000 description 1
- 102100028904 Serine/threonine-protein kinase MARK2 Human genes 0.000 description 1
- 108010003723 Single-Domain Antibodies Proteins 0.000 description 1
- 238000003639 Student–Newman–Keuls (SNK) method Methods 0.000 description 1
- 108010092262 T-Cell Antigen Receptors Proteins 0.000 description 1
- 102100035794 T-cell surface glycoprotein CD3 epsilon chain Human genes 0.000 description 1
- 102100025244 T-cell surface glycoprotein CD5 Human genes 0.000 description 1
- 241001420369 Thosea Species 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 235000010724 Wisteria floribunda Nutrition 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- ZVQOOHYFBIDMTQ-UHFFFAOYSA-N [methyl(oxido){1-[6-(trifluoromethyl)pyridin-3-yl]ethyl}-lambda(6)-sulfanylidene]cyanamide Chemical compound N#CN=S(C)(=O)C(C)C1=CC=C(C(F)(F)F)N=C1 ZVQOOHYFBIDMTQ-UHFFFAOYSA-N 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 239000003070 absorption delaying agent Substances 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 229960000548 alemtuzumab Drugs 0.000 description 1
- 230000000735 allogeneic effect Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- 239000003429 antifungal agent Substances 0.000 description 1
- 229960004405 aprotinin Drugs 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 229960004669 basiliximab Drugs 0.000 description 1
- 229950002903 bivatuzumab Drugs 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 101150058049 car gene Proteins 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 238000002659 cell therapy Methods 0.000 description 1
- 229960005395 cetuximab Drugs 0.000 description 1
- 229950002334 clenoliximab Drugs 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 208000023963 corpus uteri neoplasm Diseases 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000001461 cytolytic effect Effects 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 238000002784 cytotoxicity assay Methods 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 229960002806 daclizumab Drugs 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 239000002612 dispersion medium Substances 0.000 description 1
- 229960001776 edrecolomab Drugs 0.000 description 1
- 229960000284 efalizumab Drugs 0.000 description 1
- 206010014599 encephalitis Diseases 0.000 description 1
- 108010078428 env Gene Products Proteins 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 229950009760 epratuzumab Drugs 0.000 description 1
- 230000017188 evasion or tolerance of host immune response Effects 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 238000012239 gene modification Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 208000014951 hematologic disease Diseases 0.000 description 1
- 208000018706 hematopoietic system disease Diseases 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 125000001165 hydrophobic group Chemical group 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 229960000598 infliximab Drugs 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- ZPNFWUPYTFPOJU-LPYSRVMUSA-N iniprol Chemical compound C([C@H]1C(=O)NCC(=O)NCC(=O)N[C@H]2CSSC[C@H]3C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(N[C@H](C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC=4C=CC=CC=4)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC=4C=CC=CC=4)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC2=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H]2N(CCC2)C(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N2[C@@H](CCC2)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N2[C@@H](CCC2)C(=O)N3)C(=O)NCC(=O)NCC(=O)N[C@@H](C)C(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@H](C(=O)N1)C(C)C)[C@@H](C)O)[C@@H](C)CC)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 ZPNFWUPYTFPOJU-LPYSRVMUSA-N 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 229950010828 keliximab Drugs 0.000 description 1
- 229950000518 labetuzumab Drugs 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
- 210000001165 lymph node Anatomy 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 229960005027 natalizumab Drugs 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 229960000470 omalizumab Drugs 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 238000007427 paired t-test Methods 0.000 description 1
- 229960000402 palivizumab Drugs 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- 229960005570 pemtumomab Drugs 0.000 description 1
- 230000001124 posttranscriptional effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000001566 pro-viral effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 210000002307 prostate Anatomy 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 230000000306 recurrent effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 230000001177 retroviral effect Effects 0.000 description 1
- 206010039083 rhinitis Diseases 0.000 description 1
- 229960004641 rituximab Drugs 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 229950004951 sevirumab Drugs 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 230000009131 signaling function Effects 0.000 description 1
- 229950003804 siplizumab Drugs 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 208000017572 squamous cell neoplasm Diseases 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000004797 therapeutic response Effects 0.000 description 1
- 210000001541 thymus gland Anatomy 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 230000010474 transient expression Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 102000035160 transmembrane proteins Human genes 0.000 description 1
- 108091005703 transmembrane proteins Proteins 0.000 description 1
- 229960000575 trastuzumab Drugs 0.000 description 1
- 230000004565 tumor cell growth Effects 0.000 description 1
- 210000003171 tumor-infiltrating lymphocyte Anatomy 0.000 description 1
- 229950005082 tuvirumab Drugs 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 230000029812 viral genome replication Effects 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
- 239000012130 whole-cell lysate Substances 0.000 description 1
Images
Classifications
-
- 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
- A61K39/0005—Vertebrate antigens
- A61K39/0011—Cancer antigens
- A61K39/001102—Receptors, cell surface antigens or cell surface determinants
- A61K39/001103—Receptors for growth factors
-
- 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
-
- 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
- A61K39/0005—Vertebrate antigens
- A61K39/0011—Cancer antigens
- A61K39/001102—Receptors, cell surface antigens or cell surface determinants
-
- 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
- A61K39/0005—Vertebrate antigens
- A61K39/0011—Cancer antigens
- A61K39/001102—Receptors, cell surface antigens or cell surface determinants
- A61K39/001111—Immunoglobulin superfamily
-
- 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
- A61K39/46—Cellular immunotherapy
- A61K39/461—Cellular immunotherapy characterised by the cell type used
- A61K39/4611—T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
-
- 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
- A61K39/46—Cellular immunotherapy
- A61K39/463—Cellular immunotherapy characterised by recombinant expression
- A61K39/4631—Chimeric Antigen Receptors [CAR]
-
- 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
- A61K39/46—Cellular immunotherapy
- A61K39/463—Cellular immunotherapy characterised by recombinant expression
- A61K39/4637—Other peptides or polypeptides
-
- 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
- A61K39/46—Cellular immunotherapy
- A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
- A61K39/4643—Vertebrate antigens
- A61K39/4644—Cancer antigens
- A61K39/464402—Receptors, cell surface antigens or cell surface determinants
- A61K39/464411—Immunoglobulin superfamily
- A61K39/464412—CD19 or B4
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
-
- 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
-
- 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
- C07K14/70578—NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
-
- 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
- C07K14/71—Receptors; Cell surface antigens; Cell surface determinants for growth factors; for growth regulators
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2803—Immunoglobulins [IGs], 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
-
- 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
- C12N15/86—Viral vectors
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/52—Constant or Fc region; Isotype
- C07K2317/53—Hinge
-
- 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
- C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
-
- 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
- C07K2319/33—Fusion polypeptide fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
-
- 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
- C12N2501/10—Growth factors
- C12N2501/15—Transforming growth factor beta (TGF-β)
-
- 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
- C12N2510/00—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
- C12N2740/00—Reverse transcribing RNA viruses
- C12N2740/00011—Details
- C12N2740/10011—Retroviridae
- C12N2740/15011—Lentivirus, not HIV, e.g. FIV, SIV
- C12N2740/15041—Use of virus, viral particle or viral elements as a vector
- C12N2740/15043—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
-
- 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
- C12N2740/00—Reverse transcribing RNA viruses
- C12N2740/00011—Details
- C12N2740/10011—Retroviridae
- C12N2740/16011—Human Immunodeficiency Virus, HIV
- C12N2740/16041—Use of virus, viral particle or viral elements as a vector
- C12N2740/16043—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
-
- 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
- C12N2800/00—Nucleic acids vectors
- C12N2800/10—Plasmid DNA
- C12N2800/106—Plasmid DNA for vertebrates
- C12N2800/107—Plasmid DNA for vertebrates for mammalian
-
- 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
- C12N2830/00—Vector systems having a special element relevant for transcription
- C12N2830/15—Vector systems having a special element relevant for transcription chimeric enhancer/promoter combination
-
- 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
- C12N2830/00—Vector systems having a special element relevant for transcription
- C12N2830/36—Vector systems having a special element relevant for transcription being a transcription termination element
-
- 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
- C12N2830/00—Vector systems having a special element relevant for transcription
- C12N2830/48—Vector systems having a special element relevant for transcription regulating transport or export of RNA, e.g. RRE, PRE, WPRE, CTE
Definitions
- the invention belongs to the field of biomedicine.
- the present invention relates to improved therapeutic T cells and methods of making the same.
- the present invention relates to the preparation of an improved therapeutic T cell by co-expressing a foreign antigen-specific receptor protein and a dominant negative TGF- ⁇ type II receptor in T cells through transduction by a lentiviral vector.
- T cells are the key immune cells that perform tumor cell killing and virus infection cell killing in vivo.
- T cells have been used, including antigen-specific T cells derived from in vitro induced or tumor infiltrating lymphocytes, genetically modified chimeric antigen receptor T cells (CAR-T cells), and genetically modified T cell receptor T cells (TCR-T cells) for the treatment of malignant tumors, has shown significant tumor clearance and control in some clinical patients.
- CAR-T cells genetically modified chimeric antigen receptor T cells
- TCR-T cells genetically modified T cell receptor T cells
- TGF- ⁇ is an important T cell inhibitory factor, which causes the killing effect of T cells on target cells to weaken or disappear.
- TGF- ⁇ is widely expressed in a variety of tumor tissues, and significantly inhibits the killing activity of tumor-specific T cells on tumor cells, which is an important reason for the failure of immunotherapy.
- the dominant negative TGF- ⁇ type II receptor (dominant TGF- ⁇ receptor type II (DNRII)) is a negative regulator of TGF- ⁇ , which can inhibit the inhibitory effect of TGF- ⁇ on T cells.
- tumor cells also express TGF- ⁇ , which leads to the problem that CAR-T cells and TCR-T cell functions are inhibited. It is also desirable in the art to introduce DNRII into CAR-T cells or TCR-T cells.
- CAR / TCR and DNRII are co-expressed on the same T cell for the treatment of tumors. This may be due to the low co-expression efficiency of the two proteins, which is difficult to meet the clinical needs.
- Figure 1 shows the lentiviral vector genome and integrity identification strategy for CAR-19 expression.
- A Old vector pPVLV1 containing P EF1 ⁇ - L (long promoter, 531 bp);
- B New vector pPVLV2 including P EF1 ⁇ -S (short promoter, 212 bp).
- the expected PCR products (F1-F5: PCR fragments) were generated from cDNA reverse transcribed using random hexamer primers to identify the integrity of the viral vector genome.
- Figure 2 shows the difference between pPVLV1 and pPVLV2.
- A The expected DNA fragment was amplified from the reverse transcription cDNA of the viral genome. Defective loci were observed in the viral gene fragment containing PEF1 ⁇ - L. DNA fragments with unexpected sizes are indicated by arrows (left panel).
- B Comparison of the percentage of CAR-19-expressing cells and
- C Titration of each vector 48 hours after transduction of 293T cells.
- Figure 3 shows the structure and luciferase activity of CAR-19-Fluc.
- A and (B) used a bicistronic construct encoding CAR-19 cloned upstream of the P2A-Fluc box.
- C Schematic representation of the expressed CAR-19 and Fluc molecules.
- D Luciferase activity measured 48 hours after lentiviral vector transduction of 293T cells.
- Figure 4 shows the structure of CAR-19-DNRII and the viral vector.
- A) and (B) show vector maps of CAR-19 co-expressing truncated TGFBRII (DNRII).
- C Schematic diagram of the co-expressed CAR-19 and DNRII molecules.
- Figure 5 shows the transduction efficiency of CAR-19 and DNRII expression in transduced 293T cells.
- the numbers in the figure indicate the percentage of positive cells of CAR-19 (top) or DNRII (bottom) relative to the negative control of untransduced 293T cells. Results from representative experiments from ten independent experiments are given.
- Figure 6 shows the expression of CAR-19 and DNRII in transduced T cells.
- Activated T cells were transduced with a lentiviral vector to express CAR-19 or CAR-19-DNRII and evaluated by flow cytometry.
- the numbers in the figure indicate the percentage of positive cells of CAR-19 (top) or DNRII (bottom) relative to the negative control of untransduced T cells. The results represent three independent experiments.
- Figure 7 shows cell viability and counts after transduction of CAR-19 or CAR-19-DNRII vectors. Data are expressed as mean ⁇ SD.
- Figure 8 shows that DNRII reduces TGF- ⁇ 1-induced SMAD2 phosphorylation.
- Figure 9 shows mRNA levels of IFN- ⁇ and TNF- ⁇ in CAR-T-19 and CAR-T-19-DNRII cells. Data are expressed as mean ⁇ SEM.
- FIG. 10 shows the antigen-specific killing of CD19 + tumor cells by CAR-T-19 and CAR-T-19-DNRII cells in the presence of TGF- ⁇ 1.
- EuTDA cytotoxicity assay measures cell lysis activity. T cells were taken 3 days before the measurement and cultured with rhTGF- ⁇ 1 (10 ng / ml) for 72 hours. Target cells were labeled with BATDA reagent for 15 minutes, and then transduced T cells were added as effector cells at the specified E: T ratio. After 4 hours of incubation, lysis was determined.
- the present invention provides a method for preparing a therapeutic T cell that specifically targets a cancer-associated antigen, comprising co-expressing in a T cell a foreign cancer-associated antigen-specific receptor protein and a dominant negative TGF- ⁇ II Receptor.
- the cancer-related antigens include, but are not limited to, CD16, CD64, CD78, CD96, CLL1, CD116, CD117, CD71, CD45, CD71, CD123, CD138, ErbB2 (HER2 / neu), carcinoembryonic antigen (CEA), epithelial cell adhesion Epidermal molecule (EpCAM), epidermal growth factor receptor (EGFR), EGFR variant III (EGFRvIII), CD19, CD20, CD30, CD40, disialoganglioside GD2, ductal epithelial mucin, gp36, TAG-72 , Glycosphingolipid, glioma-associated antigen, ⁇ -human chorionic gonadotropin, alpha fetal globulin (AFP), exogenous lectin-reactive AFP, thyroglobulin, RAGE-1, MN-CA IX , Human telomerase reverse transcriptase, RU1, RU2 (AS), intestinal
- the dominant negative TGF- ⁇ II receptor refers to TGF that can compete with TGF- ⁇ RII for binding to a TGF- ⁇ ligand (such as TGF- ⁇ 1), but cannot perform the signaling function of TGF- ⁇ RII -a variant of the ⁇ II receptor.
- the intracellular signaling domain of the dominant negative TGF- ⁇ type II receptor is mutated, thereby losing intracellular signaling capabilities.
- the dominant negative TGF- ⁇ type II receptor lacks the intracellular signaling domain of the TGF- ⁇ type II receptor.
- the dominant negative TGF- ⁇ type II receptor comprises the amino acid sequence shown in SEQ ID NO: 18.
- the "foreign cancer-associated antigen-specific receptor protein" described in the present invention may be an exogenous T cell receptor (TCR) or a chimeric antigen receptor (CAR).
- TCR T cell receptor
- CAR chimeric antigen receptor
- T cell receptor is also called T cell antigen receptor. It is a molecular structure that T cells specifically recognize and bind to the antigen peptide-MHC molecule. It usually exists on the surface of T cells in the form of a complex with CD3 molecules.
- the TCR of most T cells consists of ⁇ and ⁇ peptide chains, and the TCR of a few T cells consists of ⁇ and ⁇ peptide chains.
- CAR Chimeric antigen receptor
- the TCR is a TCR that specifically binds a cancer-associated antigen (usually comprising a chain that typically contains alpha and beta).
- the CAR may comprise an extracellular antigen-binding domain directed against a cancer-associated antigen.
- the extracellular antigen-binding domain may be, for example, a monoclonal antibody, a synthetic antibody, a human antibody, a humanized antibody, a single-domain antibody, an antibody single-chain variable region (scFV), and an antigen-binding fragment thereof.
- the extracellular antigen-binding domain may be derived from one or more known antibodies including any commercially available antibody, such as FMC63, rituximab, alemtuzumab, elixir Epratuzumab, trastuzumab, bivatuzumab, cetuximab, labetuzumab, parizumab ( palivizumab), sevirumab, tuvirumab, basiliximab, daclizumab, infliximab, omalimumab ( (omalizumab), efalizumab, keliximab, siplizumab, natalizumab, clenoliximab, pemalimumab ( pemtumomab), Edrecolomab, Cantuzumab, and the like.
- FMC63 FMC63
- rituximab alemtuzumab
- the CAR further comprises a transmembrane domain and an intracellular signal transduction domain.
- the intracellular signal transduction domain of the CAR according to the present invention is responsible for intracellular signal transduction after the extracellular ligand binding domain binds to the target, leading to activation of immune cells and immune responses.
- the intracellular signal transduction domain has the ability to activate at least one normal effector function of a CAR-expressing immune cell.
- the effector function of a T cell may be cytolytic activity or auxiliary activity, including secretion of cytokines.
- the intracellular signal transduction domain for CAR can be a cytoplasmic sequence, such as, but not limited to, a cytoplasmic sequence of a T cell receptor and a co-receptor (they work in concert to initiate signal transduction after antigen receptor junction), As well as derivatives or variants of any of these sequences and any synthetic sequences having the same functional capacity.
- the intracellular signal transduction domain includes two different types of cytoplasmic signal transduction sequences: those that initiate antigen-dependent primary activation, and those that act in an antigen-independent manner to provide secondary or costimulatory signals.
- the primary cytoplasmic signal transduction sequence may include a signal transduction motif called an immune receptor tyrosine activation motif called ITAM.
- Non-limiting examples of ITAM used in the present invention may include those derived from TCR ⁇ , FcR ⁇ , FcR ⁇ , FcR ⁇ , CD3 ⁇ , CD3 ⁇ , CD5, CD22, CD79a, CD79b, and CD66d.
- the intracellular signal transduction domain of the CAR may include a CD3 ⁇ signal transduction domain.
- the intracellular signal transduction domain of the CAR of the invention further comprises a co-stimulatory domain.
- the costimulatory domain is selected from a 41BB costimulatory domain or a CD28 costimulatory domain.
- a CAR is expressed on the surface of cells. Therefore, a CAR can include a transmembrane domain.
- Suitable transmembrane domains of the CARs of the present invention have the ability to: (a) be expressed on the cell surface, preferably immune cells, such as but not limited to lymphocytes or natural killer (NK) cells, and (b) a ligand binding domain It interacts with intracellular signal transduction domains to direct the immune cells to respond to predetermined target cells.
- Transmembrane domains can be derived from natural or synthetic sources. The transmembrane domain can be derived from any membrane-bound protein or transmembrane protein.
- a transmembrane domain can be derived from a subunit of a T cell receptor, such as an ⁇ subunit, a ⁇ subunit, a ⁇ , or a ⁇ subunit, a polypeptide that makes up the CD3 complex, and p55 ( ⁇ chain), p75 ( ⁇ chain) or ⁇ , a subunit chain of the Fc receptor, especially the Fc ⁇ receptor III or CD protein.
- the transmembrane domain may be synthetic and may include mainly hydrophobic residues such as leucine and valine.
- the transmembrane domain is derived from a human CD8 ⁇ chain.
- the transmembrane domain may further include a hinge region between an extracellular ligand-binding domain and the transmembrane domain.
- the hinge region is, for example, from the extracellular region of CD8, CD4 or CD28. In some embodiments, the hinge region is part of a human CD8 ⁇ chain.
- the CAR used in the present invention may include extracellular antigen-binding domains such as scFv, CD8 hinge and transmembrane domain, CD3 ⁇ signal transduction domain, and 4-1BB that specifically bind to cancer-associated antigens. Costimulatory domain.
- the CAR comprises an extracellular antigen-binding domain directed against CD19. In some specific embodiments, the CAR comprises the amino acid sequence shown in SEQ ID NO: 16.
- the method comprises transducing the T cells with a lentiviral particle comprising a lentiviral vector, the lentiviral vector comprising a nucleotide sequence encoding a fusion polypeptide comprising a self-cleaving peptide
- the exogenous antigen-specific receptor protein and the dominant negative TGF- ⁇ type II receptor are linked, thereby co-expressing the exogenous antigen-specific receptor protein and the dominant negative TGF- in the T cells.
- ⁇ II receptor a lentiviral particle comprising a lentiviral vector, the lentiviral vector comprising a nucleotide sequence encoding a fusion polypeptide comprising a self-cleaving peptide
- a "lentiviral vector” refers to a non-replicating vector that is used to transduce a transgene comprising a cis-acting lentiviral RNA or DNA sequence to a host cell, and requires the lentiviral protein to be provided in trans form ( (Eg Gag, Pol and / or Env).
- Lentiviral vectors lack coding sequences for functional Gag, Pol, and Env proteins. Lentiviral vectors can exist in the form of RNA or DNA molecules, depending on the stage of production or development of the retroviral vector.
- Lentiviral vectors can be in the form of a recombinant DNA molecule, such as a plasmid (eg, a transfer plasmid vector).
- Lentiviral vectors can be in the form of lentiviral particle vectors, such as RNA molecules in a complex of lentivirus and other proteins.
- a lentiviral vector corresponding to a modified or recombinant lentiviral particle comprises a genome consisting of two copies of single-stranded RNA.
- RNA sequences can be obtained by transcription from a double-stranded DNA sequence (proviral vector DNA) inserted into the genome of a host cell, or can be obtained by transient expression of plasmid DNA (plasmid vector DNA) in a transformed host cell.
- a lentiviral vector may also refer to a DNA sequence integrated into a host cell.
- Lentiviral vectors can be derived from lentiviruses, especially human immunodeficiency virus (HIV-1 or HIV-2), simian immunodeficiency virus (SIV), equine infectious encephalitis virus (EIAV), goat arthritis encephalitis virus ( CAEV), bovine immunodeficiency virus (BIV), and feline immunodeficiency virus (FIV), which are modified to remove genetic determinants involved in pathogenicity and introduced into foreign expression cassettes.
- HIV-1 or HIV-2 human immunodeficiency virus
- SIV simian immunodeficiency virus
- EIAV equine infectious encephalitis virus
- CAEV goat arthritis encephalitis virus
- BIV bovine immunodeficiency virus
- FV feline immunodeficiency virus
- Self-cleaving peptide as used herein means a peptide that can achieve self-cleaving within a cell.
- the self-cleaving peptide may contain a protease recognition site so that it is recognized and specifically cleaved by a protease in the cell.
- the self-cleaving peptide may be a 2A polypeptide.
- 2A polypeptides are a class of short peptides derived from viruses that undergo self-cleaving during translation. When 2A polypeptide is used to link two different proteins of interest in the same reading frame, the two proteins of interest are generated at a ratio of almost 1: 1.
- Commonly used 2A polypeptides can be P2A from porcine techovirus-1, T2A from Thosea asnagna virus, and E2A from equine rhinitis virus And F2A from foot-and-mouth disease virus. Among them, P2A has the highest cutting efficiency and is therefore preferred.
- a variety of functional variants of these 2A polypeptides are also known in the art, and these variants can also be used in the present invention.
- Transduced cells express both proteins. Because if the two proteins are transduced to cells in different vectors, some cells may only express exogenous cancer-associated antigen-specific receptor proteins, and some cells only express dominant negative TGF- ⁇ type II receptors. The proportion of cells expressing both proteins will be low. In addition, if the expression of two proteins is driven by different promoters in the same vector, due to the difference in promoter efficiency, the proportion of cells that simultaneously express the two proteins will also be reduced.
- the nucleotide sequence encoding the fusion polypeptide is operably linked to a truncated EF1 ⁇ promoter.
- the inventors have surprisingly found that transduction of cells, such as T cells, using a lentiviral vector containing a long EF1 ⁇ promoter (such as SEQ ID NO 7), an abnormality in this promoter region can occur, resulting in the introduction of foreign genes into the cell ( In particular, a gene encoding a CAR or a fusion protein thereof) has a low expression rate.
- the use of a truncated EF1 ⁇ promoter can avoid this phenomenon and significantly increase the expression rate of the introduced foreign gene.
- the truncated EF1 ⁇ promoter is an EF1 ⁇ core promoter comprising the nucleotide sequence shown in SEQ ID NO: 13.
- the lentiviral vector further comprises at least one element selected from the group consisting of 5'LTR, ⁇ sequence, RRE sequence, cPPT / CTS sequence, WPRE sequence, and 3'LTR.
- the 5 ' LTR may be a truncated 5 ' LTR from HIV-1, which is essential for viral transcription, reverse transcription and integration.
- the ⁇ element is a packaging signal for HIV-1 and is essential for lentiviral vector packaging.
- RRE is required for Rev-dependent export of the mRNA of the virus transcript from the nucleus to the cytoplasm.
- the cPPT / CTS sequence can be cPPT / CTS of HIV1, which can improve the efficiency of vector integration and transduction.
- WPRE post-transcriptional regulatory element from marmot hepatitis virus
- the 3 ' LTR may be a self-inactivating 3 ' LTR from HIV-1, which is essential for viral transcription, reverse transcription and integration, and contains safety measures to prevent viral replication.
- the lentiviral vector comprises an operably linked 5'LTR, ⁇ element, RRE element, cPPT / CTS element, the truncated EF1 ⁇ promoter, and the nucleotide sequence encoding the fusion polypeptide , WPRE element and 3'LTR.
- the 5 ′ LTR comprises the nucleotide sequence shown in SEQ ID NO: 3 or 11; the ⁇ element comprises the nucleotide sequence shown in SEQ ID NO: 4 or 12; the RRE element Contains the nucleotide sequence shown in SEQ ID NO: 5; the cPPT / CTS element contains the nucleotide sequence shown in SEQ ID NO: 6; the WPRE element contains the nucleotide sequence shown in SEQ ID NO: 9 or 14 ; Wherein the 3'LTR comprises the nucleotide sequence shown in SEQ ID NO: 10 or 15.
- the lentiviral vector comprises an operably linked 5'LTR comprising the nucleotide sequence shown in SEQ ID NO: 11, a ⁇ element comprising the nucleotide sequence shown in SEQ ID NO: 12, comprising
- the RRE element of the nucleotide sequence shown in SEQ ID NO: 5 includes the cPPT / CTS element of the nucleotide sequence shown in SEQ ID NO: 6 and the truncated EF1 ⁇ of the nucleotide sequence shown in SEQ ID NO: 13
- the lentiviral vector is derived from SEQ ID NO: 2, wherein the nucleotide sequence encoding CAR-19 at position 2,042-3,499 of SEQ ID NO: 2 may be the nucleoside encoding the fusion polypeptide Acid sequence substitution.
- the T cells of the invention can be obtained from a number of non-limiting sources by a variety of non-limiting methods, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, umbilical cord blood, thymus tissue, ascites, pleural effusion, spleen tissue, and tumors.
- the cells can be derived from a healthy donor or from a patient diagnosed with cancer.
- the cells can be part of a mixed population of cells that exhibit different phenotypic characteristics.
- T cells can be obtained by isolating peripheral blood mononuclear cells (PBMC), and then activating and expanding with specific antibodies.
- PBMC peripheral blood mononuclear cells
- the T cell is derived from an autologous cell of a subject.
- autologous means that the cell, cell line, or cell population used to treat a subject is derived from the subject.
- the T cells are derived from an allogeneic cell, for example, derived from a donor compatible with the subject's human leukocyte antigen (HLA). Donor-derived cells can be transformed into non-allogene-reactive cells using standard protocols and replicated as needed to produce cells that can be administered to one or more patients.
- HLA human leukocyte antigen
- the present invention provides a therapeutic T cell that specifically targets a cancer-associated antigen, which is produced by the method described above of the present invention.
- the present invention provides a therapeutic T cell that specifically targets a cancer-associated antigen, which co-expresses an exogenous cancer-associated antigen-specific receptor protein and a dominant negative TGF- ⁇ type II receptor
- said Therapeutic T cells comprise a lentiviral vector (eg, a lentiviral vector integrated into the genome of a cell), said lentiviral vector comprising a nucleotide sequence encoding a fusion polypeptide, said fusion polypeptide comprising said exogenous molecule linked by a self-cleaving peptide Cancer-associated antigen-specific receptor protein and the dominant negative TGF- ⁇ type II receptor.
- the dominant negative TGF- ⁇ type II receptor in the therapeutic T cell lacks an intracellular signaling domain of a TGF- ⁇ type II receptor.
- the dominant negative TGF- ⁇ type II receptor comprises the amino acid sequence shown in SEQ ID NO: 18.
- the exogenous cancer-associated antigen-specific receptor protein in the therapeutic T cell is selected from the group consisting of a T cell receptor (TCR) and a chimeric antigen receptor (CAR).
- TCR T cell receptor
- CAR chimeric antigen receptor
- the TCR specifically binds a cancer-associated antigen
- the CAR comprises an extracellular antigen-binding domain against a cancer-associated antigen.
- the CAR includes an extracellular antigen-binding domain such as scFv, a CD8 hinge and a transmembrane domain, a CD3 ⁇ signal transduction domain, and a 4-1BB costimulatory domain that specifically binds a cancer-associated antigen.
- an extracellular antigen-binding domain such as scFv, a CD8 hinge and a transmembrane domain, a CD3 ⁇ signal transduction domain, and a 4-1BB costimulatory domain that specifically binds a cancer-associated antigen.
- the cancer-associated antigen is selected from the group consisting of CD16, CD64, CD78, CD96, CLL1, CD116, CD117, CD71, CD45, CD71, CD123, CD138, ErbB2 (HER2 / neu), Carcinoembryonic Antigen (CEA) , Epithelial cell adhesion molecule (EpCAM), epidermal growth factor receptor (EGFR), EGFR variant III (EGFRvIII), CD19, CD20, CD30, CD40, disialoganglioside GD2, ductal epithelial mucin, gp36 , TAG-72, glycosphingolipid, glioma-associated antigen, ⁇ -human chorionic gonadotropin, alpha fetal globulin (AFP), exogenous lectin-reactive AFP, thyroglobulin, RAGE-1, MN-CA IX, human telomerase reverse transcriptase, RU1, RU2 (AS), intestinal carb
- the CAR comprises an extracellular antigen-binding domain directed against CD19, for example, the CAR comprises the amino acid sequence shown in SEQ ID NO: 16.
- the self-cleaving peptide is a 2A polypeptide, for example, the self-cleaving peptide is selected from a P2A, F2A, E2A, or T2A polypeptide, or a functional variant thereof.
- the nucleotide sequence encoding the fusion polypeptide is operably linked to a truncated EF1 ⁇ promoter, for example, the truncated EF1 ⁇ promoter comprises a nucleotide represented by SEQ ID NO: 13 Sequenced EF1 ⁇ core promoter.
- the lentiviral vector further comprises at least one element selected from the group consisting of 5'LTR, ⁇ element, RRE element, cPPT / CTS element, WPRE element, and 3'LTR.
- the lentiviral vector comprises an operably linked 5'LTR, ⁇ element, RRE element, cPPT / CTS element, the truncated EF1 ⁇ promoter, and the nucleotide sequence encoding the fusion polypeptide , WPRE element and 3'LTR.
- the 5'LTR comprises the nucleotide sequence shown in SEQ ID NO: 3 or 11; the ⁇ element comprises the nucleotide sequence shown in SEQ ID NO: 4 or 12; and the RRE element comprises The nucleotide sequence shown in SEQ ID NO: 5; the cPPT / CTS element contains the nucleotide sequence shown in SEQ ID NO: 6; the WPRE element contains the nucleotide sequence shown in SEQ ID NO: 9 or 14; Wherein, the 3'LTR comprises the nucleotide sequence shown in SEQ ID NO: 10 or 15.
- the lentiviral vector comprises an operably linked 5'LTR comprising the nucleotide sequence shown in SEQ ID NO: 11, a ⁇ element comprising the nucleotide sequence shown in SEQ ID NO: 12, comprising
- the RRE element of the nucleotide sequence shown in SEQ ID NO: 5 includes the cPPT / CTS element of the nucleotide sequence shown in SEQ ID NO: 6 and the truncated EF1 ⁇ of the nucleotide sequence shown in SEQ ID NO: 13
- the present invention provides a pharmaceutical composition
- a pharmaceutical composition comprising a therapeutic T cell of the present invention that specifically targets a cancer-associated antigen, and a pharmaceutically acceptable carrier.
- pharmaceutically acceptable carrier includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible.
- the carrier is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (such as by injection or infusion).
- the present invention provides the use of a therapeutic T cell of the present invention that specifically targets a cancer-associated antigen or a pharmaceutical composition of the present invention in the manufacture of a medicament for treating cancer in a subject.
- subject refers to an organism that has or is susceptible to a disease (such as cancer) that can be treated by the cells, methods, or pharmaceutical compositions of the invention.
- a disease such as cancer
- Non-limiting examples include humans, cattle, rats, mice, dogs, monkeys, goats, sheep, cows, deer, and other non-mammals.
- the subject is a human.
- the present invention provides a method of treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of a therapeutic T cell of the present invention that specifically targets a cancer-associated antigen or a medicament of the present invention combination.
- a “therapeutically effective amount” or “therapeutically effective dose” or “effective amount” refers to an amount of a substance, compound, material, or cell that is at least sufficient to produce a therapeutic effect after administration to a subject. Therefore, it is an amount necessary to prevent, cure, ameliorate, block or partially block the symptoms of a disease or disorder.
- an "effective amount" of a cell or pharmaceutical composition of the invention preferably results in a reduction in the severity of the symptoms of the disease, an increase in the frequency and duration of the asymptomatic phase of the disease, or the prevention of injury or disability due to the pain of the disease.
- an "effective amount" of a cell or pharmaceutical composition of the present invention preferably inhibits tumor cell growth or tumor growth by at least about 10%, preferably by at least about 20%, and more by comparison with an untreated subject. It is preferably at least about 30%, more preferably at least about 40%, more preferably at least about 50%, more preferably at least about 60%, more preferably at least about 70%, and still more preferably at least about 80%.
- the ability to inhibit tumor growth can be evaluated in animal model systems that predict efficacy in human tumors. Alternatively, it can also be evaluated by examining the ability to inhibit the growth of tumor cells, and this inhibition can be determined in vitro by tests known to those skilled in the art.
- the dosage level of cells in the pharmaceutical composition of the present invention may be changed to obtain an amount of active ingredient that can effectively achieve a desired therapeutic response to a particular patient, composition, and mode of administration, but is non-toxic to the patient.
- the selected dosage level depends on a variety of pharmacokinetic factors, including the activity of the particular composition of the invention applied, the route of administration, the time of administration, the excretion rate of the particular compound applied, the duration of the treatment, and the particular application
- Other drugs, compounds and / or materials in combination with the composition the age, sex, weight, condition, general health and medical history of the patient being treated, and similar factors well known in the medical field.
- the cancer is selected from the group consisting of lung cancer, ovarian cancer, colon cancer, rectal cancer, melanoma, kidney cancer, bladder cancer, breast cancer, liver cancer, lymphoma, malignant hematological disease, head and neck cancer, Glioma, gastric cancer, nasopharyngeal cancer, laryngeal cancer, cervical cancer, uterine body tumor, osteosarcoma, bone cancer, pancreatic cancer, skin cancer, prostate cancer, uterine cancer, anal cancer, testicular cancer, fallopian tube cancer, intrauterine Membrane cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin's lymphoma, esophageal cancer, small bowel cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer Chronic or acute leukemia (including acute myeloid leukemia, chronic mye
- Lentiviral vectors used to transduce CAR should contain the desired CAR transgene and be capable of being expressed in cells.
- Two third-generation lentiviral vectors for CAR expression were designed, the old vector pPVLV1 ( Figure 1A) and the new vector pPVLV2 ( Figure 1B).
- pPVLV1 contains a 531 bp long human elongation factor 1 ⁇ (EF1 ⁇ ) promoter and pPVLV1 contains a 212 bp truncated human EF1 ⁇ promoter.
- Table 1 The various elements contained in the two vectors and their descriptions are shown in Table 1 below.
- the CAR to be expressed in the examples of the present application includes a scFv targeting CD19, a hinge and transmembrane domain of human CD8, an intracellular domain 4-1BB, and CD3 ⁇ .
- the amino acid of the CD19-targeting CAR is shown in SEQ ID NO: 16, and the nucleotide sequence is shown in SEQ ID NO: 8.
- pPVLV1 vector including EF1 ⁇ long promoter (5,320bp, SEQ ID NO: 1)
- pPVLV2 vector including EF1 ⁇ short promoter (4,417bp, SEQ ID NO: 2)
- Lentiviral supernatants were generated by transfecting 293T cells with a gag / pol packaging plasmid, a VSV-G envelope plasmid, and a transfer construct containing a lentiviral vector sequence as described above. Briefly, the DNA mixture was mixed in Opti-MEM (Life Technologies, Gaithersburg, MD, USA) and mixed with an equal volume of Opti-MEM containing Lipofectamine 3000 (Life Technologies). After 15 minutes of incubation at room temperature, the resulting mixture was applied to 293T cells. Lentivirus-containing medium was collected 24 hours after transfection. After each collection, the supernatant was filtered through a PVDF membrane (0.45 ⁇ m). The lentiviral harvests were combined and stored at 4 ° C, followed by ultracentrifugation at 20,000 xg for 1 hour and 30 minutes. The lentiviral particles were resuspended in PBS.
- Figure 1 shows a schematic diagram of the structure of two lentiviral vectors and a strategy for verifying the integrity of the lentivirus by overlapping PCR products.
- Appropriate primers were designed to amplify overlapping fragments F1-F5 from cDNA reverse transcribed using random primers.
- a PCR product of the expected size can demonstrate the integrity of the lentivirus.
- FIG. 2A shows each DNA fragment amplified from the cDNA reversely transcribed from the viral genome.
- defective gene loci were observed in viral gene fragments containing P EF1 ⁇ -L (long promoter). Arrows indicate unexpected DNA fragments (left). This phenomenon was not observed in viral gene fragments containing P EF1 ⁇ -S (short promoter). Such defective viral genomes may affect titer and transduction efficiency.
- lentiviral titration 2 ⁇ 10 6 293T cells were seeded into each well of a 6-well plate and transduced with a series of volumes of concentrated lentivirus. 48 hours after transduction, 293T cells were isolated from the plate. The presence of CAR was detected by flow cytometry using Alexa Fluor 488 labeled goat anti-human IgG F (ab) 2 . Viral genomic RNA from 5'LTR to 3'LTR was examined using conventional PCR.
- the results are shown in Figures 2B and C.
- the transduction efficiency (proportion of CAR-expressing cells) of the virus with P EF1 ⁇ - L (based on pPVLV1 vector) was only 9.95%, which was far lower than 70.4% of the virus with P EF1 ⁇ - S (based on pPVLV2 vector).
- the titer of the virus with P EF1 ⁇ - L (based on the pPVLV1 vector) was also significantly lower than that of the virus with P EF1 ⁇ - S (based on the pPVLV2 vector). It is shown that pPVLV2 vector is superior to pPVLV1 vector, and this may be caused by EF1 ⁇ promoters of different lengths.
- FIG. 3D shows the luciferase activity measured 48 hours after transduction of two lentiviral vectors into 293T cells. The results showed that cells transduced with the lentiviral vector with P EF1 ⁇ - S had significantly stronger fluorescence than cells transduced with the lentiviral vector with P EF1 ⁇ - L. It was further proved that P EF1 ⁇ -S significantly improved the expression of the transgene in cells.
- TGF- ⁇ is an important T cell inhibitory factor, which may cause the killing effect of therapeutic T cells on target cells to weaken or disappear.
- TGF- ⁇ is widely expressed in a variety of tumor tissues, and significantly inhibits the killing activity of tumor-specific T cells on tumor cells, which is an important reason for the failure of immunotherapy.
- the dominant negative TGF- ⁇ type II receptor (dominant TGF- ⁇ receptor type II (DNRII)) is a negative regulator of TGF- ⁇ , which can inhibit the inhibitory effect of TGF- ⁇ on T cells.
- DNRII dominant negative TGF- ⁇ type II receptor
- the following examples investigate the effect of co-expression of CAR and DNRII in T cells.
- the amino acid sequence of DNRII is shown in SEQ ID NO: 17, and its nucleotide sequence is shown in SEQ ID NO: 18.
- FIG. 4 shows a schematic diagram of the structure of the CAR-19 and DNRII molecules.
- DNRII lacks the intracellular serine / threonine kinase domain of TGFBRII and cannot transmit signals downstream.
- CAR-19 and DNRII coding sequences are separated by 2A coding sequences, placed in the same open reading frame, and driven by the same promoter, which can ensure that the obtained transduced cells express CAR-19 and DNRII at the same time. Because if CAR-19 and DNRII are transduced to cells in different vectors, some cells may only express CAR-19, some cells only express DNRII, and the proportion of cells co-expressing the two proteins will be very low. In addition, if the expression of two proteins is driven by different promoters in the same vector, due to the difference in promoter efficiency, the proportion of cells that simultaneously express the two proteins will also be reduced.
- the two CAR-19-DNRII lentiviral vectors were transduced to 293T cells with equal MOI (multiple infections).
- CAR or DNRII expression was detected by flow cytometry using a labeled goat anti-human IgG F (ab) 2 or anti-DNRII antibody using a MACSQuant analyzer 10 and the data was analyzed with FlowJo software.
- PBMCs Human peripheral blood mononuclear cells
- PBMCs Human peripheral blood mononuclear cells
- rhIL- 2 200 IU / mL
- IMSF100 serum-free medium LONZA, Belgium
- CAR-19 and CAR-19-DNRII lentiviral supernatants were added for transduction. It was then centrifuged at 1,200 ⁇ g for 2 hours at 32 ° C. After 24 hours, the supernatant containing the viral vector was removed.
- Cells were suspended at 3 ⁇ 10 5 cells / ml in a medium containing rhIL-2 (200IU / mL), fresh medium was replaced every 2 to 3 days, and cultured for 12 days to obtain a CAR expressing a CAR-19 molecule.
- -T-19 cells and CAR-T-19-DNRII cells co-expressing CAR-19 molecules and DNRII molecules.
- PBMC cultured under the same culture conditions but without gene transduction was used as a control (NC).
- Flow cytometry was used to detect the expression of each protein molecule of CAR-T cells obtained after transduction.
- Cells were stained with propidium iodide (PI) every 2-3 days, and cell viability was measured by flow cytometry. During the cell culture, trypan blue staining was performed every 2-3 days (three replicates per sample), and the number of cells was calculated (mean ⁇ SD).
- PI propidium iodide
- this example determines that the backbone of the pPVLV2 vector (including P EF1 ⁇ -S) is particularly suitable for CAR expression in cells, such as T cells, and is particularly suitable for co-expression of CAR and other proteins such as DNRII.
- placing the CAR and DNRII coding sequences in the same open reading frame can achieve a high co-expression rate of the two molecules.
- TFG- ⁇ The inhibitory effect of TFG- ⁇ on T cells is achieved through the phosphorylation of SMAD2 molecules after TFG- ⁇ binds to its receptor.
- CAR-T-19 cells and CAR-T-19-DNRII cells 9 days after transduction were incubated with recombinant human TFG- ⁇ 1 (10ng / ml) for 24 hours to perform expression of phosphorylated SMAD2 (pSMAD2). analysis.
- pSMAD2 phosphorylated SMAD2
- the Bradford assay kit (Sigma-Aldrich) was used to measure the protein concentration of whole cell lysates. An equal amount of protein was loaded into the wells of an SDS-PAGE gel, and the separated protein was transferred to a PVDF membrane (Thermo Scientific). The membrane was blocked with 10% (w / v) skim milk in TBST and then incubated with primary antibodies (anti-pSMAD2 and anti-SMAD2; from Cellsignaling Technologies, Danvers, MA, USA; all diluted 1: 1000) overnight at 4 ° C. .
- the membrane was then washed with TBST and incubated with HRP-conjugated goat anti-rabbit IgG (1: 2000 dilution; Cellular Signaling Technologies) at room temperature for 2 hours.
- the membrane was then exposed to ECL reagent (Thermo Scientific) and the resulting signal was detected using a luminescence image analyzer (LAS-4000, Fuji Film, Tokyo, Japan).
- Fig. 8 The level of pSAMD2 in CAR-T-19-DNRII cells was significantly lower than that of CAR-T-19 cells. This indicates that the expression of DNRII inhibits the phosphorylation of SMAD2, a key signaling molecule in the TGF- ⁇ signaling pathway.
- Example 5 Expression of IFN- ⁇ and TNF- ⁇ in CAR-T-19-DNRII cells and CAR-T-19 cells treated with recombinant human TGF- ⁇ 1
- IFN- ⁇ and TNF- ⁇ are the hallmark cytokines that T cells kill target cells.
- the high expression levels of these two cytokines indicate that T cells have high killing potential for target cells, and conversely, low killing potential.
- RNA samples were analyzed using specific primers and One-step SensiFAST SYBR Low-ROX kit (Bioline, Maryland, USA)
- Real-time quantitative RT-PCR analysis was performed using a QuantStudio3 real-time PCR detection system (Applied Biosystems, Foster City, CA, USA). 18s rRNA was amplified as an internal control. The expression level was calculated by the ⁇ Ct method, and the expression multiple was obtained using the formula 2- ⁇ Ct. All experiments were performed in triplicate.
- Example 6 CAR-T-19-DNRII cells and CAR-T-19 cells treated with recombinant human TGF- ⁇ 1 specifically kill tumor target cells
- Target cell killing experiments were performed using CAR-T-19 cells and CAR-T-19-DNRII cells transduced for 12 days.
- a TDA release assay was performed to determine the cytotoxic activity of CAR-T-19 cells and CAR-T-19-DNRII cells to K562 or CD19 + -K562 in the presence of TGF- ⁇ 1.
- CAR-T-19 cells and CAR-T-19-DNRII cells were incubated with recombinant human TGF- ⁇ 1 (10ng / ml) for 72 hours, respectively.
- Target cells were labeled with BA-TDA (Perkin Elmer, Norwalk, Connecticut, USA) for 15 minutes, and the effector cells (T cells) were 20: 1, 10: 1, 5: 1, and 2.5: 1: target cells (tumor Cells were mixed with effector cells, and TDA release (target cell lysis) was detected after 4 hours of incubation.
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)
- Cell Biology (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Epidemiology (AREA)
- Biomedical Technology (AREA)
- Mycology (AREA)
- Gastroenterology & Hepatology (AREA)
- Toxicology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Oncology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Hematology (AREA)
- Virology (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
Description
Claims (32)
- 一种制备特异性靶向癌症相关抗原的治疗性T细胞的方法,包括在T细胞中共表达外源癌症相关抗原特异性受体蛋白和显性负性TGF-βII型受体。
- 权利要求1的方法,其中所述显性负性TGF-βII型受体缺失TGF-βII型受体的胞内信号传导结构域,例如,所述显性负性TGF-βII型受体包含SEQ ID NO:18所示的氨基酸序列。
- 权利要求1或2的方法,其中所述外源癌症相关抗原特异性受体蛋白选自T细胞受体(TCR)和嵌合抗原受体(CAR)。
- 权利要求3的方法,所述TCR特异性结合癌症相关抗原,所述CAR包含针对癌症相关抗原的细胞外抗原结合结构域。
- 权利要求4的方法,所述CAR包括特异性结合癌症相关抗原的细胞外抗原结合结构域如scFv、CD8铰链和跨膜结构域、CD3ζ信号转导结构域、和4-1BB共刺激结构域。
- 权利要求1-5任一项的方法,其中所述癌症相关抗原选自CD16、CD64、CD78、CD96、CLL1、CD116、CD117、CD71、CD45、CD71、CD123、CD138、ErbB2(HER2/neu)、癌胚抗原(CEA)、上皮细胞粘附分子(EpCAM)、表皮生长因子受体(EGFR)、EGFR变体III(EGFRvIII)、CD19、CD20、CD30、CD40、双唾液酸神经节苷脂GD2、导管上皮粘蛋白、gp36、TAG-72、鞘糖脂、神经胶质瘤相关的抗原、β-人绒毛膜促性腺激素、α胎儿球蛋白(AFP)、外源凝集素反应性AFP、甲状腺球蛋白、RAGE-1、MN-CA IX、人端粒酶逆转录酶、RU1、RU2(AS)、肠羧基酯酶、mut hsp70-2、M-CSF、前列腺酶(prostase)、前列腺酶特异性抗原(PSA)、PAP、NY-ESO-1、LAGA-1a、p53、Prostein、PSMA、存活和端粒酶、前列腺癌肿瘤抗原-1(PCTA-1)、MAGE、ELF2M、嗜中性粒细胞弹性蛋白酶、肝配蛋白B2、CD22、胰岛素生长因子(IGF1)-I、IGF-II、IGFI受体、间皮素、呈递肿瘤特异性肽表位的主要组织相容性复合体(MHC)分子、5T4、ROR1、Nkp30、NKG2D、肿瘤基质抗原、纤维连接蛋白的额外结构域A(EDA)和额外结构域B(EDB)、腱生蛋白-C的A1结构域(TnC A1)、成纤维细胞相关蛋白(fap)、CD3、CD4、CD8、CD24、CD25、CD33、CD34、CD133、CD138、Foxp3、B7-1(CD80)、B7-2(CD86)、GM-CSF、细胞因子受体、内皮因子、BCMA(CD269、TNFRSF17)、TNFRSF17(UNIPROT Q02223)、SLAMF7(UNIPROT Q9NQ25)、GPRC5D(UNIPROT Q9NZD1)、FKBP11(UNIPROT Q9NYL4)、KAMP3、ITGA8(UNIPROT P53708)和FCRL5(UNIPROT Q68SN8)。
- 权利要求6的方法,其中所述CAR包含针对CD19的细胞外抗原结合结构域,例如,所述CAR包含SEQ ID NO:16所示氨基酸序列。
- 权利要求1-7中任一项的方法,其包括用包含慢病毒载体的慢病毒颗粒转导所述T细胞,所述慢病毒载体包含编码融合多肽的核苷酸序列,所述融合多肽包含通过自裂解肽连接的所述外源抗原特异性受体蛋白和所述显性负性TGF-βII型受体,从而在所述 T细胞中共表达外源抗原特异性受体蛋白和显性负性TGF-βII型受体。
- 权利要求8的方法,其中所述自裂解肽是2A多肽,例如,所述自裂解肽选自P2A、F2A、E2A或T2A多肽,或其功能性变体。
- 权利要求8或9的方法,其中所述编码融合多肽的核苷酸序列与截短的EF1α启动子可操作地连接,例如,所述截短的EF1α启动子包含SEQ ID NO:13所示核苷酸序列。
- 权利要求8-10中任一项的方法,其中所述慢病毒载体还包含选自以下的至少一种元件:5’LTR、ψ元件、RRE元件、cPPT/CTS元件、WPRE元件和3’LTR。
- 权利要求11的方法,其中所述慢病毒载体包含可操作连接的5’LTR、ψ元件、RRE元件、cPPT/CTS元件、所述截短的EF1α启动子、所述编码融合多肽的核苷酸序列、WPRE元件和3’LTR。
- 权利要求11或12的方法,其中所述5’LTR包含SEQ ID NO:3或11所示核苷酸序列;所述ψ元件包含SEQ ID NO:4或12所示核苷酸序列;所述RRE元件包含SEQ ID NO:5所示核苷酸序列;所述cPPT/CTS元件包含SEQ ID NO:6所示核苷酸序列;所述WPRE元件包含SEQ ID NO:9或14所示核苷酸序列;其中所述3’LTR包含SEQ ID NO:10或15所示核苷酸序列。
- 权利要求13的方法,其中所述慢病毒载体包含可操作连接的包含SEQ ID NO:11所示核苷酸序列的5’LTR,包含SEQ ID NO:12所示核苷酸序列的ψ元件,包含SEQ ID NO:5所示核苷酸序列的RRE元件,包含SEQ ID NO:6所示核苷酸序列的cPPT/CTS元件,包含SEQ ID NO:13所示核苷酸序列的截短的EF1α启动子,所述编码融合多肽的核苷酸序列、包含SEQ ID NO:14所示核苷酸序列的WPRE元件,包含SEQ ID NO:15所示核苷酸序列的3’LTR。
- 一种特异性靶向癌症相关抗原的治疗性T细胞,其通过权利要求1-14中任一项的方法产生。
- 一种特异性靶向癌症相关抗原的治疗性T细胞,其共表达外源癌症相关抗原特异性受体蛋白和显性负性TGF-βII型受体,所述治疗性T细胞包含慢病毒载体,所述慢病毒载体包含编码融合多肽的核苷酸序列,所述融合多肽包含通过自裂解肽连接的所述外源癌症相关抗原特异性受体蛋白和所述显性负性TGF-βII型受体。
- 权利要求16的特异性靶向癌症相关抗原的治疗性T细胞,其中所述显性负性TGF-βII型受体缺失TGF-βII型受体的胞内信号传导结构域,例如,所述显性负性TGF-βII型受体包含SEQ ID NO:18所示的氨基酸序列。
- 权利要求16或17的特异性靶向癌症相关抗原的治疗性T细胞,其中所述外源癌症相关抗原特异性受体蛋白选自T细胞受体(TCR)和嵌合抗原受体(CAR)。
- 权利要求18的特异性靶向癌症相关抗原的治疗性T细胞,所述TCR特异性结合癌症相关抗原,所述CAR包含针对癌症相关抗原的细胞外抗原结合结构域。
- 权利要求19的特异性靶向癌症相关抗原的治疗性T细胞,所述CAR包括特异 性结合癌症相关抗原的细胞外抗原结合结构域如scFv、CD8铰链和跨膜结构域、CD3ζ信号转导结构域、和4-1BB共刺激结构域。
- 权利要求16-20任一项的特异性靶向癌症相关抗原的治疗性T细胞,其中所述癌症相关抗原选自CD16、CD64、CD78、CD96、CLL1、CD116、CD117、CD71、CD45、CD71、CD123、CD138、ErbB2(HER2/neu)、癌胚抗原(CEA)、上皮细胞粘附分子(EpCAM)、表皮生长因子受体(EGFR)、EGFR变体III(EGFRvIII)、CD19、CD20、CD30、CD40、双唾液酸神经节苷脂GD2、导管上皮粘蛋白、gp36、TAG-72、鞘糖脂、神经胶质瘤相关的抗原、β-人绒毛膜促性腺激素、α胎儿球蛋白(AFP)、外源凝集素反应性AFP、甲状腺球蛋白、RAGE-1、MN-CA IX、人端粒酶逆转录酶、RU1、RU2(AS)、肠羧基酯酶、mut hsp70-2、M-CSF、前列腺酶(prostase)、前列腺酶特异性抗原(PSA)、PAP、NY-ESO-1、LAGA-1a、p53、Prostein、PSMA、存活和端粒酶、前列腺癌肿瘤抗原-1(PCTA-1)、MAGE、ELF2M、嗜中性粒细胞弹性蛋白酶、肝配蛋白B2、CD22、胰岛素生长因子(IGF1)-I、IGF-II、IGFI受体、间皮素、呈递肿瘤特异性肽表位的主要组织相容性复合体(MHC)分子、5T4、ROR1、Nkp30、NKG2D、肿瘤基质抗原、纤维连接蛋白的额外结构域A(EDA)和额外结构域B(EDB)、腱生蛋白-C的A1结构域(TnC A1)、成纤维细胞相关蛋白(fap)、CD3、CD4、CD8、CD24、CD25、CD33、CD34、CD133、CD138、Foxp3、B7-1(CD80)、B7-2(CD86)、GM-CSF、细胞因子受体、内皮因子、BCMA(CD269、TNFRSF17)、TNFRSF17(UNIPROT Q02223)、SLAMF7(UNIPROT Q9NQ25)、GPRC5D(UNIPROT Q9NZD1)、FKBP11(UNIPROT Q9NYL4)、KAMP3、ITGA8(UNIPROT P53708)和FCRL5(UNIPROT Q68SN8)。
- 权利要求21的特异性靶向癌症相关抗原的治疗性T细胞,其中所述CAR包含针对CD19的细胞外抗原结合结构域,例如,所述CAR包含SEQ ID NO:16所示氨基酸序列。
- 权利要求16-22中任一项的特异性靶向癌症相关抗原的治疗性T细胞,其中所述自裂解肽是2A多肽,例如,所述自裂解肽选自P2A、F2A、E2A或T2A多肽,或其功能性变体。
- 权利要求16-23中任一项的特异性靶向癌症相关抗原的治疗性T细胞,其中所述编码融合多肽的核苷酸序列与截短的EF1α启动子可操作地连接,例如,所述截短的EF1α启动子是包含SEQ ID NO:13所示核苷酸序列的EF1α核心启动子。
- 权利要求16-24中任一项的特异性靶向癌症相关抗原的治疗性T细胞,其中所述慢病毒载体还包含选自以下的至少一种元件:5’LTR、ψ元件、RRE元件、cPPT/CTS序列、WPRE元件和3’LTR。
- 权利要求25的特异性靶向癌症相关抗原的治疗性T细胞,其中所述慢病毒载体包含可操作连接的5’LTR、ψ元件、RRE元件、cPPT/CTS元件、所述截短的EF1α启动子、所述编码融合多肽的核苷酸序列、WPRE元件和3’LTR。
- 权利要求25或26的特异性靶向癌症相关抗原的治疗性T细胞,其中所述5’LTR 包含SEQ ID NO:3或11所示核苷酸序列;所述ψ元件包含SEQ ID NO:4或12所示核苷酸序列;所述RRE元件包含SEQ ID NO:5所示核苷酸序列;所述cPPT/CTS元件包含SEQ ID NO:6所示核苷酸序列;所述WPRE元件包含SEQ ID NO:9或14所示核苷酸序列;其中所述3’LTR包含SEQ ID NO:10或15所示核苷酸序列。
- 权利要求27的特异性靶向癌症相关抗原的治疗性T细胞,其中所述慢病毒载体包含可操作连接的包含SEQ ID NO:11所示核苷酸序列的5’LTR,包含SEQ ID NO:12所示核苷酸序列的ψ元件,包含SEQ ID NO:5所示核苷酸序列的RRE元件,包含SEQ ID NO:6所示核苷酸序列的cPPT/CTS元件,包含SEQ ID NO:13所示核苷酸序列的截短的EF1α启动子,所述编码融合多肽的核苷酸序列、包含SEQ ID NO:14所示核苷酸序列的WPRE元件,包含SEQ ID NO:15所示核苷酸序列的3’LTR。
- 药物组合物,其包含权利要求15-28中任一项的特异性靶向癌症相关抗原的治疗性T细胞,和药物可接受的载体。
- 权利要求15-28中任一项的特异性靶向癌症相关抗原的治疗性T细胞或权利要求29的药物组合物在制备用于在对象中治疗癌症的药物中的用途。
- 一种在对象中治疗癌症的方法,包括给所述对象施用治疗有效量的权利要求15-28中任一项的特异性靶向癌症相关抗原的治疗性T细胞或权利要求29的药物组合物。
- 前述权利要求中任一项的方法、治疗性T细胞、药物组合物或用途,其中所述癌症选自肺癌、卵巢癌、结肠癌、直肠癌、黑色素瘤、肾癌、膀胱癌、乳腺癌、肝癌、淋巴瘤、恶性血液病、头颈癌、胶质瘤、胃癌、鼻咽癌、喉癌、宫颈癌、子宫体瘤、骨肉瘤、骨癌、胰腺癌、皮肤癌、前列腺癌、子宫癌、肛区癌、睾丸癌、输卵管癌、子宫内膜癌、阴道癌、阴户癌、霍奇金病、非霍奇金淋巴瘤、食道癌、小肠癌、内分泌系统癌、甲状腺癌、甲状旁腺癌、肾上腺癌、软组织肉瘤、尿道癌、阴茎癌、慢性或急性白血病(包括急性髓细胞样白血病、慢性髓细胞样白血病、急性淋巴细胞性白血病、慢性淋巴细胞性白血病)、儿童实体瘤、淋巴细胞性淋巴瘤、膀胱癌、肾或输尿管癌、肾盂癌、中枢神经系统(CNS)肿瘤、原发性CNS淋巴瘤、肿瘤血管发生、脊柱肿瘤、脑干神经胶质瘤、垂体腺瘤、卡波西肉瘤、表皮状癌、鳞状细胞癌、T细胞淋巴瘤、环境诱发的癌症,包括石棉诱发的癌症,以及所述癌症的组合,例如,所述癌症是B细胞急性淋巴细胞白血病(B-ALL)。
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2021510781A JP7399157B2 (ja) | 2018-08-28 | 2019-04-28 | 改良された治療用t細胞 |
US17/272,176 US20210213119A1 (en) | 2018-08-28 | 2019-04-28 | Improved therapeutic t cell |
CA3110922A CA3110922A1 (en) | 2018-08-28 | 2019-04-28 | Improved therapeutic t cell |
KR1020217008992A KR20210063348A (ko) | 2018-08-28 | 2019-04-28 | 개선된 치료용 t 세포 |
EP19854222.7A EP3845564A4 (en) | 2018-08-28 | 2019-04-28 | IMPROVED THERAPEUTIC T-CELL |
SG11202101996QA SG11202101996QA (en) | 2018-08-28 | 2019-04-28 | Improved therapeutic t cell |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810988074.9 | 2018-08-28 | ||
CN201810988074 | 2018-08-28 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2020042647A1 true WO2020042647A1 (zh) | 2020-03-05 |
Family
ID=69643363
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/CN2019/084805 WO2020042647A1 (zh) | 2018-08-28 | 2019-04-28 | 改进的治疗性t细胞 |
Country Status (8)
Country | Link |
---|---|
US (1) | US20210213119A1 (zh) |
EP (1) | EP3845564A4 (zh) |
JP (1) | JP7399157B2 (zh) |
KR (1) | KR20210063348A (zh) |
CN (1) | CN110863013A (zh) |
CA (1) | CA3110922A1 (zh) |
SG (1) | SG11202101996QA (zh) |
WO (1) | WO2020042647A1 (zh) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20210332105A1 (en) * | 2020-04-24 | 2021-10-28 | Astrazeneca Ab | Compositions and methods of treating cancer with chimeric antigen receptors |
CN115427055A (zh) * | 2020-03-09 | 2022-12-02 | 四川大学华西医院 | IFN-γ在制备抗肿瘤辅助药物中的应用 |
US11667723B2 (en) | 2020-08-17 | 2023-06-06 | Utc Therapeutics (Shanghai) Co., Ltd. | Lymphocytes-antigen presenting cells co-stimulators and uses thereof |
Families Citing this family (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DK3959235T3 (da) * | 2019-04-26 | 2023-10-16 | Allogene Therapeutics Inc | Rituximab-resistente kimære antigenreceptorer og anvendelser deraf |
CA3176697A1 (en) * | 2020-05-11 | 2021-11-18 | Doughlas J. JOLLY | Vectors and methods for in vivo transduction |
AU2021286676A1 (en) * | 2020-06-11 | 2022-09-08 | Bioheng Therapeutics Limited | Engineered immune cell expressing NK inhibitory molecule and use thereof |
CN112210543B (zh) * | 2020-07-30 | 2023-01-03 | 山东省成体细胞产业技术研究院有限公司 | 一种针对实体瘤克服TGF-β免疫抑制的CAR-T细胞 |
WO2022033483A1 (zh) * | 2020-08-10 | 2022-02-17 | 佧珐药业有限公司 | 多功能的免疫效应细胞及其应用 |
WO2022061837A1 (en) * | 2020-09-27 | 2022-03-31 | Jiangsu Cell Tech Medical Research Institute Co., Ltd. | Fibronectin extra domain b (edb) -specific car-t for cancer |
US20220202862A1 (en) * | 2020-12-31 | 2022-06-30 | Immatics US, Inc. | Cd8 polypeptides, compositions, and methods of using thereof |
CA3234494A1 (en) * | 2021-10-15 | 2023-04-20 | Astrazeneca Ab | Anti-steap2 chimeric antigen receptors and uses thereof |
AU2022387845A1 (en) * | 2021-11-15 | 2024-05-23 | Neogene Therapeutics B.V. | Engineered t cells with reduced tgf-beta receptor signaling |
CN114606237A (zh) * | 2022-05-11 | 2022-06-10 | 上海优替济生生物医药有限公司 | Gm-csf抑制剂及其用途 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20160151491A1 (en) * | 2007-02-02 | 2016-06-02 | Yale University | Cells prepared by transient transfection and methods of use thereof |
US20160228547A1 (en) * | 2015-02-06 | 2016-08-11 | Batu Biologics, Inc. | Chimeric antigen receptor targeting of tumor endothelium |
CN107949400A (zh) * | 2015-06-24 | 2018-04-20 | 西格马-奥尔德里奇有限责任公司 | 细胞周期依赖性基因组调控和修饰 |
Family Cites Families (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20180147271A1 (en) * | 2015-05-18 | 2018-05-31 | Bluebird Bio, Inc. | Anti-ror1 chimeric antigen receptors |
WO2017103613A1 (en) * | 2015-12-18 | 2017-06-22 | Ucl Business Plc | Treatment |
SG11201807820PA (en) * | 2016-03-11 | 2018-10-30 | Bluebird Bio Inc | Genome edited immune effector cells |
CN105602992B (zh) * | 2016-03-17 | 2019-06-21 | 上海优卡迪生物医药科技有限公司 | 一种基于复制缺陷性重组慢病毒的car-t转基因载体及其构建方法和应用 |
KR20190003550A (ko) * | 2016-04-15 | 2019-01-09 | 더 트러스티스 오브 더 유니버시티 오브 펜실바니아 | 키메라 동종항원 수용체 t 세포의 조성물 및 방법 |
GB201609597D0 (en) * | 2016-06-01 | 2016-07-13 | Univ Sheffield | Therapy |
EP3487991B1 (en) * | 2016-07-25 | 2022-09-07 | The United States of America, as represented by The Secretary, Department of Health and Human Services | Methods of producing modified natural killer cells and methods of use |
WO2018071576A1 (en) * | 2016-10-14 | 2018-04-19 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Treatment of tumors by inhibition of cd300f |
US11408005B2 (en) * | 2016-12-12 | 2022-08-09 | Seattle Children's Hospital | Chimeric transcription factor variants with augmented sensitivity to drug ligand induction of transgene expression in mammalian cells |
CN108203720A (zh) * | 2016-12-20 | 2018-06-26 | 上海生博生物医药科技有限公司 | 能靶向Her2并封闭PD-L1降低肿瘤免疫逃逸的CAR-T载体及其构建方法和应用 |
-
2019
- 2019-04-28 US US17/272,176 patent/US20210213119A1/en active Pending
- 2019-04-28 EP EP19854222.7A patent/EP3845564A4/en active Pending
- 2019-04-28 CN CN201910350044.XA patent/CN110863013A/zh active Pending
- 2019-04-28 SG SG11202101996QA patent/SG11202101996QA/en unknown
- 2019-04-28 WO PCT/CN2019/084805 patent/WO2020042647A1/zh unknown
- 2019-04-28 JP JP2021510781A patent/JP7399157B2/ja active Active
- 2019-04-28 KR KR1020217008992A patent/KR20210063348A/ko not_active Application Discontinuation
- 2019-04-28 CA CA3110922A patent/CA3110922A1/en active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20160151491A1 (en) * | 2007-02-02 | 2016-06-02 | Yale University | Cells prepared by transient transfection and methods of use thereof |
US20160228547A1 (en) * | 2015-02-06 | 2016-08-11 | Batu Biologics, Inc. | Chimeric antigen receptor targeting of tumor endothelium |
CN107949400A (zh) * | 2015-06-24 | 2018-04-20 | 西格马-奥尔德里奇有限责任公司 | 细胞周期依赖性基因组调控和修饰 |
Non-Patent Citations (5)
Title |
---|
KLOSS, C, C ET AL.: "Dominant-Negative TGF-beta Receptor Enhances PSMA-Targeted Human CAR T Cell Proliferation And Augments Prostate Cancer Eradication", MOLECULAR THERAPY, vol. 26, no. 7, 5 July 2018 (2018-07-05), pages 1855 - 1866, XP055649123, DOI: 10.1016/j.ymthe.2018.05.003 * |
LEWIN, GENES VIII |
ROITT ET AL., IMMUNOLOGY |
SAMBROOK ET AL., MOLECULAR CLONING: A LABORATORY MANUAL |
See also references of EP3845564A4 |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115427055A (zh) * | 2020-03-09 | 2022-12-02 | 四川大学华西医院 | IFN-γ在制备抗肿瘤辅助药物中的应用 |
US20210332105A1 (en) * | 2020-04-24 | 2021-10-28 | Astrazeneca Ab | Compositions and methods of treating cancer with chimeric antigen receptors |
WO2021214270A1 (en) * | 2020-04-24 | 2021-10-28 | Astrazeneca Ab | Compositions and methods of treating cancer with chimeric antigen receptors |
WO2021214269A3 (en) * | 2020-04-24 | 2021-12-09 | Astrazeneca Ab | Compositions and methods of treating cancer with chimeric antigen receptors |
US11667723B2 (en) | 2020-08-17 | 2023-06-06 | Utc Therapeutics (Shanghai) Co., Ltd. | Lymphocytes-antigen presenting cells co-stimulators and uses thereof |
Also Published As
Publication number | Publication date |
---|---|
JP7399157B2 (ja) | 2023-12-15 |
CN110863013A (zh) | 2020-03-06 |
KR20210063348A (ko) | 2021-06-01 |
EP3845564A4 (en) | 2022-05-18 |
SG11202101996QA (en) | 2021-03-30 |
JP2021536240A (ja) | 2021-12-27 |
EP3845564A1 (en) | 2021-07-07 |
US20210213119A1 (en) | 2021-07-15 |
CA3110922A1 (en) | 2020-03-05 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP7399157B2 (ja) | 改良された治療用t細胞 | |
AU2019203823A1 (en) | CS1-specific chimeric antigen receptor engineered immune effector cells | |
JP7158075B2 (ja) | Gd2に基づくキメラ抗原受容体及びその利用 | |
WO2020108645A1 (en) | Cd19-and bcma-based combined car-t immunotherapy | |
WO2020108646A1 (en) | Cd19-and psma-based combined car-t immunotherapy | |
WO2019223226A1 (zh) | 改良型抗cd19 car-t细胞 | |
EP3250695A1 (en) | Universal immune cells for cancer immunotherapy | |
WO2017028374A1 (en) | Construct, genetically modified lymphocyte, preparation method and usage thereof | |
CN111733186A (zh) | 靶向cd19的人源化嵌合抗原受体制备及其应用 | |
WO2017120998A1 (zh) | 治疗脑胶质母细胞瘤的治疗组合物 | |
KR20230018376A (ko) | 키메라 항원 수용체를 발현하는 바이러스 특이적 면역세포를 사용한 암의 치료 및 예방 | |
JP7386848B2 (ja) | 改良されたレンチウイルスベクター | |
WO2018137294A1 (zh) | 共表达抗msln嵌合抗原受体和无功能egfr的转基因淋巴细胞及其用途 | |
WO2022057725A1 (zh) | 分离的能够用于表达外源基因的溶瘤腺病毒、载体、治疗剂及其用途 | |
CN111944053A (zh) | 抗bcma的car及其表达载体和应用 | |
WO2017120997A1 (zh) | 共表达抗EGFRvIII嵌合抗原受体和无功能EGFR的转基因淋巴细胞及其用途 | |
TW201927812A (zh) | 醫藥重組受體組成物及方法 | |
CN111826353B (zh) | 调节t细胞功能和反应的方法 | |
WO2021135178A1 (zh) | 一种增强型t细胞受体star及其应用 | |
JP2023541694A (ja) | がん治療剤としての二重特異的抗原結合分子を有するcar発現ナチュラルキラー細胞の生成方法及び組成物 | |
CN115960257B (zh) | 靶向IL13Rα2的经优化的嵌合抗原受体及其用途 | |
EP4357369A1 (en) | Preparation and application of chimeric antigen receptor immune cell constructed on basis of granzyme b | |
WO2023273762A1 (zh) | 介导细胞内有效滞留cd3的空间构象表位及其应用 | |
JP2023518578A (ja) | Hpv感染細胞を標的とするための組成物及び方法 | |
TW202339777A (zh) | Cd30陽性癌症之治療 |
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: 19854222 Country of ref document: EP Kind code of ref document: A1 |
|
ENP | Entry into the national phase |
Ref document number: 3110922 Country of ref document: CA Ref document number: 2021510781 Country of ref document: JP Kind code of ref document: A |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
ENP | Entry into the national phase |
Ref document number: 2019854222 Country of ref document: EP Effective date: 20210329 |