WO2020079162A1 - Procédés d'induction de l'ablation complète de l'hématopoïèse - Google Patents

Procédés d'induction de l'ablation complète de l'hématopoïèse Download PDF

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WO2020079162A1
WO2020079162A1 PCT/EP2019/078241 EP2019078241W WO2020079162A1 WO 2020079162 A1 WO2020079162 A1 WO 2020079162A1 EP 2019078241 W EP2019078241 W EP 2019078241W WO 2020079162 A1 WO2020079162 A1 WO 2020079162A1
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Prior art keywords
rac2
inhibitor
cells
hematopoiesis
cell
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PCT/EP2019/078241
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Marina Cavazzana
Isabelle Andre
Chantal LAGRESLE-PEYROU
Hanem SADEK-ROCK
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INSERM (Institut National de la Santé et de la Recherche Médicale)
Fondation Imagine
Université Paris Descartes
Assistance Publique-Hôpitaux De Paris (Aphp)
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Publication of WO2020079162A1 publication Critical patent/WO2020079162A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides
    • 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
    • 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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1137—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against enzymes

Definitions

  • the present invention relates to methods for inducting full ablation of hematopoiesis and uses thereof in the conditioning for bone marrow transplantation and the treatment of hematopoietic cell malignancies.
  • Hematopoietic stem cell transplantation is a potentially curative therapeutic approach for a variety of malignant and non-malignant hematopoietic diseases.
  • HSCT Hematopoietic stem cell transplantation
  • preparative or conditioning regimens are administered as part of the procedure to achieve 3 goals: make space in the bone-marrow (myeloablation), provide sufficient immunoablation to prevent graft rejection and reduce the tumor burden.
  • the intensity of conditioning regimens can vary substantially, and when selecting the optimal conditioning regimen for any given patient, disease-related factors such as diagnosis and remission status, as well as patient-related factors including age, donor availability, and presence of comorbid conditions, need to be considered.
  • conditioning regimens have been classified as high-dose (myelo ablative), reduced-intensity, and non-myeloablative.
  • myeloablative, or“high-dose” regimens consisting of alkylating agents (single or multiple) with or without total body irradiation, are expected to ablate hematopoiesis, not allowing autologous hematologic recovery.
  • the administration of conditioning regimens is associated with immediate and delayed toxicities. For instance, nausea, vomiting, transient acute parotiditis, xerostomia, mucositis, and diarrhea are commonly observed acute complications.
  • Interstitial pneumonitis Interstitial pneumonitis, idiopathic pulmonary fibrosis, and reduced lung pulmonary function can also be observed.
  • the occurrence of sinusoidal obstruction syndrome also known as veno- occlusive disease of the liver
  • Long-term side effects include infertility, cataract formation, hypothyroidism and thyroiditis, and secondary malignancies. Accordingly, there is a need for new method that will allow full ablation of hematopoiesis with a minimal occurrence of short and long term severe adverse side effects.
  • the present invention relates to methods for inducting full ablation of hematopoiesis and uses thereof in the conditioning for bone marrow transplantation and the treatment of hematopoietic cell malignancies.
  • the present invention is defined by the claims.
  • the first object of the present invention relates to a method of full ablating hematopoiesis in a patient in need thereof comprising administering to the patient a therapeutically effective amount of a Rac2 inhibitor.
  • a further object of the present invention relates to a method for inhibiting proliferation and differentiation of a population of hematopoietic stem cells comprising contacting said population with an effective amount of a Rac2 inhibitor.
  • a further object of the present invention relates to a method of inducing cell death of a population of hematopoietic cells comprising contacting said population with an effective amount of a Rac2 inhibitor.
  • hematopoietic cell has its general meaning in the art and refers to any type of cell of the hematopoietic system, including, but not limited to, undifferentiated cells such as hematopoietic stem cells and progenitor cells, and differentiated cells e.g. leukocytes (for example granulocytes, monocytes and lymphocytes) platelets and red blood cells.
  • undifferentiated cells such as hematopoietic stem cells and progenitor cells
  • differentiated cells e.g. leukocytes (for example granulocytes, monocytes and lymphocytes) platelets and red blood cells.
  • the term“hematopoietic stem cell” or“HSC” refers to blood cells that have the capacity to self-renew and to differentiate into precursors of circulating mature blood cells. These precursor cells are immature blood cells that cannot self-renew and differentiate into circulating mature blood cells. Within the bone marrow microenvironment, the stem cells self-renew and maintain continuous production of hematopoietic cells that give rise to all mature blood cells throughout life. In some embodiments, the hematopoietic progenitor cells or hematopoietic stem cells are isolated form peripheral blood cells.
  • hematopoiesis refers to the formation and development of hematopoietic cells involving proliferation and/or differentiation from stem cells.
  • the patient is selected from the group consisting of children, young adults, middle aged adults, and the elderly adults.
  • the method of the present invention is particularly suitable for preparing the patient to bone marrow transplantation (i.e. conditioning treatment).
  • the method of the present invention is thus particularly suitable for avoiding use of chemotherapy and radiotherapy.
  • the method of the present invention will help bone marrow for new blood stem cells to grow, helps prevent the patient's body from rejecting the transplanted cells, and helps kill any cancer cells that could be present in the body.
  • the administration of the Rac2 inhibitor is performed before bone marrow transplantation.
  • the term “bone marrow transplantation” or “hematopoietic stem cell transplantation” used herein should be considered as interchangeable, referring to the transplantation of hematopoietic stem cells in some form to a recipient.
  • the hematopoietic stem cells do not necessarily have to be derived from bone marrow, but could also be derived from other sources such as umbilical cord blood or mobilized PBMC.
  • the terms “hematopoietic stem cell transplantation” or“HSCT” refer to a component of the treatment of a wide array of hematologic disorders. Generally, there are two types of HSCTs: autologous and allogeneic transplantation.
  • the term“allogeneic” refers to deriving from, originating in, or being members of the same species, where the members are genetically related or genetically unrelated but genetically similar.
  • An“allogeneic transplant” refers to transfer of cells or organs from a donor to a recipient, where the recipient is the same species as the donor. Allogeneic transplantation involves infusion of donor stem cells, typically using a donor that matches the recipient's MHC.
  • the term“autologous” refers to deriving from or originating in the same patient.
  • An“autologous transplant” refers to collection and retransplant of a patient's own cells or organs.
  • the method of the present invention is particular suitable for the treatment of a hematopoietic cell malignancy.
  • hematopoietic cell malignancies that are cancers include leukemias, lymphomas and multiple myelomas.
  • leukemias include acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML) and chronic myelogenous leukemia (CML).
  • lymphomas include Elodgkin's disease and its subtypes; non-Hodgkin lymphomas and its subtypes including chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), hairy cell leukemia (HCL), marginal zone lymphoma (MZL), Burkitt's lymphoma (BL), Post-transplant lymphoproliferative disorder (PTLD), T-cell pro lymphocytic leukemia (T-PLL), B-cell pro lymphocytic leukemia (B-PLL), Waldenstrom's macroglobulinemia/Lymphoplasmacytic lymphoma and other natural killer cell (NK-cell) or T-cell lymphomas.
  • CLL chronic lymphocytic leukemia
  • SLL small lymphocytic lymphoma
  • DLBCL diffuse
  • MDS myelodysplastic syndrome
  • myeloproliferative diseases such as polycythemia vera (i.e., PV, PCV or polycythemia rubra vera (PRY)), essential thrombocytosis (ET), myelofibrosis
  • diseases with features of both myelodysplastic syndromes and myeloproliferative diseases such as chronic myelomonocytic leukemia (CMML), juvenile myelomonocytic leukemia (JMML), atypical chronic myeloid leukemia (aCML) and myelodysplastic/myeloproliferative disease.
  • CMML chronic myelomonocytic leukemia
  • JMML juvenile myelomonocytic leukemia
  • aCML atypical chronic myeloid leukemia
  • treatment refers to both prophylactic or preventive treatment as well as curative or disease modifying treatment, including treatment of patient at risk of contracting the disease or suspected to have contracted the disease as well as patients who are ill or have been diagnosed as suffering from a disease or medical condition, and includes suppression of clinical relapse.
  • the treatment may be administered to a patient having a medical disorder or who ultimately may acquire the disorder, in order to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of a disorder or recurring disorder, or in order to prolong the survival of a patient beyond that expected in the absence of such treatment.
  • therapeutic regimen is meant the pattern of treatment of an illness, e.g., the pattern of dosing used during therapy.
  • a therapeutic regimen may include an induction regimen and a maintenance regimen.
  • the phrase “induction regimen” or “induction period” refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the initial treatment of a disease.
  • the general goal of an induction regimen is to provide a high level of drug to a patient during the initial period of a treatment regimen.
  • An induction regimen may employ (in part or in whole) a "loading regimen", which may include administering a greater dose of the drug than a physician would employ during a maintenance regimen, administering a drug more frequently than a physician would administer the drug during a maintenance regimen, or both.
  • maintenance regimen refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the maintenance of a patient during treatment of an illness, e.g., to keep the patient in remission for long periods of time (months or years).
  • a maintenance regimen may employ continuous therapy (e.g., administering a drug at regular intervals, e.g., weekly, monthly, yearly, etc.) or intermittent therapy (e.g., interrupted treatment, intermittent treatment, treatment at relapse, or treatment upon achievement of a particular predetermined criteria [e.g., disease manifestation, etc.]).
  • Rh2 has its general meaning in the art and refers to Ras- related C3 botulinum toxin substrate 2.
  • An exemplary amino acid sequence for the human Rac2 is represented by SEQ ID NO: 1.
  • a“Rac2 inhibitor” refers to any compound natural or not which is capable of inhibiting the activity or expression of Rac2.
  • the term encompasses any Rac2 inhibitor that is currently known in the art or that will be identified in the future, and includes any chemical entity that, upon administration to a patient, results in inhibition or down- regulation of a biological activity associated with activation of Rac2.
  • the Rac2 inhibitor is selective over the other Rho GTPases especially over RAC1.
  • selective it is meant that the inhibition of the selected compound is at least 5 fold, more preferably 10-fold, most preferably 25-fold, even more preferably lOO-fold, and still preferably 300-fold higher than the inhibition of the other Rho GTPases.
  • Rho GTPase refers to a subfamily of Ras superfamily and are small, membrane-bound, Ras-related GTP-binding proteins that function by binding and hydrolyzing GTP.
  • Rho GTPases function as molecular switches, cycling between an inactive GDP-bound conformation and an active GTP-bound conformation and include RhoA, RhoB, RhoC, Cdc42, Racl, Rac2, Rac3, TC10, RhoG, RhoD, Chp, WRCH1, TCL, and RIF.
  • Rho GTPases function as molecular switches, cycling between an inactive GDP-bound conformation and an active GTP-bound conformation and include RhoA, RhoB, RhoC, Cdc42, Racl, Rac2, Rac3, TC10, RhoG, RhoD, Chp, WRCH1, TCL, and RIF.
  • the Rac2 inhibition of the compounds may be determined using various methods well known in the art.
  • the Rac2 inhibitor is a small organic molecule.
  • small organic molecule refers to a molecule of a size comparable to those organic molecules generally used in pharmaceuticals.
  • Preferred small organic molecules range in size up to about 5000 Da, more preferably up to 2000 Da, and most preferably up to about 1000 Da.
  • the Rac2 inhibitor is an antibody specific for Rac2, more particularly an intrabody.
  • the term“intrabody” is an art-recognized term that includes intracellularly expressed antibodies.
  • the antibody is a single domain antibody.
  • the term“single domain antibody” has its general meaning in the art and refers to the single heavy chain variable domain of antibodies of the type that can be found in Camelid mammals which are naturally devoid of light chains. Such single domain antibody are also“Nanobody®”.
  • (single) domain antibodies reference is also made to the prior art cited above, as well as to EP 0 368 684, Ward et al. (Nature 1989 Oct.
  • the amino acid sequence and structure of a single domain antibody can be considered to be comprised of four framework regions or“FRs” which are referred to in the art and herein as“Framework region 1” or“FR1”; as“Framework region 2” or“FR2”; as“Framework region 3” or“FR3”; and as“Framework region 4” or“FR4” respectively; which framework regions are interrupted by three complementary determining regions or“CDRs”, which are referred to in the art as“Complementarity Determining Region for “CDR1”; as “Complementarity Determining Region 2” or “CDR2” and as “Complementarity Determining Region 3” or“CDR3”, respectively.
  • the single domain antibody can be defined as an amino acid sequence with the general structure: FR1- CDR1-FR2-CDR2-FR3-CDR3-FR4 in which FR1 to FR4 refer to framework regions 1 to 4 respectively, and in which CDR1 to CDR3 refer to the complementarity determining regions 1 to 3.
  • the single domain antibody is fused to a domain of an ubiquitin ligase, such as E3 ubiquitin ligase.
  • E3 ubiquitin ligase domains examples include RING, HECT, U-box, RIBRR, F-box domain, DCAF domain, DDS2, HIF-mimetic peptides, IkB-mimetic sequences, BTB domain, or combination thereof. These E3 ligase domains facilitate ubiquitination, and when fused with the single domain antibody of the present allows for the degradation of the antigen-antibody complex by the proteasome. Any E3 ligase domains including E2 binding domains known or later discovered or developed can be used. Recombinant E3 ligase domains can be used.
  • the heterologous polypeptide is a F-box domain.
  • the F-box domain is typically a protein motif of approximately 50 amino acids.
  • the F-box domain tethers the F-box protein to other components of the SCF complex by binding the core SCF component, Skpl .
  • the Rac2 inhibitor is an inhibitor of Rac2 expression respectively.
  • An“inhibitor of expression” refers to a natural or synthetic compound that has a biological effect to inhibit the expression of a gene.
  • said inhibitor of gene expression is a siRNA, an antisense oligonucleotide or a ribozyme.
  • anti-sense oligonucleotides including anti-sense RNA molecules and anti-sense DNA molecules, would act to directly block the translation of Rac2 mRNA by binding thereto and thus preventing protein translation or increasing mRNA degradation, thus decreasing the level ofRac2, and thus activity, in a cell.
  • antisense oligonucleotides of at least about 15 bases and complementary to unique regions of the mRNA transcript sequence encoding Rac2 can be synthesized, e.g., by conventional phosphodiester techniques.
  • Methods for using antisense techniques for specifically inhibiting gene expression of genes whose sequence is known are well known in the art (e.g. see U.S. Pat. Nos. 6,566,135; 6,566,131; 6,365,354; 6,410,323; 6,107,091; 6,046,321; and 5,981,732).
  • Small inhibitory RNAs siRNAs
  • siRNAs can also function as inhibitors of expression for use in the present invention.
  • Rac2 gene expression can be reduced by contacting a patient or cell with a small double stranded RNA (dsRNA), or a vector or construct causing the production of a small double stranded RNA, such that Rac2 gene expression is specifically inhibited (i.e. RNA interference or RNAi).
  • dsRNA small double stranded RNA
  • RNAi RNA interference or RNAi
  • Antisense oligonucleotides, siRNAs, shRNAs and ribozymes of the invention may be delivered in vivo alone or in association with a vector.
  • a "vector” is any vehicle capable of facilitating the transfer of the antisense oligonucleotide, siRNA, shRNA or ribozyme nucleic acid to the cells and typically cells expressing Rac2.
  • the vector transports the nucleic acid to cells with reduced degradation relative to the extent of degradation that would result in the absence of the vector.
  • the vectors useful in the invention include, but are not limited to, plasmids, phagemids, viruses, other vehicles derived from viral or bacterial sources that have been manipulated by the insertion or incorporation of the antisense oligonucleotide, siRNA, shRNA or ribozyme nucleic acid sequences.
  • Viral vectors are a preferred type of vector and include, but are not limited to nucleic acid sequences from the following viruses: retrovirus, such as moloney murine leukemia virus, harvey murine sarcoma virus, murine mammary tumor virus, and rous sarcoma vims; adenovirus, adeno-associated vims; SV40-type vimses; polyoma vimses; Epstein-Barr vimses; papilloma vimses; herpes vims; vaccinia vims; polio vims; and RNA vims such as a retro vims.
  • retrovirus such as moloney murine leukemia virus, harvey murine sarcoma virus, murine mammary tumor virus, and rous sarcoma vims
  • adenovirus adeno-associated vims
  • SV40-type vimses polyoma vimses
  • the inhibitor of expression is an endonuclease.
  • the term “endonuclease” refers to enzymes that cleave the phosphodiester bond within a polynucleotide chain. Some, such as Deoxyribonuclease I, cut DNA relatively nonspecifically (without regard to sequence), while many, typically called restriction endonucleases or restriction enzymes, and cleave only at very specific nucleotide sequences.
  • endonuclease-based genome inactivating generally requires a first step of DNA single or double strand break, which can then trigger two distinct cellular mechanisms for DNA repair, which can be exploited for DNA inactivating: the errorprone nonhomologous end-joining (NITEJ) and the high-fidelity homology-directed repair (HDR).
  • the endonuclease is CRISPR- cas.
  • CRISPR-cas has its general meaning in the art and refers to clustered regularly interspaced short palindromic repeats associated which are the segments of prokaryotic DNA containing short repetitions of base sequences.
  • the endonuclease is CRISPR-cas9 which is from Streptococcus pyogenes.
  • the CRISPR/Cas9 system has been described in IJS 8697359 Bl and US 2014/0068797.
  • the endonuclease is CRISPR-Cpfl which is the more recently characterized CRISPR from Provote lla and Francisella 1 (Cpfl) in Zetsche et al. (“Cpfl is a Single RNA-guided Endonuclease of a Class 2 CRISPR-Cas System (2015); Cell; 163, 1-13).
  • a “therapeutically effective amount” is meant a sufficient amount of the active ingredient for treating or reducing the symptoms at reasonable benefit/risk ratio applicable to any medical treatment. It will be understood that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment.
  • the specific therapeutically effective dose level for any particular subject will depend upon a variety of factors including the disorder being treated and the severity of the disorder; activity of the specific compound employed; the specific composition employed, the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination with the active ingredients; and like factors well known in the medical arts.
  • the daily dosage of the products may be varied over a wide range from 0.01 to 1,000 mg per adult per day.
  • the compositions contain 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 250 and 500 mg of the active ingredient for the symptomatic adjustment of the dosage to the subject to be treated.
  • a medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, typically from 1 mg to about 100 mg of the active ingredient.
  • An effective amount of the drug is ordinarily supplied at a dosage level from 0.0002 mg/kg to about 20 mg/kg of body weight per day, especially from about 0.001 mg/kg to 7 mg/kg of body weight per day.
  • the active ingredient of the present invention e.g. Rac2 inhibitor
  • pharmaceutically acceptable excipients e.g. Rac2 inhibitor
  • sustained-release matrices such as biodegradable polymers
  • pharmaceutically acceptable carrier or excipient refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type.
  • the carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetables oils.
  • a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetables oils.
  • the proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
  • the prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like ln many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride.
  • Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminium monostearate and gelatin.
  • the active ingredients of the invention can be administered in a unit administration form, as a mixture with conventional pharmaceutical supports.
  • Suitable unit administration forms comprise oral-route forms such as tablets, gel capsules, powders, granules and oral suspensions or solutions, sublingual and buccal administration forms, aerosols, implants, subcutaneous, transdermal, topical, intraperitoneal, intramuscular, intravenous, subdermal, transdermal, intrathecal and intranasal administration forms and rectal administration forms.
  • FIGURE
  • FIG. 1 RAC2 mutation impaired CB CD34+ HSPC proliferation and differentiation toward lymphocyte and neutrophil lineages.
  • the representative graphs are all gated on the alive GFP+ population.
  • Statistical analysis was performed using a two tailed unpaired T- test *** ⁇ 0,001 ** ⁇ 0,01
  • RAC2 regulates survival, proliferation and differentiation of hematopoietic stem and progenitor cell (HSPC).
  • Severe Combined Immune Deficiencies are inherited disorders characterized by a blockade in T lymphoid differentiation associated with an absence or a functional defect of B NK-cell or neutrophil lineages 1 .
  • RAC2 protein belongs to the Rac family small GTPase 2 involved in various signalling pathways. In an inactive stage, the protein is localized into the cytosol, bought to GDP and upon activation, the RAC2 protein is translocated to the plasma membrane in association with GTP. RAC2 protein is highly expressed in the bone-marrow, especially in the hematopoietic stem and progenitor cell (HSPC) population unlike other member of the family as RAC 1 expressed in many tissues 2 .
  • HSPC hematopoietic stem and progenitor cell
  • CD34+ HSPC progenitors with a lentiviral vector containing either the wild-type form of RAC2 (WT) or the mutated form (G12R), both associated with the GFP reporter gene.
  • WT wild-type form of RAC2
  • G12R mutated form
  • Transduced cells were also seeded on Delta4 coated plate in order to analyse their capacity to differentiate toward T cell lineage.
  • CD7+ T-cell progenitors were highly reduced in the G12R condition as compared to the WT condition ( Figure IB).
  • the number of alive differentiated cells characterized by the co-expression of CDl lb and CD15 (G-CSF condition) or CDl lb and CD14 (M-CSF condition) was nearly absent in the GFP+ subset from the G12R condition as compared to the GFP+ subset from the WT condition.

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Abstract

La présente invention concerne des schémas thérapeutiques de conditionnement qui visent à préparer un sujet à la transplantation de cellules souches hématopoïétiques et qui sont associés à des toxicités immédiates et retardées. Par conséquent, il est nécessaire de trouver un nouveau procédé qui permettrait l'ablation complète de l'hématopoïèse. Les inventeurs ont identifié une telle nouvelle cible. En effet, les inventeurs ont été intéressés par trois patients présentant une forme grave de SCID à la naissance, associée à la septicémie. Les études génétiques des fibroblastes des patients les ont conduits à identifier la même mutation dans le gène de substrat de la toxine botulinum C3 lié à Ras 2 (RAC2). Plus particulièrement, ils ont montré que l'introduction de la mutation dans la cellule souche hématopoïétique et progénitrice (HSPC) conduit à une baisse importante de la prolifération et à la différenciation vers des lignées différentes (cellules T, granulocytes et monocytes). Ainsi, ces résultats suggèrent que l'inhibition de RAC2 pourrait convenir pour induire l'ablation de l'hématopoïèse.
PCT/EP2019/078241 2018-10-18 2019-10-17 Procédés d'induction de l'ablation complète de l'hématopoïèse WO2020079162A1 (fr)

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