127 results on '"X-Linked Combined Immunodeficiency Diseases therapy"'
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2. Restoring T and B cell generation in X-linked severe combined immunodeficiency mice through hematopoietic stem cells adenine base editing.
- Author
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Zhang L, Li K, Liu Z, An L, Wei H, Pang S, Cao Z, Huang X, Jin X, and Ma X
- Subjects
- Animals, Mice, Humans, Mice, SCID, Genetic Therapy methods, CRISPR-Cas Systems, RNA, Guide, CRISPR-Cas Systems, Gene Editing, Hematopoietic Stem Cells metabolism, Disease Models, Animal, X-Linked Combined Immunodeficiency Diseases therapy, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases immunology, Hematopoietic Stem Cell Transplantation methods, T-Lymphocytes immunology, T-Lymphocytes metabolism, Adenine analogs & derivatives, B-Lymphocytes immunology, B-Lymphocytes metabolism
- Abstract
Base editing of hematopoietic stem/progenitor cells (HSPCs) is an attractive strategy for treating immunohematologic diseases. However, the feasibility of using adenine-base-edited HSPCs for treating X-linked severe combined immunodeficiency (SCID-X1), the influence of dose-response relationships on immune cell generation, and the potential risks have not been demonstrated in vivo. Here, a humanized SCID-X1 mouse model was established, and 86.67% ± 2.52% (n = 3) of mouse hematopoietic stem cell (HSC) pathogenic mutations were corrected, with no single-guide-RNA (sgRNA)-dependent off-target effects detected. Analysis of peripheral blood over 16 weeks post-transplantation in mice with different immunodeficiency backgrounds revealed efficient immune cell generation following transplantation of different amounts of modified HSCs. Therefore, a large-scale infusion of gene-corrected HSCs within a safe range can achieve rapid, stable, and durable immune cell regeneration. Tissue-section staining further demonstrated the restoration of immune organ tissue structures, with no tumor formation in multiple organs. Collectively, these data suggest that base-edited HSCs are a potential therapeutic approach for SCID-X1 and that a threshold infusion dose of gene-corrected cells is required for immune cell regeneration. This study lays a theoretical foundation for the clinical application of base-edited HSCs in treating SCID-X1., Competing Interests: Declaration of interests The authors declare that they have no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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3. Integrome signatures of lentiviral gene therapy for SCID-X1 patients.
- Author
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Yan KK, Condori J, Ma Z, Metais JY, Ju B, Ding L, Dhungana Y, Palmer LE, Langfitt DM, Ferrara F, Throm R, Shi H, Risch I, Bhatara S, Shaner B, Lockey TD, Talleur AC, Easton J, Meagher MM, Puck JM, Cowan MJ, Zhou S, Mamcarz E, Gottschalk S, and Yu J
- Subjects
- Humans, Genetic Therapy, Retroviridae genetics, T-Lymphocytes, Genetic Vectors genetics, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Lentiviral vector (LV)-based gene therapy holds promise for a broad range of diseases. Analyzing more than 280,000 vector integration sites (VISs) in 273 samples from 10 patients with X-linked severe combined immunodeficiency (SCID-X1), we discovered shared LV integrome signatures in 9 of 10 patients in relation to the genomics, epigenomics, and 3D structure of the human genome. VISs were enriched in the nuclear subcompartment A1 and integrated into super-enhancers close to nuclear pore complexes. These signatures were validated in T cells transduced with an LV encoding a CD19-specific chimeric antigen receptor. Intriguingly, the one patient whose VISs deviated from the identified integrome signatures had a distinct clinical course. Comparison of LV and gamma retrovirus integromes regarding their 3D genome signatures identified differences that might explain the lower risk of insertional mutagenesis in LV-based gene therapy. Our findings suggest that LV integrome signatures, shaped by common features such as genome organization, may affect the efficacy of LV-based cellular therapies.
- Published
- 2023
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4. CRISPR-Cas9-AAV versus lentivector transduction for genome modification of X-linked severe combined immunodeficiency hematopoietic stem cells.
- Author
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Brault J, Liu T, Liu S, Lawson A, Choi U, Kozhushko N, Bzhilyanskaya V, Pavel-Dinu M, Meis RJ, Eckhaus MA, Burkett SS, Bosticardo M, Kleinstiver BP, Notarangelo LD, Lazzarotto CR, Tsai SQ, Wu X, Dahl GA, Porteus MH, Malech HL, and De Ravin SS
- Subjects
- Animals, Mice, Dependovirus, CRISPR-Cas Systems, Mice, SCID, Hematopoietic Stem Cells, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Introduction: Ex vivo gene therapy for treatment of Inborn errors of Immunity (IEIs) have demonstrated significant clinical benefit in multiple Phase I/II clinical trials. Current approaches rely on engineered retroviral vectors to randomly integrate copy(s) of gene-of-interest in autologous hematopoietic stem/progenitor cells (HSPCs) genome permanently to provide gene function in transduced HSPCs and their progenies. To circumvent concerns related to potential genotoxicities due to the random vector integrations in HSPCs, targeted correction with CRISPR-Cas9-based genome editing offers improved precision for functional correction of multiple IEIs., Methods: We compare the two approaches for integration of IL2RG transgene for functional correction of HSPCs from patients with X-linked Severe Combined Immunodeficiency (SCID-X1 or XSCID); delivery via current clinical lentivector (LV)- IL2RG versus targeted insertion (TI) of IL2RG via homology-directed repair (HDR) when using an adeno-associated virus (AAV)- IL2RG donor following double-strand DNA break at the endogenous IL2RG locus., Results and Discussion: In vitro differentiation of LV- or TI-treated XSCID HSPCs similarly overcome differentiation block into Pre-T-I and Pre-T-II lymphocytes but we observed significantly superior development of NK cells when corrected by TI (40.7% versus 4.1%, p = 0.0099). Transplants into immunodeficient mice demonstrated robust engraftment (8.1% and 23.3% in bone marrow) for LV- and TI- IL2RG HSPCs with efficient T cell development following TI- IL2RG in all four patients' HSPCs. Extensive specificity analysis of CRISPR-Cas9 editing with rhAmpSeq covering 82 predicted off-target sites found no evidence of indels in edited cells before ( in vitro ) or following transplant, in stark contrast to LV's non-targeted vector integration sites. Together, the improved efficiency and safety of IL2RG correction via CRISPR-Cas9-based TI approach provides a strong rationale for a clinical trial for treatment of XSCID patients., Competing Interests: BK is an inventor on patents and patent applications filed by Mass General Brigham that describe genome engineering technologies. BK is a consultant for Avectas Inc., EcoR1 capital, and ElevateBio, and is an advisor to Acrigen Biosciences and Life Edit Therapeutics. RM and GD are employees of CELLSCRIPT, LLC Madison, WI. Authors SL and XW were employed by Leidos Biomedical Research Inc. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Brault, Liu, Liu, Lawson, Choi, Kozhushko, Bzhilyanskaya, Pavel-Dinu, Meis, Eckhaus, Burkett, Bosticardo, Kleinstiver, Notarangelo, Lazzarotto, Tsai, Wu, Dahl, Porteus, Malech and De Ravin.)
- Published
- 2023
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5. Challenges in Gene Therapy for Somatic Reverted Mosaicism in X-Linked Combined Immunodeficiency by CRISPR/Cas9 and Prime Editing.
- Author
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Hou Y, Ureña-Bailén G, Mohammadian Gol T, Gratz PG, Gratz HP, Roig-Merino A, Antony JS, Lamsfus-Calle A, Daniel-Moreno A, Handgretinger R, and Mezger M
- Subjects
- Humans, CRISPR-Cas Systems genetics, Mosaicism, Gene Editing methods, Genetic Therapy methods, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
X-linked severe combined immunodeficiency (X-SCID) is a primary immunodeficiency that is caused by mutations in the interleukin-2 receptor gamma (IL2RG) gene. Some patients present atypical X-SCID with mild clinical symptoms due to somatic revertant mosaicism. CRISPR/Cas9 and prime editing are two advanced genome editing tools that paved the way for treating immune deficiency diseases. Prime editing overcomes the limitations of the CRISPR/Cas9 system, as it does not need to induce double-strand breaks (DSBs) or exogenous donor DNA templates to modify the genome. Here, we applied CRISPR/Cas9 with single-stranded oligodeoxynucleotides (ssODNs) and prime editing methods to generate an in vitro model of the disease in K-562 cells and healthy donors' T cells for the c. 458T>C point mutation in the IL2RG gene, which also resulted in a useful way to optimize the gene correction approach for subsequent experiments in patients' cells. Both methods proved to be successful and were able to induce the mutation of up to 31% of treated K-562 cells and 26% of treated T cells. We also applied similar strategies to correct the IL2RG c. 458T>C mutation in patient T cells that carry the mutation with revertant somatic mosaicism. However, both methods failed to increase the frequency of the wild-type sequence in the mosaic T cells of patients due to limited in vitro proliferation of mutant cells and the presence of somatic reversion. To the best of our knowledge, this is the first attempt to treat mosaic cells from atypical X-SCID patients employing CRISPR/Cas9 and prime editing. We showed that prime editing can be applied to the formation of specific-point IL2RG mutations without inducing nonspecific on-target modifications. We hypothesize that the feasibility of the nucleotide substitution of the IL2RG gene using gene therapy, especially prime editing, could provide an alternative strategy to treat X-SCID patients without revertant mutations, and further technological improvements need to be developed to correct somatic mosaicism mutations.
- Published
- 2022
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6. Lentivector cryptic splicing mediates increase in CD34+ clones expressing truncated HMGA2 in human X-linked severe combined immunodeficiency.
- Author
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De Ravin SS, Liu S, Sweeney CL, Brault J, Whiting-Theobald N, Ma M, Liu T, Choi U, Lee J, O'Brien SA, Quackenbush P, Estwick T, Karra A, Docking E, Kwatemaa N, Guo S, Su L, Sun Z, Zhou S, Puck J, Cowan MJ, Notarangelo LD, Kang E, Malech HL, and Wu X
- Subjects
- Antigens, CD34 genetics, Clone Cells, Genetic Therapy, Genetic Vectors genetics, Humans, Lentivirus genetics, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
X-linked Severe Combined Immunodeficiency (SCID-X1) due to IL2RG mutations is potentially fatal in infancy where 'emergency' life-saving stem cell transplant may only achieve incomplete immune reconstitution following transplant. Salvage therapy SCID-X1 patients over 2 years old (NCT01306019) is a non-randomized, open-label, phase I/II clinical trial for administration of lentiviral-transduced autologous hematopoietic stem cells following busulfan (6 mg/kg total) conditioning. The primary and secondary objectives assess efficacy in restoring immunity and safety by vector insertion site analysis (VISA). In this ongoing study (19 patients treated), we report VISA in blood lineages from first eight treated patients with longer follow up found a > 60-fold increase in frequency of forward-orientated VIS within intron 3 of the High Mobility Group AT-hook 2 gene. All eight patients demonstrated emergence of dominant HMGA2 VIS clones in progenitor and myeloid lineages, but without disturbance of hematopoiesis. Our molecular analysis demonstrated a cryptic splice site within the chicken β-globin hypersensitivity 4 insulator element in the vector generating truncated mRNA transcripts from many transcriptionally active gene containing forward-oriented intronic vector insert. A two base-pair change at the splice site within the lentiviral vector eliminated splicing activity while retaining vector functional capability. This highlights the importance of functional analysis of lentivectors for cryptic splicing for preclinical safety assessment and a redesign of clinical vectors to improve safety., (© 2022. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)
- Published
- 2022
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7. CRISPR-targeted MAGT1 insertion restores XMEN patient hematopoietic stem cells and lymphocytes.
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Brault J, Liu T, Bello E, Liu S, Sweeney CL, Meis RJ, Koontz S, Corsino C, Choi U, Vayssiere G, Bosticardo M, Dowdell K, Lazzarotto CR, Clark AB, Notarangelo LD, Ravell JC, Lenardo MJ, Kleinstiver BP, Tsai SQ, Wu X, Dahl GA, Malech HL, and De Ravin SS
- Subjects
- Animals, CRISPR-Cas Systems, Cation Transport Proteins deficiency, Cells, Cultured, Female, Genetic Therapy, Hematopoietic Stem Cell Transplantation, Hematopoietic Stem Cells pathology, Humans, Lymphocytes pathology, Male, Mice, Inbred NOD, X-Linked Combined Immunodeficiency Diseases pathology, X-Linked Combined Immunodeficiency Diseases therapy, Mice, Cation Transport Proteins genetics, Gene Editing methods, Hematopoietic Stem Cells metabolism, Lymphocytes metabolism, X-Linked Combined Immunodeficiency Diseases genetics
- Abstract
XMEN disease, defined as "X-linked MAGT1 deficiency with increased susceptibility to Epstein-Barr virus infection and N-linked glycosylation defect," is a recently described primary immunodeficiency marked by defective T cells and natural killer (NK) cells. Unfortunately, a potentially curative hematopoietic stem cell transplantation is associated with high mortality rates. We sought to develop an ex vivo targeted gene therapy approach for patients with XMEN using a CRISPR/Cas9 adeno-associated vector (AAV) to insert a therapeutic MAGT1 gene at the constitutive locus under the regulation of the endogenous promoter. Clinical translation of CRISPR/Cas9 AAV-targeted gene editing (GE) is hampered by low engraftable gene-edited hematopoietic stem and progenitor cells (HSPCs). Here, we optimized GE conditions by transient enhancement of homology-directed repair while suppressing AAV-associated DNA damage response to achieve highly efficient (>60%) genetic correction in engrafting XMEN HSPCs in transplanted mice. Restored MAGT1 glycosylation function in human NK and CD8+ T cells restored NK group 2 member D (NKG2D) expression and function in XMEN lymphocytes for potential treatment of infections, and it corrected HSPCs for long-term gene therapy, thus offering 2 efficient therapeutic options for XMEN poised for clinical translation., (© 2021 by The American Society of Hematology.)
- Published
- 2021
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8. Successful living donor liver transplantation for liver failure due to maternal T cell engraftment following cord blood transplantation in X-linked severe combined immunodeficiency disease: Case report.
- Author
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Okada N, Kawahara Y, Sanada Y, Hirata Y, Otomo S, Niijima H, Tanaka A, Morimoto A, Lefor AK, Urahashi T, Yasuda Y, Mizuta K, Sakuma Y, and Sata N
- Subjects
- Female, Humans, Infant, Living Donors, Male, Pregnancy, T-Lymphocytes, Cord Blood Stem Cell Transplantation adverse effects, Graft vs Host Disease, Hematopoietic Stem Cell Transplantation, Liver Failure surgery, Liver Transplantation adverse effects, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Maternal T cells from perinatal transplacental passage have been identified in up to 40% of patients with severe combined immunodeficiency (SCID). Although engrafted maternal T cells sometimes injure newborn tissue, liver failure due to maternal T cells has not been reported. We rescued a boy with X-linked SCID who developed liver failure due to engrafted maternal T cell invasion following living donor liver transplantation (LDLT) following unrelated umbilical cord blood transplantation (UCBT). After developing respiratory failure 3 weeks postpartum, he was diagnosed with X-linked SCID. Pathological findings showed maternal T cells engrafted in his liver and hepatic fibrosis gradually progressed. He underwent UCBT at 6 months, but hepatic function did not recover and liver failure progressed. Therefore, he underwent LDLT using an S2 monosegment graft at age 1.3 years. The patient had a leak at the Roux-en-Y anastomosis, which was repaired. Despite occasional episodes of pneumonia and otitis media, he is generally doing well 6 years after LDLT with continued immunosuppression agents. In conclusion, the combination of hematopoietic stem cell transplantation (HSCT) and liver transplantation may be efficacious, and HSCT should precede liver transplantation for children with X-linked SCID and liver failure., (© 2021 The American Society of Transplantation and the American Society of Transplant Surgeons.)
- Published
- 2021
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9. Case Report: A Novel IL2RG Frame-Restoring Rescue Mutation Mimics Early T Cell Engraftment Following Haploidentical Hematopoietic Stem Cell Transplantation in a Patient With X-SCID.
- Author
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Steininger J, Leiss-Piller A, Geier CB, Rossmanith R, Elfeky R, Bra D, Pichler H, Lawitschka A, Zubarovskaya N, Artacker G, Matthes-Leodolter S, Eibl MM, and Wolf HM
- Subjects
- Allografts, Humans, Infant, Interleukin Receptor Common gamma Subunit genetics, Interleukin Receptor Common gamma Subunit immunology, Male, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Hematopoietic Stem Cell Transplantation, Mutation, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases immunology, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Mutations of the interleukin 2 receptor γ chain (IL2RG) result in the most common form of severe combined immunodeficiency (SCID), which is characterized by severe and persistent infections starting in early life with an absence of T cells and natural killer cells, normal or elevated B cell counts and hypogammaglobulinemia. SCID is commonly fatal within the first year of life, unless the immune system is reconstituted by hematopoietic stem cell transplantation (HSCT) or gene therapy. We herein describe a male infant with X-linked severe combined immunodeficiency (X-SCID) diagnosed at 5 months of age. Genetic testing revealed a novel C to G missense mutation in exon 1 resulting in a 3' splice site disruption with premature stop codon and aberrant IL2 receptor signaling. Following the diagnosis of X-SCID, the patient subsequently underwent a TCRαβ/CD19-depleted haploidentical HSCT. Post transplantation the patient presented with early CD8
+ T cell recovery with the majority of T cells (>99%) being non-donor T cells. Genetic analysis of CD4+ and CD8+ T cells revealed a spontaneous 14 nucleotide insertion at the mutation site resulting in a novel splice site and restoring the reading frame although defective IL2RG function was still demonstrated. In conclusion, our findings describe a spontaneous second-site mutation in IL2RG as a novel cause of somatic mosaicism and early T cell recovery following haploidentical HSCT., Competing Interests: Author ME was employed by company Biomedizinische Forschungs GmbH. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Steininger, Leiss-Piller, Geier, Rossmanith, Elfeky, Bra, Pichler, Lawitschka, Zubarovskaya, Artacker, Matthes-Leodolter, Eibl and Wolf.)- Published
- 2021
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10. Gene Therapies for Primary Immune Deficiencies.
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Kohn LA and Kohn DB
- Subjects
- Genetic Therapy trends, Granulomatous Disease, Chronic genetics, Granulomatous Disease, Chronic therapy, Hematopoietic Stem Cells cytology, Humans, Primary Immunodeficiency Diseases genetics, Severe Combined Immunodeficiency genetics, Severe Combined Immunodeficiency therapy, Treatment Outcome, Wiskott-Aldrich Syndrome genetics, Wiskott-Aldrich Syndrome therapy, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases therapy, Genetic Therapy methods, Hematopoietic Stem Cell Transplantation methods, Hematopoietic Stem Cells metabolism, Primary Immunodeficiency Diseases therapy
- Abstract
Gene therapy is an innovative treatment for Primary Immune Deficiencies (PIDs) that uses autologous hematopoietic stem cell transplantation to deliver stem cells with added or edited versions of the missing or malfunctioning gene that causes the PID. Initial studies of gene therapy for PIDs in the 1990-2000's used integrating murine gamma-retroviral vectors. While these studies showed clinical efficacy in many cases, especially with the administration of marrow cytoreductive conditioning before cell re-infusion, these vectors caused genotoxicity and development of leukoproliferative disorders in several patients. More recent studies used lentiviral vectors in which the enhancer elements of the long terminal repeats self-inactivate during reverse transcription ("SIN" vectors). These SIN vectors have excellent safety profiles and have not been reported to cause any clinically significant genotoxicity. Gene therapy has successfully treated several PIDs including Adenosine Deaminase Severe Combined Immunodeficiency (SCID), X-linked SCID, Artemis SCID, Wiskott-Aldrich Syndrome, X-linked Chronic Granulomatous Disease and Leukocyte Adhesion Deficiency-I. In all, gene therapy for PIDs has progressed over the recent decades to be equal or better than allogeneic HSCT in terms of efficacy and safety. Further improvements in methods should lead to more consistent and reliable efficacy from gene therapy for a growing list of PIDs., Competing Interests: LK is the recipient of an A. P. Giannini Foundation Postdoctoral Research Fellowship. DK is a member of the Scientific Advisory Board for Orchard Therapeutics that markets Strimvelis and has licensed a lentiviral vector for gene therapy of ADA SCID from the University of California, Los Angeles for which he is an inventor. DK is also the principal investigator for clinical trials discussed in this article on gene therapy for XCGD (California Institute for Regenerative Medicine CLIN2-08231 and Orchard Therapeutics) and LAD-I (California Institute for Regenerative Medicine CLIN2-11480 and Rocket Pharmaceuticals). He is the Chair of the Data Safety Monitoring Board for Leadiant Biosciences for Revcovi PEG-ADA ERT., (Copyright © 2021 Kohn and Kohn.)
- Published
- 2021
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11. In Vivo Gene Therapy for Canine SCID-X1 Using Cocal-Pseudotyped Lentiviral Vector.
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Rajawat YS, Humbert O, Cook SM, Radtke S, Pande D, Enstrom M, Wohlfahrt ME, and Kiem HP
- Subjects
- Animals, Dogs, Genetic Therapy, Genetic Vectors genetics, Hematopoietic Stem Cells, Humans, Lentivirus genetics, Transduction, Genetic, Spumavirus, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Hematopoietic stem and progenitor cell (HSPC)-based ex vivo gene therapy has demonstrated clinical success for X-linked severe combined immunodeficiency (SCID-X1) patients who lack a suitable donor for HSPC transplantation. Nevertheless, this form of treatment is associated with an increased risk of infectious disease complications and genotoxicity mainly due to the conditioning regimen. In addition, ex vivo gene therapy approaches require sophisticated facilities to manufacture gene-modified cells and to care for the patients after chemotherapy. Considering these impediments, we have developed an in vivo gene therapy approach to treat canine SCID-X1 after HSPC mobilization and systemic delivery of the therapeutic vector. Here, we investigated the use of the cocal envelope to pseudotype a lentiviral (LV) vector expressing a functional gammaC gene. The cocal envelope is resistant to serum inactivation compared with the commonly used vesicular stomatitis virus envelope glycoprotein (VSV-G) envelope and thus well suited for systemic delivery. Two SCID-X1 neonatal canines treated with this approach achieved long-term therapeutic immune reconstitution with no prior conditioning. Therapeutic levels of gene-corrected CD3
+ T cells were demonstrated for at least 16 months, and all other correlates of T cell functionality were within normal range. Retroviral integration-site analysis demonstrated polyclonal T cell reconstitution. Comparative analysis of integration profiles of foamy viral (FV) vector and cocal LV vector after in vivo gene therapy found distinct integration-site patterns. These data demonstrate that clinically relevant and durable correction of canine SCID-X1 can be achieved with in vivo delivery of cocal LV. Since manufacturing of cocal LV is similar to VSV-G LV, this approach is easily translatable to a clinical setting, thus providing for a highly portable and accessible gene therapy platform for SCID-X1.- Published
- 2021
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12. Gene therapy for X-linked severe combined immunodeficiency: Historical outcomes and current status.
- Author
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Pai SY and Thrasher AJ
- Subjects
- Animals, Genetic Therapy trends, Humans, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases immunology, B-Lymphocytes immunology, Chromosomes, Human, X genetics, Genetic Therapy methods, T-Lymphocytes immunology, X-Linked Combined Immunodeficiency Diseases therapy
- Published
- 2020
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13. T-cell-replete haploidentical bone marrow transplantation for X-linked severe combined immunodeficiency.
- Author
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Ariffin H, Chew KS, Jawin V, and Thavagnanam S
- Subjects
- Humans, Infant, Interleukin Receptor Common gamma Subunit genetics, Malaysia, Male, T-Lymphocytes, X-Linked Combined Immunodeficiency Diseases genetics, Bone Marrow Transplantation methods, Transplantation, Haploidentical methods, X-Linked Combined Immunodeficiency Diseases therapy
- Published
- 2020
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14. Gene therapy for severe combined immunodeficiencies and beyond.
- Author
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Fischer A and Hacein-Bey-Abina S
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- Animals, Gene Editing, Gene Transfer Techniques, Genetic Therapy adverse effects, Genetic Vectors genetics, Humans, Interleukin Receptor Common gamma Subunit genetics, Leukemia etiology, Mutagenesis, Insertional methods, Mutation, Missense, Retroviridae genetics, T-Lymphocytes immunology, Adenosine Deaminase deficiency, Agammaglobulinemia therapy, Genetic Therapy methods, Severe Combined Immunodeficiency therapy, Wiskott-Aldrich Syndrome therapy, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Ex vivo retrovirally mediated gene therapy has been shown within the last 20 yr to correct the T cell immunodeficiency caused by γc-deficiency (SCID X1) and adenosine deaminase (ADA) deficiency. The rationale was brought up by the observation of the revertant of SCIDX1 and ADA deficiency as a kind of natural gene therapy. Nevertheless, the first attempts of gene therapy for SCID X1 were associated with insertional mutagenesis causing leukemia, because the viral enhancer induced transactivation of oncogenes. Removal of this element and use of a promoter instead led to safer but still efficacious gene therapy. It was observed that a fully diversified T cell repertoire could be generated by a limited set (<1,000) of progenitor cells. Further advances in gene transfer technology, including the use of lentiviral vectors, has led to success in the treatment of Wiskott-Aldrich syndrome, while further applications are pending. Genome editing of the mutated gene may be envisaged as an alternative strategy to treat SCID diseases., (© 2019 Fischer and Hacein-Bey-Abina.)
- Published
- 2020
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15. Elucidation of the Effects of a Current X-SCID Therapy on Intestinal Lymphoid Organogenesis Using an In Vivo Animal Model.
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Nochi T, Suzuki S, Ito S, Morita S, Furukawa M, Fuchimoto D, Sasahara Y, Usami K, Niimi K, Itano O, Kitago M, Matsuda S, Matsuo A, Suyama Y, Sakai Y, Wu G, Bazer FW, Watanabe K, Onishi A, and Aso H
- Subjects
- Adolescent, Adult, Animals, Animals, Genetically Modified, Child, Child, Preschool, Disease Models, Animal, Female, Gastrointestinal Microbiome immunology, Gene Knockout Techniques, Humans, Immunity, Mucosal, Immunoglobulin G blood, Immunoglobulin G immunology, Intestinal Mucosa immunology, Intestinal Mucosa microbiology, Male, Organogenesis genetics, Organogenesis immunology, Peyer's Patches immunology, Swine, Treatment Outcome, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases immunology, X-Linked Combined Immunodeficiency Diseases pathology, Bone Marrow Transplantation, Interleukin Receptor Common gamma Subunit genetics, Intestinal Mucosa growth & development, Peyer's Patches growth & development, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Background & Aims: Organ-level research using an animal model lacking Il2rg, the gene responsible for X-linked severe combined immunodeficiency (X-SCID), is clinically unavailable and would be a powerful tool to gain deeper insights into the symptoms of patients with X-SCID., Methods: We used an X-SCID animal model, which was first established in our group by the deletion of Il2rg gene in pigs, to understand the clinical signs from multiple perspectives based on pathology, immunology, microbiology, and nutrition. We also treated the X-SCID pigs with bone marrow transplantation (BMT) for mimicking a current therapeutic treatment for patients with X-SCID and investigated the effect at the organ-level. Moreover, the results were confirmed using serum and fecal samples collected from patients with X-SCID., Results: We demonstrated that X-SCID pigs completely lacked Peyer's patches (PPs) and IgA production in the small intestine, but possessed some dysfunctional intestinal T and B cells. Another novel discovery was that X-SCID pigs developed a heterogeneous intestinal microflora and possessed abnormal plasma metabolites, indicating that X-SCID could be an immune disorder that affects various in vivo functions. Importantly, the organogenesis of PPs in X-SCID pigs was not promoted by BMT. Although a few isolated lymphoid follicles developed in the small intestine of BMT-treated X-SCID pigs, there was no evidence that they contributed to IgA production and microflora formation. Consistently, most patients with X-SCID who received BMT possessed abnormal intestinal immune and microbial environments regardless of the presence of sufficient serum IgG., Conclusions: These results indicate that the current BMT therapies for patients with X-SCID may be insufficient to induce the organogenesis of intestinal lymphoid tissues that are associated with numerous functions in vivo., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2020
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16. An update on X-Linked agammaglobulinaemia: clinical manifestations and management.
- Author
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Shillitoe BMJ and Gennery AR
- Subjects
- Agammaglobulinemia therapy, Animals, Delayed Diagnosis, Genetic Therapy, Humans, Mice, X-Linked Combined Immunodeficiency Diseases therapy, Agammaglobulinemia diagnosis, Immunoglobulins, Intravenous therapeutic use, X-Linked Combined Immunodeficiency Diseases diagnosis
- Abstract
Purpose of Review: X-linked agammaglobulinaemia (XLA) is a congenital defect of development of B lymphocytes leading to agammaglobulinaemia. It was one of the first primary immunodeficiencies described, but treatment has remained relatively unchanged over the last 60 years. This summary aims to outline the current outcomes, treatments and future research areas for XLA., Recent Findings: Immunoglobulin therapy lacks IgA and IgM, placing patients at theoretical risk of experiencing recurrent respiratory tract infections and developing bronchiectasis despite best current therapy. Recent cohort studies from Italy and the USA conform that bronchiectasis remains a major burden for this group despite best current efforts. However, gene therapy offers a potential cure for these patients with proven proof of concept murine models., Summary: The potential limitations of current immunoglobulin therapy appear to be confirmed by recent cohort studies, and therefore further work in the development of gene therapy is warranted. Until this is available, clinicians should strive to reduce the diagnostic delay, regularly monitor for lung disease and individualize target immunoglobulin doses to reduce infection rates for their patients.
- Published
- 2019
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17. In-Vivo Gene Therapy with Foamy Virus Vectors.
- Author
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Rajawat YS, Humbert O, and Kiem HP
- Subjects
- Animals, Cats, Cattle, Disease Models, Animal, Dogs, Hematopoietic Stem Cells physiology, Humans, Stem Cells physiology, Transduction, Genetic veterinary, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases therapy, Genetic Therapy, Genetic Vectors, Spumavirus genetics, X-Linked Combined Immunodeficiency Diseases veterinary
- Abstract
Foamy viruses (FVs) are nonpathogenic retroviruses that infect various animals including bovines, felines, nonhuman primates (NHPs), and can be transmitted to humans through zoonotic infection. Due to their non-pathogenic nature, broad tissue tropism and relatively safe integration profile, FVs have been engineered as novel vectors (foamy virus vector, FVV) for stable gene transfer into different cells and tissues. FVVs have emerged as an alternative platform to contemporary viral vectors (e.g., adeno associated and lentiviral vectors) for experimental and therapeutic gene therapy of a variety of monogenetic diseases. Some of the important features of FVVs include the ability to efficiently transduce hematopoietic stem and progenitor cells (HSPCs) from humans, NHPs, canines and rodents. We have successfully used FVV for proof of concept studies to demonstrate safety and efficacy following in-vivo delivery in large animal models. In this review, we will comprehensively discuss FVV based in-vivo gene therapy approaches established in the X-linked severe combined immunodeficiency (SCID-X1) canine model.
- Published
- 2019
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18. New Gene Therapy Potential Cure for "Bubble Boy Disease": An experimental gene therapy has allowed children with SCID-1X to develop fully functioning immune systems.
- Subjects
- Busulfan therapeutic use, Child, Child, Preschool, Female, Gene Expression, Genetic Vectors chemistry, Genetic Vectors metabolism, Humans, Immune System drug effects, Immune System immunology, Immune System pathology, Immunosuppressive Agents therapeutic use, Infant, Interleukin Receptor Common gamma Subunit deficiency, Interleukin Receptor Common gamma Subunit immunology, Lentivirus metabolism, Male, T-Lymphocytes drug effects, T-Lymphocytes immunology, T-Lymphocytes pathology, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases immunology, X-Linked Combined Immunodeficiency Diseases pathology, Genetic Therapy methods, Interleukin Receptor Common gamma Subunit genetics, Lentivirus genetics, Therapies, Investigational methods, X-Linked Combined Immunodeficiency Diseases therapy
- Published
- 2019
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19. Hematopoietic stem cell transplantation for CD40 ligand deficiency: Results from an EBMT/ESID-IEWP-SCETIDE-PIDTC study.
- Author
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Ferrua F, Galimberti S, Courteille V, Slatter MA, Booth C, Moshous D, Neven B, Blanche S, Cavazzana M, Laberko A, Shcherbina A, Balashov D, Soncini E, Porta F, Al-Mousa H, Al-Saud B, Al-Dhekri H, Arnaout R, Formankova R, Bertrand Y, Lange A, Smart J, Wolska-Kusnierz B, Aquino VM, Dvorak CC, Fasth A, Fouyssac F, Heilmann C, Hoenig M, Schuetz C, Kelečić J, Bredius RGM, Lankester AC, Lindemans CA, Suarez F, Sullivan KE, Albert MH, Kałwak K, Barlogis V, Bhatia M, Bordon V, Czogala W, Alonso L, Dogu F, Gozdzik J, Ikinciogullari A, Kriván G, Ljungman P, Meyts I, Mustillo P, Smith AR, Speckmann C, Sundin M, Keogh SJ, Shaw PJ, Boelens JJ, Schulz AS, Sedlacek P, Veys P, Mahlaoui N, Janda A, Davies EG, Fischer A, Cowan MJ, and Gennery AR
- Subjects
- Child, Child, Preschool, Humans, Infant, Infant, Newborn, Treatment Outcome, X-Linked Combined Immunodeficiency Diseases mortality, CD40 Ligand deficiency, Hematopoietic Stem Cell Transplantation, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Background: CD40 ligand (CD40L) deficiency, an X-linked primary immunodeficiency, causes recurrent sinopulmonary, Pneumocystis and Cryptosporidium species infections. Long-term survival with supportive therapy is poor. Currently, the only curative treatment is hematopoietic stem cell transplantation (HSCT)., Objective: We performed an international collaborative study to improve patients' management, aiming to individualize risk factors and determine optimal HSCT characteristics., Methods: We retrospectively collected data on 130 patients who underwent HSCT for CD40L deficiency between 1993-2015. We analyzed outcome and variables' relevance with respect to survival and cure., Results: Overall survival (OS), event-free survival (EFS), and disease-free survival (DFS) were 78.2%, 58.1%, and 72.3% 5 years after HSCT. Results were better in transplantations performed in 2000 or later and in children less than 10 years old at the time of HSCT. Pre-existing organ damage negatively influenced outcome. Sclerosing cholangitis was the most important risk factor. After 2000, superior OS was achieved with matched donors. Use of myeloablative regimens and HSCT at 2 years or less from diagnosis associated with higher OS and DFS. EFS was best with matched sibling donors, myeloablative conditioning (MAC), and bone marrow-derived stem cells. Most rejections occurred after reduced-intensity or nonmyeloablative conditioning, which associated with poor donor cell engraftment. Mortality occurred mainly early after HSCT, predominantly from infections. Among survivors who ceased immunoglobulin replacement, T-lymphocyte chimerism was 50% or greater donor in 85.2%., Conclusion: HSCT is curative in patients with CD40L deficiency, with improved outcome if performed before organ damage development. MAC is associated with better OS, EFS, and DFS. Prospective studies are required to compare the risks of HSCT with those of lifelong supportive therapy., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2019
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20. Lentiviral Gene Therapy Combined with Low-Dose Busulfan in Infants with SCID-X1.
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Mamcarz E, Zhou S, Lockey T, Abdelsamed H, Cross SJ, Kang G, Ma Z, Condori J, Dowdy J, Triplett B, Li C, Maron G, Aldave Becerra JC, Church JA, Dokmeci E, Love JT, da Matta Ain AC, van der Watt H, Tang X, Janssen W, Ryu BY, De Ravin SS, Weiss MJ, Youngblood B, Long-Boyle JR, Gottschalk S, Meagher MM, Malech HL, Puck JM, Cowan MJ, and Sorrentino BP
- Subjects
- Antigens, Differentiation, T-Lymphocyte blood, B-Lymphocytes physiology, Hematopoietic Stem Cell Transplantation, Humans, Immunoglobulin M blood, Infant, Killer Cells, Natural, Lymphocyte Count, Male, T-Lymphocytes, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases immunology, Busulfan administration & dosage, Genetic Therapy, Genetic Vectors, Interleukin Receptor Common gamma Subunit genetics, Lentivirus, Transplantation Conditioning, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Background: Allogeneic hematopoietic stem-cell transplantation for X-linked severe combined immunodeficiency (SCID-X1) often fails to reconstitute immunity associated with T cells, B cells, and natural killer (NK) cells when matched sibling donors are unavailable unless high-dose chemotherapy is given. In previous studies, autologous gene therapy with γ-retroviral vectors failed to reconstitute B-cell and NK-cell immunity and was complicated by vector-related leukemia., Methods: We performed a dual-center, phase 1-2 safety and efficacy study of a lentiviral vector to transfer IL2RG complementary DNA to bone marrow stem cells after low-exposure, targeted busulfan conditioning in eight infants with newly diagnosed SCID-X1., Results: Eight infants with SCID-X1 were followed for a median of 16.4 months. Bone marrow harvest, busulfan conditioning, and cell infusion had no unexpected side effects. In seven infants, the numbers of CD3+, CD4+, and naive CD4+ T cells and NK cells normalized by 3 to 4 months after infusion and were accompanied by vector marking in T cells, B cells, NK cells, myeloid cells, and bone marrow progenitors. The eighth infant had an insufficient T-cell count initially, but T cells developed in this infant after a boost of gene-corrected cells without busulfan conditioning. Previous infections cleared in all infants, and all continued to grow normally. IgM levels normalized in seven of the eight infants, of whom four discontinued intravenous immune globulin supplementation; three of these four infants had a response to vaccines. Vector insertion-site analysis was performed in seven infants and showed polyclonal patterns without clonal dominance in all seven., Conclusions: Lentiviral vector gene therapy combined with low-exposure, targeted busulfan conditioning in infants with newly diagnosed SCID-X1 had low-grade acute toxic effects and resulted in multilineage engraftment of transduced cells, reconstitution of functional T cells and B cells, and normalization of NK-cell counts during a median follow-up of 16 months. (Funded by the American Lebanese Syrian Associated Charities and others; LVXSCID-ND ClinicalTrials.gov number, NCT01512888.)., (Copyright © 2019 Massachusetts Medical Society.)
- Published
- 2019
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21. The γ c Family of Cytokines: Basic Biology to Therapeutic Ramifications.
- Author
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Leonard WJ, Lin JX, and O'Shea JJ
- Subjects
- Animals, Autoimmune Diseases genetics, Autoimmune Diseases immunology, Cytokines genetics, Cytokines immunology, Evolution, Molecular, Gene Expression Regulation, Genetic Therapy, Humans, Immunologic Deficiency Syndromes immunology, Immunologic Deficiency Syndromes therapy, Janus Kinase 3 antagonists & inhibitors, Janus Kinases antagonists & inhibitors, Janus Kinases physiology, Lymphocyte Subsets immunology, Mice, Molecular Targeted Therapy, Multigene Family genetics, Neoplasms genetics, Neoplasms immunology, Protein Subunits, STAT Transcription Factors physiology, Signal Transduction, Translational Research, Biomedical, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases immunology, X-Linked Combined Immunodeficiency Diseases therapy, Cytokines classification, Interleukin Receptor Common gamma Subunit genetics, Multigene Family immunology
- Abstract
The common cytokine receptor γ chain, γ
c , is a component of the receptors for interleukin-2 (IL-2), IL-4, IL-7, IL-9, IL-15, and IL-21. Mutation of the gene encoding γc results in X-linked severe combined immunodeficiency in humans, and γc family cytokines collectively regulate development, proliferation, survival, and differentiation of immune cells. Here, we review the basic biology of these cytokines, highlighting mechanisms of signaling and gene regulation that have provided insights for immunodeficiency, autoimmunity, allergic diseases, and cancer. Moreover, we discuss how studies of this family stimulated the development of JAK3 inhibitors and present an overview of current strategies targeting these pathways in the clinic, including novel antibodies, antagonists, and partial agonists. The diverse roles of these cytokines on a range of immune cells have important therapeutic implications., (Copyright © 2019. Published by Elsevier Inc.)- Published
- 2019
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22. Gene correction for SCID-X1 in long-term hematopoietic stem cells.
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Pavel-Dinu M, Wiebking V, Dejene BT, Srifa W, Mantri S, Nicolas CE, Lee C, Bao G, Kildebeck EJ, Punjya N, Sindhu C, Inlay MA, Saxena N, DeRavin SS, Malech H, Roncarolo MG, Weinberg KI, and Porteus MH
- Subjects
- Animals, Antigens, CD34 metabolism, CRISPR-Cas Systems genetics, Cell Line, Codon, Initiator genetics, Dependovirus, Exons genetics, Fetal Blood cytology, Genetic Vectors genetics, Healthy Volunteers, Hematopoietic Stem Cells metabolism, Humans, Male, Mice, Mutation, Parvovirinae genetics, Primary Cell Culture, Time Factors, Transduction, Genetic methods, Transplantation Chimera genetics, Transplantation, Heterologous methods, X-Linked Combined Immunodeficiency Diseases genetics, DNA, Complementary genetics, Gene Editing methods, Hematopoietic Stem Cell Transplantation, Interleukin Receptor Common gamma Subunit genetics, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Gene correction in human long-term hematopoietic stem cells (LT-HSCs) could be an effective therapy for monogenic diseases of the blood and immune system. Here we describe an approach for X-linked sSevere cCombined iImmunodeficiency (SCID-X1) using targeted integration of a cDNA into the endogenous start codon to functionally correct disease-causing mutations throughout the gene. Using a CRISPR-Cas9/AAV6 based strategy, we achieve up to 20% targeted integration frequencies in LT-HSCs. As measures of the lack of toxicity we observe no evidence of abnormal hematopoiesis following transplantation and no evidence of off-target mutations using a high-fidelity Cas9 as a ribonucleoprotein complex. We achieve high levels of targeting frequencies (median 45%) in CD34
+ HSPCs from six SCID-X1 patients and demonstrate rescue of lymphopoietic defect in a patient derived HSPC population in vitro and in vivo. In sum, our study provides specificity, toxicity and efficacy data supportive of clinical development of genome editing to treat SCID-Xl.- Published
- 2019
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23. Successful in utero stem cell transplantation in X-linked severe combined immunodeficiency.
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Magnani A, Jouannic JM, Rosain J, Gabrion A, Touzot F, Roudaut C, Kracker S, Mahlaoui N, Toubert A, Clave E, Macintyre EA, Radford-Weiss I, Alcantara M, Magrin E, Ternaux B, Nisoy J, Caccavelli L, Darras AM, Picard C, Blanche S, and Cavazzana M
- Subjects
- Child, Preschool, Female, Humans, X-Linked Combined Immunodeficiency Diseases pathology, Hematopoietic Stem Cell Transplantation methods, Transplantation Conditioning methods, X-Linked Combined Immunodeficiency Diseases therapy
- Published
- 2019
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24. Impact of Day 14 Peripheral Blood Chimerism after Allogeneic Hematopoietic Stem Cell Bone Transplantation on the Treatment Outcome of Non-Malignant Disease.
- Author
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Choi YB, Lee JW, Sung KW, Koo HH, Kim HJ, and Yoo KH
- Subjects
- Adolescent, Adult, Anemia, Aplastic mortality, Anemia, Aplastic pathology, Anemia, Aplastic therapy, Child, Child, Preschool, Disease-Free Survival, Female, Graft vs Host Disease, Humans, Infant, Infant, Newborn, Male, Red-Cell Aplasia, Pure mortality, Red-Cell Aplasia, Pure pathology, Red-Cell Aplasia, Pure therapy, Retrospective Studies, Survival Rate, Transplantation Chimera blood, Transplantation, Homologous, X-Linked Combined Immunodeficiency Diseases mortality, X-Linked Combined Immunodeficiency Diseases pathology, X-Linked Combined Immunodeficiency Diseases therapy, Young Adult, Hematopoietic Stem Cell Transplantation, Transplantation Chimera genetics
- Abstract
Background: The impact of early peripheral blood chimerism on the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) is unclear. We aimed to determine whether day 14 peripheral blood chimerism after allo-HSCT predicts outcomes in patients with non-malignant diseases., Methods: Data from 56 patients who received allo-HSCT between April 2007 and March 2016 were retrospectively analyzed. Chimerism was evaluated using short-tandem repeat polymerase chain reaction, with mixed chimerism (MC) defined as greater than 1% recipient cells which was further categorized into low-level MC (> 1% and < 15% of recipient-derived cells) and high-level MC (≥ 15% of the recipient-derived cells)., Results: Thirty-six patients showed complete donor chimerism (CC), 14 low-level MC, and 6 high-level MC at day 14 post-transplant. The estimated 5-year event-free survival (EFS) was higher in the CC or low-level MC groups than in the high-level MC group (86.1% vs. 71.4% vs. 33.3%; P = 0.001). In BM or peripheral blood stem cell (BM/PBSC) transplants, the 5-year EFS was higher in the CC or low-level MC group than in the high-level MC group (93.1% vs. 66.7% vs. 0%; P < 0.001). However, in cord blood transplants, the 5-year OS and EFS according to the day 14 peripheral blood chimerism did not reach statistical significance., Conclusion: Although CC is not always necessary after allo-HSCT for non-malignant diseases, our data suggest that day 14 peripheral blood chimerism may predict outcomes in patients with non-malignant diseases who underwent BM/PBSC transplants., Competing Interests: Disclosure: The authors have no potential conflicts of interests to disclose.
- Published
- 2019
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25. Successful Bone Marrow Transplantation for XMEN: Hemorrhagic Risk Uncovered.
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Dimitrova D, Rose JJ, Uzel G, Cohen JI, Rao KV, Bleesing JH, Kanakry CG, and Kanakry JA
- Subjects
- Adolescent, Adult, Humans, Male, Young Adult, Bone Marrow Transplantation adverse effects, Epstein-Barr Virus Infections therapy, Hemorrhage etiology, Magnesium Deficiency therapy, Neoplasms therapy, X-Linked Combined Immunodeficiency Diseases therapy
- Published
- 2019
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26. Advances in site-specific gene editing for primary immune deficiencies.
- Author
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Kuo CY
- Subjects
- Animals, Humans, Agammaglobulinemia genetics, Agammaglobulinemia immunology, Agammaglobulinemia therapy, Gene Editing methods, Genetic Diseases, X-Linked genetics, Genetic Diseases, X-Linked immunology, Genetic Diseases, X-Linked therapy, Genetic Therapy methods, Wiskott-Aldrich Syndrome genetics, Wiskott-Aldrich Syndrome immunology, Wiskott-Aldrich Syndrome therapy, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases immunology, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Purpose of Review: Conventional gene therapy has been a successful, curative treatment modality for many primary immune deficiencies with significant improvements in the last decade. However, the risk of leukemic transformation with viral-mediated gene addition still remains, and unregulated gene addition is not an option for certain diseases in which the target gene is closely controlled. The recent bloom in genome modification platforms has created the opportunity to site-specifically correct mutated DNA base pairs or insert a corrective cDNA minigene while maintaining gene expression under control of endogenous regulatory elements., Recent Findings: There is an abundance of ongoing research utilizing programmable nucleases to facilitate site-specific gene correction of many primary immune deficiencies including X-linked severe combined immune deficiency, X-linked chronic granulomatous disease, Wiskott-Aldrich syndrome, X-linked hyper-IgM syndrome, X-linked agammaglobulinemia, and immune dysregulation, polyendocrinopathy, enteropathy, X-linked. In all, these studies have demonstrated the ability to integrate corrective DNA sequences at a precise location in the genome at rates likely to either cure or ameliorate disease., Summary: Gene editing for primary immune deficiency (PID) has advanced to the point to that translation to clinical trials is likely to occur in the next several years. At the current pace of research in DNA repair mechanisms, stem cell biology, and genome-editing technology, targeted genome modification represents the next chapter of gene therapy for PID.
- Published
- 2018
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27. T cell dynamics and response of the microbiota after gene therapy to treat X-linked severe combined immunodeficiency.
- Author
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Clarke EL, Connell AJ, Six E, Kadry NA, Abbas AA, Hwang Y, Everett JK, Hofstaedter CE, Marsh R, Armant M, Kelsen J, Notarangelo LD, Collman RG, Hacein-Bey-Abina S, Kohn DB, Cavazzana M, Fischer A, Williams DA, Pai SY, and Bushman FD
- Subjects
- Cell Division, Child, Preschool, Complementarity Determining Regions genetics, Humans, Receptors, Antigen, T-Cell, alpha-beta metabolism, X-Linked Combined Immunodeficiency Diseases microbiology, X-Linked Combined Immunodeficiency Diseases virology, Genetic Therapy, Microbiota, T-Lymphocytes immunology, X-Linked Combined Immunodeficiency Diseases immunology, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Background: Mutation of the IL2RG gene results in a form of severe combined immune deficiency (SCID-X1), which has been treated successfully with hematopoietic stem cell gene therapy. SCID-X1 gene therapy results in reconstitution of the previously lacking T cell compartment, allowing analysis of the roles of T cell immunity in humans by comparing before and after gene correction., Methods: Here we interrogate T cell reconstitution using four forms of high throughput analysis. (1) Estimation of the numbers of transduced progenitor cells by monitoring unique positions of integration of the therapeutic gene transfer vector. (2) Estimation of T cell population structure by sequencing of the recombined T cell receptor (TCR) beta locus. (3) Metagenomic analysis of microbial populations in oropharyngeal, nasopharyngeal, and gut samples. (4) Metagenomic analysis of viral populations in gut samples., Results: Comparison of progenitor and mature T cell populations allowed estimation of a minimum number of cell divisions needed to generate the observed populations. Analysis of microbial populations showed the effects of immune reconstitution, including normalization of gut microbiota and clearance of viral infections. Metagenomic analysis revealed enrichment of genes for antibiotic resistance in gene-corrected subjects relative to healthy controls, likely a result of higher healthcare exposure., Conclusions: This multi-omic approach enables the characterization of multiple effects of SCID-X1 gene therapy, including T cell repertoire reconstitution, estimation of numbers of cell divisions between progenitors and daughter T cells, normalization of the microbiome, clearance of microbial pathogens, and modulations in antibiotic resistance gene levels. Together, these results quantify several aspects of the long-term efficacy of gene therapy for SCID-X1. This study includes data from ClinicalTrials.gov numbers NCT01410019, NCT01175239, and NCT01129544.
- Published
- 2018
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28. Rapid immune reconstitution of SCID-X1 canines after G-CSF/AMD3100 mobilization and in vivo gene therapy.
- Author
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Humbert O, Chan F, Rajawat YS, Torgerson TR, Burtner CR, Hubbard NW, Humphrys D, Norgaard ZK, O'Donnell P, Adair JE, Trobridge GD, Scharenberg AM, Felsburg PJ, Rawlings DJ, and Kiem HP
- Subjects
- Animals, Benzylamines, CD4-CD8 Ratio, Cyclams, Disease Models, Animal, Dogs, Humans, Phosphoglycerate Kinase genetics, Dog Diseases blood, Dog Diseases genetics, Dog Diseases therapy, Genetic Therapy, Genetic Vectors pharmacology, Granulocyte Colony-Stimulating Factor pharmacology, Hematopoietic Stem Cell Mobilization, Heterocyclic Compounds pharmacology, Spumavirus, X-Linked Combined Immunodeficiency Diseases blood, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases therapy, X-Linked Combined Immunodeficiency Diseases veterinary
- Abstract
Hematopoietic stem-cell gene therapy is a promising treatment of X-linked severe combined immunodeficiency disease (SCID-X1), but currently, it requires recipient conditioning, extensive cell manipulation, and sophisticated facilities. With these limitations in mind, we explored a simpler therapeutic approach to SCID-X1 treatment by direct IV administration of foamy virus (FV) vectors in the canine model. FV vectors were used because they have a favorable integration site profile and are resistant to serum inactivation. Here, we show improved efficacy of our in vivo gene therapy platform by mobilization with granulocyte colony-stimulating factor (G-CSF) and AMD3100 before injection of an optimized FV vector incorporating the human phosphoglycerate kinase enhancerless promoter. G-CSF/AMD3100 mobilization before FV vector delivery accelerated kinetics of CD3
+ lymphocyte recovery, promoted thymopoiesis, and increased immune clonal diversity. Gene-corrected T lymphocytes exhibited a normal CD4:CD8 ratio and a broad T-cell receptor repertoire and showed restored γC-dependent signaling function. Treated animals showed normal primary and secondary antibody responses to bacteriophage immunization and evidence for immunoglobulin class switching. These results demonstrate safety and efficacy of an accessible, portable, and translatable platform with no conditioning regimen for the treatment of SCID-X1 and other genetic diseases., (© 2018 by The American Society of Hematology.)- Published
- 2018
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29. Nuclease-free Adeno-Associated Virus-Mediated Il2rg Gene Editing in X-SCID Mice.
- Author
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Hiramoto T, Li LB, Funk SE, Hirata RK, and Russell DW
- Subjects
- Alleles, Animals, Bone Marrow Cells immunology, Bone Marrow Cells metabolism, Gene Order, Genetic Therapy, Hematopoietic Stem Cells metabolism, Humans, Immunotherapy, Adoptive, Mice, Receptors, Antigen, T-Cell metabolism, T-Lymphocytes immunology, T-Lymphocytes metabolism, X-Linked Combined Immunodeficiency Diseases immunology, X-Linked Combined Immunodeficiency Diseases therapy, Dependovirus genetics, Gene Editing, Genetic Vectors genetics, Interleukin Receptor Common gamma Subunit genetics, X-Linked Combined Immunodeficiency Diseases genetics
- Abstract
X-linked severe combined immunodeficiency (X-SCID) has been successfully treated by hematopoietic stem cell (HSC) transduction with retroviral vectors expressing the interleukin-2 receptor subunit gamma gene (IL2RG), but several patients developed malignancies due to vector integration near cellular oncogenes. This adverse side effect could in principle be avoided by accurate IL2RG gene editing with a vector that does not contain a functional promoter or IL2RG gene. Here, we show that adeno-associated virus (AAV) gene editing vectors can insert a partial Il2rg cDNA at the endogenous Il2rg locus in X-SCID murine bone marrow cells and that these ex vivo-edited cells repopulate transplant recipients and produce CD4
+ and CD8+ T cells. Circulating, edited lymphocytes increased over time and appeared in secondary transplant recipients, demonstrating successful editing in long-term repopulating cells. Random vector integration events were nearly undetectable, and malignant transformation of the transplanted cells was not observed. Similar editing frequencies were observed in human hematopoietic cells. Our results demonstrate that therapeutically relevant HSC gene editing can be achieved by AAV vectors in the absence of site-specific nucleases and suggest that this may be a safe and effective therapy for hematopoietic diseases where in vivo selection can increase edited cell numbers., (Copyright © 2018 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.)- Published
- 2018
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30. Towards safe therapy for immunodeficiency.
- Author
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Tsai SQ
- Subjects
- Animals, Disease Models, Animal, Humans, Interleukin Receptor Common gamma Subunit genetics, Mice, Mice, Knockout, Mice, SCID, Gene Editing, Hematopoietic Stem Cell Transplantation, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases therapy
- Published
- 2017
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31. Acute Myeloid Leukemia in a Patient With X-linked Severe Combined Immunodeficiency.
- Author
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Shigemura T, Motobayashi M, Matsuda K, Shimodaira T, Kurata T, Kobayashi N, Agematsu K, and Nakazawa Y
- Subjects
- Chromosome Deletion, Chromosomes, Human, Pair 6, Cord Blood Stem Cell Transplantation adverse effects, Cord Blood Stem Cell Transplantation methods, Graft vs Host Disease etiology, Graft vs Host Disease prevention & control, Humans, Immunosuppressive Agents therapeutic use, In Situ Hybridization, Fluorescence, Infant, Interleukin Receptor Common gamma Subunit genetics, Loss of Heterozygosity, Male, Mutation, X-Linked Combined Immunodeficiency Diseases diagnosis, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases therapy, Leukemia, Myeloid, Acute diagnosis, Leukemia, Myeloid, Acute etiology, X-Linked Combined Immunodeficiency Diseases complications
- Abstract
Severe combined immunodeficiency (SCID) is a defect in the differentiation and function of T cells. An increased malignancy risk, mainly lymphatic malignancy, has been described in patients with SCID. We report a patient with X-linked SCID who developed acute myeloid leukemia, derived from the recipient with somatic NRAS mutation 4 months after cord blood transplantation (CBT). Loss of heterozygosity phenomenon of the recipient at 6q14 locus was observed at 2 months post-CBT and progressed to 6q deletion (6q-) chromosome abnormality. Somatic NRAS mutation was detected at 3 months post-CBT. Thus, 6q- and NRAS mutation were strongly associated with the leukemic transformation in our patient.
- Published
- 2017
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32. Targeted genome editing restores T cell differentiation in a humanized X-SCID pluripotent stem cell disease model.
- Author
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Alzubi J, Pallant C, Mussolino C, Howe SJ, Thrasher AJ, and Cathomen T
- Subjects
- Amino Acid Substitution, Animals, Cell Differentiation, Disease Models, Animal, Gene Expression, Hematopoietic Stem Cells drug effects, Hematopoietic Stem Cells pathology, Humans, Interleukin Receptor Common gamma Subunit deficiency, Interleukin Receptor Common gamma Subunit immunology, Interleukin-2 genetics, Interleukin-2 immunology, Interleukin-2 pharmacology, Interleukin-7 genetics, Interleukin-7 immunology, Interleukin-7 pharmacology, Killer Cells, Natural drug effects, Killer Cells, Natural immunology, Killer Cells, Natural pathology, Mice, Mice, SCID, Mice, Transgenic, Molecular Targeted Therapy, Mouse Embryonic Stem Cells drug effects, Mouse Embryonic Stem Cells pathology, Mutation, T-Lymphocytes drug effects, T-Lymphocytes immunology, T-Lymphocytes pathology, Transcription Activator-Like Effector Nucleases genetics, Transcription Activator-Like Effector Nucleases immunology, Transgenes, X-Linked Combined Immunodeficiency Diseases immunology, X-Linked Combined Immunodeficiency Diseases pathology, Gene Editing methods, Hematopoietic Stem Cells immunology, Interleukin Receptor Common gamma Subunit genetics, Mouse Embryonic Stem Cells immunology, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
The generation of T cells from pluripotent stem cells (PSCs) is attractive for investigating T cell development and validating genome editing strategies in vitro. X-linked severe combined immunodeficiency (X-SCID) is an immune disorder caused by mutations in the IL2RG gene and characterised by the absence of T and NK cells in patients. IL2RG encodes the common gamma chain, which is part of several interleukin receptors, including IL-2 and IL-7 receptors. To model X-SCID in vitro, we generated a mouse embryonic stem cell (ESC) line in which a disease-causing human IL2RG gene variant replaces the endogenous Il2rg locus. We developed a stage-specific T cell differentiation protocol to validate genetic correction of the common G691A mutation with transcription activator-like effector nucleases. While all ESC clones could be differentiated to hematopoietic precursor cells, stage-specific analysis of T cell maturation confirmed early arrest of T cell differentiation at the T cell progenitor stage in X-SCID cells. In contrast, genetically corrected ESCs differentiated to CD4 + or CD8 + single-positive T cells, confirming correction of the cellular X-SCID phenotype. This study emphasises the value of PSCs for disease modelling and underlines the significance of in vitro models as tools to validate genome editing strategies before clinical application.
- Published
- 2017
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33. Incident cancers and late mortality in Australian children treated by allogeneic stem cell transplantation for non-malignant diseases.
- Author
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Nelson AS, Vajdic CM, Ashton LJ, Le Marsney RE, Nivison-Smith I, Wilcox L, Dodds AJ, and O'Brien TA
- Subjects
- Adolescent, Anemia, Aplastic complications, Australia epidemiology, Bone Marrow Diseases complications, Bone Marrow Failure Disorders, Child, Child, Preschool, Female, Follow-Up Studies, Graft vs Host Disease epidemiology, Graft vs Host Disease etiology, Hemoglobinuria, Paroxysmal complications, Humans, Incidence, Infant, Infant, Newborn, Male, Metabolism, Inborn Errors complications, Neoplasms epidemiology, Neoplasms etiology, Prognosis, Risk Factors, Survival Rate, Transplantation, Homologous, X-Linked Combined Immunodeficiency Diseases complications, Anemia, Aplastic therapy, Bone Marrow Diseases therapy, Graft vs Host Disease mortality, Hematopoietic Stem Cell Transplantation adverse effects, Hemoglobinuria, Paroxysmal therapy, Metabolism, Inborn Errors therapy, Neoplasms mortality, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Background: Hematopoietic stem cell transplantation (HSCT) is a life-saving procedure for children with a variety of non-malignant conditions. However, these children face an increased risk of late death and incident cancers after HSCT, which may occur many years after their initial HSCT., Procedure: We examined cancer occurrence and late mortality in a population-based cohort of 318 Australian children who underwent allogeneic HSCT for non-malignant disease. Standardized incident ratios (SIRs) and standardized mortality ratios (SMRs) were calculated and compared with population controls., Results: We identified six (1.9%) cancers at a median 9.2 years post-HSCT. Cancer occurred 15 times more frequently than in the general population (SIR 15.4, 95% CI = 6.9-34.2). Of the 198 patients who survived for at least 2 years post-HSCT, 11 (5.6%) died at a median 7.5 years post-HSCT. The mortality rate was 17 times higher than in the general population (SMR 17.5, 95% CI = 9.7-31.2)., Discussion: Children transplanted for non-malignant conditions require evidence-based survivorship programs to reduce excess morbidity and mortality., (© 2016 Wiley Periodicals, Inc.)
- Published
- 2017
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34. Long-Term Safety Follow-Up of Subjects Previously Treated with Non-Replicating Retroviral Vector-Based Gene Therapies.
- Author
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Mohanlal R, Qiu Y, Zheng M, Mirkou A, Sridharan K, and Keir C
- Subjects
- Adult, Female, Follow-Up Studies, Genetic Vectors adverse effects, HIV Infections therapy, Hemophilia A therapy, Humans, Male, Middle Aged, Neoplasms therapy, Retroviridae isolation & purification, X-Linked Combined Immunodeficiency Diseases therapy, Young Adult, Genetic Therapy, Genetic Vectors genetics, Retroviridae genetics
- Abstract
Objective: Our objective was to evaluate the life-long safety profile of gene therapy using retroviral (non-replicating) vectors (nRCR), or cell products in 127 subjects with hemophilia, human immunodeficiency virus (HIV), or cancer, previously treated with such gene therapy., Methods: We assessed the occurrence of serious adverse events (SAEs), deaths and presence of replication competent retrovirus (RCR)., Results: A total of 23 subjects remained until the data cut-off date of 31 July 2013 and provided safety information of up to 18 years. Of the 104 subjects who discontinued, the primary reason was loss to follow-up (47.2 %; n = 60). The follow-up period for the 60 subjects lost to follow-up was 7-10 years. A total of 41 subjects experienced at least one SAE, and 15 subjects died. We reviewed SAEs and cause of death (none related to the active therapy), but no evidence was found for safety signals related to new malignancy or neurologic, rheumatological, autoimmune, or hematologic disorder. RCR results were negative, indicating no evidence for in vivo vector persistence., Conclusion: Despite the loss of follow-up, which is the limiting factor in this long-term safety trial, the findings from this long-term follow-up study are encouraging., Competing Interests: and disclosures Yuhong Qiu (YQ), Ming Zheng (MZ), Asmae Mirkou (AM), Kanaka Sridharan (KS), and Christopher Keir (CK) are employees of Novartis. At the time the study was conducted and during development of the publication, Ramon Mohanlal (RM) was an employee at Novartis. Currently, RM is an independent consultant. Funding The study was supported by Novartis Oncology. Ethical approval and informed consent This study was conducted in accordance with the ethical principles of the Declaration of Helsinki and approved by the institutional review board or ethics committee at each participating site. All subjects provided written informed consent before enrolment (Protocol identification: Chiron Corporation protocol number CS-GT005/Novartis protocol number CRV789A2201).
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- 2016
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35. Twelfth Annual Meeting of the French Society of Cell and Gene Therapy.
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Verhoeyen E, Gomez S, Galy A, Ayuso E, Midoux P, Pucéat M, Vassaux G, and Cordelier P
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- CRISPR-Cas Systems genetics, Cell- and Tissue-Based Therapy methods, Congresses as Topic, France, Genetic Therapy methods, Humans, Neoplasms genetics, Neoplasms therapy, Societies, Scientific, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases therapy, Cell- and Tissue-Based Therapy trends, Genetic Therapy trends
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- 2016
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36. Genomics of Immune Diseases and New Therapies.
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Lenardo M, Lo B, and Lucas CL
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- Animals, High-Throughput Nucleotide Sequencing, Humans, Immune System Diseases therapy, Male, Molecular Targeted Therapy, X-Linked Combined Immunodeficiency Diseases therapy, CTLA-4 Antigen genetics, Cation Transport Proteins genetics, Genomics, Immune System Diseases genetics, Mutation genetics, Phosphatidylinositol 3-Kinases genetics, X-Linked Combined Immunodeficiency Diseases genetics
- Abstract
Genomic DNA sequencing technologies have been one of the great advances of the 21st century, having decreased in cost by seven orders of magnitude and opening up new fields of investigation throughout research and clinical medicine. Genomics coupled with biochemical investigation has allowed the molecular definition of a growing number of new genetic diseases that reveal new concepts of immune regulation. Also, defining the genetic pathogenesis of these diseases has led to improved diagnosis, prognosis, genetic counseling, and, most importantly, new therapies. We highlight the investigational journey from patient phenotype to treatment using the newly defined XMEN disease, caused by the genetic loss of the MAGT1 magnesium transporter, as an example. This disease illustrates how genomics yields new fundamental immunoregulatory insights as well as how research genomics is integrated into clinical immunology. At the end, we discuss two other recently described diseases, CHAI/LATAIE (CTLA-4 deficiency) and PASLI (PI3K dysregulation), as additional examples of the journey from unknown immunological diseases to new precision medicine treatments using genomics.
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- 2016
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37. Lentiviral hematopoietic stem cell gene therapy for X-linked severe combined immunodeficiency.
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De Ravin SS, Wu X, Moir S, Anaya-O'Brien S, Kwatemaa N, Littel P, Theobald N, Choi U, Su L, Marquesen M, Hilligoss D, Lee J, Buckner CM, Zarember KA, O'Connor G, McVicar D, Kuhns D, Throm RE, Zhou S, Notarangelo LD, Hanson IC, Cowan MJ, Kang E, Hadigan C, Meagher M, Gray JT, Sorrentino BP, Malech HL, and Kardava
- Subjects
- Adolescent, Adult, B-Lymphocytes metabolism, Child, Genetic Vectors genetics, Humans, Interleukin Receptor Common gamma Subunit genetics, Killer Cells, Natural metabolism, Male, T-Lymphocytes metabolism, X-Linked Combined Immunodeficiency Diseases genetics, Young Adult, Genetic Therapy methods, Hematopoietic Stem Cells metabolism, Lentivirus genetics, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
X-linked severe combined immunodeficiency (SCID-X1) is a profound deficiency of T, B, and natural killer (NK) cell immunity caused by mutations inIL2RGencoding the common chain (γc) of several interleukin receptors. Gamma-retroviral (γRV) gene therapy of SCID-X1 infants without conditioning restores T cell immunity without B or NK cell correction, but similar treatment fails in older SCID-X1 children. We used a lentiviral gene therapy approach to treat five SCID-X1 patients with persistent immune dysfunction despite haploidentical hematopoietic stem cell (HSC) transplant in infancy. Follow-up data from two older patients demonstrate that lentiviral vector γc transduced autologous HSC gene therapy after nonmyeloablative busulfan conditioning achieves selective expansion of gene-marked T, NK, and B cells, which is associated with sustained restoration of humoral responses to immunization and clinical improvement at 2 to 3 years after treatment. Similar gene marking levels have been achieved in three younger patients, albeit with only 6 to 9 months of follow-up. Lentiviral gene therapy with reduced-intensity conditioning appears safe and can restore humoral immune function to posthaploidentical transplant older patients with SCID-X1., (Copyright © 2016, American Association for the Advancement of Science.)
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- 2016
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38. Gene Therapy for X-Linked Severe Combined Immunodeficiency: Where Do We Stand?
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Cavazzana M, Six E, Lagresle-Peyrou C, André-Schmutz I, and Hacein-Bey-Abina S
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- B-Lymphocytes immunology, B-Lymphocytes pathology, Clinical Trials as Topic, Gammaretrovirus immunology, Genetic Vectors immunology, Hematopoietic Stem Cell Transplantation, Hematopoietic Stem Cells cytology, Hematopoietic Stem Cells immunology, Humans, Killer Cells, Natural immunology, Killer Cells, Natural pathology, Lentivirus immunology, Patient Safety, Retroviridae immunology, T-Lymphocytes immunology, T-Lymphocytes pathology, Virus Integration genetics, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases immunology, X-Linked Combined Immunodeficiency Diseases pathology, Gammaretrovirus genetics, Genetic Therapy methods, Genetic Vectors chemistry, Lentivirus genetics, Retroviridae genetics, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
More than 20 years ago, X-linked severe combined immunodeficiency (SCID-X1) appeared to be the best condition to test the feasibility of hematopoietic stem cell gene therapy. The seminal SCID-X1 clinical studies, based on first-generation gammaretroviral vectors, demonstrated good long-term immune reconstitution in most treated patients despite the occurrence of vector-related leukemia in a few of them. This gene therapy has successfully enabled correction of the T cell defect. Natural killer and B cell defects were only partially restored, most likely due to the absence of a conditioning regimen. The success of these pioneering trials paved the way for the extension of gene-based treatment to many other diseases of the hematopoietic system, but the unfortunate serious adverse events led to extensive investigations to define the retrovirus integration profiles. This review puts into perspective the clinical experience of gene therapy for SCID-X1, with the development and implementation of new generations of safer vectors such as self-inactivating gammaretroviral or lentiviral vectors as well as major advances in integrome knowledge.
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- 2016
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39. Faster T-cell development following gene therapy compared with haploidentical HSCT in the treatment of SCID-X1.
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Touzot F, Moshous D, Creidy R, Neven B, Frange P, Cros G, Caccavelli L, Blondeau J, Magnani A, Luby JM, Ternaux B, Picard C, Blanche S, Fischer A, Hacein-Bey-Abina S, and Cavazzana M
- Subjects
- Adolescent, Allografts, Child, Child, Preschool, Female, Graft vs Host Disease etiology, Graft vs Host Disease genetics, Graft vs Host Disease immunology, Graft vs Host Disease pathology, Humans, Infant, Interleukin Receptor Common gamma Subunit genetics, Male, Mutation, Prospective Studies, Retrospective Studies, T-Lymphocytes pathology, Thymus Gland pathology, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases immunology, X-Linked Combined Immunodeficiency Diseases pathology, Genetic Therapy, Hematopoietic Stem Cell Transplantation, Interleukin Receptor Common gamma Subunit immunology, T-Lymphocytes immunology, Thymus Gland immunology, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
During the last decade, gene therapy via ex vivo gene transfer into autologous hematopoietic stem cells has emerged as a convincing therapy for severe combined immunodeficiency caused by ILR2G mutation (SCID-X1) despite the occurrence of genotoxicity caused by the integration of first-generation retroviral vectors. However, the place of gene therapy among the therapeutic armamentarium remains to be defined. We retrospectively analyze and compare clinical outcomes and immune reconstitution in 13 consecutive SCID-X1 patients having undergone haploidentical hematopoietic stem cell transplantation (HSCT) and 14 SCID-X1 patients treated with gene therapy over the same period at a single center level: the Necker Children's Hospital (Paris, France). Our results show a clear advantage in terms of T-cell development of gene therapy over HSCT with a mismatched donor. Patients treated with gene therapy display a faster T-cell reconstitution and a better long-term thymic output. Interestingly, this advantage of gene therapy vs haploidentical HSCT seems to be independent of the existence of clinical graft-versus-host disease in the latter condition. If data of safety are confirmed over the long term, gene therapy for SCID-X1 appears to be an equal, if not superior, alternative to haploidentical HSCT., (© 2015 by The American Society of Hematology.)
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- 2015
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40. Gene therapy outpaces haplo for SCID-X1.
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Kohn DB
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- Female, Humans, Male, Genetic Therapy, Hematopoietic Stem Cell Transplantation, Interleukin Receptor Common gamma Subunit immunology, T-Lymphocytes immunology, Thymus Gland immunology, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
In this issue of Blood, Touzot et al report that autologous gene therapy/hematopoietic stem cell transplantation (HSCT) for infants with X-linked severe combined immune deficiency (SCID-X1) lacking a matched sibling donor may have better outcomes than haploidentical (haplo) HSCT. Because gene therapy represents an autologous transplant, it obviates immune suppression before and after transplant, eliminates risks of graft versus host disease (GVHD), and, as the authors report, led to faster immunological reconstitution after transplant than did haplo transplant.
- Published
- 2015
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41. Toward Clinical Translation of New Gene Targeting Technologies for Correcting Inherited Mutations and Empowering Adoptive Immunotherapy of Cancer (SUPERSIST).
- Subjects
- Gene Targeting, Germ-Line Mutation, Hematopoietic Stem Cell Transplantation, Humans, Interleukin Receptor Common gamma Subunit genetics, T-Lymphocytes transplantation, X-Linked Combined Immunodeficiency Diseases genetics, Genetic Therapy, Immunotherapy, Adoptive, Neoplasms therapy, X-Linked Combined Immunodeficiency Diseases therapy
- Published
- 2015
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- View/download PDF
42. Clinical translation of TALENS: Treating SCID-X1 by gene editing in iPSCs.
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Biffi A
- Subjects
- Humans, Genetic Therapy methods, Immunotherapy, Adoptive, Induced Pluripotent Stem Cells physiology, Killer Cells, Natural physiology, Precursor Cells, T-Lymphoid physiology, Regeneration, Regenerative Medicine, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Mutations causing X-linked severe combined immunodeficiency (SCID-X1) reduce immune cell populations and function and may be amenable to targeted gene correction strategies. Now in Cell Stem Cell, Menon et al. (2015) correct SCID-X1-related blood differentiation defects by TALEN-mediated genome editing in patient-derived iPSCs, suggesting a possible strategy for autologous cell therapy of SCID-X1., (Copyright © 2015 Elsevier Inc. All rights reserved.)
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- 2015
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43. Lymphoid regeneration from gene-corrected SCID-X1 subject-derived iPSCs.
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Menon T, Firth AL, Scripture-Adams DD, Galic Z, Qualls SJ, Gilmore WB, Ke E, Singer O, Anderson LS, Bornzin AR, Alexander IE, Zack JA, and Verma IM
- Subjects
- Antigens, CD metabolism, Bacterial Proteins metabolism, Cell Differentiation genetics, Cell Line, DNA Repair, DNA Repair Enzymes metabolism, Humans, Induced Pluripotent Stem Cells transplantation, Infant, Interleukin Receptor Common gamma Subunit genetics, Killer Cells, Natural transplantation, Mutation genetics, Precursor Cells, T-Lymphoid transplantation, X-Linked Combined Immunodeficiency Diseases genetics, Genetic Therapy methods, Immunotherapy, Adoptive, Induced Pluripotent Stem Cells physiology, Killer Cells, Natural physiology, Precursor Cells, T-Lymphoid physiology, Regeneration, Regenerative Medicine, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
X-linked Severe Combined Immunodeficiency (SCID-X1) is a genetic disease that leaves newborns at high risk of serious infection and a predicted life span of less than 1 year in the absence of a matched bone marrow donor. The disease pathogenesis is due to mutations in the gene encoding the Interleukin-2 receptor gamma chain (IL-2Rγ), leading to a lack of functional lymphocytes. With the leukemogenic concerns of viral gene therapy there is a need to explore alternative therapeutic options. We have utilized induced pluripotent stem cell (iPSC) technology and genome editing mediated by TALENs to generate isogenic subject-specific mutant and gene-corrected iPSC lines. While the subject-derived mutant iPSCs have the capacity to generate hematopoietic precursors and myeloid cells, only wild-type and gene-corrected iPSCs can additionally generate mature NK cells and T cell precursors expressing the correctly spliced IL-2Rγ. This study highlights the potential for the development of autologous cell therapy for SCID-X1 subjects., (Copyright © 2015 Elsevier Inc. All rights reserved.)
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- 2015
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44. Gene therapy studies in a canine model of X-linked severe combined immunodeficiency.
- Author
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Felsburg PJ, De Ravin SS, Malech HL, Sorrentino BP, Burtner C, and Kiem HP
- Subjects
- Animals, Disease Models, Animal, Dogs, Humans, X-Linked Combined Immunodeficiency Diseases immunology, Genetic Therapy, Retroviridae genetics, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Since the occurrence of T cell leukemias in the original human γ-retroviral gene therapy trials for X-linked severe combined immunodeficiency (XSCID), considerable effort has been devoted to developing safer vectors. This review summarizes gene therapy studies performed in a canine model of XSCID to evaluate the efficacy of γ-retroviral, lentiviral, and foamy viral vectors for treating XSCID and a novel method of vector delivery. These studies demonstrate that durable T cell reconstitution and thymopoiesis with no evidence of any serious adverse events and, in contrast to the human XSCID patients, sustained marking in myeloid cells and B cells with reconstitution of normal humoral immune function can be achieved for up to 5 years without any pretreatment conditioning. The presence of sustained levels of gene-marked T cells, B cells, and more importantly myeloid cells for almost 5 years is highly suggestive of transduction of either multipotent hematopoietic stem cells or very primitive committed progenitors.
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- 2015
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45. Quantitative shearing linear amplification polymerase chain reaction: an improved method for quantifying lentiviral vector insertion sites in transplanted hematopoietic cell systems.
- Author
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Zhou S, Bonner MA, Wang YD, Rapp S, De Ravin SS, Malech HL, and Sorrentino BP
- Subjects
- Animals, Genetic Vectors genetics, HEK293 Cells, Humans, Jurkat Cells, Macaca nemestrina, Young Adult, Gene Targeting methods, Genetic Therapy methods, Hematopoietic Stem Cell Transplantation methods, Lentivirus genetics, Polymerase Chain Reaction methods, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
In gene therapy trials targeting blood disorders, it is important to detect dominance of transduced hematopoietic stem cell (HSC) clones arising from vector insertion site (VIS) effects. Current methods for VIS analysis often do not have defined levels of quantitative accuracy and therefore can fail to detect early clonal dominance. We have developed a rapid and inexpensive method for measuring clone size based on random shearing of genomic DNA, minimal exponential PCR amplification, and shear site counts as a quantitative endpoint. This quantitative shearing linear amplification PCR (qsLAM PCR) assay utilizes an internal control sample containing 19 lentiviral insertion sites per cell that is mixed with polyclonal samples derived from transduced human CD34+ cells. Samples were analyzed from transplanted pigtail macaques and from a participant in our X-linked severe combined immunodeficiency (XSCID) lentiviral vector trial and yielded controlled and quantitative results in all cases. One case of early clonal dominance was detected in a monkey transplanted with limiting numbers of transduced HSCs, while the clinical samples from the XSCID trial participant showed highly diverse clonal representation. These studies demonstrate that qsLAM PCR is a facile and quantitative assay for measuring clonal repertoires in subjects enrolled in human gene therapy trials using lentiviral-transduced HSCs.
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- 2015
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46. Trial watch: Self-inactivating gene-therapy vector alleviates safety concerns.
- Author
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Cully M
- Subjects
- Clinical Trials as Topic adverse effects, Genetic Vectors administration & dosage, Genetic Vectors adverse effects, Humans, Leukemia chemically induced, Leukemia genetics, Clinical Trials as Topic standards, Genetic Vectors genetics, Retroviridae genetics, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases therapy
- Published
- 2014
- Full Text
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47. Progress in gene therapy for primary immunodeficiencies using lentiviral vectors.
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Sauer AV, Di Lorenzo B, Carriglio N, and Aiuti A
- Subjects
- Clinical Trials as Topic, Common Variable Immunodeficiency genetics, Common Variable Immunodeficiency immunology, Humans, Wiskott-Aldrich Syndrome genetics, Wiskott-Aldrich Syndrome immunology, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases immunology, Common Variable Immunodeficiency therapy, Genetic Therapy, Genetic Vectors, Lentivirus, Wiskott-Aldrich Syndrome therapy, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Purpose of Review: This review gives an overview over the most recent progress in the field of lentiviral gene therapy for primary immunodeficiencies (PIDs). The history and state-of-the-art of lentiviral vector development are summarized and the recent advancements for a number of selected diseases are reviewed in detail. Past retroviral vector trials for these diseases, the most recent improvements of lentiviral vector platforms and their application in preclinical development as well as ongoing clinical trials are discussed., Recent Findings: Main focus is on the preclinical studies and clinical trials for the treatment of Wiskott-Aldrich syndrome, chronic granulomatous disease, adenosine deaminase deficient severe combined immunodeficiency (ADA-SCID) and X-linked severe combined immunodeficiency with lentiviral gene therapy., Summary: Gene therapy for PIDs is an effective treatment, providing potential long-term clinical benefit for affected patients. Substantial progress has been made to make lentiviral gene therapy platforms available for a number of rare genetic diseases. Although many ongoing gene therapy trials are based on ex-vivo approaches with autologous hematopoietic stem cells, other approaches such as in-vivo gene therapy or gene-repair platforms might provide further advancement for certain PIDs.
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- 2014
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48. X-linked immunodeficiency with magnesium defect, Epstein-Barr virus infection, and neoplasia disease: a combined immune deficiency with magnesium defect.
- Author
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Ravell J, Chaigne-Delalande B, and Lenardo M
- Subjects
- Cation Transport Proteins, Child, Epstein-Barr Virus Infections diagnosis, Epstein-Barr Virus Infections therapy, Humans, Magnesium Deficiency diagnosis, Magnesium Deficiency therapy, Male, Neoplasms diagnosis, Neoplasms therapy, Syndrome, X-Linked Combined Immunodeficiency Diseases diagnosis, X-Linked Combined Immunodeficiency Diseases therapy, Epstein-Barr Virus Infections complications, Magnesium Deficiency complications, Neoplasms complications, X-Linked Combined Immunodeficiency Diseases complications
- Abstract
Purpose of Review: To describe the role of the magnesium transporter 1 (MAGT1) in the pathogenesis of 'X-linked immunodeficiency with magnesium defect, Epstein-Barr virus (EBV) infection, and neoplasia' (XMEN) disease and its clinical implications., Recent Findings: The magnesium transporter protein MAGT1 participates in the intracellular magnesium ion (Mg) homeostasis and facilitates a transient Mg influx induced by the activation of the T-cell receptor. Loss-of-function mutations in MAGT1 cause an immunodeficiency named 'XMEN syndrome', characterized by CD4 lymphopenia, chronic EBV infection, and EBV-related lymphoproliferative disorders. Patients with XMEN disease have impaired T-cell activation and decreased cytolytic function of natural killer (NK) and CD8 T cells because of decreased expression of the NK stimulatory receptor 'natural-killer group 2, member D' (NKG2D). Patients may have defective specific antibody responses secondary to T cell dysfunction, but B cells have not been shown to be directly affected by mutations in MAGT1., Summary: XMEN disease has revealed a novel role for free intracellular magnesium in the immune system. Further understanding of the MAGT1 signaling pathway may lead to new diagnostic and therapeutic approaches.
- Published
- 2014
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- View/download PDF
49. [Autumn 2014, new chapter of gene therapy for "bubble baby"].
- Author
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Nau JY
- Subjects
- Humans, Infant, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases therapy, Genetic Therapy methods
- Published
- 2014
50. A modified γ-retrovirus vector for X-linked severe combined immunodeficiency.
- Author
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Hacein-Bey-Abina S, Pai SY, Gaspar HB, Armant M, Berry CC, Blanche S, Bleesing J, Blondeau J, de Boer H, Buckland KF, Caccavelli L, Cros G, De Oliveira S, Fernández KS, Guo D, Harris CE, Hopkins G, Lehmann LE, Lim A, London WB, van der Loo JC, Malani N, Male F, Malik P, Marinovic MA, McNicol AM, Moshous D, Neven B, Oleastro M, Picard C, Ritz J, Rivat C, Schambach A, Shaw KL, Sherman EA, Silberstein LE, Six E, Touzot F, Tsytsykova A, Xu-Bayford J, Baum C, Bushman FD, Fischer A, Kohn DB, Filipovich AH, Notarangelo LD, Cavazzana M, Williams DA, and Thrasher AJ
- Subjects
- Animals, Antigens, CD34, DNA, Complementary therapeutic use, Gene Expression, Gene Silencing, Humans, Infant, Interleukin Receptor Common gamma Subunit genetics, Male, Mice, Mutation, T-Lymphocytes immunology, Transduction, Genetic, Transgenes physiology, X-Linked Combined Immunodeficiency Diseases genetics, X-Linked Combined Immunodeficiency Diseases immunology, Gammaretrovirus genetics, Genetic Therapy adverse effects, Genetic Vectors, X-Linked Combined Immunodeficiency Diseases therapy
- Abstract
Background: In previous clinical trials involving children with X-linked severe combined immunodeficiency (SCID-X1), a Moloney murine leukemia virus-based γ-retrovirus vector expressing interleukin-2 receptor γ-chain (γc) complementary DNA successfully restored immunity in most patients but resulted in vector-induced leukemia through enhancer-mediated mutagenesis in 25% of patients. We assessed the efficacy and safety of a self-inactivating retrovirus for the treatment of SCID-X1., Methods: We enrolled nine boys with SCID-X1 in parallel trials in Europe and the United States to evaluate treatment with a self-inactivating (SIN) γ-retrovirus vector containing deletions in viral enhancer sequences expressing γc (SIN-γc)., Results: All patients received bone marrow-derived CD34+ cells transduced with the SIN-γc vector, without preparative conditioning. After 12.1 to 38.7 months of follow-up, eight of the nine children were still alive. One patient died from an overwhelming adenoviral infection before reconstitution with genetically modified T cells. Of the remaining eight patients, seven had recovery of peripheral-blood T cells that were functional and led to resolution of infections. The patients remained healthy thereafter. The kinetics of CD3+ T-cell recovery was not significantly different from that observed in previous trials. Assessment of insertion sites in peripheral blood from patients in the current trial as compared with those in previous trials revealed significantly less clustering of insertion sites within LMO2, MECOM, and other lymphoid proto-oncogenes in our patients., Conclusions: This modified γ-retrovirus vector was found to retain efficacy in the treatment of SCID-X1. The long-term effect of this therapy on leukemogenesis remains unknown. (Funded by the National Institutes of Health and others; ClinicalTrials.gov numbers, NCT01410019, NCT01175239, and NCT01129544.).
- Published
- 2014
- Full Text
- View/download PDF
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