Gene Therapy and Hematopoietic Stem Cell Research to Treat Inherited Primary Immune Deficiencies

National Institute Of Allergy And Infectious Diseases

Investigators

Linked publications & trials

Abstract

This project is focused on developing curative allogeneic HSC transplants and autologous HSC gene therapies for primary immune deficiencies (PIDs) and primary immune regulatory disorders (PIRDs). Our laboratory program is designed to facilitate the translation of our laboratory efforts into therapeutic clinical trials of transplant and gene therapy. The project also must include clinical studies to understand the basic physiology, genetic defects, clinical problems, and management issues affecting the patient groups for which we are developing gene therapies. We use a variety of cell lines, primary patient cells, and animal models to develop these gene and cell therapy treatments; and a variety of tools including integrating and non-integrating gene transfer vectors, as well as gene editing reagents/approaches; as well as methods to transiently correct function in mature immune cells by transfection with mRNA. We also study means of enhancing HSC engraftment and preventing GVHD. Our 13 ongoing therapeutic, lab and clinic facilitating, and natural history clinical trials listed elsewhere in this report are a central element to progress of our research initiatives. A summary of research accomplishments published during the reporting period for this report and deriving from studies in our lab and clinic, and in collaboration with other investigative teams follows below. The numbering refers to the number of the citation included in the Bibliography section associated with this report: Reference # 1. First demonstration of the use of lentivector gene therapy to achieve successful long term correction of the oxidase defect in the neutrophils of patients with X-linked chronic granulomatous disease. This is an important clinical proof of principle breakthrough gene therapy employing lentivector transduction of patient autologous CD34+ HSC. 2. A new clinical lentivector to treat the autosomal recessive p47phox deficient CGD has been developed that appears to be safe and effective and that will be used in a clinical trial in development that will open for recruitment in the next year. 3. First demonstration of high level correction of X-linked CGD by gene editing using a AAV donor encoding the CYBB gene cDNA. 4. This is a report of development of a method of optimum conditions for CRISPR/Cas9 gene editing of human hematopoietic stem cells. 5. This represents a definitive description of clinical trials results of lentivector gene therapy to achieve functional cure of a very large cohort of patients with ADA-SCID. 6. Definitive report of the long term outcomes of gene therapy for ADA SCID using murine gamma retrovirus vectors indicating a broad range of degree of immune correction. 7. Case report demonstration that lentivector gene therapy for infants with X-linked SCID can result in resolution of disseminated BCGosis. 8. Important gene editing methods patent application applying the methods described in reference #4 just above. 9. Improved methods of gene editing used to demonstrate functional correction of T cells and hematopoietic stem cells from patients with MAGT1 deficiency. 10. A novel artificial thymic organoid system combined with included pluripotent stem cells a an extraordinarily informative system for examining both the immune defect in RAG-2 deficiency and as a model to test methods to correct this immune deficiency. 11. Correction to reference #10 above 12. Editorial providing some history of gene editing as applied to treatment of hemoglobinopathies. 13. First demonstration of efficient gene editing correction of the sickle mutation in human patient blood stem cells in vitro and in a mouse/human xenograft model; and use of the same gene editing methods targeting the homologous genetic site to create the sickle trait in non-human primates. 14. A first demonstration that CGD neutrophils can achieve oxidase activity correction at clinical scale using mRNA transfection. A clinical trial will shortly proceed using this methodology to provide autologous corrected granulocyte transfusions to patients with CGD. 15. Similar to reference #14 using mRNA transfection to correct CGD neutrophils, this demonstrates that the same methodology of mRNA transfection can correct T lymphocyte function in T cells from patients with MAGT1 deficiency (XMEN syndrome). 16. Report of the largest series of haploidentical transplants for chronic granulomatous disease employing post-transplant Cytoxan, demonstrating high rates of engraftment but also unacceptably high rates of graft versus host disease including two deaths from GVHD among the 7 transplanted patients. A follow on study adding campath to the regimen to prevent GVHD is in progress suggesting an reduction in the rate and severity of GVHD. 17. First report of multicenter review of outcomes of transplant for chronic granulomatous disease from the Primary Immune Deficiency Treatment Consortium showing that bone marrow transplant for CGD patients who have CGD related significant inflammatory bowel disease will fully resolve the IBD by one year after successful transplant. It is the first demonstration that transplant can correct CGD colitis. 18. Demonstration of successful outcome of allogeneic transplant of patients with a variety of immune deficiencies using a radiation-free reduced intensity regimen. This is an important advance in transplant for this patient population. 19. A novel ex vivo method of culture to recapitulate thymus differentiation of lymphocytes was developed and shows to be useful in delineating the step in T cell differentiation that is impaired in patients with inherited defects in T lymphocytes. This is important to delineating a therapeutic plan for these patients 20. A definite report of the range of diagnostics findings using computerized tomography to delineate GI disease occurring in patients with CGD. 21. This is a report of the safety and efficacy of ustekinumab to treat severe inflammatory bowel disease in patients with CGD showing that some patients safely achieve significant benefit, but others either may not benefit or the improvement can be transient. 22. This is a report of the safety and efficacy of vedolizumab to treat severe inflammatory bowel disease in patients with CGD showing that some patients safely achieve significant benefit, but others either may not benefit or the improvement can be transient. 23. This is a definitive report of showing that mutation of the IL37 gene can be a cause of infantile onset severe inflammatory bowel disease. 24. This is a case report demonstrating that a homozygous variant p. Arg90His in NCF1 is associated with early-onset interferonopathy autoinflammatory disease. 25. Plasma gelsolin (pGSN) is reduced in several disease states, including severe trauma, burns, and sepsis. This study demonstrated low pGSN in patients with CGD and this is normalized after successful transplant. 26. This is a case report of a patient with X-linked severe combined immunodeficiency (X-SCID) who survived for over 20 years without hematopoietic stem cell transplantation (HSCT) because of a somatic reversion mutation, but who had progressive loss of B cells over that period of time. This phenomenon of progressive B cell loss is increasingly being seen in long term X-SCID survivors. 27. Granulibacter bethesdensis demonstrate presence of a unique cell wall glycolipid that is hypostimulatory to neutrophils and may explain how this organism evades host immunosurveillance. 28. This is an editorial commenting on the importance of the discovery of a new genetic form of severe chronic neutropenia. 29. This is an updated report of newly reported mutations in the CYBB gene causing X-linked chronic granulomatous disease. Knowing new mutations informs studies of gene editing correction of CGD and assists with genetic diagnosis.