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Principal Investigator: Harry L Malech
Organization: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
Fiscal Year: 2020
Award: $669,910
Funding agency: National Institute of Allergy and Infectious Diseases
This project is focused on developing curative allogentic 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:
1. Bosticardo M, et al, (2020) Blood Adv 4, 2611; 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
2. Dimitrova D, et al. (2020) Biol Blood Marrow Transplant 26, 94; 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.
3. Espinoza DA, et al. (2019) Mol Ther 27, 1074; Reported markedly abnormal dysplastic clonal hematopoiesis in a rhesus macaque transplanted with hematopoietic stem cells that were transduced with a lentivector containing a strong retroviral murine stem cell virus (MSCV) constitutive promoter-enhancer in the long terminal repeat, suggesting that that strong constitutive promoters should not be included in lentivectors.
4. Jones K, et al. (2019) Biol Blood Marrow Transplant 25, 193; Demonstration of a novel pregnancy related immunosuppressive protein used as a treatment to prevent graft versus host disease in a mouse model of transplant.
5. Kohn DB, et al (2020) Nat Med 26, 200; 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.
6. Kuhns DB, et al. (2019) Blood Adv 3, 136; First pcr based genetic analysis of the NCF1 gene and its pseudogenes NCF1B and NCF1C quantifying the exon 2 GT deletion used as diagnostic tool to diagnose patients with the p47phox deficient autosomal recessive form of CGD; demonstration that there is extraordinarily high rates of recombination related loss or gain of pseudogene or gene alleles; and demonstration that carriers of p47phox deficient AR CGD have less than normal amounts of p47phox protein in their myeloid cells.
7. Lin CH, et al. (2020) J Clin Immunol, in press; 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.
8. Malech HL, et al. (2019) Methods Mol Biol 2087, 3; An introductory overview of advances across the field of study of biology and function of human neutrophils is provided.
9. Mamcarz E, et al. (2019) N Engl J Med 380, 1525; Reported first in human restoration of both cellular and humoral immunity in 8 infants newly diagnosed with X-linked severe combined immune deficiency treated with lentivector transduced autologous stem cells following low dose targeted busulfan myeloid conditioning.
10. Marsh RA, et al. (2020) J Clin Immunol 39, 653; 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.
11. Parta M, et al (2020) J Clin Immunol 40, 619; 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. However, among the surviving 5 are 3 who entered the study with severe likely fatal fungal infections not responding to conventional therapy. Thus the haploidentical transplant was a life-saving successful salvage therapy for these patients. A follow on study adding campath to the regimen to prevent GVHD is in progress with successful transplant and no GVHD in the first patient treated in this follow on study.
12. Pavel-Dinu M, et al, (2019) Nat Commun 10, 1634; First demonstration ex vivo and in vivo with xenograft in the immune deficient mouse of correction of human patient derived X-SCID CD34+ HSC using a novel approach to CRISPR/Cas9 gene editing. This is an important first pre-clinical demonstration that will lead to use of gene editing in a future clinical trial of gene editing planned for treatment of X-SCID.
13. Author correction to 12.
14. Author correction to 12.
15. Ravell JC, et al. (2019) J Clin Invest 130, 507; Demonstration that the MagT1 deficiency immune defect is mediated by the requirement for this magnesium transporter for functionally essential glycosylation of a large range of essential receptor molecules on immune cells leading to multisystem abnormalities. This is an important pathophysiology component of XMEN that will be important to developing gene editing and other gene therapy corrective approaches to treat this PID.
16. Sweeney CL, et al. Methods Mol Biol 1982, 623; A detailed review of novel methods to achieve functional correction of chronic granulomatous disease in CD34+ HSC and iPSC. This will be an important reference document to assist in advancing research in the field.
Terms: <19S Gamma Globulin><2019 novel coronavirus><2019-nCoV><Acute><Affect><Aldrich Syndrome><Alkylating Agents><Alkylators><Alleles><Allelomorphs><Allogeneic Transplantation><Allogenic><Animal Model><Animal Models and Related Studies><Antibodies><Antiviral Agents><Antiviral Drugs><Antivirals><Autoimmune Status><Autoimmunity><Autologous><B blood cells><B cell><B cells><B-Cells><B-Lymphocytes><B-cell><Bibliography><Biological><Biology><Blood><Blood Diseases><Blood Neutrophil><Blood Polymorphonuclear Neutrophil><Blood Precursor Cell><Blood Reticuloendothelial System><Bone Marrow Grafting><Bone Marrow Transplant><Bone Marrow Transplantation><Bussulfam><Busulfan><Busulfanum><CAR T cells><CD154 Antigens><CD34><CD34 gene><CD40 Ligand><CD40-L><COVID-19><COVID19><CRISPR><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/Cas method><CRISPR/Cas system><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><CTX><CYCLO-cell><Campath><Carloxan><Cas nuclease technology><Case Study><Cell Body><Cell Function><Cell Line><Cell Mediated Immunology><Cell Process><Cell Therapy><Cell model><Cell physiology><Cell-Mediated Immunity><CellLine><Cells><Cellular Function><Cellular Immune Function><Cellular Immunity><Cellular Physiology><Cellular Process><Cellular model><Cessation of life><Chronic GVHD><Chronic Granulomatous Disease><Ciclofosfamida><Ciclofosfamide><Cicloxal><Clafen><Claphene><Clinic><Clinical><Clinical Research><Clinical Study><Clinical Trials><Clustered Regularly Interspaced Short Palindromic Repeats><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Colitis><Collaborations><Cycloblastin><Cycloblastine><Cyclophospham><Cyclophosphamide><Cyclophosphamidum><Cyclophosphan><Cyclophosphane><Cyclophosphanum><Cyclostin><Cyclostine><Cytophosphan><Cytophosphane><Cytoxan><DNA Recombination><DNA Therapy><Death><Defect><Development><Diagnosis><Diagnostic><Disease><Disorder><Dose><Drugs><Dysfunction><Elements><Endoxan><Endoxana><Enduxan><Engineering><Engraftment><Enhancers><Epidemiology><Exons><Failure><Fosfaseron><Functional disorder><Fungus Diseases><Future><Gene Transfer Clinical><Genes><Genetic><Genetic Alteration><Genetic Change><Genetic Intervention><Genetic Recombination><Genetic analyses><Genetic defect><Genoxal><Genuxal><Gestation><Goals><Graft Enhancements><GvHD><HIGM1><HPCA1><HSC differentiation><HSC transplantation><Hematologic Diseases><Hematological Disease><Hematological Disorder><Hematopoiesis><Hematopoietic Cellular Control Mechanisms><Hematopoietic Progenitor Cells><Hematopoietic Stem Cell Research><Hematopoietic 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Disorder><Inherited><Institutes><Institution><Investigators><Laboratories><Ledoxina><Lentiviral Vector><Lentivirus Vector><Life><Link><Long Terminal Repeats><Lymphocyte><Lymphocytic><M mulatta><M. mulatta><MabCampath><Macaca mulatta><Magnesium><Marrow><Marrow Neutrophil><Marrow Transplantation><Mediating><Medical><Medication><Messenger RNA><Metabolic Glycosylation><Methods><Mg element><Mice><Mice Mammals><Mitoxan><Modern Man><Murine><Mus><Mutation><Mycoses><Myelogenous><Myeloid><Myeloid Cells><NHGRI><NHLBI><NIAID><NIH><National Center for Human Genome Research><National Heart, Lung, and Blood Institute><National Human Genome Research Institute><National Institute of Allergy and Infectious Disease><National Institutes of Health><Natural History><Neosar><Neutropenia><Neutrophilic Granulocyte><Neutrophilic Leukocyte><Newly Diagnosed><Other Genetics><Outcome><Oxidases><Patients><Pharmaceutic Preparations><Pharmaceutical Preparations><Physiology><Physiopathology><Polymorphonuclear 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