Primary immune deficiencies comprise a group of inherited genetic disorders caused by interruption of normal lymphoid development. These diseases are considered prime candidates for gene therapy by introduction of the missing gene into hematopoietic stem cells that can then differentiate into lymphoid cells, and thus restore immunity. X-linked severe combined immunodeficiency (X-SCID), caused by a mutation in the gene encoding the common ( chain gene ((c) of the receptors for IL-2, IL-4, IL-7, IL-9, IL-15, and IL-213, is one of the more common of the primary immunodeficiencies. Clinical gene therapy trials for X-SCID have focused on the use of retroviruses as an integrating viral vector for introduction of the (c gene, resulting in restoration of immunity in patients that have engrafted with transduced, autologous hematopoietic stem cells. However, serious adverse events have emerged in these clinical trials, namely a T-cell leukemia-like syndrome developing in 5 of 21 treated patients. Therefore, safer, alterative therapies are needed. At Discovery Genomics, Inc. (DGI), we are working on developing the Sleeping Beauty (SB) transposon system for gene therapy, including the targeting of hematopoietic stem cells for treatment of primary immunodeficiencies. The two-component SB system consists of a transposon (inverted repeats (IR's) flanking a therapeutic gene of interest) and a transposase that catalyzes excision of the transposon at the ends of the IRs and then integration into host cell chromosomal sequence. As a lead immunodeficiency, here we propose that X-SCID may be treatable without the use of a virus by combining the power of electroporation for introduction of DNA into cells, along with use of DGI's Sleeping Beauty transposon system to achieve integration and long-term (c gene expression. In this Phase I application, the overall goal is to establish conditions for SB-mediated gene therapy for X-SCID. In this regard, there are two key questions that will need to be addressed: (i) Will SB-mediated transposition of the (c gene allow functional correction of lymphoid cells? (ii) What is the effectiveness of SB-mediated (c gene insertion in the treatment of an animal model of X-SCID? There are two Specific Aims: Aim 1. Evaluate the effectiveness of Sleeping Beauty transposons for correction of (c chain deficiency in a lymphoblastoid cell line derived from an X-linked SCID patient. Aim 2. Evaluate Sleeping Beauty-mediated transposition and long term expression of (c gene in hematopoietic stem cells derived of X-SCID mice, as a model for SB-mediated gene therapy of X-SCID. Successful accomplishment of these goals will provide evidence for the effectiveness of the SB system to mediate non-viral gene transfer in murine hematopoietic stem cells (previously undemonstrated), as well as provide key preclinical data for the development of SB transposons for X-SCID as a lead primary immunodeficiency.
Public Health Relevance: Results from these experiments will provide the technical basis for achieving transposon-mediated integration and long-term expression in hematopoietic stem cells, the cellular target inr gene therapy of primary immunodeficiencies. In addition, it will provide the basis for the development of the SB transposon system for treatment of other hematologic diseases.
Thesaurus Terms: Abscission; Address; Allogenic; Animal Model; Animal Models And Related Studies; Autologous; B Blood Cells; B Cell Activating Factor; B Cell Growth Factor; B-Cell Differentiation Factor-1; B-Cell Growth Factor-1; B-Cell Growth Factor-I; B-Cell Proliferating Factor; B-Cell Stimulating Factor; B-Cell Stimulating Factor-1; B-Cell Stimulation Factor-1; B-Cell Stimulatory Factor-1; B-Cells; B-Lymphocytes; Baf; Bcdf-1; Bcgf; Bcgf-1; Bcsf 1; Bsf-1; Bsf1; Bsf1 (B Cell Stimulating Factor 1); Binetrakin; Blood Diseases; Blood Precursor Cell; Bone Marrow; Bursa-Dependent Lymphocytes; Bursa-Equivalent Lymphocyte; Cd34; Cd34 Gene; Candidate Disease Gene; Candidate Gene; Cell Communication And Signaling; Cell Signaling; Cells; Clinical Trials; Clinical Trials, Unspecified; Common Cytokine Receptor Gamma Chain; Common Gamma Chain; Cytofluorometry, Flow; Cytokine Signal Transduction; Cytokine Signaling; Dna; Data; Deoxyribonucleic Acid; Development; Disease; Disorder; Effectiveness; Electroporation; Engraftment; Evaluation; Excision; Extirpation; Fetal Liver; Flow Cytofluorometries; Flow Cytometry; Flow Microfluorimetry; Gvhd; Gene Expression; Gene Transfer; Gene Transfer Clinical; Gene Transfer Procedure; Gene-Tx; Genes; Genetic Alteration; Genetic Change; Genetic Condition; Genetic Diseases; Genetic Intervention; Genetic Defect; Genomics; Goals; Graft Rejection; Graft-Versus-Host Disease; Graft-Vs-Host Disease; Hp40; Hpca1; Hematologic Diseases; Hematological Disease; Hematological Disorder; Hematopoietic; Hematopoietic Stem Cells; Hereditary; Hereditary Disease; High Affinity Interleukin-2 Receptor; Homolog Of Mouse T Cell And Mast Cell Growth Factor 40; Homologous Wasting Disease; Human; Human, General; Il-15; Il-4; Il-7; Il-9; Il15; Il15 Protein; Il2 Receptors; Il4; Il4 Protein; Il7 Protein; Il9 Protein; Immune; Immune Function, Cellular; Immunity; Immunodeficiency Disorder; Immunodeficiency Syndrome; Immunologic Deficiency Syndromes; Immunological Deficiency Syndromes; Inherited; Interleukin 2 Receptor; Interleukin 2 Receptor Gamma; Interleukin 2 Receptor, Gamma Chain; Interleukin 7 Precursor; Interleukin 9 Precursor; Interleukin-15; Interleukin-15 Precursor; Interleukin-4; Interleukin-4 Precursor; Interleukin-7; Interleukin-9; Interruption; Intervention, Genetic; Intracellular Communication And Signaling; Investigators; Laboratories; Lead; Leukemia, T-Cell; Link; Lymphocyte Stimulatory Factor 1; Lymphocytic Leukemia, T-Cell; Lymphoid; Lymphoid Cell; Lymphopoietin-1; Mcgf-2; Mgc9721; Mammals, Mice; Man (Taxonomy); Man, Modern; Marrow; Mast Cell Growth Factor-2; Mediating; Mice; Microfluorometry, Flow; Modeling; Molecular; Molecular Biology, Gene Therapy; Molecular Disease; Monitor; Murine; Mus; Mutation; Pathway Interactions; Patients; Pb Element; Phase; Physiologic Pulse; Plasmids; Preclinical Testing; Progenitor Cell Transplantation; Progenitor Cells, Hematopoietic; Proteins; Pulse; Receptors, Il-2; Recovery; Removal; Reporting; Research Personnel; Researchers; Reticuloendothelial System, Bone Marrow; Retroviridae; Retroviruses; Runt Disease; Sae; Sbir; Sbirs (R43/44); Serious Adverse Event; Signal Transduction; Signal Transduction Systems; Signaling; Site; Sleeping Beauty; Small Business Innovation Research; Small Business Innovation Research Grant; Source; Stem Cell Transplantation; Stem Cell Transplant; Surgical Removal; Syndrome; System; System, Loinc Axis 4; T-Cell Growth Factor 2; T-Cell Growth Factor P40; T-Cell Growth Factor Receptors; T-Cell Leukemia; T-Cell/Mast Cell Growth Factor P40; T-Lymphocytic Leukemia; Tcgf Receptors; Testing; Therapy, Dna; Transplant Rejection; Transplantation; Transplantation Rejection; Transposase; Viral; Viral Genes; Viral Vector; Virus; Virus-Retrovirus; Viruses, General; Work; X-Linked Severe Combined Immunodeficiency; Allotransplant; Base; Biological Signal Transduction; Blood Disorder; Cellular Targeting; Clinical Investigation; Disease/Disorder; Experiment; Experimental Research; Experimental Study; Flow Cytophotometry; Gene Product; Gene Therapy; Gene Therapy Clinical Trial; Genetic Disorder; Genetic Therapy; Genome Mutation; Genotoxicity; Heavy Metal Pb; Heavy Metal Lead; Hereditary Disorder; Hypoimmunity; Immune Deficiency Disorder; Immune Function; Immunodeficiency; Interest; Lymphoblastoid Cell Line; Model Organism; P40 Cytokine; P40 Protein; Pathway; Pre-Clinical; Preclinical; Public Health Relevance; Research Study; Resection; Restoration; Therapeutic Gene; Transfer Of A Gene; Transplant
