Mucopolysaccharidosis type II (MPS II, Hunter syndrome) is an X-linked recessive inherited disorder caused by absence of iduronate-2-sulfatase, resulting in systemic accumulation of glycosaminoglycans heparan sulphate and dermatan sulphate. Affected individuals suffer from skeletal abnormalities, organomegaly, life- threatening obstructive airway disease, and, in the severely enzyme deficient form, neurologic degeneration and death by age 15. While transplantation and engraftment of hematopoietic stem cells has shown efficacy in the treatment of some MPS diseases, allografted MPSII patients have thus far not exhibited improved neurologic outcomes. In this project, we hypothesize that allotransplant for MPSII is ineffective due to insufficient generation of IDS enzyme from engrafted cells, and that genetic engineering of donor cells to express high levels of IDS will overcome this insufficiency and provide effective metabolic cross- correction that includes neurologic manifestations of the disease. Lentigen is a leading company in the development of lentiviral vectors for treatment of human disease. In this Phase I STTR project, we propose to combine Lentigen's lentiviral vector technology with the University of Minnesota's experience in cellular therapies for lysosomal storage diseases by developing an ex vivo transduction approach for the treatment of Hunter syndrome, MPS II. The Specific Aims of the proposal are: (i) To construct and test lentiviral vectors for transduction of the human IDS gene along with green fluorescent protein as a cellular marker. Vector constructs will be generated based on dual promoter, bicistronic and fusion protein strategies, and packaged using Lentigen's proprietary LentiMax platform. (ii) Correction of metabolic and neurologic disease by ex vivo lentiviral transduction of the human IDS gene into hematopoietic stem cells of MSPII mice. Marrow from IDS deficient mice will be transduced with lentiviral vector carrying the IDS gene and transplanted into IDS deficient recipients as a model for ex vivo gene therapy of Hunter syndrome targeting hematopoietic stem cells. Treated animals will be tested for engraftment and transduction of donor cells, IDS enzyme expression in plasma and tissues, clearing of storage materials in urine and in tissues, and improved performance in neurobehavioral tests of learning and motor function. The overall goal of the proposed studies is to provide preclinical data to support the most straightforward and feasible approach for implementation of gene therapy for MPS II, with implications for the development of lentiviral gene therapies for other lysosomal storage diseases in the future.
Public Health Relevance: Lysosomal storage disorders are a rare group of inherited diseases caused by genetic deficiency in which patients suffer from skeletal abnormalities, heart and breathing problems, mental retardation and death. It is envisioned in this grant application that one way to treat these diseases would be to restore the missing gene in patients cells so that the cells can be returned to the patient. The specific research proposed in this application will use a mouse model for a lysosomal storage disease to establish and test conditions for introduction of the correcting gene and its effectiveness when reintroduced into the animals.
Thesaurus Terms: Adrgnd; Aids Virus; Acquired Immune Deficiency Syndrome Virus; Acquired Immunodeficiency Syndrome Virus; Adrenal Glands; Adrenals; Affect; Age; Allogenic; Allografting; Ammon Horn; Animal Experimental Use; Animal Experimentation; Animal Research; Animal Testing; Animals; Applications Grants; Aspiration, Respiratory; Assay; Bioassay; Biologic Assays; Biological Assay; Biotechnology, Genetic Engineering; Blood Coagulation Disorders; Blood Plasma; Blood Precursor Cell; Blood Leukocyte; Body Tissues; Bone Marrow Transplant; Bone Marrow Transplantation; Brain; Breathing; Cells; Central Nervous System; Cerebellum; Cessation Of Life; Chimera Protein; Chimeric Proteins; Chondroitin Sulfate B; Clinical Trials; Clinical Trials, Unspecified; Coagulation Disorder; Coagulopathy; Cornu Ammonis; Corpus Striatum; Corpus Striatum Structure; Cytofluorometry, Flow; Data; Death; Dermatan Sulfate; Dermatan, 4-(Hydrogen Sulfate); Development; Disease; Disease Model; Disorder; Effectiveness; Encephalon; Encephalons; Engineering; Engineerings; Engraftment; Enzymes; Exhibits; Fibroblasts; Flow Cytofluorometries; Flow Cytometry; Flow Microfluorimetry; Fusion Protein; Future; Gag; Gag Gene; Gfp; Gargoylism, Hunter Syndrome; Gene Transfer; Gene Transfer Clinical; Gene Transfer Procedure; Gene-Tx; Generations; Genes; Genetic; Genetic Diseases, Inborn; Genetic Engineering; Genetic Intervention; Glycosaminoglycans; Goals; Grafting, Bone Marrow; Grant Proposals; Grants, Applications; Green Fluorescent Proteins; Hiv; Hsc Transplantation; Htlv-Iii; Harvest; Heart; Hematopoietic Stem Cell Transplantation; Hematopoietic Stem Cells; Hemoglobinopathies; Hemoglobinopathies / Iron Metabolism; Heparan Sulfate; Heparitin Sulfate; Hereditary; Hippocampus; Hippocampus (Brain); Histologic; Histologically; Human; Human Immunodeficiency Viruses; Human T-Cell Leukemia Virus Type Iii; Human T-Cell Lymphotropic Virus Type Iii; Human T-Lymphotropic Virus Type Iii; Human, General; Hunter Corrective Factor; Hunter Syndrome Gargoylisms; Hunter Syndrome; Hunter's Syndrome; Hunter-Fraser Syndrome; Hunter-Mcalpine Syndrome; Iduronate Sulfatase; Iduronate Sulfate Sulfatase; Iduronatesulfate Sulfohydrolase; Immune; In Vitro; Inborn Genetic Diseases; Individual; Inhalation; Inhaling; Inherited; Inherited Disorder; Inspiration, Respiratory; Intervention, Genetic; Kidney; L-Iduronate-2-Sulfate 2-Sulfohydrolase; Lav-Htlv-Iii; Lentiviral Vector; Lentivirus Vector; Leukocytes; Life; Link; Liver; Lung; Lung Diseases, Obstructive; Lymphadenopathy-Associated Virus; Lysosomal Enzyme Disorders; Lysosomal Storage Diseases; Lysosomes; Mps Ii; Malignant; Malignant - Descriptor; Mammals, Mice; Man (Taxonomy); Man, Modern; Marrow; Marrow Transplantation; Marrow Leukocyte; Mediating; Mental Retardation; Metabolic; Mice; Microfluorometry, Flow; Minnesota; Modeling; Molecular Biology, Gene Therapy; Molecular Biology, Genetic Engineering; Mucopolysaccharides; Mucopolysaccharidoses; Mucopolysaccharidosis; Mucopolysaccharidosis 2; Mucopolysaccharidosis Ii; Murine; Mus; Nervous System, Brain; Nervous System, Cns; Neuraxis; Neurologic; Neurologic Manifestations; Neurologic Signs And Symptoms; Neurologic Outcome; Neurological; Neurological Manifestations; Neurological Outcome; Obstructive Lung Diseases; Patients; Performance; Peripheral; Phase; Plasma; Population; Prevention; Procedures; Progenitor Cells, Hematopoietic; Programs (Pt); Programs [publication Type]; Promoter; Promoter Regions; Promoter Regions (Genetics); Promoters (Genetics); Promotor; Promotor (Genetics); Promotor Regions; Promotor Regions (Genetics); Protocol; Protocols Documentation; Recombinant Dna Technology; Research; Respiratory System, Lung; Reticuloendothelial System, Leukocytes; Reticuloendothelial System, Serum, Plasma; Reticuloendothelial System, Spleen; Risk; Rotarod Assay; Rotarod Method; Rotarod Performance Test; Rotarod Test; Sttr; Safety; Serum, Plasma; Small Business Technology Transfer Research; Spleen; Striate Body; Striatum; Sulfoiduronate Sulfatase; T-Cells; T-Lymphocyte; Technology; Testing; Therapeutic; Therapy, Dna; Thymus-Dependent Lymphocytes; Time; Tissues; Transplantation; Universities; Urinary System, Kidney; Urinary System, Urine; Urine; Virus-Hiv; Visceromegaly; White Blood Cells; White Cell; X-Linked Hurler Syndrome; Allotransplant; Base; Beta-Heparin; Body System, Hepatic; Clinical Investigation; Clotting Disorder; Disease/Disorder; Disorder Model; Enzyme Activity; Enzyme Deficiency; Enzyme Therapy; Experience; Flow Cytophotometry; Gene Correction; Gene Therapy; Gene-Corrected; Genetic Promoter Element; Genetic Therapy; Hippocampal; Human Disease; Iduronate Sulfatase (Ids) Deficiency; Iduronate Sulfatase Deficiency; Iduronate-2-Sulfatase; Iduronate-2-Sulfatase Deficiency; Improved; In Vivo; Inborn Error; Inborn Lysosomal Enzyme Disorder; Inspiration; Leukodystrophy; Locomotor Learning; Lymphoblastoid Cell Line; Morris Water Maze; Morris Watermaze; Motor Learning; Mouse Model; Mucopolysaccharide Storage Disease Ii; Mucopolysaccharidosis (Mps) Ii; Mucopolysaccharidosis Type Ii; Neural Manifestation; Neurobehavioral Test; Obstructive Airway Disease; Obstructive Lung Disease (Generalized); Organ System, Hepatic; Organomegaly; Peripheral Blood; Pre-Clinical; Preclinical; Prevent; Preventing; Programs; Public Health Relevance; Pulmonary; Renal; Respiratory Airway Obstruction; Safety Testing; Skeletal Abnormality; Splanchnomegaly; Striatal; Sulfo-Iduronate Sulfatase (Sids) Deficiency; Sulfo-Iduronate Sulfatase Deficiency; Suprarenal Gland; Thymus Derived Lymphocyte; Transfer Of A Gene; Transplant; Vector; White Blood Cell; White Blood Corpuscle
