SBIR-STTR Award

Mucopolysaccharidosis type I (MPS I) is an autosomal recessive storage disease caused by the absence of - L-iduronidase (IDUA), resulting in systemic accumulation of glycosaminoglycan (GAG) storage materials, hepatosplenomegaly, skeletal dysplasias, cardiopulmonary obstruction, progressive neurologic impairment and death by age 15. MPS I is currently treated by enzyme replacement therapy and by allogeneic hematopoietic stem cell transplantation, but these treatments are extraordinarily expensive and do not fully address the skeletal, cardiac and neurologic manifestations of the disease. Here we propose an entirely novel approach to the treatment of MPS I. Immusoft Corp. is developing genetically engineered autologous human B cells for production of therapeutic proteins upon infusion into patients. Discovery Genomics, Inc. (DGI) focuses on clinical development of the Sleeping Beauty (SB) transposon system for integrative gene transfer and expression. Here we propose to combine Immusoft's novel B cell expression platform with DGI's advanced DNA-mediated cellular genetic engineering technology for the purpose of expressing human IDUA from B cells in vivo as an approach to achieve systemic expression of IDUA as a treatment for MPS I. This collaborative project is further strengthened by the extensive experience of DGI's investigative team and colleagues at the University of Minnesota in the conduct of preclinical studies and clinical trials of new treatments for lysosomal storage diseases, in particular MPS I. For this Phase I study, the Specific Aims are; (i) Sleeping Beauty mediated human iduronidase gene transfer and expression in primary human B cells cultured in vitro. B cells will be isolated from human blood and expanded in culture using Immusoft's Immune System Programming technology. Early in the expansion process, cells will be collected and nucleofected with SB transposon DNA encoding human IDUA along with a plasmid encoding SB transposase to mediate integration into host B cell chromosomes. (ii) These cells will then be infused into immunodeficient (NOD-SCID), IDUA deficient mice, evaluating recipient animals for engraftment, expression and distribution of IDUA activity, and correction of metabolic and neurologic disease. Results from these studies will be directly applicable to the development of a clinical protocol for treatment of human MPS I by infusion of B cells genetically engineered using the SB transposon system.

Public Health Relevance Statement:


Public Health Relevance:
Lysosomal storage disorders are a rare group of inherited diseases in which patients suffer from skeletal abnormalities, heart and breathing problems, mental retardation and death. While some of these diseases can be treated by protein therapy or by bone marrow transplantation, these treatments are expensive and incompletely effective. It is envisioned in this grant application that one way to treat these diseases would be to restore the missing gene in patients' white blood cells as a way of providing the missing protein and restoring function in the different organs.

Project Terms:
Address; Affect; Age; Allogenic; Animals; Applications Grants; Autologous; B-Lymphocytes; Biological Assay; Blood; Blood Circulation; bone; Bone Marrow Transplantation; Breathing; Cardiac; Cardiopulmonary; Cell Culture Techniques; Cell physiology; cell type; Cells; cellular engineering; Cessation of life; Chromosomes; Clinical; Clinical Protocols; Clinical Trials; Congenital Heart Defects; Cornea; Dermatan Sulfate; Development; Disease; DNA; DNA Transposons; Effectiveness; Engineering; Engraftment; enzyme activity; enzyme replacement therapy; Enzymes; experience; functional restoration; Gene Expression; Gene Transfer; Generations; Genes; Genetic Engineering; Genomics; Glycosaminoglycans; Goals; Heart; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Heparitin Sulfate; Hepatosplenomegaly; Human; Immune system; Impaired cognition; Impairment; improved; In Vitro; in vivo; Individual; Infusion procedures; Inherited; L-Iduronidase; Leukocytes; Lysosomal Storage Diseases; Mediating; Mental Retardation; Metabolic; Minnesota; Modeling; Morbidity - disease rate; Mortality Vital Statistics; mouse model; Mucopolysaccharidoses; Mucopolysaccharidosis I; Mucopolysaccharidosis I H; Mus; Nervous System Physiology; Nervous system structure; Neurologic; Neurologic Manifestations; novel; novel strategies; Obstruction; Organ; Patients; Penetration; peripheral blood; phase 1 study; Physiological; Plasma Cells; plasmid DNA; Plasmids; Population; preclinical study; Production; programs; protein function; Proteins; public health relevance; restoration; Risk; skeletal; skeletal abnormality; skeletal dysplasia; Sleeping Beauty; standard of care; Stem cell transplant; synergism; System; Technology; Testing; therapeutic protein; Tissues; Translations; Transposase; Universities

Mucopolysaccharidosis type I (MPS I) is an autosomal recessive storage disease caused by the absence of ?- L-iduronidase (IDUA), resulting in systemic accumulation of glycosaminoglycan (GAG) storage materials, hepatosplenomegaly, skeletal dysplasias, cardiopulmonary obstruction, progressive neurologic impairment and death by age 10. MPS I is currently treated by enzyme replacement therapy and by allogeneic hematopoietic stem cell transplantation, but these treatments are extraordinarily expensive and do not fully address the skeletal, cardiac and neurologic manifestations of the disease. Here we propose an entirely novel approach to the treatment of MPS I. Immusoft Corp. is developing genetically engineered autologous human B cells for production of iduronidase protein upon infusion into patients. This Phase II SBIR application is based on a Phase I award to Immusoft and Discovery Genomics, Inc. (DGI) to apply DGI's Sleeping Beauty (SB) transposon system for non-viral genetic engineering and expression of human iduronidase in B cells as a novel cellular therapy for MPS I. Through this collaboration, we have demonstrated; (i) reliable SB-mediated IDUA transposition and expression in primary human B cells in large scale culture, and (ii) significant reduction of metabolic disease when these B cells are adoptively transferred into immunodeficient NSG-MPS I mice. In this Phase II proposal, we describe plans to carry out all necessary activities supporting IND submission and initiation of clinical testing of Immusoft's B cell product in MPS I patients. In Aim 1 we will conduct complete proof-of-concept studies testing the therapeutic efficacy of IDUA-expressing B cell product adoptively transferred into immunodeficient NSG-MPSI mice, including dose-ranging studies, multiple dose studies, and thorough analysis of physiological outcomes including cardiac, skeletal, and neurologic assessment. In Aim 2 we will test the neurologic effectiveness of B cells expressing IDUA that has been modified for enhanced transit across the blood-brain barrier into the central nervous system, a key target for treating MPS I. In Aim 3 we will conduct a one-year adoptive transfer study to assess the persistence of IDUA expressing B cells in NSG-MPSI mice. In Aim 4 we will conduct complete toxicologic studies under Good Laboratory Practice conditions to determine the safety of Immusoft's B cell product. These studies will provide IND-enabling results essential to the development and clinical testing of Immusoft's IDUA-expressing B cell product for the treatment of MPS I. Effectiveness of SB-engineered B cells for treatment of MPS I will have substantial ramifications for future treatment of other metabolic diseases by infusion of B cells genetically engineered to express secretable protein for system-wide distribution.

Public Health Relevance Statement:
Project Narrative Lysosomal storage disorders are a rare group of inherited diseases in which patients suffer from skeletal abnormalities, heart and breathing problems, mental retardation and death. While some of these diseases can be treated by protein therapy or by bone marrow transplantation, these treatments are expensive and incompletely effective. It is envisioned in this grant application that one way to treat these diseases would be to restore the missing gene in patients' white blood cells as a way of providing the missing protein and restoring function in the different organs.

Project Terms:
Address; Adoptive Cell Transfers; Adoptive Transfer; Affect; Age; Allogenic; Animals; Apolipoprotein E; Applications Grants; Autologous; Award; B-Lymphocytes; base; Blood - brain barrier anatomy; bone; Bone Marrow Transplantation; Breathing; Cardiac; Cardiopulmonary; Cells; cellular engineering; Cessation of life; Clinical; Clinical Chemistry; Clinical Trials; Collaborations; compare effectiveness; Complement; Cornea; Dermatan Sulfate; Development; Disease; Dose; Effectiveness; Engineering; enzyme replacement therapy; Enzymes; Extraordinary Treatments; Feedback; functional restoration; Future; Genes; Genetic Engineering; genetically modified cells; Genomics; Glycosaminoglycans; Goals; good laboratory practice; Heart; Heart Abnormalities; Heart Diseases; Hematology; Hematopoietic Stem Cell Transplantation; Heparitin Sulfate; Hepatosplenomegaly; Hereditary Disease; Human; Impaired cognition; Impairment; improved; in vivo; Individual; Infusion procedures; L-Iduronidase; Leukocytes; Longitudinal Studies; Mediating; Mental Retardation; Metabolic; Metabolic Diseases; Monitor; Morbidity - disease rate; mortality; Mucopolysaccharidoses; Mucopolysaccharidosis I; Mucopolysaccharidosis I H; Mus; Nerve Tissue; nervous system disorder; Nervous system structure; Neuraxis; Neurocognitive; Neurologic; Neurologic Symptoms; novel; novel strategies; novel therapeutics; Obstruction; Organ; Outcome; overexpression; Patients; Penetration; Phase; phase 1 study; phase 2 study; Physiological; Production; protein expression; protein function; Proteins; research clinical testing; Research Design; Risk; Safety; safety testing; skeletal; skeletal abnormality; skeletal disorder; skeletal dysplasia; Sleeping Beauty; Small Business Innovation Research Grant; standard of care; System; Teleconferences; Testing; Therapeutic; therapeutic evaluation; therapeutic protein; Time; Tissues; Toxicology; Treatment Efficacy