Lentiviral gene therapy for mucopolysaccharidosis

粘多糖贮积症的慢病毒基因治疗

• 批准号: 7805078
• 负责人: R. Scott McIvor
• 金额: $ 36.48万
• 依托单位: LENTIGEN CORPORATION
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7805078
• 关键词: Adrenal Glands; Affect; Age; Allogenic; Allografting; Animal Testing; Animals; Applications Grants; Biological Assay; Blood Coagulation Disorders; Bone Marrow Transplantation; Brain; Breathing; Cells; Cerebellum; Cessation of life; Chimeric Proteins; Clinical Trials; Corpus striatum structure; Data; Dermatan Sulfate; Development; Disease; Disease model; Effectiveness; Engineering; Engraftment; Enzymes; Exhibits; Fibroblasts; Flow Cytometry; Future; GAG Gene; Gene Transfer; Generations; Genes; Genetic; Genetic Engineering; Glycosaminoglycans; Goals; Green Fluorescent Proteins; HIV; Harvest; Heart; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hemoglobinopathies; Heparitin Sulfate; Hippocampus (Brain); Histologic; Human; Immune; In Vitro; Inborn Genetic Diseases; Individual; Inherited; Kidney; Lentivirus Vector; Leukocytes; Life; Link; Liver; Lung; Lysosomal Storage Diseases; Lysosomes; Malignant - descriptor; Marrow; Mediating; Mental Retardation; Metabolic; Minnesota; Modeling; Mucopolysaccharidoses; Mucopolysaccharidosis II; Mus; Neuraxis; Neurologic; Neurologic Manifestations; Neurological outcome; Obstructive Lung Diseases; Patients; Performance; Peripheral; Phase; Plasma; Population; Prevention; Procedures; Promoter Regions; Protocols documentation; Research; Risk; Rotarod Performance Test; Safety; Small Business Technology Transfer Research; Spleen; T-Lymphocyte; Technology; Testing; Therapeutic; Time; Tissues; Transplantation; Universities; Urine; Visceromegaly; allotransplant; base; enzyme activity; enzyme deficiency; enzyme therapy; experience; gene correction; gene therapy; human disease; iduronate-2-sulfatase; improved; in vivo; leukodystrophy; lymphoblastoid cell line; morris water maze; motor learning; mouse model; neurobehavioral test; peripheral blood; pre-clinical; prevent; programs; promoter; public health relevance; safety testing; skeletal abnormality; vector

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：粘多糖病II型（MPS II， Hunter综合征）是一种x连锁隐性遗传疾病，由伊杜醛酸-2-硫酸酯酶缺乏引起，导致糖胺聚糖硫酸肝素和硫酸皮聚糖的系统性积累。受影响的个体患有骨骼异常、器官肿大、危及生命的阻塞性气道疾病，并且，在严重缺乏酶的情况下，神经系统退化并在15岁之前死亡。虽然造血干细胞的移植和植入在治疗一些MPS疾病方面显示出疗效，但同种异体移植的MPSII患者迄今尚未表现出改善的神经系统预后。在这个项目中，我们假设同种异体移植治疗MPSII是无效的，因为从移植的细胞中产生的IDS酶不足，而供体细胞的基因工程表达高水平的IDS将克服这种不足，并提供有效的代谢交叉校正，包括疾病的神经系统表现。Lentigen是开发用于治疗人类疾病的慢病毒载体的领先公司。在这个I期STTR项目中，我们建议将Lentigen的慢病毒载体技术与明尼苏达大学在溶酶体贮积病细胞治疗方面的经验结合起来，开发一种体外转导方法来治疗亨特综合征（MPS II）。该提案的具体目的是：(i)构建和测试慢病毒载体，用于人类IDS基因的转导以及绿色荧光蛋白作为细胞标记。载体构建将基于双启动子、双顺子和融合蛋白策略生成，并使用Lentigen专有的LentiMax平台进行包装。（ii）通过体外慢病毒将人IDS基因转导到MSPII小鼠的造血干细胞中来纠正代谢和神经系统疾病。将携带IDS基因的慢病毒载体转导IDS缺陷小鼠的骨髓，并将其移植到IDS缺陷受体中，作为针对造血干细胞的亨特综合征体外基因治疗的模型。接受治疗的动物将测试供体细胞的移植和转导、血浆和组织中的IDS酶表达、尿液和组织中储存物质的清除，以及学习和运动功能神经行为测试中的表现改善。拟议研究的总体目标是提供临床前数据，以支持实施MPS II基因治疗的最直接和可行的方法，这对未来开发其他溶酶体贮积病的慢病毒基因治疗具有重要意义。