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）：II 型粘多糖贮积症（MPS II，亨特综合征）是一种 X 连锁隐性遗传性疾病，由艾杜糖醛酸-2-硫酸酯酶缺失引起，导致糖胺聚糖硫酸乙酰肝素和硫酸皮肤素的全身积累。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，并且在酶严重缺乏的情况下，会出现神经退行性变，并在 15 岁时死亡。虽然造血干细胞移植和植入在治疗某些 MPS 方面已显示出疗效疾病，同种异体移植的 MPSII 患者迄今为止尚未表现出神经系统结果的改善。在这个项目中，我们假设 MPSII 的同种异体移植是无效的，因为移植细胞产生的 IDS 酶不足，而表达高水平 IDS 的供体细胞的基因工程将克服这种不足，并提供有效的代谢交叉校正，包括神经系统的代谢交叉校正。疾病的表现。 Lentigen 是开发用于治疗人类疾病的慢病毒载体的领先公司。在这个第一期 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相结合，开发一种治疗亨特综合征（MPS II）的离体转导方法。该提案的具体目标是： (i) 构建并测试慢病毒载体，用于转导人类 IDS 基因以及作为细胞标记的绿色荧光蛋白。载体构建体将基于双启动子、双顺反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)通过将人类IDS基因离体慢病毒转导至MSPII小鼠的造血干细胞来纠正代谢和神经系统疾病。来自IDS缺陷小鼠的骨髓将被携带IDS基因的慢病毒载体转导并移植到IDS缺陷受体中，作为针对造血干细胞的亨特综合征离体基因治疗的模型。将测试接受治疗的动物的供体细胞的植入和转导、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的神经行为测试中表现的改善。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、可行的方法，并对未来其他溶酶体贮积病的慢病毒基因治疗的开发具有影响。