GENE THERAPY OF INHERITED ENZYME DEFICIENCIES

遗传性酶缺陷的基因治疗

• 批准号: 6111859
• 负责人: R O BRADY
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6111859
• 关键词: CD34 molecule; Fabry's disease; Gaucher's disease; cell transplantation; clinical research; clinical trial phase I; enzyme activity; enzyme deficiency; gene expression; gene mutation; gene targeting; gene therapy; genetic disorder; genetic transduction; glucosylceramidase; hematopoietic stem cells; hematopoietic tissue transplantation; human subject; human therapy evaluation; inborn metabolism disorder; laboratory mouse; macrophage; molecular cloning; tissue /cell culture; transfection /expression vector

We have completed a Phase 1 safety and gene trial in patients with Type 1 Gaucher disease. CD34+ stem and progenitor cells were obtained from peripheral blood by apheresis after stimulating patients with G-CSF (NIH) or from bone marrow biopsies (Children's Hospital, Los Angeles). The first recipient was a 21-year-old male with Type 1 Gaucher disease where only bone marrow stromal cells were used in the expansion of his CD34+ cells. Barely detectable and short-lasting marking of granulocytes was observed. The second patient was a 22-year-old female. Autologous marrow stromal cells along with IL-3, IL-6 and SCF were employed to expand the transduced cells. More significant and longer lasting granulocyte marking was observed in this patient. We are developing more efficient procedures to infect CD34+ cells and to segregate the transduced cells. We are also investigating methods to provide a growth advantage to the transduced cells. We have carried out a series of pre-clinical investigations concerning gene therapy for Fabry disease. A recombinant retroviral gene transfer vector was designed that engineers efficient traduction of cells and expression of human alpha-Gal A activity. Enzymatic correction was observed in cultured skin fibroblasts and B cell lines derived from patients with Fabry disease. Corrected cells secreted significant quantities of alpha-Gal A into the culture medium, and the enzyme is taken up by uncorrected bystander cells. We have also achieved enzymatic correction in CD34+ and progenitor colony cells obtained from bone-marrow aspirates from patients with Fabry disease. We are producing a high-titer clinical grade vector under GMP conditions for gene therapy trials in Fabry disease. We shall also explore gene therapy in the transgenic alpha-Gal A knock-out mouse model of Fabry disease that we created. In addition, we have made gene transfer vector constructs based on HIV-1 that are highly effective in the transduction of quiescent cells including rat cerebellar neurons. This investigation has particular relevance to gene therapy since most stem cells in the bone marrow are non-dividing and cells within the nervous system are in the post- mitotic state.

我们已经完成了第一阶段的安全性和基因试验 1型戈谢病患者中。cd 34+干细胞和 通过单采血液成分术从外周血中获得祖细胞 在用G-CSF（NIH）或从骨髓刺激患者后， 活组织检查（儿童医院，洛杉矶）。第一位获奖者是 一名21岁的男性，患有1型戈谢病， 骨髓基质细胞用于扩增his CD 34 + 细胞粒细胞标记几乎不可检测且持续时间短 观察了第二名患者是一名22岁的女性。 自体骨髓基质细胞沿着IL-3、IL-6和SCF 用于扩增转导的细胞。更显著和 在该患者中观察到更持久的粒细胞标记。我们 正在开发更有效的程序来感染CD 34+细胞， 以分离转导的细胞。我们也在调查 为转导细胞提供生长优势的方法。我们 进行了一系列临床前研究， 法布里病的基因治疗重组逆转录病毒基因 设计了一种有效的传递载体， 细胞和表达人α-Gal A活性。酶 在培养的皮肤成纤维细胞和B细胞系中观察到校正 来源于法布里病患者。分泌的校正细胞 培养基中加入大量的α-Gal A，以及 该酶被未校正的旁观者细胞摄取。我们有 还实现了CD 34+和祖细胞的酶校正 从患者骨髓抽吸物中获得的集落细胞 Fabry病我们正在生产高滴度的临床级 用于法布里病基因治疗试验的GMP条件下的载体 疾病我们还将探索转基因小鼠的基因治疗。 α-Gal A基因敲除小鼠法布里病模型， 创造此外，我们还构建了基因转移载体， 基于HIV-1的基因， 静止细胞包括大鼠小脑神经元。这次调查 与基因治疗特别相关，因为大多数干细胞在 骨髓不分裂，神经系统内的细胞 处于有丝分裂后的状态。