Enhancement of Gene Therapy Outcomes for Fabry Disease

提高法布里病的基因治疗效果

• 批准号: 6797751
• 负责人: JEFFREY A MEDIN
• 金额: $ 20万
• 依托单位: ONTARIO CANCER INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-29 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6797751
• 关键词: Fabry' s disease; alpha galactosidase; carbohydrate receptor; cell line; cofactor; enzyme mechanism; gene therapy; hematopoietic tissue transplantation; laboratory mouse; nonhuman therapy evaluation; receptor mediated endocytosis; recombinant proteins; transfection /expression vector

DESCRIPTION (provided by applicant): Fabry disease is the 2nd-most prevalent lysosomal storage disorder (LSD) in humans. It is X-linked and pan-ethnic with a frequency of about1:40,000 males. In Fabry disease the deficiency of the lysosomal enzyme alpha-galactosidase A (a-gal A) are mainly manifested in the vascular endothelium. These cells build up excessive lipids with terminal galactose residues (mainly ceramide trihexoside; CTH) leading to vessel occlusions. A major source of CTH in Fabry disease is from the hematopoietic system-specifically from the breakdown of RBCs by macrophages. Patients succumb to renal, cardiovascular, or cerebrovascular disease in mid life. Current treatment for the disorder is only palliative. Unlike many other LSDs, limited primary nervous system involvement is observed in Fabry disease and even 5 percent of normal enzyme activity may improve the clinical course. An a-gal A deficient mouse has been created that offers a model for studies on the pathophysiology and the development of therapeutic strategies. Fabry disease and this model offer compelling systems to develop and test methods for the improvement of gene therapy. A phenomenon called metabolic cooperativity exists wherein genetically corrected cells secrete the hydrolase-which can be taken up and used by bystander cells. This allows for correction of a lower number of cells to impact therapy. We have previously created retroviral vectors and corrected a variety of Fabry patient and a-gal A-deficient mouse cells. We have also demonstrated that metabolic cooperativity occurs in vivo. The Specific Aims of this application are designed to further this therapeutic approach in order to improve outcomes for Fabry disease-and possibly for other LSDs. Hypothesis 1: In the a-gal A-deficient mouse pre-selection of retrovirally transduced cells co-expressing a selectable marker and the therapeutic gene will improve metabolic cooperativity over that seen with non-enriched cells, as measured systemically by increased levels of a-gal A activity and decreased CTH levels. Hypothesis 2: Retroviral vectors (including novel recombinant lentiviruses) can be generated that encode marking or the therapeutic a-gal A gene that lead to the specific amplification of transduced cells relevant to Fabry disease upon the addition of selectively active co-factors both in vitro and in vivo. Hypothesis: Co-overexpression of the prosaposin gene, a protein co-factor for the a-gal A enzyme, leads to higher catalytic activity in genetically corrected cells than overexpression of a-gal A alone.

描述（由申请人提供）：Fabry病是第二大流行 人类的溶酶体存储障碍（LSD）。它是X连锁和泛种族的 大约1：40,000名男性的频率。在法布里病中 溶酶体酶α-半乳糖苷酶A（A-GAL A）主要体现在 血管内皮。这些细胞与终端产生过多的脂质 半乳糖残留物（主要是神经酰胺三己糖苷； CTH）导致血管 闭塞。 Fabry病中CTH的主要来源是造血 特定于巨噬细胞的RBC分解。患者屈服 中期肾脏，心血管或脑血管疾病。当前的 该疾病的治疗仅是姑息治疗。与许多其他LSD不同，有限 原发性神经系统在Fabry疾病中观察到，甚至5个 正常酶活性的百分比可以改善临床过程。一个a-gal a 已经创建了不足的鼠标，为研究提供了一个模型 病理生理学和治疗策略的发展。法布里病 该模型提供了令人信服的系统来开发和测试 改善基因疗法。存在一种称为代谢合作的现象 其中遗传校正的细胞分泌可以吸收的水解酶 并由旁观者细胞使用。这允许校正较低的数量 细胞影响治疗。我们以前创建了逆转录病毒载体， 纠正了各种Fabry患者和A-GAL A缺陷小鼠细胞。我们有 还证明代谢合作发生在体内。具体 该应用的目的旨在进一步进一步 为了改善Fabry病的预后，并可能用于其他LSD。 假设1：在A-GAL A缺陷小鼠逆转录病毒预选中 转导的细胞共表达可选标记和治疗基因 将改善代谢合作，而不是富含富集的细胞，因为 通过a-gal A活性的水平增加并降低CTH，从系统地测量 水平。 假设2：逆转录病毒载体（包括新的重组慢病毒）可以 生成编码标记或导致的治疗A-GAL A基因 与Fabry疾病相关的转导细胞的特定扩增 在体外和体内添加选择性活跃的共同因素。 假设：prosaposin Gene的共透表达，一种蛋白质的共同因素 A-GAL A酶导致遗传校正的催化活性较高 细胞比单独的A-GAL A的过表达。