Pre-clinical Modeling of Foamy Viral gene Therapy for Murine and Human SCID-X1

小鼠和人类 SCID-X1 泡沫病毒基因治疗的临床前模型

• 批准号: 8278864
• 负责人: David J Rawlings
• 金额: $ 25.01万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-07 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8278864
• 关键词: Adverse event; Allogenic; Animal Model; Animals; Biological Assay; CD34 gene; Cells; Clinical Research; Cytokine Receptors; Data; Disease; Elements; Enhancers; Exhibits; Family; Gene Delivery; Generations; Genes; Grant; Hematopoietic stem cells; Human; Immunologic Deficiency Syndromes; In Vitro; Interleukin 2 Receptor Gamma; Lead; Left; Life; Link; Molecular Analysis; Multipotent Stem Cells; Mus; Mutation; Natural Killer Cells; Outcome; Patients; Pattern; Pre-Clinical Model; Recombinants; SCID Mice; Safety; Siblings; Signal Transduction; Source; Spumavirus; Stem cell transplant; Stem cells; T-Lymphocyte; Testing; Therapeutic; Transactivation; Viral; Viral Genes; Viral Vector; base; design; gene replacement; gene therapy; genotoxicity; improved; in vivo; leukemogenesis; pre-clinical; programs; transgene expression; vector

SCID-Xl is catastrophic immunodeficiency disorder caused by mutations within the common gamma chain (yc) gene. While stem cell transplantation using a matched sibling donor can be curative, most patients lack optimal donors leading to poorer outcomes. Gene replacement has many theoretical advantages as an alternative therapeutic approach for SCID-Xl; and pioneering clinical studies using gammaretroviral yc delivery lead to both significant benefit as well as unanticipated adverse events due to viral enhancer triggered leukemogenesis. The overarching hypothesis of this PPG is that both the efficacy and safety of yc gene delivery can be significantly improved using recombinant foamy virus (FV) based vectors. Studies in Project 1 are designed to test the hypotheses that yc FV vectors devoid of viral enhancers (with or without additional enhancer blocking elements flanking the transcriptional cassette) will exhibit levels of transgene expression sufficient for functional rescue in vivo while concurrently showing reduced genotoxicity. The aims of Project 1 are designed to test these hypotheses via detailed phenotypic, functional, and molecular analysis in both: 1) a small animal model of SCID-Xl and 2) hematopoietic stem cells (HSC) derived from SCID-Xl patients. Our specific studies will include efficacy and safety assessment of 1) EFIa-hu-yc FV vectors in vivo in myeloablated vs. non-myeloablated murine SCID-Xl recipients; and in alternative in vitro transactivation assays; 2) Preclinical and GMP-grade 1^' generation yc FV in transduced SCID-Xl patient CD34* BM cells; and 3) Candidate insulated 2

SCID-Xl 是由共同伽马链内突变引起的灾难性免疫缺陷病 (yc) 基因。虽然使用匹配的兄弟姐妹捐赠者的干细胞移植可以治愈，但大多数患者缺乏 最佳捐助者会导致较差的结果。基因替代作为一种方法具有许多理论优势 SCID-Xl 的替代治疗方法；以及使用γ逆转录病毒yc的开创性临床研究 由于病毒增强剂，递送既带来显着的益处，也带来意想不到的不良事件 引发白血病发生。该 PPG 的总体假设是 yc 的有效性和安全性 使用基于重组泡沫病毒（FV）的载体可以显着改善基因递送。研究于 项目 1 旨在测试 yc FV 载体缺乏病毒增强子（有或没有）的假设 转录盒侧翼的附加增强子阻断元件）将表现出转基因水平 表达足以进行体内功能性救援，同时显示出降低的遗传毒性。目标 项目 1 的设计旨在通过详细的表型、功能和分子特征来检验这些假设 两种分析：1) SCID-Xl 的小动物模型和 2) 源自的造血干细胞 (HSC) SCID-X1 患者。我们的具体研究将包括 1) EFIa-hu-yc FV 的功效和安全性评估 清髓性与非清髓性小鼠 SCID-X1 受体体内的载体；或者在体外 反式激活测定； 2) 转导 SCID-Xl 患者的临床前和 GMP 级 1^' 代 yc FV CD34* 骨髓细胞； 3) 候选人绝缘 2