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

SCID-X1是由常见γ链内的突变引起的灾难性免疫缺陷病症 (yc)基因虽然使用匹配的同胞供体进行干细胞移植可以治愈，但大多数患者缺乏 最佳捐助者导致较差的结果。基因替代作为一种治疗癌症的方法有许多理论上的优势。 SCID-X1的替代治疗方法;以及使用γ逆转录病毒γ c的开创性临床研究 递送导致显著益处以及由于病毒增强剂引起的意外不良事件 引发了白血病该PPG的总体假设是，YC的有效性和安全性均 使用基于重组泡沫病毒（FV）的载体可以显著改善基因递送。研究 设计项目1以检验缺乏病毒增强子（有或没有）的γ c FV载体在体外表达的假设。 转录盒侧翼的另外的增强子阻断元件）将表现出转基因水平 表达足以在体内进行功能拯救，同时显示降低的遗传毒性。目标 的项目1的目的是通过详细的表型，功能和分子生物学测试这些假设， 在以下两者中的分析：1）SCID-X1的小动物模型和2）来源于SCID-X1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的疗效和安全性评估 在骨髓清除的鼠SCID-X1接受者与非骨髓清除的鼠SCID-X1接受者中的体内载体;以及 反式激活测定; 2）转导的SCID-X1患者中的临床前和GMP级第1代γ c FV ⑶ 34 * BM细胞;和3）候选绝缘2

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