Gene Therapy for SCID-X1 with Low Dose Busulfan and a SIN-lentiviral Vector

使用低剂量白消安和 SIN 慢病毒载体对 SCID-X1 进行基因治疗

• 批准号: 10827632
• 负责人: DAVID A WILLIAMS
• 金额: $ 128.56万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-07 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10827632
• 关键词: Gene; Therapy; SCID; X1; Low; Gene; Therapy; SCID; X1; Low

Gene therapy using autologous C034+ cells is a promising treatment for primary immunodeficiency, particularly for individuals without optimal allogeneic donors. SCIO-X1 is caused by mutations in IL2RG, which encodes the common gamma chain (ye) of multiple cytokine receptors. Boys with SCIO-X1 lack T and NK cells, and their B cells fail to produce antibodies due to the lack of IL-7, IL-15 and IL-21 function respectively. This project seeks to test the efficacy and safety of a new self-inactivating lentiviral (LV) vector to treat SCIO- X1. We hypothesize that this trial will improve immune reconstitution through the introduction of low dose busulfan conditioning (Aim 1) and improve safety through the change from a gammaretroviral (yRV) vector used in previous trials to the LV vector in this trial (Aim 2). Previous trials of gene therapy for SCIO-X1 have infused cells without chemotherapy conditioning, which resulted in robust T cell recovery and gene marking, but negligible gene marking in B cells and failure of humoral immune reconstitution. Initial development and marking in NK cells was not sustained. In Aim 1 we will examine the impact of low dose busulfan conditioning on 1) cell type specific engraftment and gene marking, 2) in vivo T cell reconstitution, T cell phenotype and TRB repertoire by deep sequencing, 3) in vivo humoral immune reconstitution, B cell number, phenotype, IL-21 dependent function and IGH repertoire by deep sequencing, 4) NK cell number, phenotype and function. Previous trials of gene therapy for SCIO-X1 have used a yRV vector with intact viral promoters/enhancers, which resulted in 5/20 patients developing T cell leukemia due to insertional oncogenesis. Gene therapy using a self-inactivating yRV vector in which viral enhancers have been deleted shows encouraging evidence of reduced insertion sites near lymphoid oncogenes, but an initial insertion site pattern that is still risky. The proposed trial in this application will further improve safety by using a self-inactivating LV vector. In Aim 2 we will investigate the initial insertion site pattern in the patients' C034+ transduced cells and compare samples from the proposed trial to historical trials using yRV, analyze insertion site profile in peripheral blood after gene therapy to perform lineage tracing and compare clustering with samples from previous trials.

使用自体C034+细胞的基因疗法是用于原发性免疫缺陷的有前途的治疗方法 特别是对于没有最佳同种异体供体的个人。 SCIO-X1是由IL2RG突变引起的， 编码多个细胞因子受体的常见伽马链（YE）。 Scio-x1的男孩缺乏T NK细胞以及其B细胞由于缺乏IL-7，IL-15和IL-21功能而无法产生抗体 分别。该项目旨在测试新的自动慢病毒（LV）的功效和安全性 处理scio-x1的矢量。我们假设该试验将通过 引入低剂量的Busulfan调节（AIM 1），并通过更改从 在此试验中，LV载体的先前试验中使用的γ逆转录病毒（YRV）载体（AIM 2）。 先前对SCIO-X1基因治疗的试验已注入细胞，没有化学疗法调节，这 导致稳健的T细胞恢复和基因标记，但在B细胞中可忽略不计 体液免疫重建。 NK细胞中的初始开发和标记不持续。在目标1中 我们将研究低剂量的busulfan调节对1）特定细胞类型的植入和 基因标记， 2）体内T细胞重建，T细胞表型和TRB曲目通过深度测序，3）体内 体液免疫重建，B细胞数，表型，IL-21依赖性功能和IGH曲目 通过深度测序，4）NK细胞数，表型和功能。 先前对SCIO-X1基因治疗的试验使用了具有完整病毒的YRV载体 启动子/增强剂，导致5/20例由于插入而产生T细胞白血病的患者 肿瘤发生。使用病毒增强剂的自动灭活YRV载体的基因疗法 删除显示的令人鼓舞的证据表明淋巴癌基因附近插入位点减少，但初始 仍然有风险的插入站点模式。该应用程序中拟议的试验将进一步提高安全性 使用自动化的LV矢量。在AIM 2中，我们将研究最初的插入位点模式 患者的C034+转导细胞，并使用拟议试验的样品与历史试验进行了比较 YRV，分析基因治疗后外周血中的插入部位谱图进行谱系跟踪和 将聚类与以前试验的样本进行比较。