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

基本信息

批准号：
9143841
负责人：
SUNG-YUN PAI
金额：
$ 113.23万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-07 至 2021-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9143841
关键词：
Acute T Cell Leukemia Address Allogenic Antibodies Antibody Response Autologous B-Lymphocytes Biological Assay Birth Busulfan CD34 gene Cell Count Cells Cessation of life Characteristics Clinical Trials Cytokine Receptors Cytokine Signaling Development Dose Elongation Factor Engraftment Enhancers Failure Gene therapy trial Genes HIV Hematopoietic stem cells Hereditary Disease IGH@ gene cluster IL2RG gene Immune Immune system Immunoglobulin Class Switching Immunoglobulin Somatic Hypermutation Immunoglobulins In Vitro Individual Infant Infection Interleukin 2 Receptor Gamma Interleukin-15 Interleukin-7 Lentivirus Vector Life Link Long Terminal Repeats Lymphoid Measurement Moloney Leukemia Virus Mutation Natural Killer Cells Neonatal Screening Oncogenes Output Patients Pattern Phase Phenotype Plasmablast Population Sizes Proto-Oncogenes Recovery Reporting Safety Sampling Severe Combined Immunodeficiency Site T-Cell Leukemia T-Lymphocyte Testing Transgenes Transplantation Vertebral column Viral adenosine deaminase base boys cell type cellular transduction chemotherapy conditioning congenital immunodeficiency deep sequencing efficacy testing gene therapy graft vs host disease granulocyte hematopoietic cell transplantation improved in vivo integration site leukemia next generation sequencing overexpression peripheral blood progenitor promoter reconstitution response tumorigenesis vector

项目摘要

Project Summary Gene therapy using autologous CD34+ cells is a promising treatment for primary immunodeficiency, particularly for individuals without optimal allogeneic donors. SCID-X1 is caused by mutations in IL2RG, which encodes the common gamma chain (γc) of multiple cytokine receptors. Boys with SCID-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 SCID- 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 (γRV) vector used in previous trials to the LV vector in this trial (Aim 2). Previous trials of gene therapy for SCID-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 SCID-X1 have used a γRV 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 γRV 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’ CD34+ transduced cells and compare samples from the proposed trial to historical trials using γRV, analyze insertion site profile in peripheral blood after gene therapy to perform lineage tracing and compare clustering with samples from previous trials.

项目概要使用自体 CD34+ 细胞的基因治疗是治疗原发性免疫缺陷的一种有前途的治疗方法，特别是对于没有最佳同种异体供体的个体来说，SCID-X1 是由 IL2RG 突变引起的。编码多种细胞因子受体的共同伽马链 (γc) 患有 SCID-X1 的男孩缺乏 T 和 NK。细胞及其 B 细胞分别由于缺乏 IL-7、IL-15 和 IL-21 功能而无法产生抗体。该项目旨在测试一种新的自失活慢病毒 (LV) 载体治疗 SCID 的有效性和安全性 X1.我们追求该试验将通过引入低剂量来改善免疫重建。白消安调节（目标 1）并通过改变伽玛逆转录病毒 (γRV) 载体来提高安全性之前的试验中使用了本次试验中的 LV 载体（目标 2）。之前针对 SCID-X1 的基因治疗试验是在没有化疗条件下输注细胞，这导致了 T 细胞的强劲恢复和基因标记，但 B 细胞中的基因标记可以忽略不计并且失败在目标 1 中，我们将无法维持 NK 细胞的初始发育和标记。检查低剂量白消安调理对 1) 细胞类型特异性植入和基因标记的影响， 2) 通过深度测序进行体内 T 细胞重建、T 细胞表型和 TRB 库，3) 体内体液免疫重建、B 细胞数量、表型、IL-21 依赖性功能和 IGH 库测序，4) NK 细胞数量、表型和功能。之前针对 SCID-X1 的基因治疗试验使用了具有完整病毒启动子/增强子的 γRV 载体，这导致 5/20 的患者由于使用插入性肿瘤发生而患上 T 细胞白血病。删除病毒增强子的自失活 γRV 载体显示出令人鼓舞的证据淋巴癌基因附近的插入位点减少，但初始插入位点模式仍然存在风险。在该应用中提出的试验将通过使用自失活 LV 载体进一步提高安全性。将研究患者 CD34+ 转导细胞中的初始插入位点模式并比较样本从拟议的试验到使用 γRV 的历史试验，分析基因后外周血中的插入位点概况疗法进行谱系追踪并将聚类与之前试验的样本进行比较。