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: 9977108
• 负责人: DAVID A WILLIAMS
• 金额: $ 98.23万
• 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 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9977108
• 关键词: Acute T Cell Leukemia; Address; Allogenic; Antibodies; Antibody Response; Autologous; Automobile Driving; B-Lymphocytes; Biological Assay; Birth; Busulfan; CD34 gene; Cell Count; Cells; Cessation of life; Characteristics; Cytokine Receptors; Cytokine Signaling; Development; Dose; Elongation Factor; Engraftment; Enhancers; Failure; Gene therapy trial; Genes; Genetic Diseases; HIV; Hematopoietic stem cells; IGH@ gene cluster; IL2RG gene; Immune system; Immunoglobulin Class Switching; Immunoglobulin Somatic Hypermutation; Immunoglobulins; In Vitro; Individual; Infant; Infection; Interleukin 2 Receptor Gamma; Interleukin-15; Interleukin-7; Lentivirus Vector; Link; Long Terminal Repeats; Lymphoid; Measurement; Moloney Leukemia Virus; Mutation; Natural Killer Cells; Neonatal Screening; Oncogenes; Opportunistic Infections; Output; Patients; Pattern; Phase I/II Clinical Trial; Phenotype; Plasmablast; Population Sizes; Proto-Oncogenes; Recovery; Reporting; Safety; Sampling; Severe Combined Immunodeficiency; Site; T cell reconstitution; T-Cell Development; T-Cell Leukemia; T-Lymphocyte; T-cell receptor repertoire; Testing; Thymus Gland; Transcription Initiation Site; Transgenes; Transplantation; Vaccination; 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; immune reconstitution; improved; in vivo; integration site; leukemia; leukemogenesis; next generation sequencing; overexpression; peripheral blood; progenitor; promoter; reconstitution; response; tumorigenesis; vector

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.

项目摘要 使用自体CD34+细胞的基因治疗是一种原发性免疫缺陷的有望治疗 特别是对于没有最佳同种异体供体的个人。 SCID-X1是由IL2RG突变引起的，该突变是 编码多个细胞因子受体的常见γ链（γc）。患有SCID-X1的男孩缺乏T和NK 细胞，其B细胞分别缺乏IL-7，IL-15和IL-21功能，无法产生抗体。 该项目旨在测试新的自动慢病毒（LV）向量的效率和安全性，以治疗SCID- x1。我们假设该试验将通过引入低剂量来改善免疫重建 Busulfan条件（AIM 1）并通过从伽马甲（γRV）载体的变化来提高安全性 在此试验中，在先前的LV载体试验中使用（AIM 2）。 先前对SCID-X1基因治疗的试验已感染细胞，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 检查低剂量的busulfan调节对1）细胞类型特异性植入和基因标记的影响， 2）体内T细胞重建，T细胞表型和TRB曲目通过深度测序，3）体内体内 免疫重建，B细胞数，表型，IL-21依赖性功能和IGH曲目 测序，4）NK细胞数，表型和功能。 先前对SCID-X1基因治疗的试验已将γRV载体与完整的病毒启动子/增强子一起使用， 由于插入肿瘤发生，导致5/20例患者患有T细胞白血病。基因治疗使用 删除病毒增强子的自动灭活γRV载体显示令人鼓舞的证据 淋巴癌基因附近的插入部位减少了，但仍有风险的初始插入位点模式。 在本应用中提议的试验将通过使用自动化的LV载体进一步提高安全性。在目标2中我们 将研究患者CD34+翻译细胞中的初始插入位点模式，并比较样品 从拟议的试验到使用γRV的历史试验，分析基因后血液中的插入位点谱。 治疗以执行谱系跟踪并将聚类与以前试验的样品进行比较。