In vivo HSC gene therapy using a multi-modular HDAd vector for HIV cure

使用多模块 HDAd 载体进行体内 HSC 基因治疗以治愈 HIV

基本信息

批准号：
10599503
负责人：
HANS-PETER KIEM
金额：
$ 68.59万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-01-10 至 2027-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10599503
关键词：
AIDS therapy Address Adenoviruses Ambulatory Care Animals B-Lymphocytes Berlin Biological Biological Models Blood Blood Cells Bone Marrow Bone Marrow Purging Brain CCR5 gene CD46 Antigen CXCR4 gene Carmustine Cell Lineage Cells Clinical Complex Developing Countries Disease model Drug Kinetics Effector Cell Erythroid Cells Escape Mutant Gene Transfer Genes Goals HIV HIV therapy HIV/AIDS Hematopoiesis Hematopoietic Stem Cell Mobilization Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Hemoglobinopathies Hemophilia A High Dose Chemotherapy Homing Human Hyperactivity IL8RB gene Immune Immunotoxins In Vitro Infection Intravenous Knock-out London Macaca mulatta Macrophage Methylation Microglia Modeling Mus New York Patients Pharmaceutical Preparations Preparation Prophylactic treatment Protocols documentation Regimen Resistance Risk SIV Safety Sleeping Beauty Spleen Stream System T-Lymphocyte Terminator Codon Testing Tissues Toxic effect Transgenes Transgenic Organisms Transposase Viral reservoir Virulent Virus anti-viral efficacy base editor cellular transduction chimeric antigen receptor conditioning cost cost efficient expression vector gene therapy gene transfer vector genome editing gutless adenoviral vector humanized mouse immunogenicity in vivo intravenous injection knockout gene mouse model nonhuman primate peripheral blood portability prevent prime editor promoter receptor safety testing side effect simian human immunodeficiency virus stem cell gene therapy targeted treatment trafficking transgene expression vector

项目摘要

Hematopoietic stem cell (HSC) transplantation can provide durable HIV elimination as exemplified in the “Berlin” patient, the “London” patient, and recently, in a third (“New York”) patient. This gives a strong rationale for HSC gene therapy of HIV/AIDS. Current clinical HSC gene therapy protocols (e.g. for hemoglobinopathies) involve high-dose chemotherapy to make space in the bone marrow, and the transplantation of HSCs after ex vivo gene transfer. Because of the risk, cost, and technical complexity, it is unlikely that ex vivo protocols will be widely applicable, specifically in developing countries where the greatest demand for HIV/AIDS therapy lies. We have developed an in vivo HSC transduction approach that requires only intravenous injections and could be provided as an outpatient treatment. In this approach, HSCs are mobilized from the bone marrow into the peripheral blood stream and transduced with intravenously injected in vivo gene transfer vectors (helper-dependent adenovirus vectors) that target receptors present on primitive HSCs. HSCs transduced in the periphery return to the bone marrow, persist there long-term, and contribute to all blood cell lineages. The central goal of this application is to further develop our in vivo approach toward HIV prophylaxis and therapy with persistent eradication of HIV in target/reservoir cells. The Specific aims are. 1. Optimize HSC mobilization regimens, HSC homing, and HDAd vectors/expression systems to achieve i) efficient bone marrow homing of mobilized HSCs, ii) efficient trafficking of transduced HSC progeny cells, specifically to the brain, a main HIV reservoir tissue that is difficult to target by therapeutics, and iii) increase the level and safety of transgene expression. 2. Prevent HIV/SIV escape mutants and eliminate virus from reservoirs by a multi-modular in vivo HSC gene therapy approach. Modules will exert anti-HIV activity based on different mechanisms (e.g. opsonization of virus in blood by eCD4-Ig, protection of target cells by co-receptor knockout through in vivo genome editing, and killing of infected cells independently of MHC-I presentation by a CD4 chimeric antigen receptor (CD4-CAR) expressed on immune effector cells. 3. Demonstrate in NHPs that the optimized in vivo HSC gene therapy approach will allow for i) complete protection against SIV challenges (absence of escape mutants) and ii) SIV elimination in infected animals (including the brain). Model systems to test the safety and antiviral efficacy of the approaches will include primary HSCs/HSC- derived cells, transgenic and humanized mouse models (with and without SIV infection), as well as NHPs (in prophylaxis and therapy setting). Our efforts will address important biological obstacles in HIV therapy in the context of a technically simple, cost-efficient, and portable approach.

造血干细胞（HSC）移植可以持久消除艾滋病毒，“柏林”就是例证患者、“伦敦”患者以及最近的第三名患者（“纽约”）这为 HSC 提供了强有力的理由。 HIV/AIDS 的基因治疗目前的临床 HSC 基因治疗方案（例如血红蛋白病）涉及高剂量化疗在骨髓中腾出空间，离体基因后移植造血干细胞由于风险、成本和技术复杂性，离体方案不太可能被广泛使用。适用，特别是在对艾滋病毒/艾滋病治疗需求最大的发展中国家。开发了一种体内 HSC 转导方法，只需静脉注射即可提供作为门诊治疗，HSC 从骨髓转移到外周血中。流并用静脉注射的体内基因转移载体（辅助依赖性腺病毒载体），其目标是在外周转导的原始 HSC 上的受体返回骨。骨髓，长期存在，并有助于所有血细胞谱系。该应用的中心目标是。进一步发展我们的体内艾滋病毒预防和治疗方法，并在体内持续根除艾滋病毒具体目标是 1. 优化 HSC 动员方案、HSC 归巢和 HDAd。载体/表达系统，以实现 i) 动员的 HSC 的有效骨髓归巢，ii) 有效运输转导的 HSC 子代细胞，特别是大脑，这是 HIV 的主要储存组织，很难通过治疗，以及 iii) 提高转基因表达的水平和安全性 2. 防止 HIV/SIV 逃逸突变体。并通过多模块体内 HSC 基因治疗方法消除病毒库。基于不同机制的抗 HIV 活性（例如 eCD4-Ig 对血液中病毒的调理作用、保护通过体内基因组编辑通过共受体敲除来靶向细胞，并独立地杀死受感染的细胞由免疫效应细胞上表达的 CD4 嵌合抗原受体 (CD4-CAR) 呈递 MHC-I 3。在 NHP 中证明优化的体内 HSC 基因治疗方法将实现 i) 完全保护对抗 SIV 挑战（不存在逃逸突变体）和 ii) 消除受感染动物（包括测试这些方法的安全性和抗病毒功效的模型系统将包括初级 HSC/HSC-。衍生细胞、转基因和人源化小鼠模型（有或没有 SIV 感染）以及 NHP（在预防和治疗环境）。我们的努力将解决艾滋病毒治疗中的重要生物学障碍。技术上简单、成本效益高且可移植的方法。