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）移植可以提供耐用的HIV消除，如“柏林”的例子患者，“伦敦”患者，最近在第三名（“纽约”）患者中。这为HSC提供了强大的理由艾滋病毒/艾滋病的基因治疗。当前临床HSC基因治疗方案（例如，用于血红蛋白病）高剂量化疗以在骨髓中腾出空间，并在体内基因后移植HSC 转移。由于风险，成本和技术复杂性，离体协议不太可能广泛适用，特别是对艾滋病毒/艾滋病治疗需求最大的发展中国家。我们有开发了一种仅需要静脉注射的体内HSC转移方法，可以提供作为门诊治疗。在这种方法中，HSC从骨髓动员到外周血流并用静脉注射的体内基因转移载体（依赖辅助腺病毒病毒）转移载体）靶向接收器存在于原始HSC上。 HSC在外围返回到骨骼中转移骨髓，长期持续存在，并为所有血细胞谱系做出了贡献。此应用的核心目标是为了进一步发展我们的体内预防艾滋病毒和治疗的体内方法，并持续目标/储层细胞。具体目标是。 1。优化HSC动员方案，HSC HOUNT和HDAD 实现i）动员HSC的有效骨髓归巢的载体/表达系统，ii）有效的贩运翻译的HSC后代细胞，特别是大脑，这是一种主要的HIV储层组织，很难通过治疗和iii）提高转化表达的水平和安全性。 2。防止艾滋病毒/SIV逃脱突变体并通过多模块化体内HSC基因治疗方法从储层中消除病毒。模块将执行基于不同机制的抗HIV活性（例如，通过ECD4-Ig对血液的病毒调整，保护通过体内基因组编辑的共受体敲除靶细胞，并独立于杀死感染细胞 MHC-1通过在免疫效应细胞上表达的CD4嵌合抗原受体（CD4-CAR）表示。 3。在NHP中证明，经过优化的体内HSC基因治疗方法将允许i）完全保护反对SIV挑战（缺乏逃生突变体）和II）被感染动物消除SIV（包括脑）。测试方法的安全性和抗病毒效率的模型系统将包括主要的HSC/HSC- 衍生的细胞，转基因和人源化的小鼠模型（有和没有SIV感染）以及NHP（在预防和治疗设置）。我们的努力将解决艾滋病毒疗法中重要的生物障碍技术上简单，成本效益和便携式方法的背景。