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

• 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
• 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

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)移植可以提供持久的艾滋病毒消除，就像“柏林”所证明的那样 患者，“伦敦”患者，最近，第三名(“纽约”)患者。这为HSC提供了一个强有力的理由 艾滋病毒/艾滋病的基因治疗。目前的临床HSC基因治疗方案(例如，针对血红蛋白疾病)包括 大剂量化疗为骨髓腾出空间，体外基因后HSCs移植 调职。由于风险、成本和技术的复杂性，体外方案不太可能广泛应用 适用，特别是在艾滋病毒/艾滋病治疗需求最大的发展中国家。我们有 开发了一种体内HSC转导方法，只需静脉注射即可提供 作为门诊治疗。在这种方法中，造血干细胞被从骨髓动员到外周血中。 并通过静脉注射体内基因转移载体(辅助依赖腺病毒)进行转导 载体)，靶向原始造血干细胞上存在的受体。在外周转导的HSCs返回到骨骼 骨髓，在那里长期存在，并对所有血细胞谱系做出贡献。这个应用程序的中心目标是 进一步发展我们体内预防和治疗艾滋病毒的方法，持续根除艾滋病毒 目标/储集层单元。具体目标是。1.优化HSC动员方案、HSC归巢、HDAd 载体/表达系统实现i)动员的HSCs的高效骨髓归巢，ii)高效的运输 转导的HSC子代细胞，特别是大脑，这是一种主要的HIV储存组织，很难被 3)提高转基因表达的水平和安全性。2.防止HIV/SIV逃逸突变 并通过体内多模块HSC基因治疗方法从储存库中清除病毒。模块将发挥作用 基于不同机制的抗艾滋病毒活性(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，通过共受体敲除靶细胞，并独立于 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基因治疗方法将允许i)完全保护 对抗SIV挑战(没有逃逸突变体)和II)在受感染动物中消除SIV(包括 大脑)。用于测试这些方法的安全性和抗病毒效果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(感染和不感染SIV)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障碍 技术简单、经济高效且可移植的方法的背景。