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HIV消除，如在“柏林”中举例说明的。 病人，“伦敦”病人，最近，在第三个（“纽约”）病人。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（例如用于血红蛋白病）涉及 大剂量化疗使骨髓中产生空间，并移植造血干细胞后， 转移由于风险、成本和技术复杂性，离体方案不太可能被广泛应用。 这是一个非常普遍的问题，特别是在对艾滋病毒/艾滋病治疗需求最大的发展中国家。我们有 开发了一种体内HSC转导方法，该方法仅需要静脉注射， 作为门诊治疗。在这种方法中，HSC从骨髓动员到外周血中， 流并用静脉内注射体内基因转移载体（辅助细胞依赖性腺病毒）转导 载体）靶向存在于原始HSC上的受体。在外周转导的HSC返回到骨 骨髓，长期存在，并有助于所有血细胞谱系。此应用程序的中心目标是 进一步发展我们的体内方法，以持续根除艾滋病毒， 靶/储库细胞。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后代细胞，特别是大脑，一个主要的HIV储库组织，很难通过靶向 iii）增加转基因表达的水平和安全性。2.预防HIV/SIV逃逸突变体 并通过多模块体内HSC基因治疗方法从宿主中清除病毒。模块将发挥 基于不同机制的抗HIV活性（例如，eCD 4-IG对血液中病毒的调理作用， 通过体内基因组编辑通过共受体敲除靶细胞，并独立于 通过在免疫效应细胞上表达的CD 4嵌合抗原受体（CD 4-CAR）的MHC-I呈递。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攻击（不存在逃逸突变体）和ii）在感染动物（包括 脑）。用于测试这些方法的安全性和抗病毒功效的模型系统将包括原代HSC/HSC- 衍生的细胞、转基因和人源化小鼠模型（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治疗中的重要生物学障碍， 这是一种技术上简单、成本效益高、便携式的方法。