Engineering T-cells with optimized anti-HIV chimeric antigen receptors and CCR5 disruption as a strategy to target HIV-infected cells

使用优化的抗 HIV 嵌合抗原受体和 CCR5 破坏改造 T 细胞，作为针对 HIV 感染细胞的策略

• 批准号: 9276567
• 负责人: Thor Andrew Wagner
• 金额: $ 48.9万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-15 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9276567
• 关键词: Adoptive Transfer; Affect; Affinity; Alpha Cell; Amino Acids; Animal Model; Animals; Anti-Retroviral Agents; Antibodies; Autopsy; Binding; Blood - brain barrier anatomy; Bone Marrow; CCR5 gene; Cell Line; Cell physiology; Cells; Clinical; Clinical Trials; Control Animal; Cytotoxic T-Lymphocytes; DNA; Data; Effectiveness; Engineering; Epitopes; Future; Goals; HIV; HIV Antibodies; HIV Infections; HIV therapy; Human; Human immunodeficiency virus test; IgG4; Immune response; Immune system; Immunoglobulin Variable Region; Individual; Length; Ligands; Link; Liver; Lymphocyte; Major Histocompatibility Complex; Malignant Neoplasms; Methods; Mus; Mutation; Peripheral Blood Mononuclear Cell; Pharmacology; Plasma; Proteins; RNA; Research; Residual state; T-Lymphocyte; Testing; Therapeutic Intervention; Thymus Gland; Time; Toxic effect; Transmembrane Domain; Variant; Viral; Virus; antiretroviral therapy; autologous lymphocytes; autoreactivity; base; cellular engineering; chimeric antigen receptor; efficacy testing; humanized mouse; improved; in vivo; insight; killings; neutralizing antibody; novel therapeutic intervention; public health relevance; screening

 DESCRIPTION (provided by applicant): A cure for HIV is an important goal of HIV treatment. Various "kick and kill" strategies have been envisioned to eradicate the HIV-infected cells that persist during antiretroviral therapy. However, there are few therapeutic interventions that specifically kill HIV-infected cells. Our preliminary data demonstrates that T-cells engineered to express anti-HIV chimeric antigen receptors (CAR) can effectively kill HIV infected cells. CAR were first described 25 years ago, and early clinical trials of adoptive transfer of CAR-expressing T-cells for a variety of malignancies, as well as for HIV, showed promise but their efficacy was limited by insufficient function and/or off-target toxicity. Current trials of newer CR, engineered to incorporate intracellular co-stimulatory domains, have shown dramatic clinical benefit against specific malignancies. CAR-based strategies are attractive for HIV because CAR T-cells function independently of major histocompatibility complexes, and therefore can target HIV-infected cells that have escaped the host immune response. CAR-expressing T-cells have also been shown to persist for years, providing a potential long-term strategy to target HIV-infected cells that reactivate in the future. Most previous anti-HIV CAR used CD4 as a broadly reactive ligand for HIV. However in recent years the breadth and potency of available HIV antibodies has improved significantly. We hypothesize that newer anti-HIV CAR, which include intracellular co-stimulatory domains, linked to the single chain variable fragment (scFv) of potent HIV antibodies, will be an effective strategy for targeting the residual HIV reservoir. We expect these CAR to be more specific for HIV-expressing cells than anti-HIV CAR that utilize CD4 as the ligand for HIV. A concern with any CAR-based approach for HIV is that the CAR+ T-cells can be infected with HIV. Our data indicates that protecting the CAR+ T-cells from HIV infection by disrupting CCR5 increases the efficacy of anti-HIV CAR+ T-cells. This project has two specific aims: 1. Optimize the potency and breadth of an antibody-based anti-HIV CAR by screening scFv from several different antibodies; optimizing the length of the linker between the variable region of the antibody and the transmembrane domain; and testing combinations of CAR targeting different epitopes. 3. Test the efficacy of the optimized CAR(s) in a small animal model of HIV latency. Together these results will help optimize an antibody-based anti-HIV CAR and provide important insight into the feasibility and effectiveness of this strategy.

 描述（由适用提供）：艾滋病毒治愈是艾滋病毒治疗的重要目标。已经设想了各种“踢和杀戮”策略，可以在抗逆转录病毒治疗期间持续存在的HIV感染细胞。但是，很少有治疗性干预措施专门杀死了HIV感染的细胞。我们的初步数据表明，用于表达抗HIV嵌合抗原受体（CAR）的T细胞有效地杀死了HIV感染的细胞。 CAR是25年前首次描述的，对表达各种恶性肿瘤和HIV的表达T细胞的自适应转移的早期临床试验表现出了希望，但其有效性受功能不足和/或脱离目标毒性的限制。当前针对结合细胞内共刺激域的新型CR的试验显示出针对特定恶性肿瘤的巨大临床益处。基于CAR的策略对HIV具有吸引力，因为CAR T细胞独立于主要的组织相容性复合物的作用，因此可以靶向已逃脱宿主免疫反应的HIV感染细胞。表达汽车的T细胞也已显示多年来持续存在，为未来重新激活的HIV感染细胞提供了潜在的长期策略。大多数以前的抗HIV汽车都使用CD4作为HIV的广泛反应性配体。然而，近年来，可用的HIV抗体的广度和效力显着提高。我们假设包括细胞内共刺激域，与有效的单链可变片段（SCFV）相关的新抗HIV汽车 HIV抗体将是针对残留HIV储层的有效策略。我们期望这些汽车比使用CD4作为HIV配体的抗HIV CAR更为特异。任何基于汽车的HIV方法的关注点是CAR+ T细胞可以感染HIV。我们的数据表明，通过破坏CCR5来保护CAR+ T细胞免受HIV感染的影响，可以提高抗HIV CAR+ T细胞的有效性。该项目有两个具体的目标：1。通过从几种不同的抗体中筛选SCFV，优化基于抗体的抗HIV汽车的效力和广度；优化抗体的变量区域和跨膜结构域之间的连接器长度；以及针对不同表位的汽车的测试组合。 3。在艾滋病毒潜伏期的小动物模型中测试优化汽车的效率。这些结果共同有助于优化基于抗体的抗HIV汽车，并为该策略的可行性和有效性提供重要的见解。