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（由申请人提供）：治愈艾滋病毒是艾滋病毒治疗的一个重要目标，人们设想了各种“踢杀”策略来根除抗逆转录病毒治疗期间持续存在的艾滋病毒感染细胞。我们的初步数据表明，经过改造后表达抗 HIV 嵌合抗原受体 (CAR) 的 T 细胞可以有效地杀死 HIV 感染细胞，而且早在 25 年前就已被描述。表达 CAR 的 T 细胞过继转移治疗多种恶性肿瘤以及 HIV 的临床试验显示出希望，但其功效因功能不足和/或脱靶毒性而受到限制。结合细胞内共刺激结构域，已显示出针对特定恶性肿瘤的显着临床益处，基于 CAR 的策略对 HIV 很有吸引力，因为 CAR T 细胞的功能独立于主要组织相容性复合物，因此可以靶向逃脱 HIV 感染的细胞。表达 CAR 的 T 细胞也被证明可以持续存在多年，这为未来重新激活的 HIV 感染细胞提供了一种潜在的长期策略。然而，近年来，可用的 HIV 抗体的广度和效力已显着提高，我们捕获了新型抗 HIV CAR，其中包括与有效的单链可变片段 (scFv) 连接的细胞内共刺激结构域。 HIV 抗体将成为针对残留 HIV 储存库的有效策略，我们预计这些 CAR 比利用 CD4 作为 HIV 配体的抗 HIV CAR 对 HIV 表达细胞更具特异性。对于 HIV 来说，CAR+ T 细胞可能会被 HIV 感染。我们的数据表明，通过破坏 CCR5 来保护 CAR+ T 细胞免受 HIV 感染，可以提高抗 HIV CAR+ T 细胞的功效。 1. 通过筛选多种不同抗体的scFv，优化基于抗体的抗HIV CAR的效力和广度；优化抗体可变区和跨膜结构域之间的接头长度，并测试针对不同表位的CAR组合； 3. 在 HIV 潜伏期小动物模型中测试优化的 CAR 的功效，这些结果将有助于优化基于抗体的抗 HIV CAR，并为该方案的可行性和有效性提供重要见解。战略。