Development of Gamma delta CAR-T cells to target CNS HIV reservoir

开发针对 CNS HIV 储存库的 Gamma delta CAR-T 细胞

• 批准号: 10620021
• 负责人: Namita Rout
• 金额: $ 28.46万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10620021
• 关键词: Address; Adoptive Transfer; Autopsy; Bar Codes; Biodistribution; Brain; CAR T cell therapy; CD4 Positive T Lymphocytes; Cell model; Cells; Cellular Stress; Central Nervous System; Data; Development; Engineering; Engraftment; Foundations; Functional disorder; Future; Genes; Genetic Engineering; Goals; Granzyme; HIV; HIV-associated neurocognitive disorder; Hematologic Neoplasms; Immune; Impaired cognition; In Vitro; Infection; Inflammation; Innovative Therapy; Lentivirus Vector; Location; Macaca; Macaca mulatta; Macrophage; Microglia; Modeling; Modernization; Mutation; Myelogenous; Myeloid Cells; Nervous System Trauma; Neurocognitive; Neurocognitive Deficit; Neurons; Pathogenesis; Patients; Persons; Pharmaceutical Preparations; Property; Research; Resistance; Retroviral Vector; Rhesus; Risk; SIV; Safety; Site; Specificity; Stress; T-Lymphocyte; Testing; Therapeutic; Tissues; Viral; Virus; Virus Replication; antiretroviral therapy; brain tissue; cancer immunotherapy; cell transformation; chimeric antigen receptor; chimeric antigen receptor T cells; clinically relevant; cytotoxicity; design; feasibility testing; improved; in vitro Assay; in vivo; in vivo evaluation; insight; latent HIV reservoir; lymph nodes; migration; monocyte; neuroAIDS; neuroinflammation; novel; novel strategies; prevent; purge; simian human immunodeficiency virus; transmission process; γδ T cells

PROJECT SUMMARY HIV persistence in reservoir tissues remains a major barrier to achieving a therapeutic cure despite effective antiretroviral therapy (ART). Eradicating HIV-infected cells in the central nervous system (CNS) is particularly challenging since it is a site of immune privilege and is poorly accessible to ART drugs. While cognitive impairment among people living with HIV has significantly reduced in the era of modern HIV treatment, about 30% of ART-suppressed HIV patients still display conditions of HIV-associated neurocognitive disorder (HAND), suggesting that even low levels of HIV persisting in the brain can cause neurological damage. Thus, functional cure strategies that can safely eradicate replicating virus the CNS reservoir are urgently needed. Emerging evidence suggests that myeloid cells, including brain macrophages (Mf), are sanctuaries of HIV persistence in the CNS, and that HIV-infected macrophages are more resistant to CTLs than CD4 T cells. The goal of this project is to develop a universal gamma delta (γδ) CAR-T platform with dual HIVenv/Mf-targeting for HIV- infected myeloid cells in the CNS reservoir. Our central hypothesis is that adding Mf specific CAR to anti-HIV γδ CAR-T cells will enable efficient targeting of HIV-infected cells in the CNS reservoir. Our data in rhesus macaques show enhanced in vitro killing of HIV-infected monocytic cells by γδ CD4-CAR-T cells and greater Granzyme B cytotoxicity, which has been shown to be critical for CTL killing of HIV-infected primary macrophages. Based on the unique functional properties of γδ T cells, including (i) well-documented CTL activity in immunotherapy of cancer, (ii) ability to migrate to sites of neuroinflammation, and (iii) innate anti-HIV/SIV CTL functions; we hypothesize that the dual γδ CAR-T cells will induce more effective targeting of myeloid HIV reservoirs. In Aim 1, we will develop approaches for generating retroviral and lentiviral vector-based γδ CD4- CAR-T cells toward enhanced targeting of SHIV-infected primary Mf and microglial cells. In Aim 2, we will test the in vivo potential of the optimized γδ CAR-T cell to migrate and engraft in the CNS during viremic infection with the novel macrophage-tropic TF SHIV.D model of CNS pathogenesis. The proposed research is significant because its successful completion will lead to the development of a universal γδ CAR-T cell model with enhanced targeting of tissue macrophage reservoirs including the CNS in a clinically relevant model of SHIV.D infected rhesus macaques.

项目概要 尽管有效，但艾滋病毒在储存组织中的持续存在仍然是实现治疗治愈的主要障碍 抗逆转录病毒治疗（ART）尤其是消除中枢神经系统（CNS）中感染艾滋病毒的细胞。 具有挑战性，因为它是一个免疫特权位点，并且在认知的同时很难获得 ART 药物。 在现代艾滋病毒治疗时代，艾滋病毒感染者的损伤已显着减少，大约 30% 的 ART 抑制的 HIV 患者仍然表现出 HIV 相关神经认知障碍 (HAND) 的症状， 这表明即使大脑中持续存在低水平的艾滋病毒也会导致神经功能损伤。 迫切需要能够安全根除中枢神经系统病毒库中复制病毒的治疗策略。 有证据表明，骨髓细胞，包括脑巨噬细胞（Mf），是艾滋病毒持续存在的避难所 中枢神经系统，并且 HIV 感染的巨噬细胞比 CD4 T 细胞对 CTL 具有更强的抵抗力。 该项目旨在开发一个通用的 γ δ (γδ) CAR-T 平台，具有 HIVenv/Mf 双重靶向，用于治疗 HIV- 我们的中心假设是，将 Mf 特异性 CAR 添加到抗 HIV γδ 中。 CAR-T 细胞将能够靶向中枢神经系统储存库中感染 HIV 的细胞。 猕猴在体外表现出 γδ CD4-CAR-T 细胞对 HIV 感染的单核细胞的更强杀伤力 颗粒酶 B 的细胞毒性，已被证明对于 CTL 杀死 HIV 感染的原代细胞至关重要 基于 γδ T 细胞的独特功能特性，包括 (i) 有据可查的 CTL 活性。 在癌症免疫治疗中，(ii) 迁移至神经炎症部位的能力，以及 (iii) 先天抗 HIV/SIV CTL 我们追求双γδ CAR-T细胞能够更有效地靶向骨髓HIV 在目标 1 中，我们将开发生成基于逆转录病毒和慢病毒载体的 γδ CD4- 的方法。 CAR-T 细胞增强对感染 SHIV 的原代 Mf 和小胶质细胞的靶向作用。在目标 2 中，我们将进行测试。 优化的 γδ CAR-T 细胞在病毒血症感染期间迁移和植入中枢神经系统的体内潜力 与中枢神经系统发病机制的新型巨噬细胞趋向性 TF SHIV.D 模型相关的研究具有重要意义。 因为它的成功完成将导致通用γδ CAR-T细胞模型的开发，该模型具有增强的 在 SHIV.D 感染的临床相关模型中靶向包括中枢神经系统在内的组织巨噬细胞储库 恒河猴。