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

项目总结 HIV在储存组织中的持久性仍然是实现治疗治愈的主要障碍，尽管有效 抗逆转录病毒疗法(ART)。根除中枢神经系统(CNS)中的艾滋病毒感染细胞尤其重要 具有挑战性，因为它是一个享有免疫特权的地方，而且很难获得抗逆转录病毒药物。在认知上 在现代艾滋病毒治疗的时代，艾滋病毒携带者的损害显著减少，大约 30%的抗逆转录病毒药物抑制的艾滋病毒患者仍然表现出艾滋病毒相关神经认知障碍(HAND)的状况， 这表明，即使低水平的艾滋病毒在大脑中持续存在，也会导致神经损伤。因此，功能性的 迫切需要能够安全地根除中枢神经系统病毒宿主的治疗策略。新兴 有证据表明，髓系细胞，包括脑巨噬细胞(MF)，是艾滋病毒持续存在的避难所 感染HIV的巨噬细胞比CD4T细胞对CTL的抵抗力更强。这样做的目的是 该项目是开发一个通用伽马三角洲(γδ)CAR-T平台，具有针对艾滋病毒的双重HIV/MF靶向- 中枢神经系统储备库中受感染的髓系细胞。我们的中心假设是，将MF特异性CAR添加到抗艾滋病毒γδ中 CAR-T细胞将能够有效地靶向中枢神经系统储备库中的艾滋病毒感染细胞。我们在恒河猴中的数据 猕猴表现出增强了γδCD4CAR-T细胞对感染艾滋病毒的单核细胞的体外杀伤作用 颗粒酶B的细胞毒性，已被证明是HIV感染的原发患者CTL杀伤的关键 巨噬细胞。基于γδT细胞的独特功能特性，包括(I)有充分证据的细胞毒性T细胞活性 在癌症的免疫治疗中，(Ii)迁移到神经炎症部位的能力，以及(Iii)天生的抗HIV/SIV CTL 功能：我们假设双重γδCAR-T细胞将诱导更有效的针对髓系艾滋病毒的靶向 水库。在目标1中，我们将开发产生基于逆转录病毒和慢病毒载体的γδCD4- CAR-T细胞对SHV感染的原代MF和小胶质细胞的靶向性增强。在目标2中，我们将测试 优化的γδCAR-T细胞在病毒感染期间在中枢神经系统内迁移和植入的体内潜能 用新的巨噬细胞嗜性因子SHIV.D模型研究中枢神经系统的发病机制。这项拟议的研究具有重要意义 因为它的成功完成将导致开发具有增强功能的通用γδCAR-T细胞模型 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存库 恒河猴。

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.