A novel CAR-T cell therapy for the one-time treatment of chronic HIV infection in patients who are not ART suppressed

摘要 我们的目标是开发一种一次性治疗方法，用于持久缓解人类免疫缺陷病毒(HIV) 未被抗逆转录病毒疗法(ART)抑制的患者，包括ART幼稚或ART患者 不合规。我们的治疗是一种自体HIV特异性嵌合抗原受体(CAR)-T细胞疗法， 使用CXCR5趋化因子受体作为归巢装置，引导抗HIV杀伤T细胞进入免疫- 淋巴B细胞滤泡中受保护的“隐藏”病毒库，大多数产生病毒的细胞在 慢性感染。病毒特异性CD8T细胞对HIV-1和猿猴具有很强的抗病毒活性 免疫缺陷病毒(SIV)，艾滋病毒的动物模型。然而，尽管有丰富的CD8 T细胞反应 感染HIV-1的人类和感染SIV的猕猴，这些细胞可能不能完全抑制病毒复制 因为大多数HIV-1和SIV复制发生在集中在B细胞滤泡内的CD4+T细胞中 次级淋巴组织，很少有病毒特异性CD8 T细胞驻留。事实上，我们展示了In的比率 体内效应病毒特异性CD8 T细胞对SIV RNA+细胞的靶向是体内比体外低40倍 SIV感染猕猴淋巴组织中的B细胞滤泡。此外，大多数病毒特异性CD8 T细胞 细胞不表达毛囊归巢分子CXCR5，可能解释了病毒特异性CD8 T细胞水平低的原因 定位和监测B细胞滤泡。这些数据表明，HIV和SIV特异性CD8 T细胞 细胞完全抑制病毒复制可能是由于B细胞滤泡中缺乏病毒特异性CD8T细胞所致。 由于绝大多数病毒产生细胞是CD4T细胞，位于慢性萎缩性胃炎的次级淋巴组织中 HIV和SIV感染，我们通过自体输注抗病毒CAR(特别是 Car/CXCR5)T细胞进入慢性SIV感染猕猴体内。处理组动物显示CAR/CXCR5- 与对照组动物相比，T细胞定位于B细胞滤泡，减少了病毒在滤泡中的复制。 此外，该产品在ART抑制的siv感染的先导性临床前研究中显示出安全性和有效性。 猕猴。基于这些发现和我们对抑制抗逆转录病毒药物的猕猴的临床前研究的成功， 我们建议：(阶段1，目标1)通过产生和表征一个 优化的(OPT)CAR/CXCR5结构，包含一个三特异性HIV结合域，一种交替的共刺激 (阶段2，目标2)开发一种GMP可扩展的方法来生产CAR-T 使用optCAR/CXCR5结构的细胞和来自SIV感染的未经ART治疗的恒河猴的PBMC 用于启用IND的临床前研究；以及b)感染艾滋病毒的未接受抗逆转录病毒治疗的个人，为 第一个人类第一阶段临床试验；以及(第二阶段，目标3)评估生产的CAR-T细胞的安全性和有效性 与人optCAR/CXCR5构建成猴/人HIV(SIV)慢性感染模型。 成功完成拟议的研究将支持IND，移动optCAR/CXCR5-T细胞产品 从临床前阶段进入临床发展阶段。

ABSTRACT We aim to develop a one-time treatment for durable remission of human immunodeficiency virus (HIV) for patients who are not suppressed by antiretroviral therapy (ART), including patients who are ART naïve or ART non-compliant. Our treatment is an autologous HIV-specific chimeric antigen receptor (CAR)-T cell therapy that employs the CXCR5 chemokine receptor as a homing device to direct anti-HIV killer T cells into immune- protected “hidden” viral reservoirs in lymphoid B cell follicles, where most virus-producing cells are located during chronic infections. Virus-specific CD8 T cells exert potent antiviral activity against HIV-1 and Simian immunodeficiency virus (SIV), an animal model of HIV. Nevertheless, despite abundant CD8 T cell responses in HIV-1-infected humans and SIV-infected macaques, these cells do not fully suppress virus replication, likely because the majority of HIV-1 and SIV replication occurs in CD4+ T cells concentrated within B cell follicles in secondary lymphoid tissues where few virus-specific CD8 T cells reside. In fact, we showed that the ratio of in vivo effector virus-specific CD8 T cells to target SIV RNA+ cells is >40-fold lower inside compared to outside of B cell follicles in lymphoid tissues in SIV-infected macaques. Furthermore, the majority of virus-specific CD8 T cells fail to express the follicular homing molecule CXCR5, likely explaining low levels of virus-spcific CD8 T cells localizing to and surveilling B cell follicles. These data suggest that the inability of HIV- and SIV-specific CD8 T cells to fully suppress virus replication may be due to a deficiency of virus-specific CD8 T cells in B cell follicles. As the vast majority of virus-producing cells are CD4 T cells located in secondary lymphoid tissue during chronic HIV and SIV infections, we targeted lymphoid cells by autologously infusing anti-viral CAR (specifically CAR/CXCR5) T cells into chronically SIV-infected rhesus macaques. The treated animals showed CAR/CXCR5- T cell localization to B cell follicles and decreased virus replication in the follicles compared to control animals. In addition, this product showed safety and efficacy in a pilot preclinical study of ART-suppressed SIV-infected macaques. Based on these findings and the success of our preclinical studies in ART-suppressed macaques, we propose to: (Phase 1, Aim 1) modify our current CAR/CXCR5 construct by producing and characterizing an optimized (opt) CAR/CXCR5 construct that contains a tri-specific HIV binding domain, an alternate co-stimulatory domain, and an HIV resistance domain; (Phase 2, Aim 2) develop a GMP-scalable method to produce CAR-T cells using the optCAR/CXCR5 construct and PBMCs from a) SIV-infected non-ART-treated rhesus macaques for use in an IND-enabling preclinical study; and b) HIV-infected non-ART treated individuals to prepare for the first-in-human Phase 1 clinical trial; and (Phase 2, Aim 3) assess the safety and efficacy of CAR-T cells produced with the human optCAR/CXCR5 construct in a simian/human HIV (SHIV) model of chronic HIV infection. Successful completion of the proposed studies will be IND-enabling, moving the optCAR/CXCR5-T cell product from the preclinical stage into clinical development.