Multiomics characterization, induction, and elimination of the HIV gut reservoir

HIV 肠道储存库的多组学表征、诱导和消除

• 批准号: 10661821
• 负责人: Nancie M Archin
• 金额: $ 72.52万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10661821
• 关键词: Animal Model; Bioinformatics; Biopsy; Blood; Blood specimen; CD8-Positive T-Lymphocytes; Cells; Chimera organism; Clinical; Clonal Expansion; Engineering; Environment; Estrogens; HIV; Immune; Maintenance; Mediating; Persons; Phenotype; Postmenopause; Premenopause; Proteome; Recording of previous events; Resistance; Shock; Signal Pathway; Signal Transduction; Surface; Testing; Tissues; Viral; Woman; antiretroviral therapy; chimeric antigen receptor T cells; cytokine; design; in vivo; latent HIV reservoir; men; multiple omics; next generation; novel; novel marker; novel therapeutic intervention; peripheral blood; purge; recruit; transcriptome; treatment response

PROJECT SUMMARY Studies in animal models have suggested 98% of the HIV reservoir to reside within the gut, yet most studies of reservoir dynamics derive from peripheral blood. Unique challenges likely exist for purging the gut reservoir as an HIV cure strategy. One of these is the high concentrations of the cytokine TGF? in this tissue, as TGF? can have the combined undesired effects of reinforcing latency and dampening effector CD8+ T cells that could otherwise recognize and eliminate reactivated cells. A second challenge for the design of a universal HIV cure is latency-promoting effects of estrogen, which may necessitate different approaches for HIV cure in infected men vs. women. In this study, we will recruit, among virally-suppressed people living with HIV (PLWH), both pre- and post-menopausal women (who harbor different endogenous estrogen levels) and clinically-matched men, for donation of gut biopsy and paired blood specimens. In Aim 1, we will characterize at the single-cell level the transcriptomes, surface proteomes, and clonal expansion histories of the gut reservoir cells, to better understand the mechanisms allowing for their persistence and to try to identify novel biomarkers of these cells. In Aim 2, we will test ex vivo combinations of next-generation latency-reversal agents (LRAs) – ones that have documented in vivo latency reversal activity and are promising agents to achieve HIV cure through “shock and kill” – and use CyTOF and bioinformatics approaches to determine the phenotypes of gut reservoir cells that do vs. do not reactivate in response to these treatments. These studies will not only reveal how effective the various LRA combinations are against the gut reservoir, but also reveal features of gut reservoir cells resistant to reactivation by these treatments which can be targeted using adjunctive approaches. In Aim 3, we will develop and test “kill” approaches designed to eliminate reservoir cells in the TGF?-rich environment of the gut, by 1) blocking TGF? signaling during recognition of infected cells by HIV-specific CD8+ T cells, and 2) by transforming TGF? signal from an immunosuppressive to an activating one, via engineering HIV-specific CAR T cells co-expressing a TGF?-41BB chimera. In addition to establishing a better understanding of the gut reservoir and its maintenance (including through extensive comparison with its blood counterpart) in both men and women living with HIV, this study will test multiple novel aspects of “shock and kill” designed to eliminate the most abundant HIV reservoir in PLWH.

项目总结 在动物模型中的研究表明，98%的艾滋病毒宿主存在于肠道内，但大多数 对储集层动力学的研究源于外周血液。净化肠道可能存在独特的挑战 作为一种治疗艾滋病毒的策略。其中之一就是高浓度的细胞因子转化生长因子？在这个组织中， 作为转化生长因子？可能具有增强潜伏期和抑制效应CD8+T细胞的组合不良效果 以其他方式可以识别和消除重新激活的细胞。通用汽车设计面临的第二个挑战 HIV治愈是雌激素的潜伏期促进作用，这可能需要不同的方法来治愈HIV 受感染的男性与女性。在这项研究中，我们将在病毒抑制的艾滋病毒携带者中招募 (PLWH)，绝经前和绝经后妇女(内源性雌激素水平不同)和 临床匹配的男性，用于捐赠肠道活检和配对血液样本。在目标1中，我们将描述 在单细胞水平上，肠道的转录体、表面蛋白质组和克隆性扩张史 水库细胞，以更好地理解其持久性的机制，并试图识别新的 这些细胞的生物标志物。在目标2中，我们将测试下一代潜伏期逆转的体外组合 药物(LRA)-记录了体内潜伏期逆转活动的药物，并有望 通过“休克和杀戮”来治愈艾滋病毒--并使用细胞和生物信息学方法来确定 肠道储存库细胞的表型对这些处理不会重新激活。这些研究 不仅揭示了不同的LRA组合对肠道蓄积物的有效性，而且还揭示了 耐受这些处理的肠道储存库细胞的特征，可用于靶向 辅助性方法。在目标3中，我们将开发和测试旨在消除油层的“压井”方法。 细胞在富含转化生长因子的肠道环境中，通过1)阻断转化生长因子？识别感染细胞过程中的信号转导 通过HIV特异性CD8+T细胞，以及2)转化转化生长因子？从免疫抑制到激活的信号 其一，通过改造HIV特异性CAR T细胞，共表达一种转化生长因子？-41BB嵌合体。除了建立 更好地了解肠道储液器及其维护(包括通过广泛比较 在艾滋病毒携带者的男性和女性身上，这项研究将测试多个新的方面 “电击和杀戮”旨在消灭PLWH中最丰富的艾滋病毒宿主。