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: 10528874
• 负责人: Nancie M Archin
• 金额: $ 79.24万
• 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/10528874
• 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; Transforming Growth Factor alpha; Viral; Woman; antiretroviral therapy; chimeric antigen receptor T cells; chromosome 7 loss; 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.

项目概要 动物模型研究表明 98% 的 HIV 储存库存在于肠道内，但大多数 储库动力学研究源自外周血。净化肠道可能存在独特的挑战 储存库作为艾滋病毒治疗策略。其中之一是该组织中细胞因子 TGF 的浓度很高， 因为 TGF 可能会产生增强潜伏期和抑制效应 CD8+ T 细胞的综合不良影响 否则可以识别并消除重新激活的细胞。通用设计的第二个挑战 HIV 治疗是雌激素的潜伏促进作用，这可能需要不同的 HIV 治疗方法 受感染的男性与女性。在这项研究中，我们将招募病毒受到抑制的艾滋病毒感染者 （PLWH），绝经前和绝经后女性（具有不同的内源性雌激素水平）和 临床匹配的男性，捐献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，肠道的转录组、表面蛋白质组和克隆扩增历史 储存细胞，以更好地了解其持久存在的机制，并尝试识别新的 这些细胞的生物标志物。在目标 2 中，我们将测试下一代延迟逆转的离体组合 药物（LRAs）——已记录体内潜伏期逆转活性的药物，并且是有前途的药物 通过“休克和杀戮”实现艾滋病毒治愈——并使用 CyTOF 和生物信息学方法来确定 肠道储存细胞的表型是否会响应这些治疗而重新激活。这些研究 不仅会揭示各种 LRA 组合对肠道储存库的有效性，而且还会揭示 肠道储存细胞抵抗这些治疗重新激活的特征，可以使用这些治疗来靶向 辅助方法。在目标 3 中，我们将开发和测试旨在消除水库的“杀灭”方法 肠道内富含 TGF 的环境中的细胞，通过 1) 在识别受感染细胞期间阻断 TGF 信号传导 通过 HIV 特异性 CD8+ T 细胞，以及 2) 通过将 TGF 信号从免疫抑制转化为激活 一是通过工程化 HIV 特异性 CAR T 细胞共表达 TGF-41BB 嵌合体。除了建立 更好地了解肠道储存库及其维护（包括通过与 它的血液对应物）在男性和女性艾滋病毒感染者中，这项研究将测试多个新的方面 “休克和杀戮”旨在消除艾滋病毒感染者中最丰富的艾滋病毒储存库。