Inhibition of heat shock protein 90 for sustained remission of HIV from persistent tissue reservoirs

抑制热休克蛋白 90 使持久性组织储存库中的 HIV 持续缓解

• 批准号: 10184988
• 负责人: Cheryl Stoddart
• 金额: $ 72.35万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-10 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10184988
• 关键词: Address; Aftercare; Animal Model; Anti-Retroviral Agents; Autopsy; Blood; Bone Marrow; Brain; CD4 Positive T Lymphocytes; Cell Line; Cell division; Cells; Chronic; Clinical; Complex; DNA Sequence; Disease remission; Double Stranded DNA Virus; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Genome; Grant; HIV; HIV Infections; HSP 90 inhibition; Half-Life; Heat-Shock Proteins 90; Heat-Shock Response; Highly Active Antiretroviral Therapy; Histone Deacetylase Inhibitor; Human; Human Resources; Immune system; Infection; Infection prevention; Integration Host Factors; Interphase Cell; Interruption; Investigation; Laboratories; Lead; Length; Liver; Location; Lung; Molecular; Molecular Chaperones; Mus; NF-kappa B; Nature; Organ; Pathogenesis; Patients; Pharmaceutical Preparations; Phenotype; Plasma; Positive Transcriptional Elongation Factor B; Proviruses; Public Health; RNA; Reporting; Research; Resources; Rest; Role; Sampling; Source; Spleen; Stat5 protein; T-Lymphocyte; Therapeutic; Tissues; Viral; Viral reservoir; Viremia; Virus; Withholding Treatment; antiretroviral therapy; base; collaboratory; experience; follow-up; humanized mouse; in vivo; inhibitor/antagonist; innovation; insight; lymph nodes; macrophage; mouse model; next generation; preservation; prevent; protein complex; reproductive tract; screening; seroconversion; skills; transcription factor

PROJECT SUMMARY Persistent HIV-infected resting CD4+ T cells can remain undetected in tissue and organ reservoirs despite decades of successful antiretroviral therapy (ART). The lack of gene activity in resting CD4+ T cells enables the integrated replication-competent provirus to persist indefinitely. HIV replication in activated CD4+ T cells is cytopathic, and infected cells are cleared by the immune system. However, a small percentage of HIV-infected activated CD4+ T cells revert to the resting G0 phenotype and remain undetected in tissue reservoirs. These HIV- infected resting cells can be readily activated, and rebound viremia in patients with interrupted ART invariably originates from persistent HIV-infected tissue reservoirs. An ongoing project in my laboratory is focused on the essential role of heat shock protein 90 (Hsp90) in HIV replication. We have shown that mild heat shock (39.5°C) accelerates HIV transcription in chronically infected T-cell lines and increases HIV replication up to 30-fold in primary human cells. Accelerated HIV transcription coincides with increased cellular Hsp90 activity at 39.5°C. Hsp90 is responsible for activating cellular transcription factors, and the Hsp90 inhibitor 17-AAG significantly reduces gene expression by the NF-kB, NFAT, and STAT5 host transcription factors. Further, 39.5°C reactivates HIV replication in ART-suppressed aviremic HIV-infected patient samples and in human resting CD4+ T cells isolated from fully suppressed humanized mice. Further, we show that a more potent next-generation Hsp90 inhibitor has even greater anti-HIV activity with a highly favorable therapeutic ranking and that clinical HIV subtypes display increased viral transcription at 39.5°C in primary human T cells and macrophages. Most importantly, Hsp90 inhibition prevents HIV rebound in fully suppressed humanized mice, and we identified persistent HIV-infected human cells in the mouse spleen. Two different Hsp90 inhibitors blocked HIV transcription in this persistent tissue reservoir and sustained HIV remission up to 11 weeks after drug cessation. Further, we now have evidence that persistent HIV infection in spleen, brain, and lung reservoirs is established soon after inoculation, which produces infectious virus despite highly potent ART. HIV-infected human T cells and macrophages are also found in the mouse liver, bone marrow, and reproductive tract. We hypothesize that increased Hsp90 activity at 39.5°C activates host transcription factors that are essential for HIV transcription. This hypothesis will be addressed in the following Specific Aims: 1) Determine the gene expression profile of heat-shock activated HIV-infected human resting CD4+ T cells, 2) Identify specific Hsp90 protein complexes that regulate HIV persistence in resting CD4+ T cells, and 3) Characterize persistent HIV reservoirs in vivo and investigate the source of rebound viremia. We anticipate these studies will lead to a better understanding of how Hsp90 regulates HIV transcription, and our in vivo results will provide insight into the nature of rebound viremia from persistent tissue reservoirs.

项目概要 尽管存在持续感染 HIV 的静息 CD4+ T 细胞，但在组织和器官储存库中仍无法检测到 数十年来成功的抗逆转录病毒治疗（ART）。静息 CD4+ T 细胞中基因活性的缺乏使得 整合的具有复制能力的原病毒可以无限期地存在。 HIV 在激活的 CD4+ T 细胞中的复制是 细胞病变和受感染的细胞被免疫系统清除。然而，一小部分艾滋病毒感染者 激活的 CD4+ T 细胞恢复到静息 G0 表型，并且在组织储存库中保持未被检测到。这些艾滋病毒—— 受感染的静息细胞很容易被激活，并且中断 ART 的患者病毒血症总是会反弹 源自持续感染艾滋病毒的组织储存库。 我实验室正在进行的一个项目重点关注热休克蛋白 90 (Hsp90) 在 HIV 中的重要作用 复制。我们已经证明，轻度热休克（39.5°C）可加速慢性感染者的 HIV 转录 T 细胞系，可使人类原代细胞中的 HIV 复制增加高达 30 倍。加速 HIV 转录 与 39.5°C 时细胞 Hsp90 活性增加一致。 Hsp90 负责激活细胞 转录因子和 Hsp90 抑制剂 17-AAG 显着降低 NF-kB、NFAT、 和 STAT5 宿主转录因子。此外，39.5°C 会重新激活 ART 抑制的无病毒血症中的 HIV 复制 HIV 感染患者样本以及从完全抑制的人源化小鼠中分离出的人类静息 CD4+ T 细胞。 此外，我们还表明，一种更有效的下一代 Hsp90 抑制剂具有更强的抗 HIV 活性，并且具有 非常有利的治疗排名，临床 HIV 亚型在 39.5°C 时显示病毒转录增加 存在于原代人类 T 细胞和巨噬细胞中。 最重要的是，Hsp90 抑制可防止 HIV 在完全抑制的人源化小鼠中反弹，我们发现 小鼠脾脏中持续感染艾滋病毒的人类细胞。两种不同的 Hsp90 抑制剂可阻断 HIV 转录在这种持久的组织储存库中，并在停药后长达 11 周内持续缓解 HIV。 此外，我们现在有证据表明，脾脏、大脑和肺储存库中存在持续的艾滋病毒感染 接种后不久，尽管抗逆转录病毒疗法非常有效，但仍会产生传染性病毒。感染 HIV 的人类 T 细胞 巨噬细胞也存在于小鼠肝脏、骨髓和生殖道中。 我们假设 39.5°C 下 Hsp90 活性增加会激活宿主转录因子，这对于 HIV转录。这一假设将在以下具体目标中得到解决：1）确定基因 热休克激活的 HIV 感染人静息 CD4+ T 细胞的表达谱，2) 识别特定的 Hsp90 调节静息 CD4+ T 细胞中 HIV 持久性的蛋白质复合物，以及 3) 表征持久性 HIV 体内储存库并调查反弹病毒血症的来源。 我们预计这些研究将有助于更好地了解 Hsp90 如何调节 HIV 转录，以及 我们的体内结果将有助于深入了解持久性组织储存库的反弹病毒血症的性质。