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.

项目摘要 持续的HIV感染的静止CD4+ T细胞可能在组织和器官储层目的地仍未发现 成功的抗逆转录病毒疗法（ART）。静止CD4+ T细胞中缺乏基因活性使 综合复制能力病毒无限期地持续。激活的CD4+ T细胞中的HIV复制是 通过免疫系统清除细胞病和感染细胞。但是，一小部分艾滋病毒感染者 活化的CD4+ T细胞还原为静止的G0表型，并在组织储层中保持未发现。这些艾滋病毒 - 感染的静息细胞可以很容易地被激活，并且ART中断的患者反弹病毒血症 起源于持续的HIV感染组织储层。 我的实验室中正在进行的项目集中于热休克蛋白90（HSP90）在HIV中的重要作用 复制。我们已经表明，温和的热休克（39.5°C）加速了慢性感染的HIV转录 在原代人细胞中，T细胞系并增加了30倍的HIV复制。加速HIV转录 与39.5°C下的细胞HSP90活性增加相吻合。 HSP90负责激活细胞 转录因子和HSP90抑制剂17-AAG显着降低了NF-KB，NFAT，NFAT， 和STAT5宿主转录因子。此外，39.5°C重新激活了毒Art抑制的无毒的艾滋病毒 HIV感染的患者样品以及从完全抑制的人道抑制小鼠分离的人类静止的CD4+ T细胞中。 此外，我们表明，具有更大潜在的下一代HSP90抑制剂具有更大的抗HIV活性 高度有利的治疗排名和临床HIV亚型显示在39.5°C时的病毒转录增加 在原代人T细胞和巨噬细胞中。 最重要的是，HSP90抑制可防止完全抑制的人性化小鼠反弹，我们确定 小鼠脾脏中持续的HIV感染的人类细胞。两种不同的HSP90抑制剂阻断了HIV 该持续组织储层中的转录和持续的HIV缓解量可在药物停止后长达11周。 此外，我们现在有证据表明，建立了脾脏，大脑和肺部的持续性艾滋病毒感染 接种后不久，产生感染性病毒迫切可能是高度潜在的艺术。 HIV感染的人T细胞 在小鼠肝，骨髓和生殖道中也发现了巨噬细胞。 我们假设在39.5°C下增加HSP90活性会激活宿主转录因子，这对于 HIV转录。该假设将在以下特定目的中解决：1）确定基因 热激活的HIV感染的人静息CD4+ T细胞的表达曲线，2）确定特定的HSP90 调节静止CD4+ T细胞中HIV持续性的蛋白质复合物，3）表征持续的HIV 体内水库并研究反弹病毒血症的来源。 我们预计这些研究将使人们更好地了解HSP90如何调节艾滋病毒转录，并且 我们的体内结果将为持续组织储层的反弹病毒血症的性质提供深入的了解。