Heat shock protein 90 and HIV persistence

热休克蛋白 90 和 HIV 持久性

• 批准号: 8663192
• 负责人: Cheryl Stoddart
• 金额: $ 29.64万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-17 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8663192
• 关键词: 1-Phosphatidylinositol 3-Kinase; Achievement; Acquired Immunodeficiency Syndrome; Address; Affinity; Affinity Chromatography; Aftercare; C-terminal; CD4 Positive T Lymphocytes; Cell Death; Cell Line; Cell Nucleus; Cells; Chromatin; Client; Complex; DNA Polymerase II; Detection; Disease remission; Epigenetic Process; Family; Fever; Frequencies; Genes; Genetic Transcription; Genomics; HIV; HIV Infections; Heat-Shock Proteins 90; Heat-Shock Response; Highly Active Antiretroviral Therapy; Histone Deacetylation; Human; Hyperthermia; Immune system; Integration Host Factors; Interleukin-1; Interleukin-2; Interleukin-6; Lead; Length; Life Expectancy; Lymphocyte; Mass Spectrum Analysis; Mediating; Memory; Modeling; Molecular Chaperones; Mus; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Physiological; Plasma; Positive Transcriptional Elongation Factor B; Production; Protein Analysis; Protein Isoforms; Protein Kinase C; Proteins; Public Health; RNA; RNA chemical synthesis; Reporter; Rest; Role; Sendai virus; Signal Transduction; Stress; TNF gene; Temperature; Testing; Therapeutic; Transcript; Transcriptional Regulation; Viral; Viral Load result; Viremia; Virus; Vorinostat; bryostatin; chaperone machinery; chromatin immunoprecipitation; chromatin remodeling; cytokine; expectation; inhibitor/antagonist; innovation; mouse model; novel; novel therapeutics; prevent; promoter; protein expression; public health relevance; purge; reactivation from latency; research study; transcription factor; treatment strategy; ultraviolet irradiation; vector

DESCRIPTION: Highly active antiretroviral therapy (HAART) successfully reduces the viral load to below the limit of detection, but HIV replication resumes soon after patients cease therapy. It is believed that a small subset of persistent HIV- infected resting CD4+ T cells are activated in the absence of HAART and contribute to the rebound in patient viral load. This has led to a new treatment strategy that proposes a combination of latency-purging drugs and HAART, with the expectation that activating HIV-infected resting CD4+ T cells will eliminate the persistent viral reservoir by virus-mediated cell death, by the immune system, and by HAART. Reactivation of HIV latency in resting CD4+ T cells can also be induced nonchemically by heat shock. In ongoing experiments, we have found that heat shock protein 90 (Hsp90) is essential for HIV replication and that heat shock (39.5oC) significantly increases HIV infectivity with a simultaneous increase in Hsp90 protein expression. Similar studies by others suggest that reactivation of HIV latency by heat shock is dependent on Hsp90 expression. HIV latency is maintained in resting CD4+ T cells by suppression of the level of virus- specific proteins. Cellular activation results in production of these host factors, but these proteins need to be activated in order to induce HIV reactivation. The active P-TEFb complex and the NF-?B family of transcription factors are essential for HIV reactivation, and these cellular proteins are activated by the Hsp90 chaperone machinery. Similarly, certain chromatin-remodeling proteins that control HIV latency are activated by Hsp90 and lead to HIV reactivation. The examples mentioned above are part of a larger body of work showing that heat shock, and specifically Hsp90, influence HIV replication. Given these observations, we hypothesize that Hsp90 has a functional role in reactivating HIV latency. This hypothesis will be addressed in the experiments of the following Specific Aims: (1) to study how Hsp90 reactivates HIV latency in resting CD4+ T cells, and (2) to identify Hsp90-interacting proteins that reactivate HIV latency. These studies include expressing Hsp90 in untransformed primary human lymphocytes and analyzing the specific Hsp90-interacting proteins that induce HIV reactivation in latently infected resting CD4+ T cells. This hypothesis will be validated in the humanized NSG-BLT mouse model of HIV latency. The objective of this proposal is to identify Hsp90-interacting proteins that reactivate HIV latency, and analysis of these proteins may reveal novel targets for elimination of the latent HIV reservoir.

描述：高效抗逆转录病毒疗法 (HAART) 成功地将病毒载量降至检测限以下，但患者停止治疗后不久，HIV 复制就会恢复。据信，一小部分持续性HIV感染的静息CD4+T细胞在缺乏HAART的情况下被激活，并有助于患者病毒载量的反弹。这催生了一种新的治疗策略，提出将潜伏期清除药物和HAART相结合，期望激活HIV感染的静息CD4+T细胞将通过病毒介导的细胞死亡、免疫系统和HAART消除持久的病毒库。热休克也可以非化学方式诱导静息 CD4+ T 细胞中 HIV 潜伏期的重新激活。在正在进行的实验中，我们发现热休克蛋白 90 (Hsp90) 对于 HIV 复制至关重要，并且热休克 (39.5oC) 显着增加 HIV 感染性，同时 Hsp90 蛋白表达增加。其他人的类似研究表明，热休克导致的 HIV 潜伏期重新激活取决于 Hsp90 的表达。通过抑制病毒特异性蛋白的水平，在静息 CD4+ T 细胞中维持 HIV 潜伏期。细胞激活导致这些宿主因子的产生，但这些蛋白质需要被激活才能诱导 HIV 重新激活。活性 P-TEFb 复合物和转录因子 NF-κB 家族对于 HIV 重新激活至关重要，并且这些细胞蛋白由 Hsp90 分子伴侣机制激活。同样，某些控制 HIV 潜伏期的染色质重塑蛋白被 Hsp90 激活并导致 HIV 重新激活。上面提到的例子是大量研究工作的一部分，这些研究表明热休克，特别是 Hsp90，会影响 HIV 复制。鉴于这些观察结果，我们假设 Hsp90 在重新激活 HIV 潜伏期中具有功能性作用。这一假设将在以下具体目标的实验中得到解决：(1) 研究 Hsp90 如何重新激活静息 CD4+ T 细胞中的 HIV 潜伏期，以及 (2) 鉴定重新激活 HIV 潜伏期的 Hsp90 相互作用蛋白。这些研究包括在未转化的原代人类淋巴细胞中表达 Hsp90，并分析在潜伏感染的静息 CD4+ T 细胞中诱导 HIV 重新激活的特定 Hsp90 相互作用蛋白。这一假设将在 HIV 潜伏期人源化 NSG-BLT 小鼠模型中得到验证。该提案的目的是确定重新激活 HIV 潜伏期的 Hsp90 相互作用蛋白，对这些蛋白的分析可能揭示消除潜伏 HIV 储存库的新靶点。

项目成果

Genetically edited CD34+ cells derived from human iPS cells in vivo but not in vitro engraft and differentiate into HIV-resistant cells.

来自人类 iPS 细胞的基因编辑 CD34 细胞在体内但不在体外移植并分化为 HIV 抗性细胞。

• DOI: 10.1073/pnas.2102404118
• 发表时间: 2021
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Morvan,MaeligG;Teque,Fernando;Ye,Lin;Moreno,MaryE;Wang,Jiaming;VandenBerg,Scott;Stoddart,CherylA;Kan,YuetWai;Levy,JayA
• 通讯作者: Levy,JayA