Role of viral tropism in molecular signatures of HIV latency

病毒趋向性在 HIV 潜伏期分子特征中的作用

• 批准号: 10434386
• 负责人: Nadejda S Beliakova-Bethell
• 金额: $ 59.29万
• 依托单位: VETERANS MEDICAL RESEARCH FDN/SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-15 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10434386
• 关键词: AIDS/HIV problem; Affect; Antibodies; Biological Assay; Blood specimen; CCR5 gene; CD4 Positive T Lymphocytes; CXCR4 gene; Cell surface; Cells; Characteristics; Cytometry; DNA; Data; Development; Dyes; Event; Flow Cytometry; Frequencies; Future; Germ Cells; Goals; HIV; HIV Infections; Health; Heterogeneity; In Vitro; Individual; Infection; Knowledge; Laboratories; Membrane Proteins; Memory; Mission; Molecular; Molecular Profiling; National Institute of Allergy and Infectious Disease; Participant; Performance; Persons; Phenotype; Predisposition; Proteins; Protocols documentation; Proviruses; Public Health; RNA; Reporting; Research; Role; Testing; Tissue Sample; Tropism; United States National Institutes of Health; Viral; Virus; antiretroviral therapy; base; burden of illness; cell type; fighting; improved; innovation; liquid chromatography mass spectrometry; multidisciplinary; novel; peripheral blood; receptor; single-cell RNA sequencing; viral transmission; virus tropism

The latent reservoir remains the major obstacle to a cure for HIV/AIDS. Results from recent reports are consistent with the idea that the HIV reservoir cannot be accurately defined using expression of a single protein. The reported protein profiles also showed donor-dependent attributes. Better understanding is needed of what factors contribute to heterogeneity of the reservoir signatures. Our long-term goal is to develop strategies to target latently infected cells for elimination. The specific objective of the proposed research is to determine the contribution of viral tropism to molecular signatures of the latenly infected cells. Our central hypothesis is that cells latently infected with HIV will have both shared and unique molecular signatures when infected with CCR5- or CXCR4-tropic virus, tied to the phenotypic composition of infected cells. The rationale of the proposed research is to provide a molecular platform for the future development of improved strategies of reservoir targeting using antibody-based approaches, or optimization of latency reversal. We will test our central hypothesis by pursuing the following specific aims: 1) Determine how CCR5- and CXCR4-tropic infection in phenotypically different CD4+ T cells affects the characteristics of the latent reservoir; 2) Identify molecular signatures of CD4+ T cells infected with virus of different tropism; 3) Determine contribution of tropism-dependent cell markers to the performance of antibody panels to capture latently infected cells from persons with HIV. The novel aspect of our proposal is the focus on three major reservoir subsets: established in naïve cells, established in naïve cells that transitioned to memory, and established in memory cells, in the context on CXCR4- and CCR5-tropic infection. We will characterize integration frequencies and responsiveness of the reservoir to latency reversing agents (LRAs) in vitro, and identify phenotypic differences at the protein and RNA level between latently infected and uninfected cells individually within each of these subsets. The latest innovations in liquid chromatography mass spectrometry and single cell RNA sequencing will be used. Peripheral blood and tissue samples from persons with HIV will be used to determine the contribution of tropism-specific signatures of latency to the efficiency of reservoir capture ex vivo. We expect that using no more than 15 antibodies, at least 70-90% of the latent reservoir will be captured, but different antibody panels may be needed depending on the tropism of the virus infecting each individual. Our proposed multidisciplinary efforts will reveal the complete molecular signatures of cells of different phenotypic subsets infected with virus of different tropism. This research will be important for people with HIV, because it represents a significant step towards achieving the ultimate goal to reduce the latent reservoir size to the levels at which viral suppression can be sustained upon cessation of antiretroviral therapy. Identified molecules will serve as a platform for development of antibody-based strategies to target latently infected cells for elimination or optimization of LRA combinations to efficiently reactivate provirus of different tropism in different cell types.

潜在水库仍然是治愈艾滋病毒/艾滋病的主要障碍。最近报告的结果是 与无法使用单个表达式准确定义HIV储存库的想法一致 蛋白质。报道的蛋白质谱也显示了供体依赖性属性。需要更好的理解 哪些因素导致储层特征的异质性。我们的长期目标是发展 靶向潜在感染细胞以消除的策略。拟议研究的具体目标是 确定病毒朝托对最近感染细胞分子特征的贡献。我们的中心 假设是，当受HIV感染的细胞将具有共享和独特的分子特征时 感染了CCR5或CXCR4-纤维化病毒，与感染细胞的表型组成有关。理由 拟议的研究是为未来发展的改进策略提供分子平台 使用基于抗体的方法或优化潜伏期逆转的储层靶向。我们将测试我们的 通过追求以下特定目的：1）确定CCR5-和CXCR4-Tropic的中心假设 表型不同的CD4+ T细胞中的感染会影响潜在储层的特征。 2）识别 感染了不同卓越病毒的CD4+ T细胞的分子特征； 3）确定贡献 依赖于抗体的细胞标记抗体面板的性能，以捕获潜在的感染细胞 艾滋病毒的人。我们提案的新颖方面是关注三个主要储层子集：已建立 在幼稚的细胞中，在过渡到记忆并在记忆细胞中建立的幼稚细胞中建立 关于CXCR4和CCR5循环感染的上下文。我们将表征集成频率和 水库对潜伏期逆转剂（LRAS）的反应性，并确定表型差异 在每个蛋白质和未感染细胞之间的蛋白质和RNA水平下，每个细胞中的每一个分别 子集。液相色谱质谱和单细胞RNA测序的最新创新 将使用。来自HIV患者的外周血和组织样本将用于确定 延迟特异性特异性签名对储层捕获的效率的贡献。我们期望 使用不超过15种抗体，将捕获至少70-90％的潜在储层，但不同 可能需要根据感染每个人的病毒的向性，需要抗体面板。我们的建议 多学科的工作将揭示不同表型子集细胞的完整分子特征 感染了不同偏向主义的病毒。这项研究对艾滋病毒患者很重要，因为它 代表了实现最终目标的重要一步，将潜在储层尺寸降低到水平 停止抗逆转录病毒疗法后，可以维持病毒抑制。确定的分子会 作为开发基于抗体的策略的平台 或优化LRA组合以有效重新激活不同细胞类型中不同偏向主义的病毒。