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.

最近报告的结果显示，潜伏病毒库仍然是治愈艾滋病毒/艾滋病的主要障碍。 与 HIV 储存库不能使用单个病毒的表达来准确定义的观点相一致。 所报道的蛋白质谱还显示出依赖于供体的属性。 我们的长期目标是开发哪些因素导致储层特征的异质性。 针对潜在感染细胞进行消除的策略 本研究的具体目标是 确定病毒趋向性对潜伏感染细胞分子特征的贡献。 假设潜伏感染 HIV 的细胞在以下情况下将具有共同的和独特的分子特征： 感染了 CCR5 或 CXCR4 病毒，与感染细胞的表型组成有关。 拟议研究的目的是为未来开发改进策略提供一个分子平台 我们将测试我们的基于抗体的方法的储库靶向或潜伏期逆转的优化。 通过追求以下具体目标来得出中心假设：1) 确定 CCR5 和 CXCR4 向性如何 表型不同的 CD4+ T 细胞的感染会影响潜伏病毒库的特征；2) 识别 感染不同向性病毒的CD4+ T细胞的分子特征；3)确定 向性依赖性细胞标记物对抗体组捕获潜伏感染细胞的性能的影响 我们提案的新颖之处在于关注三个主要的储存者亚群：已确定。 在幼稚细胞中，在转变为记忆的幼稚细胞中建立，在记忆细胞中建立，在 我们将描述整合频率和 CXCR4- 和 CCR5-tropic 感染的背景。 体外储库对潜伏期逆转剂 (LRA) 的反应性，并识别表型差异 在每个这些细胞内的潜伏感染细胞和未感染细胞之间的蛋白质和 RNA 水平上 液相色谱质谱法和单细胞 RNA 测序的最新创新。 将使用艾滋病毒感染者的外周血和组织样本来确定。 我们期望潜伏期的向性特异性特征对储库离体捕获效率的贡献。 使用不超过 15 种抗体，至少 70-90% 的潜在储存库将被捕获，但不同 根据我们建议的病毒的趋向性，可能需要抗体组。 多学科的努力将揭示不同表型亚群细胞的完整分子特征 这项研究对于艾滋病毒感染者来说很重要，因为它 代表着朝着实现将潜在储层尺寸减少到水平的最终目标迈出的重要一步 在停止抗逆转录病毒治疗后，病毒抑制可以持续。 作为开发基于抗体的策略的平台，以消除潜在感染的细胞 或优化LRA组合以有效地重新激活不同细胞类型中不同向性的原病毒。