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.

潜伏的宿主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 这与HIV储库不能使用单个HIV受体的表达来准确定义的想法一致。 蛋白报告的蛋白质谱也显示出供体依赖性属性。需要更好的理解 哪些因素会导致储层特征的非均质性。我们的长期目标是发展 针对潜伏感染细胞进行清除的策略。拟议研究的具体目标是 确定病毒嗜性对潜伏感染细胞的分子特征的贡献。我们的中央 一种假设是，当细胞被HIV潜伏感染时， 感染CCR 5或CXCR 4嗜性病毒，与受感染细胞的表型组成有关。的理由 这项研究的目的是为未来开发改进的策略提供分子平台。 使用基于抗体的方法的储库靶向，或潜伏期逆转的优化。我们将测试 通过追求以下具体目标来实现中心假设：1）确定CCR 5-和CXCR 4-嗜性 表型不同的CD 4 + T细胞中的感染影响潜伏储库的特征; 2）鉴定 用不同嗜性病毒感染的CD 4 + T细胞的分子特征; 3）确定 嗜性依赖性细胞标志物对抗体组捕获潜伏感染细胞的性能的影响 艾滋病毒携带者。我们的建议的新颖之处在于重点关注三个主要的水库子集： 在幼稚细胞中建立，在幼稚细胞中建立，过渡到记忆，在记忆细胞中建立， CXCR 4和CCR 5嗜性感染的背景。我们将描述集成频率， 储库对体外潜伏期逆转剂（LRA）的反应性，并确定表型差异 在蛋白质和RNA水平上，潜伏感染和未感染的细胞之间的蛋白质和RNA水平分别在这些细胞中的每一个内， 子集液相色谱质谱和单细胞RNA测序的最新创新 将用于艾滋病毒感染者的外周血和组织样本将用于确定 潜伏期的向性特异性特征对离体储库捕获效率的贡献。我们预计 使用不超过15种抗体，至少70-90%的潜伏库将被捕获，但不同的是， 根据感染每个个体的病毒的嗜性，可能需要抗体组。我们提出的 多学科的努力将揭示不同表型亚群细胞的完整分子特征 感染了不同嗜性的病毒。这项研究对艾滋病毒感染者很重要，因为它 代表着朝着实现将潜在储层大小减少到 在该水平上，在停止抗逆转录病毒治疗后病毒抑制可以持续。识别的分子将 作为开发基于抗体的策略的平台，以靶向潜伏感染的细胞进行消除 或优化LRA组合以在不同细胞类型中有效地再活化具有不同向性的原病毒。