Loss of Y-chromosome as a driver of HIV-1 latency

Y 染色体丢失是 HIV-1 潜伏期的驱动因素

基本信息

批准号：
10882257
负责人：
OLAF KUTSCH
金额：
$ 42.43万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-16 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10882257
关键词：
Address Aging Binding Sites CD28 gene CD3 Antigens CD4 Positive T Lymphocytes Cell Line Cells Chromatin Chromosome abnormality Complex Data Development Drug Targeting Environment Event Exhibits Female Genes HIV HIV Infections HIV resistance HIV-1 Heterogeneity Histones IL8 gene Immunologics Impairment In Vitro Individual Infection Interleukin Gene Interleukin-2 Laboratories Long Terminal Repeats Mediating Memory Modeling Modification Monoclonal Antibodies Numerical Chromosomal Abnormality Patients Persons Phase Phenotype Play Population Positioning Attribute Premature aging syndrome Proteomics Proviruses RNA Polymerase II Reporting Resistance Rest Role Series Sex Chromosomes Signal Transduction Splenocyte Stress Surrogate Markers T-Lymphocyte Therapeutic Intervention Viral Viral reservoir Virus Latency Y Chromosome age effect antiretroviral therapy chromosome X loss chromosome Y loss curative treatments early onset exhaustion in vitro Model insight integration site latent HIV reservoir latent infection male member novel therapeutic intervention phenotypic biomarker phosphoproteomics promoter proteomic signature response response biomarker single cell analysis single-cell RNA sequencing targeted treatment therapeutic development therapeutic evaluation transcription factor transcriptomics

项目摘要

ABSTRACT The presence of extensive heterogeneity between individual latently HIV-1 infected T cells suggests that latent HIV-1 infection can persist under greatly differing host cell conditions. While we and others have described the biomolecular phenotype of latently infected T cells, the biomolecular driver(s) of these stable phenotypic changes remain unknown. For primary T cells, the transition to a resting memory state or T cell exhaustion effects may play a role, but latent infection is also found in naïve T cells or TfH cells suggesting additional contributing mechanisms. Leading to this application, we addressed the question whether HIV infection can trigger an irreversible modification of host cells that can explain the stability of latent infection events, and could occur independent of memory status or CD4+ T cell subtype. We show that latently infected T cell lines and a large percentage of in vitro generated latently infected primary T cells, when derived from male donors, exhibited a Loss of Y-chromosome (LOY) phenotype. Numerical chromosomal aberrations, including the loss of sex chromosomes, cause nonlinear transcriptomic changes, which would explain the observed extensive transcriptomic heterogeneity between individual latently HIV-1 infected T cells. Extensive transcriptomic changes can also result in impaired cellular signaling, and explain the widely differing reactivation response spectrum between individual cells. For primary T cells, we demonstrate that latent HIV-1 infection events in LOY cells are largely resistant to TCR/CD3-complex activation mediated reactivation, while latently infected T cells in possession of their Y-chromosome promoted HIV-1 reactivation following TCR/CD3 activation. Loss of Y- chromosome phenomena could thus explain the presence of a reactivation-resistant reservoir that has been first described by the Siliciano group. As LOY is irreversible, this is the first report of a biomolecular alteration that can mechanistically explain HIV-1 latency stability and reactivation inertness, which we consider the major hurdle to viral eradication. In this application we will extend our studies on the correlation of LOY and reactivation resistant HIV-1 latency. Our discovery provides us with the unique opportunity to separate the two phenotypic parts of the latent reservoir based on a causative mechanism (LOY) that has functional consequences (reactivation resistance), which will allow for the exploration and development of therapeutic strategies to individually target these reservoir components using single cell analysis approaches (Aim 1). In parallel we will investigate whether similar latency phenotypes can be found in T cells from female donors and whether loss of X-chromosome (LOX), the most frequent numerical chromosomal aberration in females, plays a similar role for HIV-1 latency control than LOY (Aim 2). The insights generated from this proposal should provide fundamental new insights into HIV-1 latency control and guide the development of new therapeutic strategies to target the latent HIV-1 reservoir, specifically the reactivation-resistant part of the HIV-1 reservoir.

抽象的个体潜伏感染 T 细胞之间存在广泛的异质性表明尽管我们和其他人已经描述过，潜伏的 HIV-1 感染可以在截然不同的宿主细胞条件下持续存在。潜伏感染 T 细胞的生物分子表型，这些稳定表型的生物分子驱动因素对于原代 T 细胞，向静息记忆状态或 T 细胞耗竭的转变仍然未知。效应可能发挥一定作用，但在幼稚 T 细胞或 TfH 细胞中也发现了潜伏感染，这表明额外的感染在这项应用之前，我们解决了艾滋病毒感染是否会影响的问题。触发宿主细胞的不可逆修饰，可以解释潜伏感染事件的稳定性，并且可以其发生与记忆状态或 CD4+ T 细胞亚型无关。我们表明，潜伏感染的 T 细胞系和 a 当来自男性供体时，大量体外产生的潜伏感染的原代 T 细胞表现出 a Y 染色体（LOY）表型缺失，染色体数值畸变，包括性别缺失。染色体，引起非线性转录组变化，这可以解释观察到的广泛现象个体潜伏 HIV-1 感染的 T 细胞之间的转录组异质性。也可能导致细胞信号传导受损，并解释了截然不同的重新激活反应谱对于原代 T 细胞，我们证明 LOY 细胞中的潜在 HIV-1 感染事件是在单个细胞之间进行的。很大程度上抵抗 TCR/CD3 复合物激活介导的再激活，而潜伏感染的 T 细胞 Y 染色体的拥有促进了 TCR/CD3 激活后 HIV-1 的重新激活。因此，染色体现象可以解释第一个出现的抗再激活病毒库的存在。 Siliciano 小组描述，由于 LOY 是不可逆的，这是生物分子改变的第一份报告。可以从机制上解释 HIV-1 潜伏稳定性和重新激活惰性，我们认为这是主要障碍在此应用中，我们将扩展对 LOY 和重新激活的相关性的研究。我们的发现为我们提供了分离这两种表型的独特机会。基于具有功能性后果的因果机制（LOY）的潜在储存库的部分（再激活抵抗），这将允许探索和开发治疗策略我们将同时使用单细胞分析方法单独针对这些储层成分（目标 1）。研究是否可以在女性捐赠者的 T 细胞中发现类似的潜伏表型，以及是否丢失 X 染色体 (LOX) 是女性中最常见的染色体数值畸变，在 HIV-1 潜伏期控制优于 LOY（目标 2）。该提案产生的见解应提供基础。 HIV-1潜伏期控制的新见解并指导新治疗策略的开发潜伏的 HIV-1 储存库，特别是 HIV-1 储存库的抗再激活部分。