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

摘要 潜伏感染HIV-1的个体T细胞之间存在广泛的异质性，这表明 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。虽然我们和其他人描述了 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(S) 变化仍是未知数。对于初级T细胞，向静止记忆状态或T细胞耗尽的转变 效应可能起了一定作用，但在幼稚T细胞或Tfh细胞中也发现了潜伏感染，这表明 贡献机制。导致这一应用的是，我们解决了艾滋病毒感染是否可以 引发宿主细胞的不可逆修饰，这可以解释潜伏感染事件的稳定性，并可能 与记忆状态或CD4T细胞亚型无关。我们发现潜伏感染的T细胞株和一个 来自男性捐赠者的体外产生的潜伏感染的初级T细胞中，有很大比例表现出 Y染色体(Loy)表型丢失。数量上的染色体异常，包括性丧失 染色体，导致非线性转录改变，这可以解释观察到的广泛的 个体潜伏感染HIV-1的T细胞之间的转录异质性。广泛的转录改变 也可能导致细胞信号受损，并解释了重新激活反应谱差异很大的原因 在单个细胞之间。对于原代T细胞，我们证明了Loy细胞中潜伏的HIV-1感染事件是 在很大程度上抵抗TCR/CD3复合体激活介导的重新激活，而潜伏感染T细胞在 拥有他们的Y染色体促进了TCR/CD3激活后HIV-1的重新激活。Y-损失- 因此，染色体现象可以解释为什么存在第一个抗再激活的储存库 由Siliciano小组描述。由于LOY是不可逆转的，这是第一次关于生物分子变化的报告 可以机械地解释HIV-1潜伏期的稳定性和重新激活的惰性，我们认为这是主要的障碍 为了根除病毒。在这一应用中，我们将扩展我们对LOY和再激活的相关性的研究 耐药的HIV-1潜伏期。我们的发现为我们提供了分离这两种表型的独特机会 基于具有功能后果的致因机制(LOY)的部分潜在储集层 (再激活抵抗)，这将允许探索和开发治疗策略 使用单细胞分析方法分别针对这些储集层组件(目标1)。同时，我们将 调查是否可以在女性捐献者的T细胞中发现类似的潜伏期表型，以及 X染色体(LOX)是女性最常见的数字染色体异常，在 HIV-1潜伏期控制优于Loy(目标2)。从这项提案中产生的见解应该提供基本的 对HIV-1潜伏期控制的新见解并指导开发新的治疗策略以针对 潜伏的HIV-1储存库，特别是HIV-1储存库的抗重新激活部分。

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.