The intersection of diet, cell metabolic state, and SIV reservoir transcription

项目摘要/摘要 治愈HIV-1感染的主要障碍是由静止的记忆CD4T细胞组成的蓄水池，其基因组 含有可诱导和复制能力强的HIV-1前病毒。水库的腐烂是缓慢的，这加强了 认为前病毒HIV-1DNA在转录上基本上是沉默的。最近的大量证据表明， 恰恰相反。在任何时间点，HIV-1储存库中不可忽视的一小部分仍保持转录活性，尽管 用抗逆转录病毒疗法(ART)完全抑制。此外，潜伏的HIV-1转录可以在这些之上被诱导 HIV-1刺激性化合物的表观基线水平，被认为是潜伏期反转剂(LRA)。一些LRA有 在HIV-1人类或SIV恒河猴的小型临床或临床前试验中，能够适度减少储存池的大小 猕猴(RMS)。然而，目前LRA使用的一个限制是，到目前为止，这些试剂只在非常 狭窄的患者/动物队列，不能代表HIV-1人群中存在的异质性，也不能说明 关于年龄、性别或与HIV-1相关的其他共病的流行情况。尤其是2型糖尿病是一种共同的 在PLWH中普遍存在的发病率，是饮食激素胰岛素失衡的结果。在预赛中 研究发现，CD4T细胞(它是HIV-1的主要储存库)表达胰岛素受体并响应 通过激活PI3K信号通路对胰岛素的作用。已知PI3K信号位于(1)代谢调节因子的上游。 潜伏的HIV-1转录(mTORC信号)和(2)与HIV-1启动子(Sp1)结合的转录因子 和FOXO1)。在这些拟议的研究中，我们将利用现有的完全抑制ART的强大队列 SIV RMS以探讨胰岛素信号在HIV-1/SIV生命周期中的调节作用。我们将采用尖端技术 用磷酸化蛋白质组学方法检测饮食激素胰岛素如何影响潜伏的SIV转录(AIM 1) 基于LRA的生物制剂的背景与胰岛素和(目标2)天然的 饮食摄入量引起的血糖/胰岛素波动。应用程序将寻求在基本级别(1)上定义 CD4T细胞中胰岛素“信号体”的研究(2)--可能影响潜伏期转录活性的饮食因素 HIV-1和更重要的是(3)为未来在胰岛素环境下使用LRA的研究提供了一个概念性框架 信号受损。

Project Summary/Abstract The main obstacle to curing HIV-1 infection is a reservoir that consists of resting memory CD4 T cells whose genomes contain inducible and replication-competent HIV-1 proviruses. Decay of the reservoir is slow and this has reinforced a notion that proviral HIV-1 DNA is largely transcriptionally silent. A wide body of recent evidence has pointed to the contrary. At any point in time, a non-negligible fraction of the HIV-1 reservoir remains transcriptionally active despite complete suppression with antiretroviral therapy (ART). Moreover, latent HIV-1 transcription can be induced above these apparent baseline levels with HIV-1 stimulatory compounds, deemed latency reversal agents (LRAs). Some LRAs have been able to modestly reduce reservoir size in small clinical or pre-clinical trials of HIV-1+ humans or SIV+ rhesus macaques (RMs). However, a current limitation of LRA use is that to date, these agents have been tested only in very narrow patient/animal cohorts, not representative of heterogeneity that exists in the HIV-1+ population, and not accounting for age, gender, or the prevalence of additional co-morbidities associated with HIV-1. Type 2 diabetes in particular is a co- morbidity that is prevalent in PLWH, and is the result of an imbalance of the dietary hormone insulin. In preliminary studies, we find that CD4 T cells (which serve as the principle reservoir for HIV-1) express the insulin receptor and respond to insulin by activating the PI3K signaling pathway. PI3K signaling is known to be upstream of (1) metabolic regulators of latent HIV-1 transcription (mTORC signaling) and (2) transcription factors involved in binding to the HIV-1 promoter (Sp1 and FOXO1). In these proposed studies, we will take advantage of an existing well-powered cohort of fully ART-suppressed SIV+ RMs to probe the role of insulin signaling as a regulator of the HIV-1/SIV lifecycle. We will employ cutting-edge phospho-proteomic methods to examine how the dietary hormone insulin influences transcription of latent SIV in (Aim 1) the context of LRA-based biologics that share common signaling cascades with insulin and (Aim 2) the context of natural blood glucose/insulin fluctuations that occur with dietary intake. The application will seek to define on a basic level (1) the insulin “signalome” in CD4 T cells (2) as-yet explored dietary factors that may influence transcriptional activity of latent HIV-1 and importantly (3) provide a conceptual framework for future studies on LRA use in settings in which insulin signaling is impaired.