Investigate Host Gene Isoforms Contributing to HIV Persistence in Cocaine Users

项目总结 虽然抗逆转录病毒疗法(ART)成功地阻止了艾滋病毒的活跃复制，但它并不能完全阻止 根除感染。艾滋病毒仍然持续感染，病毒载量在ART停用后可能会反弹， 这是治愈艾滋病毒/艾滋病的主要障碍。调控HIV复制的宿主机制的研究 将有助于提高对支持艾滋病毒持续感染的机制的理解。它还将提供 扰乱宿主监管因素以消除残留艾滋病毒的新战略。此主题与以下内容尤其相关 对于感染艾滋病毒的吸毒者，由于吸毒对艾滋病毒感染造成深刻影响，增加了难度 管理艾滋病毒病毒库。我们早期的工作包括识别新的寄主限制因素 特别与一群罕见的艾滋病毒感染者(1%)有关，他们被称为精英控制者(ECs)，他们 在没有抗逆转录病毒治疗的情况下，可以保持对艾滋病毒复制的长期控制。在初步研究中，我们进行了 HIV感染者ECs细胞的RNA测序分析和选择性剪接变异体的鉴定 接受抑制性ART，ART天真的HIV感染者和健康对照。差异基因 确定了针对内皮细胞并可能影响艾滋病毒耐药性的表达模式，包括 CREM/ICER基因(cAMP反应元件)的RNA选择性剪接和外显子使用变体 调节剂/诱导型cAMP早期抑制物)。特定ICER外显子的敲除和敲除导致 显著增加了艾滋病毒的感染。ICER亚型的过表达可降低HIV感染。我们也 初步表明，ICER亚型在可卡因吸毒者中存在调节失调。总之，这些早期的研究 确认CREM/ICER是一种独特且新颖的抑制HIV复制的宿主限制因子。我们建议 全面调查它们在管理艾滋病毒感染方面的作用，特别是对可卡因使用者的作用。此外， 我们还建议确定其他因可卡因使用而失调的宿主基因亚型，它们总体上促进了 艾滋病毒持续感染。我们的中心假设是，某些宿主基因，包括CREM/ICER，经历了 深刻的RNA剪接在HIV感染的可卡因使用者中产生不同的异构体，从而支持HIV 这一人群中的持续感染，可以有针对性地受益于艾滋病毒的功能性治疗和缓解艾滋病毒- 引起炎症。这些研究包括三个相互关联但独立的目标。在目标1中，我们将确定 CREM/ICER与HIV感染、炎症和可卡因使用参数的相关性。在目标2中，我们将 研究CREM/ICER在可卡因对HIV感染和炎症的影响中的作用。在目标3中，我们 将在因可卡因使用而失调的CD4+T细胞和单核细胞中发现新的基因异构体。

PROJECT SUMMARY Although antiretroviral therapy (ART) is successful to block active replication of HIV, it does not completely eradicate the infection. HIV remains persistently infected and viral load can rebound after ART withdrawal, presenting a major obstacle for cure of HIV/AIDS. Investigation of host machineries that regulate HIV replication will help to improve the understanding of the mechanisms supporting HIV persistent infection. It will also provide new strategies to perturb host regulatory factors for eliminating residual HIV. This topic is especially relevant for the HIV-infected drug users, since drug abuse creates a profound impact on HIV infection, increasing the difficulty to manage HIV viral reservoirs. Our earlier effort includes the identification of novel host restriction factors specifically associated with a rare subset of HIV-infected individuals (<1%), termed elite controllers (ECs), who can maintain long-term control over HIV replication in the absence of ART. In a preliminary study, we performed RNA sequencing analysis and identified alternative splicing variants in cells from ECs, HIV-infected individuals undergoing suppressive ART, ART-naive HIV-infected individuals, and healthy controls. Differential gene expression patterns that are specific to ECs and may influence HIV resistance were identified, including alternative RNA splicing and exon usage variants of the CREM/ICER gene (cAMP-responsive element modulator/inducible cAMP early repressors). The knockout and knockdown of specific ICER exons resulted in significantly increased HIV infection. Overexpression of ICER isoforms decreased HIV infection. We also preliminarily showed that ICER isoforms are dysregulated in cocaine users. Together, these earlier studies confirm that CREM/ICER is a unique and novel host restriction factor suppressing HIV replication. We propose to comprehensively investigate their roles in regulating HIV infection, particularly for cocaine users. Furthermore, we also propose to identify other host gene isoforms that are dysregulated by cocaine use, which overall promote HIV persistent infection. Our central hypothesis is that certain host genes, including CREM/ICER, undergo profound RNA splicing to generate distinct isoforms in HIV-infected cocaine users and thus support HIV persistent infection in this population, which can be targeted to benefit HIV functional cure and mitigate HIV- induced inflammation. These studies include three related but independent aims. In Aim 1, we will determine correlations of CREM/ICER, HIV infection, inflammation, and cocaine use parameters. In Aim 2, we will investigate roles of CREM/ICER in mediating cocaine’s effect on HIV infection and inflammation. In Aim 3, we will identify novel gene isoforms in CD4+ T cells and monocytes dysregulated by cocaine use.