Investigate Host Gene Isoforms Contributing to HIV Persistence in Cocaine Users

项目摘要 尽管抗逆转录病毒疗法（ART）成功地阻断了HIV的活跃复制，但它并不能完全阻断HIV的活跃复制。 根除感染艾滋病毒仍然持续感染，病毒载量在停止抗逆转录病毒治疗后可能反弹， 这是治疗艾滋病的主要障碍。调节HIV复制的宿主机制的研究 这将有助于提高对艾滋病毒持续感染机制的认识。它还将提供 干扰宿主调节因子以消除残留HIV的新策略。本主题特别适用于 艾滋病病毒感染的吸毒者，由于吸毒对艾滋病病毒感染产生了深刻的影响， 来管理HIV病毒库。我们早期的努力包括鉴定新的宿主限制因子 特别是与一种罕见的HIV感染者（<1%）有关，称为精英控制者（EC）， 可以在没有ART的情况下维持对HIV复制的长期控制。在一项初步研究中，我们进行了 RNA测序分析和识别的细胞中的选择性剪接变异体从EC，HIV感染的个体 接受抑制性ART、ART初治HIV感染个体和健康对照。差异基因 鉴定了EC特异性表达模式并可能影响HIV抗性，包括 CREM/ICER基因的可变RNA剪接和外显子使用变体（cAMP反应元件 调节剂/诱导型cAMP早期阻遏物）。特异性ICER外显子的敲除和敲低导致 艾滋病毒感染率大幅上升。ICER亚型的过表达降低了HIV感染。我们也 初步表明，ICER亚型在可卡因使用者中失调。这些早期的研究 证实CREM/ICER是一种独特的、新型的抑制HIV复制的宿主限制因子。我们提出 全面调查它们在调节艾滋病毒感染方面的作用，特别是可卡因使用者。此外，委员会认为， 我们还建议鉴定其他因可卡因使用而失调的宿主基因亚型，这些基因亚型总体上促进了 HIV持续感染。我们的中心假设是，某些宿主基因，包括CREM/ICER， 深刻的RNA剪接，在感染艾滋病毒的可卡因使用者中产生不同的亚型，从而支持艾滋病毒 在这一人群中持续感染，这可以有针对性地有利于艾滋病毒功能性治愈和减轻艾滋病毒- 诱发炎症。这些研究包括三个相关但独立的目标。在目标1中，我们将确定 CREM/ICER、HIV感染、炎症和可卡因使用参数的相关性。在目标2中，我们 研究CREM/ICER在介导可卡因对HIV感染和炎症作用中的作用。在目标3中， 将确定新的基因亚型在CD 4 + T细胞和单核细胞失调可卡因的使用。

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.