Investigate Host Gene Isoforms Contributing to HIV Persistence in Cocaine Users

研究导致可卡因吸食者中艾滋病毒持续存在的宿主基因亚型

• 批准号: 10788990
• 负责人: Wei Jiang
• 金额: $ 74.11万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10788990
• 关键词: Affect; Alternative Splicing; Anti-Retroviral Agents; Binding; Biological Assay; CD4 Positive T Lymphocytes; CREB1 gene; CREM protein; Cells; Chemicals; Chronic; Cocaine; Cocaine Users; Cyclic AMP; DNA Markers; Dinoprostone; Disease Progression; Dose; Drug abuse; Drug usage; Drug user; Exons; Frequencies; Gene Expression Profile; Generations; Genes; Genetic Transcription; HIV; HIV Infections; HIV resistance; HIV-1; HIV/AIDS; IRF3 gene; Immune; In Vitro; Individual; Infection; Inflammation; Inflammatory Response; Integration Host Factors; Investigation; Knock-out; Length; Measures; Mediating; Methods; Microglia; NF-kappa B; Pathogenesis; Patient Recruitments; Phase; Population; Protein Isoforms; RNA Splicing; Residual state; Role; Signal Transduction; Testing; Tissue Sample; Tissue-Specific Gene Expression; Validation; Variant; Viral; Viral Load result; Viral reservoir; Virus; Virus Replication; Withdrawal; antiretroviral therapy; brain tissue; chronic infection; clinically significant; cocaine use; digital; improved; inflammatory marker; knock-down; monocyte; mortality; non-drug; novel; nuclear factors of activated T-cells; overexpression; potential biomarker; prevent; prospective; resistance mechanism; therapeutic development; transcription factor; transcriptome sequencing

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.

项目概要 尽管抗逆转录病毒疗法 (ART) 能够成功阻止 HIV 的主动复制，但它并不能完全阻止病毒的复制。 根除感染，HIV 仍然持续感染，并且在 ART 停药后病毒载量可能会反弹， 调节艾滋病毒复制的宿主机制的研究是治疗艾滋病毒/艾滋病的主要障碍。 将有助于提高对支持艾滋病毒持续感染的机制的理解。 扰乱宿主调节因素以消除残留艾滋病毒的新策略本主题尤其与相关。 感染艾滋病毒的吸毒者，因为吸毒对艾滋病毒感染产生深远的影响，增加了难度 我们早期的努力包括识别新的宿主限制因素。 与一小部分 HIV 感染者 (<1%) 特别相关，称为精英控制者 (EC)，他们 在一项初步研究中，我们进行了在没有 ART 的情况下维持对 HIV 复制的长期控制。 RNA 测序分析并鉴定了来自 EC、HIV 感染者的细胞中的选择性剪接变体 接受抑制性 ART、未接受 ART 的 HIV 感染者和健康对照。 确定了 EC 特有且可能影响 HIV 耐药性的表达模式，包括 CREM/ICER 基因的替代 RNA 剪接和外显子使用变体（cAMP 响应元件） 特定 ICER 外显子的敲除和敲除导致了调节剂/诱导型 cAMP 早期阻遏物。 ICER 亚型的过度表达也显着增加了 HIV 感染。 这些早期研究初步表明，ICER 亚型在可卡因使用者中失调。 证实 CREM/ICER 是一种独特且新颖的抑制 HIV 复制的宿主限制因子。 全面调查它们在调节艾滋病毒感染方面的作用，特别是对于可卡因使用者。此外， 我们还建议确定因使用可卡因而失调的其他宿主基因亚型，这总体上促进了 我们的中心假设是某些宿主基因（包括 CREM/ICER）会经历 HIV 持续感染。 大量 RNA 剪接可在感染 HIV 的可卡因使用者中产生不同的亚型，从而支持 HIV 该人群的持续感染，可以有针对性地有利于艾滋病毒功能性治愈并减轻艾滋病毒- 这些研究包括三个相关但独立的目标，我们将确定目标 1。 在目标 2 中，我们将研究 CREM/ICER、HIV 感染、炎症和可卡因使用参数的相关性。 在目标 3 中，我们研究了 CREM/ICER 在调节可卡因对 HIV 感染和炎症的影响中的作用。 将鉴定可卡因使用失调的 CD4+ T 细胞和单核细胞中的新基因亚型。