Characterization of cocaine induced signaling pathways that enhances HIV transcription

可卡因诱导的增强 HIV 转录的信号通路的表征

基本信息

批准号：
9926231
负责人：
Mudit Tyagi
金额：
$ 45.31万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9926231
关键词：
Acquired Immunodeficiency Syndrome Affect Applications Grants Biological Markers Cell Line ChIP-seq Chromatin Chromatin Structure Cocaine Cocaine Dependence Complex Data Development Diagnostic Diagnostic tests Drug abuse Drug usage ELK1 gene Enzymes Epigenetic Process Euchromatin Gene Expression Gene Expression Regulation Generations Genes Genetic Transcription Genome Genomics Goals Grant HIV HIV Infections HIV-1 HIV-associated neurocognitive disorder Highly Active Antiretroviral Therapy Histone Deacetylase Histone H3 IKK alpha Illicit Drugs Infection Intervention Knowledge Lead MAP Kinase Gene MAPK8 gene Microglia Modification Molecular Morbidity - disease rate NF-kappa B Nature Nervous system structure PI3K/AKT Pathway interactions Patients Pattern Peripheral Blood Mononuclear Cell Pharmaceutical Preparations Pharmacologic Substance Phase Phosphorylation Phosphotransferases Positioning Attribute Positive Transcriptional Elongation Factor B Process Proteins Proto-Oncogene Proteins c-akt Publishing RPS6KA5 gene Role Serine Signal Pathway Signal Transduction Structure Testing Transcript Transcription Elongation Transcription Initiation Viral Work activating transcription factor addiction cocaine exposure cocaine use cofactor comorbidity design drug addict drug of abuse experimental study improved knock-down macrophage monocyte new therapeutic target novel p65 public health relevance recruit transcription factor transcriptome sequencing transmission process treatment strategy

项目摘要

DESCRIPTION: Cocaine is a significant cofactor for HIV-1 infection, transmission and morbidity. Cocaine accelerates HIV-1 gene expression by altering specific cell-signaling and epigenetic pathways and promotes HIV-associated neurocognitive disorders (HAND). Our published data demonstrates that cocaine-induced signaling pathways lead to the activation of kinases, such as MSK1 which subsequently catalyzes the phosphorylation of p65 subunit of NF- ĸB at position S276 (P-p65S276) and histone H3 at Serine 10 (P-H3S10). P-H3S10, besides facilitating the establishment of transcriptionally active chromatin structures, promotes the recruitment of P-TEFb at HIV LTR. Consequently, cocaine accelerates both the initiation and the elongation phases of HIV transcription by activating NF-kB and P-TEFb, respectively, a prerequisite to generate complete HIV genomic transcript and new viral progeny. However, our understanding of the signaling pathways that cocaine utilizes in order to promote these changes and induce enzymes, including MSK1/2, RSK 1/2, Aurora-B, ELK1 and IKK-α is highly obscure. These enzymes directly and indirectly induce different epigenetic modifications, which never been investigated in the context of HIV infection. Thus, it is imperative to have clear knowledge about the cocaine induced signaling pathways that affect HIV replication. In this grant we have proposed a systematic analysis. First, we will define and characterize the cocaine induced signaling pathways that lead to the activation of above mentioned different kinases, their target proteins and their gene regulation. Later, we will define the role of cocaine induced epigenetic modifications and the precise role of involved enzymes and their effect on HIV gene expression and replication. Subsequently, we will define the signaling pathways which cocaine induces to activate NF-kB and P-TEFb. Finally, we will utilize high throughput approaches to characterize involved epigenetic modifications and the genes which are influenced by cocaine exposure; in order to find unique biomarkers of cocaine use. All the results will be reproduced in monocytic/macrophage and microglial cell lines. Crucial findings will be confirmed using primary Monocyte derived macrophages (MDMs) from cocaine using HIV patients. Finally, to validate the direct effect of cocaine, we will perform ex vivo experiments where we will expose PBMCs from HIV infected patients to cocaine. Broader Impact: These studies will identify many novel actions of cocaine, involved pathways and enzymes which can be exploited for the development of improved diagnostic tests and may open up new avenues for better pharmaceutical interventions in drug addict HIV patients.

描述：可卡因是 HIV-1 感染、传播和发病的重要辅助因子。可卡因通过改变特定的细胞信号传导和表观遗传途径加速 HIV-1 基因表达，并促进 HIV 相关的神经认知障碍 (HAND)。 -诱导的信号通路导致激酶的激活，例如 MSK1，随后催化 NF-ĸB 的 p65 亚基磷酸化S276 (P-p65S276) 和丝氨酸 10 处的组蛋白 H3 (P-H3S10) 除了促进转录活性染色质结构的建立外，还促进 HIV LTR 处 P-TEFb 的募集，可卡因可加速这两种过程的启动。以及分别通过激活 NF-kB 和 P-TEFb 的 HIV 转录延伸阶段，这是生成完整 HIV 基因组的先决条件然而，我们对可卡因促进这些变化和诱导酶（包括 MSK1/2、RSK 1/2、Aurora-B、ELK1 和 IKK-α）的信号通路的了解非常模糊。这些酶直接和间接诱导不同的表观遗传修饰，而这些修饰从未在 HIV 感染的情况下进行过研究。因此，必须清楚地了解可卡因诱导的影响 HIV 复制的信号通路。 .首先，我们将定义和表征可卡因诱导的信号通路，这些信号通路导致上述不同激酶、其靶蛋白及其基因调控的激活，稍后，我们将定义可卡因诱导的表观遗传修饰的作用以及相关酶的精确作用及其对基因的影响。随后，我们将定义可卡因诱导激活 NF-kB 和 P-TEFb 的信号通路。最后，我们将利用高通量方法来表征所涉及的表观遗传修饰和受可卡因暴露影响的基因；为了找到独特的生物标志物所有结果将在单核细胞/巨噬细胞和小胶质细胞系中重现，使用来自使用可卡因的 HIV 患者的原代单核细胞衍生的巨噬细胞 (MDM) 来证实重要的发现。进行离体实验，我们将 HIV 感染患者的 PBMC 暴露于可卡因更广泛的影响：这些研究将确定可卡因的许多新作用，涉及的途径和酶可用于开发改进的诊断测试，并可能开辟新的途径。对吸毒成瘾的艾滋病毒患者进行更好的药物干预的新途径。