Cocaine induced selective epigenetic and signaling pathways enhance HIV replicati

说明(申请人提供)：注射和非注射毒品的使用和滥用仍然是艾滋病毒感染和传播的重要辅助因素。滥用药物，如可卡因，也与HIV-1相关的致病机制有关，如HAD、HAND。很明显，滥用药物，包括可卡因，可以改变细胞表观遗传和信号通路，最终调节几个细胞基因的表达。因此，如果整合的人类免疫缺陷病毒(HIV)前病毒的基因表达也受到这种类型的刺激的影响，也就不足为奇了。虽然到目前为止，可卡因对HIV基因的表达主要归因于几种趋化因子、细胞因子、信号通路和一些病毒蛋白(如TAT和Env)的上调，但药物滥用引起的表观遗传和信号通路的变化等分子机制可以更好地解释即使在戒毒后仍有几个基因的持续诱导。为了更好地了解滥用药物和艾滋病毒复制之间的复杂相互作用，重要的是调查滥用药物对艾滋病毒基因表达的影响，特别是在脑细胞中，因为大脑是滥用药物和艾滋病毒的靶器官。可卡因是美国滥用最广泛的药物之一，它既损害脑细胞的正常功能，又激活中枢神经系统(CNS)中HIV基因的表达。因此，与非滥用可卡因的人相比，滥用可卡因的艾滋病毒感染者经历了更严重和更快的神经艾滋病发作。在这项资助中，我们将研究可卡因对两种原代巨噬细胞--小胶质细胞(来自脑)和外周巨噬细胞(来自外周血单核细胞)--HIV基因表达和复制的调控机制。确切地说，我们将检验这一假设，即可卡因通过促进组蛋白H3在丝氨酸10个残基的磷酸化以及通过激活特定的转录因子，如NF-kB和AP-1，促进P-TEFb在HIV LTR上的招募，从而诱导HIV复制。这些修饰最终会导致免疫和神经系统更快地恶化，这在吸毒者艾滋病毒患者中相当普遍。更广泛的影响：这项工作有望导致新的潜在分子途径的发现，这可能指导新的治疗策略，以解决由于吸毒者艾滋病毒患者异常的前病毒基因表达和复制而导致的几种不良疾病后果。

DESCRIPTION (provided by applicant): Injection and non-injection drug use and abuse remain significant cofactors for HIV infection and transmission. The drugs of abuse such as cocaine has also been implicated in HIV-1-associated pathogenesis e.g. HAD, HAND. It has become clear that drugs of abuse, including cocaine can modify both cellular epigenetic and signaling pathways, which ultimately modulate the expression of several cellular genes. Therefore, it would not be surprising if the gene expressions of integrated human immunodeficiency virus (HIV) provirus's are also influenced with this type of stimuli. Although, HIV gene expression by cocaine until now is mainly attributed to up regulation of several chemokines, cytokines, signaling pathways and some of viral proteins (e.g. Tat and Env), however, the molecular mechanism such as changes in epigenetic and signaling pathways by drugs of abuse could explain better the continuous induction of several genes even after removal of drug of abuse, which is normally the case. In order to develop better understanding of the complex interplay between drugs of abuse and HIV replication, it is important to investigate the impact of drugs of abuse on HIV gene expression especially in brain cells, as brain is the target organ for both drugs of abuse and HIV. Cocaine is one of the most widely abused drugs in the United States, which both impair the normal functioning of brain cells and also activate HIV gene expression in central nervous system (CNS). As a result, HIV-infected individuals who abuse cocaine experience more severe and rapid onset of NeuroAIDS than non-abusing individuals. In this grant we will study the molecular mechanisms involved in the regulation of HIV gene expression and replication by cocaine in two primary macrophage cells, microglial (from brain) and peripheral macrophage (MDMs from PBMCs). Precisely, we will test the hypothesis that cocaine induces HIV replication by facilitating the recruitment of P-TEFb at HIV LTR via enhancing the phosphorylation of Histone H3 at serine 10 residue and through activation of specific transcription factors such as NF-kB and AP-1. These modifications ultimately contribute to more rapid deterioration of immune and nervous system, which is quite prevalent in drug addict HIV patients. Broader Impact: This work is expected to lead towards the discovery of new underlying molecular pathways, which could direct towards the new therapeutic strategies to address several adverse disease outcomes due to aberrant proviral gene expression and replication in drug addict HIV patients.