Stress response networks in HIV infection

 描述（由申请人提供）：本提案的目标是了解HIV感染中的细胞应激反应及其与循环外泌体货物的关系。在初步研究中，使用的样本和临床数据来自主要是男性非洲裔美国人受试者感染艾滋病毒的队列，可卡因的使用与炎症，氧化应激，线粒体功能障碍和改变单胺代谢的标志物增加。与HIV对照相比，HIV+可卡因使用者的血浆外泌体和外泌体相关HSP 70也增加。此外，我们在HIV+可卡因使用者中鉴定了血浆外泌体miRNA，这些miRNA也在细胞培养模型中的细胞应激反应期间诱导。基于初步数据和预测的网络模型，我们假设HIV感染诱导细胞应激反应，作为炎症，氧化应激，线粒体功能障碍和单胺代谢改变的结果。辅酶A通过对儿茶酚胺和多巴胺的作用增强这些细胞应激反应，从而增加能量需求、线粒体功能障碍和氧化应激。包括REST、PGC-1 α和sirtuins在内的转录和表观遗传调节因子预计在调节这些细胞应激反应网络和调节可卡因对细胞的相关作用中发挥关键作用。在细胞应激反应期间释放的外来体和其他微泡含有代表这些应激反应的潜在生物标志物的蛋白质和RNA货物。为了解决这一假设，我们提出了综合生物学方法，使用临床样本和数据来表征使用和不使用可卡因的种族多样性HIV+受试者队列中的外泌体和细胞应激反应，以及它们与临床数据的关系。血浆外泌体货物，基因和miRNA表达谱，炎症和氧化应激标志物，代谢组学以及病毒学和免疫学数据的综合分析沿着有针对性的实验将用于构建，测试和完善参与介导HIV感染中细胞应激反应的网络和途径的模型及其与外泌体货物的关系。

 DESCRIPTION (provided by applicant): The goal of this proposal is to understand cellular stress responses in HIV infection, and their relationship to circulating exosome cargo. In preliminary studies using samples and clinical data from cohorts of predominantly male African-American subjects with HIV infection, cocaine use was associated with increased markers of inflammation, oxidative stress, mitochondrial dysfunction, and altered monoamine metabolism. Plasma exosomes and exosome-associated HSP70 were also increased in HIV+ cocaine users compared to HIV- controls. Furthermore, we identified plasma exosome miRNAs in HIV+ cocaine users that were also induced during cell stress responses in cell culture models. Based on preliminary data and predicted network models, we hypothesize that HIV infection induces cellular stress responses as a consequence of inflammation, oxidative stress, mitochondrial dysfunction, and altered monoamine metabolism. Cocaine augments these cellular stress responses through effects on catecholamines and dopamine, which increase energy demands, mitochondrial dysfunction, and oxidative stress. Transcriptional and epigenetic regulators including REST, PGC-1alpha, and sirtuins are predicted to play key roles in regulating these cellular stress response networks and modulating cocaine-related effects on cells. Exosomes and other microvesicles released during cell stress responses contain protein and RNA cargo that represent potential biomarkers of these stress responses. To address this hypothesis, we propose integrated biology approaches using clinical samples and data to characterize exosomes and cellular stress responses in racially diverse cohorts of HIV+ subjects with and without cocaine use, and their relationship to clinical data. Integrative analysis of plasma exosome cargo, gene and miRNA expression profiles, inflammation and oxidative stress markers, metabolomics, and virological and immunological data along with targeted experimentation will be used to build, test, and refine models of networks and pathways involved in mediating cellular stress responses in HIV infection and their relationship to exosome cargo.