Elucidation of Tissue-Specific Transcriptomic Profiles in Cardiometabolic Disease

描述(由申请人提供)：最近的全基因组关联研究(GWAS)提供了对复杂心脏代谢性疾病(CMD)的遗传基础的更多洞察力，包括2型糖尿病和动脉粥样硬化性心血管疾病。然而，这些发现只解释了遗传性的一小部分，这表明需要考虑普通DNA变异以外的遗传影响。转录组的知识对于更全面地了解基因组的遗传功能元件是必不可少的。高通量RNA测序(RNA-seq)的出现，包括我们的工作，证明了存在比以前编目的更大的转录复杂性和多样性。宿主对内毒素(LPS)的反应提供了对先天免疫转录调控的基本见解，同时也是炎症性CMD的模型。在烟酸诱发反应遗传学和内毒素血症(基因)研究中，我们招募健康人(N=286)为住院方案，并在静脉注射内毒素(1 ng/kg)前后采集血液和脂肪样本。在这里，我们建议在实验性人类内毒素血症期间进行单核细胞和脂肪转录和表观基因组谱分析，以发现人类疾病的组织特异性机制。在目标1中，我们将利用基因参与者的临床炎症反应来识别极端“低”(~&lt；5%PC)和“高”(~&gt；95%PC)响应者，以检测内毒素诱导的最具临床相关性的转录反应。在目标2中，我们将复制内毒素诱导的转录改变，定义表观遗传调节，并对重复的发现进行临床翻译。在目标3中，我们假设来自基因参与者的分化的人诱导多能干细胞(HiPSC)的基础和内毒素诱导的表观/转录组反应将类似于原代细胞和组织。这一建议结合了独特的临床资源、组织特异性转录组学和HiPSC的体外图谱，以确定对CMD的遗传贡献，发现新的疾病机制和治疗机会，并推动人类疾病研究的创新。 公共卫生相关性：全基因组关联研究(GWAS)只解释了复杂心脏代谢性疾病遗传性的一小部分。转录组的知识对于更全面地了解基因组的功能元件是必不可少的。我们的翻译转录切割提案的重点是应用人类组织的深度RNA测序，识别新的转录切割变异，作为心脏代谢性疾病中组织特异性遗传功能障碍的机制。这些研究将揭示复杂心脏代谢性疾病的新的遗传机制和治疗靶点，超越DNA变异。

DESCRIPTION (provided by applicant): Recent genome wide association studies (GWAS) have provided increased insight into the genetic basis of complex cardio-metabolic diseases (CMD) including type-2 diabetes and atherosclerotic cardiovascular diseases. Such discoveries, however, only explain a small proportion of the heritability suggesting genetic influences other than common DNA variation need to be considered. Knowledge of the transcriptome is essential for a more complete understanding of the inherited functional elements of the genome. The advent of high-throughput RNA sequencing (RNA-seq), including our work, demonstrates the existence of a far greater transcriptomic complexity and diversity than previously catalogued. The host response to endotoxin (LPS) provides fundamental insights into transcriptomic regulation of innate immunity while also serving as a model for inflammatory CMD. In the Genetics of Evoked-Responses to Niacin and Endotoxemia (GENE) study, we recruited healthy individuals (N=286) to an inpatient protocol and collected blood and adipose samples before and after administration of endotoxin (1ng/kg intravenously). Here, we propose to perform monocyte and adipose transcriptomic and epigenomic profiling during experimental human endotoxemia in order to discover tissue-specific mechanisms of human disease. In Aim 1, we will utilize the clinical-inflammatory response in GENE participants to identify extreme "low" (~<5%PC) and "high" (~>95%PC) responders in order to detect endotoxin-induced transcriptomic responses of greatest clinical relevance. In Aim 2, we will replicate LPS- induced transcriptomic changes, define the epigenetic regulation, and pursue clinical translation of replicated findings. In Aim 3, we hypothesize that basal and endotoxin-evoked epigenome/transcriptome response in differentiated human induced pluripotent stem cells (hiPSC) from GENE participants will resemble that in primary cells and tissues. This proposal combines unique clinical resources, tissue-specific transcriptomics, and ex vivo profiling of hiPSC in order to identify heritable contribution to CMD, discover novel disease mechanisms and therapeutic opportunities, and advance innovation in the study of human disease. PUBLIC HEALTH RELEVANCE: Genome-wide association studies (GWAS) only explain a small proportion of the heritability of complex cardio-metabolic disease. Knowledge of the transcriptome is essential for more complete understanding of the functional elements of the genome. The focus of our translational transcriptomic proposal, applying deep RNA sequencing of human tissue, is identification of novel transcriptomic variations as mechanisms of tissue-specific genetic dysfunction in cardio- metabolic disease. These studies will reveal novel inherited mechanisms and therapeutic targets, beyond DNA variation, for complex cardio-metabolic disease.