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）为复杂心脏代谢疾病（CMD）的遗传基础提供了更多的见解，包括2型糖尿病和动脉粥样硬化性心血管疾病。然而，这些发现只解释了一小部分的遗传性，这表明除了常见的DNA变异之外，还需要考虑遗传影响。转录组的知识对于更全面地了解基因组的遗传功能元素是必不可少的。高通量RNA测序（RNA-seq）的出现，包括我们的工作，证明了比以前编目的转录组复杂性和多样性大得多的存在。宿主对内毒素（LPS）的反应为先天免疫的转录组调控提供了基本的见解，同时也作为炎症性CMD的模型。在烟酸和内毒素血症诱发反应遗传学（GENE）研究中，我们招募健康个体（N=286）进行住院治疗，并在静脉注射内毒素（1ng/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组学和表观基因组学分析，以发现人类疾病的组织特异性机制。在Aim 1中，我们将利用GENE参与者的临床炎症反应来识别极端“低”（~<5%PC）和“高”（~>95%PC）应答者，以检测具有最大临床相关性的内毒素诱导的转录组反应。在目标2中，我们将复制LPS诱导的转录组变化，定义表观遗传调控，并追求复制结果的临床翻译。在Aim 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）的基础和内毒素诱发的表观基因组/转录组反应与原代细胞和组织中的相似。本提案结合独特的临床资源、组织特异性转录组学和hiPSC的离体分析，以确定CMD的遗传贡献，发现新的疾病机制和治疗机会，并推进人类疾病研究的创新。

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.