心脏代谢疾病组织特异性转录组谱的阐明-猫眼课题宝

心脏代谢疾病组织特异性转录组谱的阐明

批准号：

10460231

负责人：

Mingyao Li

金额：

$70.59万

依托单位：

COLUMBIA UNIVERSITY HEALTH SCIENCES

依托单位国家：

美国

项目类别：

财政年份：

2012

资助国家：

美国

项目状态：

已结题

起止时间：

2012-04-16 至 2024-07-31

来源：

https://reporter.nih.gov/project-details/10460231

关键词：

Address Age Alleles Atherosclerosis Biological Models Biology CD14 gene Candidate Disease Gene Cardiometabolic Disease Cardiovascular Diseases Cells Cigarette Smoker Clinical Research Clinical Trials Complex Coupled Custom Data Data Set Disease Enrollment FCGR3B gene Flow Cytometry Functional disorder Gene Expression Genes Genetic Genetic Identity Genetic Transcription Genetic study Genomics Health Homeostasis Human Human Genetics Kinetics Knock-out Knowledge Maps Masks Membrane Proteins Messenger RNA Modeling Mus Myeloid Cells Noise Participant Patients Population Genetics Primates Procedures Property Proteins Race Recovery Regulation Regulator Genes Resolution Risk Factors Rodent Role Safety Signal Transduction Source Specificity Suggestion Surface Testing Tissues Type 2 diabetic Untranslated RNA Variant base cardiovascular disorder risk cardiovascular disorder therapy clinical translation differential expression functional genomics genetic association genetic signature genome-wide genomic signature human disease human model induced pluripotent stem cell innovation knock-down macrophage monocyte novel overexpression sex single-cell RNA sequencing targeted treatment tool transcriptome sequencing transcriptomics validation studies

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中文摘要

翻译

最近的临床试验和人类遗传学研究暗示单核细胞和巨噬细胞（M β）在动脉粥样硬化心血管疾病（CVD）。然而，对安全性和特异性的担忧限制了骨髓细胞定向移植的潜力。 CVD和复杂心脏代谢疾病（CMD）的治疗。这反映了身份方面的知识差距人类单核细胞亚群在体内平衡和疾病中的作用。在R 01-HL 113147的第一个周期中，MPI 致力于人类单核细胞-巨噬细胞的高产批量RNAseq和功能基因组学。然而，“数据批量RNAseq中的“平均”掩盖了表达变异并限制了鉴定单核细胞的能力亚群人单核细胞及其调节特征与小鼠不同。所以，尽管有一些研究啮齿动物单核细胞亚群及其“主调节”蛋白（MR）的遗传和功能，人单核细胞亚群的身份仍不清楚。基于表面标志物，人单核细胞可以分为三个子集，但新出现的数据暗示更大的多样性。我们初步的单细胞（sc）来自多个人的超过20，000个血液单核细胞中的RNAseq表明（i）至少六个单核细胞亚群和（ii）CVD危险因素状态下亚群分布的变异性。在这次更新中，我们提出对人类单核细胞进行尖端的scRNAseq分析，并结合创新的群体遗传学和功能基因组学来确定人单核细胞亚群在CMD中的作用。在目标1中，我们将招募 240名参与者（60名健康人，60名高胆固醇血症患者，60名2型糖尿病患者和60名吸烟者），在60名参与者中每个受试者约5000个单核细胞的3 '端scRNAseq，以鉴定单核细胞亚群，以及使用scRNAseq数据从所有240名受试者的批量RNAseq解卷积亚群分布。然后，我们将确定健康和心血管疾病中单核细胞亚群的差异表达基因和MR 风险国家。目的2将联合收割机scRNAseq和全基因组SNP数据结合起来，以鉴定单核细胞亚群- 特异性顺式eQTL变体和具有亚群特异性转录爆发的基因。这些基因工具将用于大型遗传数据集，以测试特定亚群与CMD的关联。在目标3中，专注于亚群特异性长基因间非编码RNA（lincRNA），我们的研究涉及的分子，组和其他在单核细胞-M β功能和人类疾病中的作用。初步数据显示，大多数lincRNA 仅在单核细胞亚群中表达。我们将使用插补程序恢复lincRNA 表达以鉴定单核细胞亚群特异性lincRNA及其顺式eQTL变体的关联以及CMD的突发特性。关键亚群mRNA和lincRNA的机制研究将使用人单核细胞源性单核细胞中的敲减和过表达以及人中用于敲除的基因编辑诱导性多能干细胞衍生的骨髓基质细胞。我们解决了人类单核细胞生物学的一个主要知识缺口，定义人类单核细胞亚群，询问它们与CMD的遗传关系，并探索亚群特异性mRNA和lincRNA在人骨髓瘤中的功能。

英文摘要

Recent clinical trials and human genetic studies implicate monocytes and macrophages (Mϕ) in atherosclerotic cardiovascular disease (CVD). Yet, concerns on safety and specificity limit the potential of myeloid cell directed therapies for CVD and complex cardiometabolic disorders (CMD). This reflects knowledge-gaps in the identity of human monocyte subpopulations in homeostasis and disease. In the first cycle of R01-HL113147, the MPIs pursued highly productive bulk RNAseq and functional genomics of human monocyte-Mϕ. However, “data averaging” in bulk RNAseq masks expression variation and limits capacity to identify monocyte subpopulations. Human monocytes and their regulatory features differ from mouse. So, despite elegant studies of rodent monocyte subpopulations and their “master regulator” proteins (MRs), the genetic and functional identities of human monocyte subpopulations remain unclear. Based on surface markers, human monocytes can be classified into three subsets, yet emerging data hint at greater diversity. Our preliminary single cell (sc)RNAseq in over 20,000 blood monocytes from multiple humans suggest (i) at least six monocyte subpopulations and (ii) variability in subpopulations distributions across CVD risk factor states. In this renewal, we propose cutting-edge scRNAseq profiling of human monocytes coupled to innovative population genetics and functional genomics to define the role of human monocyte subpopulations in CMD. In Aim 1, we will enroll 240 participants (60 healthy, 60 hypercholesterolemic, 60 type 2 diabetic and 60 cigarette smokers), perform 3’-end scRNAseq of ~5000 monocytes per subject in 60 participants to identify monocyte subpopulations, and use scRNAseq data to deconvolute subpopulation distributions from bulk RNAseq for all 240 subjects. Then, we will determine differentially expressed genes and MRs of monocyte subpopulations in health and in CVD risk states. Aim 2 will combine scRNAseq and genome wide SNP data to identify monocyte subpopulation- specific cis-eQTL variants and genes with subpopulation-specific transcriptional bursting. These genetic tools will be used in large genetic datasets to test the association of specific subpopulations with CMD. In Aim 3 we focus on subpopulation-specific long intergenic non-coding RNAs (lincRNAs), molecules implicated by our group and others in monocyte-Mϕ functions and human diseases. Preliminary data reveals that most lincRNAs are expressed only in subsets of monocytes. We will use an imputation procedure for recovery of lincRNA expression to identify monocyte subpopulation-specific lincRNAs and association of their cis-eQTL variants and bursting features with CMD. Mechanistic studies of key subpopulation mRNAs and lincRNAs will use knockdown and overexpression in human monocyte-derived Mϕ and gene-editing, for knockout, in human induced pluripotent stem cell-derived Mϕ. We address a major knowledge gap in human monocyte biology by defining human monocyte subpopulations, interrogating their genetic relationship with CMD, and exploring functions of subpopulation-specific mRNAs and lincRNAs in human Mϕs.

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