Defining the Role of FHL5, a Novel Hypertension Associated Gene in Smooth Muscle Cells

定义平滑肌细胞中新型高血压相关基因 FHL5 的作用

基本信息

批准号：
10490245
负责人：
Doris Wong
金额：
$ 2.36万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-01 至 2022-10-04
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10490245
关键词：
Actins Address Alleles Animal Model Antihypertensive Agents Arteries Binding Binding Sites Biological Blood Pressure Blood Vessels CREB1 gene CRISPR/Cas technology Calcium Candidate Disease Gene Cardiovascular Diseases Cell physiology Cellular biology Cervical Chromatin Chronic Clinic Clustered Regularly Interspaced Short Palindromic Repeats Complex Coronary Arteriosclerosis Coronary artery Cytoskeletal Modeling Data Development Disease Dissection Down-Regulation Endothelin Enhancers Epigenetic Process FHL1 gene FHL2 gene Family Family member Functional disorder Future Gene Cluster Gene Expression Genes Genetic Genetic Transcription Genetic Variation Heritability Human Hyperplasia Hypertension In Vitro Individual Intervention Studies LIM Domain Link Linkage Disequilibrium Maps Methods Migraine Molecular Muscle Muscle Contraction Myocardial Infarction Myocardium Nucleic Acid Regulatory Sequences Participant Pathogenicity Pathway interactions Pericytes Phenotype Process Proteins Public Health Quantitative Trait Loci Regulatory Element Relaxation Reporting Resolution Role Serum Response Factor Signal Transduction Single Nucleotide Polymorphism Skeletal Muscle Smooth Muscle Myocytes Specificity Stress Stress Fibers Surveys Testing Therapeutic Intervention Tibial Arteries Tissue Sample Tissues Translating Untranslated RNA Variant Vascular Diseases Vasoconstrictor Agents Veins blood pressure elevation blood pressure regulation cardiovascular risk factor causal variant cell type cofactor drug candidate familial hypertension genetic association genetic risk factor genome wide association study genome-wide in silico insight lifetime risk member novel novel therapeutics nuclease overexpression response risk variant targeted treatment trait transcription factor transcriptomics

项目摘要

ABSTRACT Hypertension, a condition defined by a chronic elevation in blood pressure is one of the most significant heritable risk factors for cardiovascular disease. Genome wide association studies (GWAS) in hypertension have provided an unbiased survey of loci relevant to blood pressure regulation, many of which harbor genes that cluster into functional pathways that regulate smooth muscle cell (SMC) phenotypes. As the most abundant cell type in the healthy vessel wall, SMCs regulate vascular tone through coordinated contraction and relaxation. In response to environmental stress, SMCs dedifferentiate and contribute to the pathogenic remodeling of the vessel that increases vascular tone. Functional characterization of these loci, may reveal new insights into SMC dysregulation and inform novel drug candidates. One such hypertension locus with a predicted role in the vessel wall, UFL1-FHL5 is also associated with multiple vascular disorders, including coronary artery disease, myocardial infarction, and migraines. Statistical fine-mapping approaches implicate FHL5 as the top candidate gene at the UFL1-FHL5 locus. FHL5 expression is enriched in artery tissues, specifically in contractile SMC and pericytes in the vessel wall. FHL5 is a member of Four and Half LIM domain family, which include proteins FHL1, FHL2, and FHL3 that function as transcriptional regulators for the transcription factors, SRF and CREB1 in skeletal and cardiac muscle tissue. FHL5, the most understudied member of this family, was implicated in vein intimal hyperplasia by activating CREB1 target genes. Thus, I hypothesize that vascular disease associated genetic variation reduces FHL5 expression and that FHL5 functions as a critical cofactor to activate contractile SMC pathways in the vessel wall. Studies in aim 1 will provide insights into the upstream regulatory mechanisms of FHL5 by identifying critical FHL5 regulatory elements. I will epigenetically activate putative enhancers harboring top candidate hypertension variants in primary SMCs and assess changes in FHL5 gene expression, SMC contractility, actin cytoskeletal organization and intracellular calcium levels. In aim 2, I will elucidate the downstream functional role of FHL5 protein by mapping its binding sites in human coronary artery tissues using the high resolution, low input Cleavage Under Target and Release Upon Nuclease (CUT&RUN) method. I will intersect FHL5 binding sites with SRF and CREB1 binding sites in matched tissue samples to determine the pathways regulated by different FHL5 transcriptional complexes. I will also determine the impact of deleting top FHL5 regulatory elements residing in hypertension loci on SMC functions. Overall, in this study I will characterize a novel hypertension associated locus and define the role of this cofactor in SMC biology. These studies may also inform the development of anti-hypertensive therapies targeting vessel wall related pathways to circumvent multiple vascular diseases and address significant gaps in the clinic.

抽象的高血压，血压慢性升高所定义的疾病是最重要的心血管疾病的遗传风险因素。高血压的基因组广泛关联研究（GWAS）已经提供了与血压调节相关的基因座的无偏调查，其中许多含有基因该聚集成调节平滑肌细胞（SMC）表型的功能途径。最多 SMC在健康容器壁中的丰富细胞类型通过协调收缩调节血管张力和放松。为了应对环境压力，SMCS去分化并有助于致病性增加血管张力的血管的重塑。这些基因座的功能表征可能会揭示对SMC失调的新见解并为新颖的候选药物提供信息。一个这样的高血压基因座预测在容器壁中的作用，UFL1-FHL5也与多种血管疾病有关，包括冠状动脉疾病，心肌梗塞和偏头痛。统计精细映射方法暗示 FHL5是UFL1-FHL5基因座的顶级候选基因。 FHL5表达富含动脉组织，特别是在容器壁上的收缩SMC和周细胞中。 FHL5是四个半域的成员包括蛋白质FHL1，FHL2和FHL3的家族作为转录调节剂的作用转录因子，骨骼肌和心脏肌肉组织中的SRF和CREB1。 FHL5，最不受限制的该家族的成员通过激活CREB1靶基因与静脉内膜增生有关。因此，我假设血管疾病相关的遗传变异降低了FHL5的表达，而FHL5 充当临界辅助因子，以激活血管壁中的收缩SMC途径。 AIM 1的研究将通过识别关键的FHL5调节性，提供有关FHL5上游调节机制的见解元素。我将表观遗传激活推定的增强剂，该增强剂具有顶级候选高血压变体初级SMC并评估FHL5基因表达，SMC收缩力，肌动蛋白细胞骨架组织的变化和细胞内钙水平。在AIM 2中，我将通过使用高分辨率，低输入裂解在人类冠状动脉组织中绘制其结合位点目标并在核酸酶（切割和运行）方法上释放。我将与SRF相交FHL5绑定位点，然后匹配的组织样品中的CREB1结合位点，以确定由不同FHL5调节的途径转录复合物。我还将确定删除位于 SMC功能上的高血压基因座。总体而言，在这项研究中，我将表征与新型高血压相关的基因座并定义了该辅因子在SMC生物学中的作用。这些研究也可能告知针对围绕血管壁相关的抗高血压疗法，以规避多种血管疾病并解决诊所中的显着差距。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Multi-ancestry genome-wide study identifies effector genes and druggable pathways for coronary artery calcification.

DOI：
10.1038/s41588-023-01518-4
发表时间：
2023-10
期刊：
NATURE GENETICS
影响因子：
30.8
作者：
Kavousi, Maryam;Bos, Maxime M.;Barnes, Hanna J.;Cardenas, Christian L. Lino;Wong, Doris;Lu, Haojie;Hodonsky, Chani J.;Landsmeer, Lennart P. L.;Turner, Adam W.;Kho, Minjung;Hasbani, Natalie R.;de Vries, Paul S.;Bowden, Donald W.;Chopade, Sandesh;Deelen, Joris;Benavente, Ernest Diez;Guo, Xiuqing;Hofer, Edith;Hwang, Shih-Jen;Lutz, Sharon M.;Lyytikainen, Leo-Pekka;Slenders, Lotte;Smith, Albert V.;Stanislawski, Maggie A.;van Setten, Jessica;Wong, Quenna;Yanek, Lisa R.;Becker, Diane M.;Beekman, Marian;Budoff, Matthew J.;Feitosa, Mary F.;Finan, Chris;Hilliard, Austin T.;Kardia, Sharon L. R.;Kovacic, Jason C.;Kral, Brian G.;Langefeld, Carl D.;Launer, Lenore J.;Malik, Shaista;Hoesein, Firdaus A. A. Mohamed;Mokry, Michal;Schmidt, Reinhold;Smith, Jennifer A.;Taylor, Kent D.;Terry, James G.;van der Grond, Jeroen;van Meurs, Joyce;Vliegenthart, Rozemarijn;Xu, Jianzhao;Young, Kendra A.;Zilhao, Nuno R.;Zweiker, Robert;Assimes, Themistocles L.;Becker, Lewis C.;Bos, Daniel;Carr, J. Jeffrey;Cupples, L. Adrienne;de Kleijn, Dominique P. V.;de Winther, Menno;den Ruijter, Hester M.;Fornage, Myriam;Freedman, Barry I.;Gudnason, Vilmundur;Hingorani, Aroon D.;Hokanson, John E.;Ikram, M. Arfan;Isgum, Ivana;Jacobs, David R., Jr.;Kahonen, Mika;Lange, Leslie A.;Lehtimaki, Terho;Pasterkamp, Gerard;Raitakari, Olli T.;Schmidt, Helena;Slagboom, P. Eline;Uitterlinden, Andre G.;Vernooij, Meike W.;Bis, Joshua C.;Franceschini, Nora;Psaty, Bruce M.;Post, Wendy S.;Rotter, Jerome I.;Bjorkegren, Johan L. M.;O'Donnell, Christopher J.;Bielak, Lawrence F.;Peyser, Patricia A.;Malhotra, Rajeev;van der Laan, Sander W.;Miller, Clint L.
通讯作者：
Miller, Clint L.

Genetic Regulation of SMC Gene Expression and Splicing Predict Causal CAD Genes