Regulation and Function of SRF in Vascular Pathiobiology

SRF 在血管病理生物学中的调节和功能

• 批准号: 10337251
• 负责人: Joseph M Miano
• 金额: $ 52.84万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-11-22 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10337251
• 关键词: Acute; Aneurysm; Apoptosis; Arterial Injury; Atherosclerosis; Automobile Driving; Base Pairing; Binding; Binding Sites; Bioinformatics; Biology; Blood Vessels; CRISPR/Cas technology; Cells; ChIP-seq; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; Color; Complement; Cre driver; DNA Binding; Data; Development; Differentiated Gene; Differentiation and Growth; Disease; Disease model; Elements; Excision; Future; Gastrointestinal tract structure; Gender; Gene Abnormality; Gene Expression; Genes; Genetic; Genetic Transcription; Genomics; Growth; Homeostasis; Human; Hyperplasia; IL6 gene; In Vitro; Inflammation; Inflammatory; Injury; Knowledge; Lesion; Lipids; LoxP-flanked allele; Luciferases; Mediating; Mediator of activation protein; Methods; Modeling; Mus; Pathogenicity; Pathologic; Phenotype; Regulation; Regulatory Element; Research; Role; Serum Response Factor; Single base substitution; Smooth Muscle Myocytes; Specificity; Tamoxifen; Testing; Text; Therapeutic; Time; Transcriptional Regulation; Untranslated RNA; Variant; Vascular Diseases; Vascular Smooth Muscle; Visceral; base; cell growth; cell type; cofactor; combat; experimental study; gain of function; gene function; genome editing; genomic data; in vivo; injured; insight; loss of function; macrophage; mouse model; novel; novel therapeutic intervention; overexpression; programs; promoter; response; tool; transcription factor; transcriptome sequencing; transdifferentiation; uptake

There is now incontrovertible evidence that vascular smooth muscle cells (VSMCs) contribute substantively to vascular diseases. The function or dysfunction of VSMCs is driven, in part, by the activity of key transcription factors (TF). Serum response factor (SRF), an abundantly expressed TF in VSMCs that binds a large cadre of CArG boxes colloquially known as the CArGome, orchestrates a number of disparate gene programs. Surprisingly, almost nothing is known about the in vivo regulation of Srf transcription and, while the function of SRF in vascular development is well understood, there is no information about its direct role in vascular diseases; and gender-based studies are not possible given the limitation of the most popular SMC Cre driver (Myh11). Moreover, the full complement of SRF target genes (notably long noncoding RNAs, lncRNAs) is not known. We have been a leading lab in SRF research and now offer fresh insights into these major scientific gaps that, collectively, form the basis of this application. First, we provide CRISPR-Cas9 genome editing results, bioinformatic predictions, ChIP-seq, and luciferase data supporting functional transcription factor bind- ing sites (TFBS) controlling Srf transcription in vivo. Second, gene expression and Srf loss-of-function (LOF) studies support SRF as an early mediator of VSMC growth, inflammation, and neointimal formation following acute vascular insult. Importantly, existing Cre driver mice limit analysis of Srf LOF to a narrow time window of only 10-14 days post-tamoxifen due to a competing, lethal phenotype of the gastrointestinal (GI) tract. How- ever, we have recently generated and validated a new Cre driver mouse with restricted Cre-mediated excision to VSMCs; little activity is observed in visceral SMCs of the GI tract, and a cross with floxed Srf mice shows extended survival providing the first ever opportunity to interrogate the function of SRF in both acute and chronic models of vascular disease without confounding phenotypes. Finally, genomic studies implicate a new SRF-dependent VSMC inflammatory gene program and CRISPR studies show a specific base substitution within the CArG box nullifies SRF-dependent gene expression in vivo. Three integrated aims will rigorously test the hypothesis that multiple TFBS control Srf expression to direct CArG-dependent homeostatic or pathogenic gene programs in the vessel wall. Aim 1 will evaluate the function of new TFBS governing Srf transcription using CRISPR editing in the mouse. Aim 2 will elucidate phenotypes associated with Srf LOF and gain-of-function in acute and chronic models of vascular disease using a novel Cre driver mouse for unparalleled VSMC specificity. Aim 3 will further utilize mice in Aim 2 for integrative VSMC ChIP-seq and RNA- seq studies to elucidate novel SRF target genes, particularly the class of lncRNAs, in the control of VSMC phenotypes; CRISPR editing of key CArG boxes are planned as a new paradigm to study gene function. These studies will vertically advance our knowledge of SRF regulation and function, paving the way towards new therapeutic approaches to combat vascular diseases while advancing new directives for further research.

现在有无可争议的证据表明血管平滑肌细胞（VSMCs）对

项目成果

CRISPR-Mediated Single Nucleotide Polymorphism Modeling in Rats Reveals Insight Into Reduced Cardiovascular Risk Associated With Mediterranean G6PD Variant.

• DOI: 10.1161/hypertensionaha.120.14772
• 发表时间: 2020-08
• 期刊: Hypertension (Dallas, Tex. : 1979)
• 作者: Kitagawa A;Kizub I;Jacob C;Michael K;D'Alessandro A;Reisz JA;Grzybowski M;Geurts AM;Rocic P;Gupte R;Miano JM;Gupte SA
• 通讯作者: Gupte SA

Fate and State of Vascular Smooth Muscle Cells in Atherosclerosis.

• DOI: 10.1161/circulationaha.120.049922
• 发表时间: 2021-05-25
• 期刊: Circulation
• 影响因子: 37.8
• 作者: Miano JM;Fisher EA;Majesky MW
• 通讯作者: Majesky MW

HDL regulates TGFß-receptor lipid raft partitioning, restoring contractile features of cholesterol-loaded vascular smooth muscle cells.

HDL 调节 TGFα 受体脂筏分配，恢复负载胆固醇的血管平滑肌细胞的收缩特征。

• DOI: 10.1101/2023.10.19.562786
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Nagesh,PrashanthThevkar;Nishi,Hitoo;Rawal,Shruti;Zahr,Tarik;Miano,JosephM;Sorci-Thomas,Mary;Xu,Hao;Akbar,Naveed;Choudhury,RobinP;Misra,Ashish;Fisher,EdwardA
• 通讯作者: Fisher,EdwardA