Nuclear FAK-mediated VSMC differentiation via epigenetic reprograming invascular diseases

通过表观遗传重编程血管疾病中核 FAK 介导的 VSMC 分化

基本信息

批准号：
10584581
负责人：
Steve Lim
金额：
$ 47.54万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-16 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10584581
关键词：
Apolipoprotein E Arterial Fatty Streak Arteries Atherosclerosis Biochemical CD47 gene Cardiovascular system Cell Cycle Cell Differentiation process Cell Nucleus Cell Proliferation Cells Complex Cyclin D1 Cytoplasm DNA Methylation DNA Modification Methylases Data Deacetylase Disease Epigenetic Process Failure Focal Adhesion Kinase 1 Gene Expression Genes Genetic Genetic Models Genetic Transcription Growth Factor Histones Human Hyperlipidemia Inflammatory Injury Integrins Intervention Intervention Studies Investments Macrophage Mediating Modeling Molecular Molecular Biology Mus Nuclear Nucleosomes Phenotype Play Process Production Proliferating Protein Tyrosine Kinase Repression Role Sampling Signal Transduction Smooth Muscle Myocytes Specimen Stimulus Testing Therapeutic Ubiquitination Vascular Smooth Muscle cell dedifferentiation cell motility chromatin remodeling epigenetic regulation histone modification insight kinase inhibitor mouse model novel oxidized lipid pharmacologic prevent promoter response tool transdifferentiation vascular injury

项目摘要

VSMCs dedifferentiate into a proliferative state upon vessel injury or transdifferentiate into macrophage-like cells (MLCs) during atherosclerosis progression. VSMC phenotypic switching are driven by multiple transcriptional and epigenetic changes that lead to increased proliferation with reduced contractile gene expression and increased matrix production, which is detrimental to atherosclerotic lesions, direct interventional studies that target this process have been lacking. Increased matrix and growth factors alter integrin signaling and leads to aberrant focal adhesion kinase (FAK) activation, which may promote VSMC phenotypic switching. We demonstrated that FAK is inactive and primarily localized in the nuclei of VSMCs of healthy arteries. However, vessel injury promoted FAK activation and cytoplasmic relocalization, which increased cyclin D1 transcription and cell cycling. While we found that inhibition of FAK activity in VSMCs induced nuclear localization of FAK and increased contractile gene transcription, the underlying mechanism by which FAK regulates the contractile genes is not known. We have identified DNA methyltransferase 3A (DNMT3A) and the nucleosome remodeling and deacetylase (NuRD) complex, two key epigenetic repression machineries, as nuclear FAK-interacting partners in VSMCs. FAK inhibition decreased DNMT3A and NuRD component expression, which was associated with decreased DNA methylation and increased active histone marks within contractile gene promoters. Using genetic FAK cytoplasmic (Cyto) restricted VSMCs, we found that nuclear FAK is required for reducing DNMT3A/NuRD and for increasing contractile gene expression. Additionally, ApoE-/-;FAK-Cyto mice showed increased atherosclerosis compared to WT mice, suggesting that active cytoplasmic FAK exacerbates atherosclerosis. Further, FAK showed increased cytoplasmic localization and activity within human atherosclerotic lesions compared to healthy specimens. Importantly, FAK inhibitor reduced advanced atherosclerotic lesions in ApoE-/- mice, which was associated with reduced DNMT3A and NuRD component expression with increased ACTA2+ cells in the fibrous cap. Our hypothesis is that FAK catalytic inhibition forces FAK nuclear localization and promotes VSMC differentiation via reduced expression of epigenetic regulators DNMT3A and NuRD complex. Aim 1 will elucidate the molecular mechanism of nuclear FAK-mediated VSMC phenotypic switching via epigenetic modulation of DNA methylation, chromatin remodeling, and histone modification. Aim 2 will investigate the role of DNMT3A and the NuRD complex in VSMC dedifferentiation upon vascular injury using both DNMT3A and NuRD genetic models. Aim 3 will evaluate the effect of FAK inhibition on blocking VSMC transdifferentiation and promoting plaque stability in early and advanced atherosclerosis. This study will provide new insights into VSMC phenotype switching via FAK-mediated epigenetic control through DNMT3A and NuRD complex stability. The therapeutic potential of FAK inhibitors in alleviating intimal thickening in vascular injury and atherosclerosis will also be assessed.

VSMC在血管损伤或转变为巨噬细胞状后将其推导成增殖状态动脉粥样硬化进展过程中的细胞（MLC）。 VSMC表型切换由多个驱动转录和表观遗传变化导致收缩基因减少的增殖增加表达并增加基质产生，这对动脉粥样硬化病变有害针对此过程的介入研究缺乏。增加基质和生长因子改变了整联蛋白信号传导并导致异常局灶性粘附激酶（FAK）激活，这可能促进VSMC 表型切换。我们证明了FAK是不活跃的，主要定位于VSMC的核健康的动脉。然而，血管损伤促进了FAK激活和细胞质重新定位，这 Cyclin D1转录和细胞循环增加。虽然我们发现VSMC中FAK活性的抑制作用诱导FAK的核定位和增加的收缩基因转录，这是基本机制 FAK调节收缩基因尚不清楚。我们已经确定了DNA甲基转移酶3A （DNMT3A）和核小体重塑和脱乙酰基酶（NURD）复合物，两个关键的表观抑制作用机器，作为VSMC中的核FAK相互作用伙伴。 FAK抑制减少了DNMT3A和NURD 成分表达，与DNA甲基化降低和活性组蛋白增加有关收缩基因启动子中的标记。使用遗传FAK细胞质（CYTO）受限的VSMC，我们发现减少DNMT3A/NURD并增加收缩基因表达所必需的核FAK。另外，与WT小鼠相比，ApoE - / - ; Fak-Cyto小鼠显示动脉粥样硬化增加，这表明主动细胞质FAK加剧了动脉粥样硬化。此外，FAK显示细胞质定位增加与健康标本相比，人动脉粥样硬化病变中的活性。重要的是，FAK抑制剂 apoE - / - 小鼠中的晚期动脉粥样硬化病变减少，与DNMT3A降低有关 NURD成分表达在纤维帽中具有增加的ACTA2+细胞。我们的假设是Fak 催化抑制力FAK核定位并通过降低表达促进VSMC分化表观遗传调节剂DNMT3A和Nurd Complex。 AIM 1将阐明核FAK介导的VSMC表型通过DNA甲基化的表观遗传调节，染色质调节重塑和组蛋白修饰。 AIM 2将调查DNMT3A和NURD综合体在使用DNMT3A和NURD遗传模型对血管损伤进行VSMC去分化。目标3意志评估FAK抑制对阻断VSMC跨分化和促进斑块稳定性的影响早期和晚期动脉粥样硬化。这项研究将为通过VSMC表型切换提供新的见解 FAK介导的表观遗传学通过DNMT3A和NURD复合稳定性。治疗潜力还将评估减轻血管损伤内膜增厚和动脉粥样硬化的FAK抑制剂。