LRP1 Repression by SNAIL Results in ECM Remodeling in Genetic Risk for Vascular Diseases.

IF 16.5 1区 医学 Q1 CARDIAC & CARDIOVASCULAR SYSTEMS
Circulation research Pub Date : 2024-11-08 Epub Date: 2024-10-02 DOI:10.1161/CIRCRESAHA.124.325269
Lu Liu, Joséphine Henry, Yingwei Liu, Charlène Jouve, Jean-Sébastien Hulot, Adrien Georges, Nabila Bouatia-Naji
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引用次数: 0

Abstract

Background: Genome-wide association studies implicate common genetic variations in the LRP1 (low-density lipoprotein receptor-related protein 1 gene) locus at risk for multiple vascular diseases and traits. However, the underlying biological mechanisms are unknown.

Methods: Fine mapping analyses included Bayesian colocalization to identify the most likely causal variant. Human induced pluripotent stem cells were genome-edited using CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR associated protein 9) to delete or modify candidate enhancer regions and generate LRP1 knockout cell lines. Cells were differentiated into smooth muscle cells through a mesodermal lineage. Transcription regulation was assessed using luciferase reporter assay, transcription factor knockdown, and chromatin immunoprecipitation. Phenotype changes in cells were conducted using cellular assays, bulk RNA sequencing, and mass spectrometry.

Results: Multitrait colocalization analyses pointed at rs11172113 as the most likely causal variant in LRP1 for fibromuscular dysplasia, migraine, pulse pressure, and spontaneous coronary artery dissection. We found the rs11172113-T allele to associate with higher LRP1 expression. Genomic deletion in induced pluripotent stem cell-derived smooth muscle cells supported rs11172113 to locate in an enhancer region regulating LRP1 expression. We found transcription factors MECP2 (methyl CpG binding protein 2) and SNAIL (Zinc Finger Protein SNAI1) to repress LRP1 expression through an allele-specific mechanism, involving SNAIL interaction with disease risk allele. LRP1 knockout decreased induced pluripotent stem cell-derived smooth muscle cell proliferation and migration. Differentially expressed genes were enriched for collagen-containing extracellular matrix and connective tissue development. LRP1 knockout and deletion of rs11172113 enhancer showed potentiated canonical TGF-β (transforming growth factor beta) signaling through enhanced phosphorylation of SMAD2/3 (Mothers against decapentaplegic homolog 2/3). Analyses of the protein content of decellularized extracts indicated partial extracellular matrix remodeling involving enhanced secretion of CYR61 (cystein rich angiogenic protein 61), a known LRP1 ligand involved in vascular integrity and TIMP3 (Metalloproteinase inhibitor 3), implicated in extracellular matrix maintenance and also known to interact with LRP1.

Conclusions: Our findings support allele-specific LRP1 expression repression by the endothelial-to-mesenchymal transition regulator SNAIL. We propose decreased LRP1 expression in smooth muscle cells to remodel the extracellular matrix enhanced by TGF-β as a potential mechanism of this pleiotropic locus for vascular diseases.

SNAIL 对 LRP1 的抑制导致血管疾病遗传风险中的 ECM 重塑。
背景:全基因组关联研究表明,LRP1(低密度脂蛋白受体相关蛋白 1)位点的常见遗传变异与多种血管疾病和性状的风险有关。然而,其潜在的生物学机制尚不清楚:方法:精细图谱分析包括贝叶斯共定位,以确定最有可能的致病变体。利用CRISPR-Cas9对人类诱导多能干细胞进行基因组编辑,删除或修改候选增强子区域,生成LRP1基因敲除细胞系。细胞通过中胚层系分化成平滑肌细胞。利用荧光素酶报告分析、转录因子敲除和染色质免疫沉淀评估转录调控。细胞表型的变化是通过细胞测定、大量 RNA 测序和质谱分析进行的:结果:多性状共定位分析表明,rs11172113是LRP1中最有可能导致纤维肌发育不良、偏头痛、脉压和肺功能性状的因果变异。我们发现 rs11172113-T 等位基因与较高的 LRP1 表达有关。诱导多能干细胞衍生的平滑肌细胞基因组缺失支持 rs11172113 位于调节 LRP1 表达的增强子区域。我们发现转录因子MECP2(甲基CpG结合蛋白2)和SNAIL通过等位基因特异性机制抑制LRP1的表达,其中SNAIL与疾病风险等位基因相互作用。LRP1基因敲除会减少诱导多能干细胞衍生的平滑肌细胞的增殖和迁移。差异表达的基因主要集中在含胶原的细胞外基质、结缔组织发育和肺发育方面。LRP1基因敲除和rs11172113增强子的缺失显示,通过增强SMAD2/3的磷酸化,TGF-β(转化生长因子β)信号传导得到加强。对脱细胞提取物蛋白质含量的分析表明,部分细胞外基质重塑涉及 CYR61 和 TIMP3 的分泌增强,CYR61 是已知的 LRP1 配体,与血管完整性有关,而 TIMP3 与细胞外基质的维持有关,也已知与 LRP1 有相互作用:我们的研究结果支持等位基因特异性 LRP1 基因受内皮到间质转化调节因子 SNAIL 的抑制。我们认为,LRP1 在平滑肌细胞中的表达减少可重塑由 TGF-β 增强的细胞外基质,这是该多效应位点导致血管疾病的潜在机制。
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来源期刊
Circulation research
Circulation research 医学-外周血管病
CiteScore
29.60
自引率
2.00%
发文量
535
审稿时长
3-6 weeks
期刊介绍: Circulation Research is a peer-reviewed journal that serves as a forum for the highest quality research in basic cardiovascular biology. The journal publishes studies that utilize state-of-the-art approaches to investigate mechanisms of human disease, as well as translational and clinical research that provide fundamental insights into the basis of disease and the mechanism of therapies. Circulation Research has a broad audience that includes clinical and academic cardiologists, basic cardiovascular scientists, physiologists, cellular and molecular biologists, and cardiovascular pharmacologists. The journal aims to advance the understanding of cardiovascular biology and disease by disseminating cutting-edge research to these diverse communities. In terms of indexing, Circulation Research is included in several prominent scientific databases, including BIOSIS, CAB Abstracts, Chemical Abstracts, Current Contents, EMBASE, and MEDLINE. This ensures that the journal's articles are easily discoverable and accessible to researchers in the field. Overall, Circulation Research is a reputable publication that attracts high-quality research and provides a platform for the dissemination of important findings in basic cardiovascular biology and its translational and clinical applications.
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