内皮染色质重塑酶调控小鼠肺发育过程中关键 ECM 成分的产生

IF 7.4 1区 医学 Q1 HEMATOLOGY Arteriosclerosis, Thrombosis, and Vascular Biology Pub Date : 2024-08-01 Epub Date: 2024-06-13 DOI:10.1161/ATVBAHA.124.320881
Meng-Ling Wu, Kate Wheeler, Robert Silasi, Florea Lupu, Courtney T Griffin
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引用次数: 0

摘要

背景:染色质重塑酶BRG1(梵天相关基因1)和CHD4(染色质链螺旋酶DNA结合蛋白4)独立调控对血管发育至关重要的基因转录,但它们在胚胎晚期对血管的协调影响尚未探究:在这项研究中,我们在混合背景小鼠(Brg1fl/fl;Chd4fl/fl;VE-Cadherin-Cre+)中遗传性地删除了内皮 Brg1 和 Chd4,并以 Cre 重组酶阴性的同窝小鼠作为对照。组织通过免疫染色、免疫印迹和流式细胞术进行分析。定量反转录聚合酶链反应用于确定基因表达,染色质免疫沉淀揭示了培养的内皮细胞中 BRG1 和 CHD4 的基因靶标:结果:我们发现Brg1/Chd4双突变体生长正常,但出生后不久即死亡,肺小而紧凑。尽管突变体肺的细胞组成正常,但其远端气囊的 ECM(细胞外基质)成分和 TGFβ(转化生长因子-β)信号转导却减少了,而 TGFβ 通常会促进 ECM 的合成。在突变的肺内皮细胞中,胶原蛋白和弹性蛋白相关基因以及 TGFβ 配体 Tgfb1 的转录物减少,但遗传性删除内皮 Tgfb1 无法再现 Brg1/Chd4 突变体中出现的小肺和 ECM 缺陷。相反,我们发现在培养的内皮细胞中,多个 ECM 基因是 BRG1 和 CHD4 的直接靶标:总之,我们的数据强调了内皮染色质重塑酶在胚胎肺发育的囊状阶段促进远端肺组织中 ECM 沉积的重要作用。
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Endothelial Chromatin-Remodeling Enzymes Regulate the Production of Critical ECM Components During Murine Lung Development.

Background: The chromatin-remodeling enzymes BRG1 (brahma-related gene 1) and CHD4 (chromodomain helicase DNA-binding protein 4) independently regulate the transcription of genes critical for vascular development, but their coordinated impact on vessels in late-stage embryos has not been explored.

Methods: In this study, we genetically deleted endothelial Brg1 and Chd4 in mixed background mice (Brg1fl/fl;Chd4fl/fl;VE-Cadherin-Cre), and littermates that were negative for Cre recombinase were used as controls. Tissues were analyzed by immunostaining, immunoblot, and flow cytometry. Quantitative reverse transcription polymerase chain reaction was used to determine gene expression, and chromatin immunoprecipitation revealed gene targets of BRG1 and CHD4 in cultured endothelial cells.

Results: We found Brg1/Chd4 double mutants grew normally but died soon after birth with small and compact lungs. Despite having normal cellular composition, distal air sacs of the mutant lungs displayed diminished ECM (extracellular matrix) components and TGFβ (transforming growth factor-β) signaling, which typically promotes ECM synthesis. Transcripts for collagen- and elastin-related genes and the TGFβ ligand Tgfb1 were decreased in mutant lung endothelial cells, but genetic deletion of endothelial Tgfb1 failed to recapitulate the small lungs and ECM defects seen in Brg1/Chd4 mutants. We instead found several ECM genes to be direct targets of BRG1 and CHD4 in cultured endothelial cells.

Conclusions: Collectively, our data highlight essential roles for endothelial chromatin-remodeling enzymes in promoting ECM deposition in the distal lung tissue during the saccular stage of embryonic lung development.

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来源期刊
CiteScore
15.60
自引率
2.30%
发文量
337
审稿时长
2-4 weeks
期刊介绍: The journal "Arteriosclerosis, Thrombosis, and Vascular Biology" (ATVB) is a scientific publication that focuses on the fields of vascular biology, atherosclerosis, and thrombosis. It is a peer-reviewed journal that publishes original research articles, reviews, and other scholarly content related to these areas. The journal is published by the American Heart Association (AHA) and the American Stroke Association (ASA). The journal was published bi-monthly until January 1992, after which it transitioned to a monthly publication schedule. The journal is aimed at a professional audience, including academic cardiologists, vascular biologists, physiologists, pharmacologists and hematologists.
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