长链非编码 RNA GAS5 通过募集 KDM5B 并促进 PDGFRα/β 启动子的 H3K4me2/3 去甲基化来抑制 TGF-β1 诱导的肺周细胞向肌成纤维细胞的转化。
LncRNA GAS5 suppresses TGF-β1-induced transformation of pulmonary pericytes into myofibroblasts by recruiting KDM5B and promoting H3K4me2/3 demethylation of the PDGFRα/β promoter.
机构信息
Department of Critical Care Medicine, Key Laboratory for Major Obstetric Diseases of Guangdong Province, The Third Affiliated Hospital of Guangzhou Medical University, No. 63 Duobao Road, Liwan District, Guangzhou, 510150, Guangdong Province, People's Republic of China.
CAS Key Laboratory of Regenerative Biology and Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510150, Guangdong Province, People's Republic of China.
出版信息
Mol Med. 2023 Mar 14;29(1):32. doi: 10.1186/s10020-023-00620-x.
BACKGROUND
Idiopathic pulmonary fibrosis (IPF) is a condition that may cause persistent pulmonary damage. The transformation of pericytes into myofibroblasts has been recognized as a key player during IPF progression. This study aimed to investigate the functions of lncRNA growth arrest-specific transcript 5 (GAS5) in myofibroblast transformation during IPF progression.
METHODS
We created a mouse model of pulmonary fibrosis (PF) via intratracheal administration of bleomycin. Pericytes were challenged with exogenous transforming growth factor-β1 (TGF-β1). To determine the expression of target molecules, we employed quantitative reverse transcription-polymerase chain reaction, Western blotting, and immunohistochemical and immunofluorescence staining. The pathological changes in the lungs were evaluated via H&E and Masson staining. Furthermore, the subcellular distribution of GAS5 was examined using FISH. Dual-luciferase reporter assay, ChIP, RNA pull-down, and RIP experiments were conducted to determine the molecular interaction.
RESULTS
GAS5 expression decreased whereas PDGFRα/β expression increased in the lungs of IPF patients and mice with bleomycin-induced PF. The in vitro overexpression of GAS5 or silencing of PDGFRα/β inhibited the TGF-β1-induced differentiation of pericytes to myofibroblasts, as evidenced by the upregulation of pericyte markers NG2 and desmin as well as downregulation of myofibroblast markers α-SMA and collagen I. Further mechanistic analysis revealed that GAS5 recruited KDM5B to promote H3K4me2/3 demethylation, thereby suppressing PDGFRα/β expression. In addition, KDM5B overexpression inhibited pericyte-myofibroblast transformation and counteracted the promotional effect of GAS5 knockdown on pericyte-myofibroblast transformation. Lung fibrosis in mice was attenuated by GAS5 overexpression but promoted by GAS5 deficiency.
CONCLUSION
GAS5 represses pericyte-myofibroblast transformation by inhibiting PDGFRα/β expression via KDM5B-mediated H3K4me2/3 demethylation in IPF, identifying GAS5 as an intervention target for IPF.
背景
特发性肺纤维化（IPF）是一种可能导致持续性肺损伤的疾病。周细胞向肌成纤维细胞的转化已被认为是 IPF 进展过程中的关键因素。本研究旨在探讨长链非编码 RNA 生长停滞特异性转录物 5（GAS5）在 IPF 进展过程中肌成纤维细胞转化中的作用。
方法
通过气管内给予博来霉素建立小鼠肺纤维化（PF）模型。用外源性转化生长因子-β1（TGF-β1）刺激周细胞。采用实时定量逆转录聚合酶链反应、Western blot、免疫组化和免疫荧光染色检测靶分子的表达。通过 H&E 和 Masson 染色评估肺的病理变化。此外，通过 FISH 检测 GAS5 的亚细胞分布。通过双荧光素酶报告基因检测、ChIP、RNA 下拉和 RIP 实验确定分子相互作用。
结果
在 IPF 患者和博来霉素诱导的 PF 小鼠的肺中，GAS5 的表达降低，而 PDGFRα/β 的表达增加。在体外过表达 GAS5 或沉默 PDGFRα/β 抑制 TGF-β1 诱导的周细胞向肌成纤维细胞的分化，表现为周细胞标志物 NG2 和 desmin 的上调以及肌成纤维细胞标志物 α-SMA 和胶原 I 的下调。进一步的机制分析表明，GAS5 募集 KDM5B 以促进 H3K4me2/3 去甲基化，从而抑制 PDGFRα/β 的表达。此外，KDM5B 的过表达抑制周细胞-肌成纤维细胞转化，并抵消了 GAS5 敲低对周细胞-肌成纤维细胞转化的促进作用。GAS5 的过表达可减轻小鼠的肺纤维化，但 GAS5 的缺乏可促进肺纤维化。
结论
GAS5 通过 KDM5B 介导的 H3K4me2/3 去甲基化抑制 PDGFRα/β 的表达，从而抑制 IPF 中的周细胞-肌成纤维细胞转化，鉴定 GAS5 为 IPF 的干预靶点。
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