ATF3 -activated accelerating effect of LINC00941/lncIAPF on fibroblast-to-myofibroblast differentiation by blocking autophagy depending on ELAVL1/HuR in pulmonary fibrosis.
ATF3 -activated accelerating effect of LINC00941/lncIAPF on fibroblast-to-myofibroblast differentiation by blocking autophagy depending on ELAVL1/HuR in pulmonary fibrosis.
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DOI：
10.1080/15548627.2022.2046448
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发表时间：
2022-11
期刊：
影响因子：
13.3
通讯作者：
Song, Xiaodong
中科院分区：
文献类型：
--
作者：
Zhang, Jinjin;Wang, Haixia;Chen, Hongbin;Li, Hongbo;Xu, Pan;Liu, Bo;Zhang, Qian;Lv, Changjun;Song, Xiaodong
关键词：
特发性肺纤维化（IPF）的特征是肺瘢痕形成，目前尚无有效的治疗方法。成纤维细胞向肌成纤维细胞分化和肌成纤维细胞增殖和迁移是这种疾病的主要临床表现;因此，阻断这些过程是一种实用的治疗策略。在此，发现高度上调的LINC 00941/lncIAPF通过促进成纤维细胞向肌成纤维细胞分化以及肌成纤维细胞增殖和迁移来加速肺纤维化。使用测序和染色质免疫沉淀实验对转座酶可接近的染色质进行的测定阐明了组蛋白3赖氨酸27乙酰化（H3 K27 ac）激活LINC 00941启动子中的染色体区域开口。结果，转录因子ATF 3（转录激活因子3）结合到该区域，并且LINC 00941转录增强。RNA亲和分离、RNA免疫沉淀（RIP）、RNase-RIP、半衰期分析和泛素化实验揭示LINC 00941与ELAVL 1/HuR（ELAV like RNA binding protein 1）形成RNA-蛋白复合物以发挥其促纤维化功能。双荧光mRFP-GFP-MAP 1 LC 3/LC 3（微管相关蛋白1轻链3）腺病毒监测技术、人自噬RT 2 profiler PCR阵列和自噬通量揭示LINC 00941-ELAVL 1轴抑制自噬体与溶酶体融合。ELAVL 1 RIP-seq、RIP-PCR、mRNA稳定性和拯救实验表明，LINC 00941-ELAVL 1复合物通过控制靶基因EZH 2（zeste 2多梳抑制复合物2亚基增强子）、STAT 1（信号转导和转录激活因子1）和FOXK 1（叉头盒K1）的稳定性来抑制自噬。最后，LINC 00941的治疗效果在小鼠模型和IPF患者中得到了证实。这项工作为IPF提供了一个与自噬相关的治疗靶点和一种新的有效治疗策略。缩略语：ACTA2/a-SMA：肌动蛋白α 2，平滑肌; ATF 3：转录激活因子3; ATG：自噬相关; Baf-A1：巴弗洛霉素A1; BLM：博来霉素; CDKN：细胞周期蛋白依赖性激酶抑制剂; CLN 3：CLN 3溶酶体/内体跨膜蛋白，battenin; COL 1A：I型胶原α; COL 3A：III型胶原α; CXCR 4：C-X-C基序趋化因子受体4; DRAM 2：DNA损伤调节自噬调节因子2; ELAVL 1/HuR：ELAV样RNA结合蛋白1; EZH 2：zeste 2多梳抑制复合物2亚基增强子; FADD：通过死亡结构域相关的Fas; FAP/FAPα：成纤维细胞活化蛋白α; FOXK 1：叉头盒K1; FVC：用力肺活量; GABARAP：GABA A型受体相关蛋白; GABARAPL 2：GABA A型受体相关蛋白样2; IGF 1：胰岛素样生长因子1; IPF：特发性肺纤维化; LAMP：溶酶体相关膜蛋白; lncRNA：长链非编码RNA; MAP 1 LC 3/LC 3：微管相关蛋白1轻链3; NPC 1：NPC细胞内胆固醇转运蛋白1; RGS：G蛋白信号调节因子; RPLP 0：核糖体蛋白侧柄亚基P0; ROC：受试者操作特征; S100 A4：S100钙结合蛋白A4; SQSTM 1/p62：隔离体1; STAT 1：信号转导和转录激活因子1; TGFB 1/TGF-β1：转化生长因子β 1; TNF：肿瘤坏死因子; UIP：普通型间质性肺炎; ULK1：unc-51样自噬激活激酶1; Vim：波形蛋白。
Idiopathic pulmonary fibrosis (IPF) is characterized by lung scarring and has no effective treatment. Fibroblast-to-myofibroblast differentiation and myofibroblast proliferation and migration are major clinical manifestations of this disease; hence, blocking these processes is a practical treatment strategy. Here, highly upregulated LINC00941/lncIAPF was found to accelerate pulmonary fibrosis by promoting fibroblast-to-myofibroblast differentiation and myofibroblast proliferation and migration. Assay for transposase-accessible chromatin using sequencing and chromatin immunoprecipitation experiments elucidated that histone 3 lysine 27 acetylation (H3K27ac) activated the chromosome region opening in the LINC00941 promoter. As a consequence, the transcription factor ATF3 (activating transcription factor 3) bound to this region, and LINC00941 transcription was enhanced. RNA affinity isolation, RNA immunoprecipitation (RIP), RNase-RIP, half-life analysis, and ubiquitination experiments unveiled that LINC00941 formed a RNA-protein complex with ELAVL1/HuR (ELAV like RNA binding protein 1) to exert its pro-fibrotic function. Dual-fluorescence mRFP-GFP-MAP1LC3/LC3 (microtubule associated protein 1 light chain 3) adenovirus monitoring technology, human autophagy RT2 profiler PCR array, and autophagic flux revealed that the LINC00941-ELAVL1 axis inhibited autophagosome fusion with a lysosome. ELAVL1 RIP-seq, RIP-PCR, mRNA stability, and rescue experiments showed that the LINC00941-ELAVL1 complex inhibited autophagy by controlling the stability of the target genes EZH2 (enhancer of zeste 2 polycomb repressive complex 2 subunit), STAT1 (signal transducer and activators of transcription 1) and FOXK1 (forkhead box K1). Finally, the therapeutic effect of LINC00941 was confirmed in a mouse model and patients with IPF. This work provides a therapeutic target and a new effective therapeutic strategy related to autophagy for IPF. Abbreviations: ACTA2/a-SMA: actin alpha 2, smooth muscle; ATF3: activating transcription factor 3; ATG: autophagy related; Baf-A1: bafilomycin A1; BLM: bleomycin; CDKN: cyclin dependent kinase inhibitor; CLN3: CLN3 lysosomal/endosomal transmembrane protein, battenin; COL1A: collagen type I alpha; COL3A: collagen type III alpha; CXCR4: C-X-C motif chemokine receptor 4; DRAM2: DNA damage regulated autophagy modulator 2; ELAVL1/HuR: ELAV like RNA binding protein 1; EZH2: enhancer of zeste 2 polycomb repressive complex 2 subunit; FADD: Fas associated via death domain; FAP/FAPα: fibroblast activation protein alpha; FOXK1: forkhead box K1; FVC: forced vital capacity; GABARAP: GABA type A receptor-associated protein; GABARAPL2: GABA type A receptor associated protein like 2; IGF1: insulin like growth factor 1; IPF: idiopathic pulmonary fibrosis; LAMP: lysosomal associated membrane protein; lncRNA: long noncoding RNA; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; NPC1: NPC intracellular cholesterol transporter 1; RGS: regulator of G protein signaling; RPLP0: ribosomal protein lateral stalk subunit P0; ROC: receiver operating characteristic; S100A4: S100 calcium binding protein A4; SQSTM1/p62: sequestosome 1; STAT1: signal transducers and activators of transcription 1; TGFB1/TGF-β1: transforming growth factor beta 1; TNF: tumor necrosis factor; UIP: usual interstitial pneumonia; ULK1: unc-51 like autophagy activating kinase 1; VIM: vimentin.
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DOI：
10.4049/jimmunol.1402277
发表时间：
2015-06-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Abdel Fattah E;Bhattacharya A;Herron A;Safdar Z;Eissa NT
通讯作者：
Eissa NT
排序Nexin 5介导病毒诱导的自噬和免疫力。
影响因子：
64.8
作者：
Dong X;Yang Y;Zou Z;Zhao Y;Ci B;Zhong L;Bhave M;Wang L;Kuo YC;Zang X;Zhong R;Aguilera ER;Richardson RB;Simonetti B;Schoggins JW;Pfeiffer JK;Yu L;Zhang X;Xie Y;Schmid SL;Xiao G;Gleeson PA;Ktistakis NT;Cullen PJ;Xavier RJ;Levine B
通讯作者：
Levine B
影响因子：
2.9
作者：
Du Y;Hao X;Liu X
通讯作者：
Liu X
影响因子：
4.7
作者：
Han, Ruhui;Ji, Xiaoming;Ni, Chunhui
通讯作者：
Ni, Chunhui
影响因子：
4.4
作者：
Kobayashi, Kenji;Araya, Jun;Kuwano, Kazuyoshi
通讯作者：
Kuwano, Kazuyoshi