Long non-coding RNA ACTA2-AS1 promotes ductular reaction by interacting with the p300/ELK1 complex.
Long non-coding RNA ACTA2-AS1 promotes ductular reaction by interacting with the p300/ELK1 complex.
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DOI：
10.1016/j.jhep.2021.12.014
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发表时间：
2022-04
影响因子：
25.7
通讯作者：
Huebert RC
中科院分区：
文献类型：
--
作者：
Navarro-Corcuera A;Sehrawat TS;Jalan-Sakrikar N;Gibbons HR;Pirius NE;Khanal S;Hamdan FH;Aseem SO;Cao S;Banales JM;Kang N;Faubion WA;LaRusso NF;Shah VH;Huebert RC
胆道疾病与增殖/纤维化导管反应（DR）有关。 p300 是一种表观遗传调节因子，可乙酰化组蛋白 3 (H3K27ac) 上的赖氨酸 27，并在纤维化过程中被激活。长非编码 RNA (lncRNA) 在胆管病中异常表达，但人们对它们如何募集表观遗传复合物并调节 DR 知之甚少。我们研究了表观遗传复合物，包括转录因子 (TF) 和 lncRNA，它们在纤维化过程中促进 p300 介导的转录。我们使用他莫昔芬诱导、胆管细胞选择性、p300 敲除 (KO) 结合胆管结扎 (BDL) 和用 SGC-CBP30 治疗的 Mdr KO 小鼠体内评估了 p300。通过 qPCR 和 RNA ISH 分析原代胆管细胞和肝组织的 Acta2-as1 lncRNA 表达。在体外，我们使用 p300 抑制剂对人胆管细胞进行了 RNA 测序。将胆管细胞暴露于脂多糖（LPS）作为损伤模型。我们通过免疫沉淀 (IP) 证实了 p300/ELK1 复合物的形成。 RNA IP 检查了 ACTA2-AS1 和 p300 之间的相互作用。染色质 IP 测定评估了 p300/ELK1 占用和 p300 介导的 H3K27ac。类器官是由 ACTA2-AS1 耗尽的胆管细胞产生的。 Krt19-CreERT/p300fl/fl 小鼠中 BDL 诱导的 DR 和纤维化减少。同样，Mdr KO 小鼠在 SGC-CBP30 治疗后也免受 DR 和纤维化的影响。在体外，去除 ACTA2-AS1 会降低增殖/纤维化标志物的表达，减少 LPS 诱导的胆管细胞增殖，并损害类器官的形成。 ACTA2-AS1 通过在 LPS 后促进 p300/ELK1 与 PDGFB 启动子结合来调节转录。相应地，LPS 诱导的 H3K27ac 由 p300/ELK1 介导，并且在 ACTA2-AS1 耗尽的胆管细胞中减少。胆管细胞选择性 p300 KO 或 p300 抑制可减轻小鼠的 DR/纤维化。 ACTA2-AS1 影响 p300/ELK1 向特定启动子的募集，以驱动 H3K27ac 和增殖/纤维化基因的表观遗传激活。这表明表观遗传共激活因子和 lncRNA 之间的合作促进了胆道疾病的 DR/纤维化。我们鉴定了一个由三部分组成的复合物，其中包含 RNA 分子、转录因子和表观遗传酶。该复合物在受损的胆管细胞中具有活性，有助于激活参与增殖和纤维化的基因。
Biliary disease is associated with a proliferative/fibrogenic ductular reaction (DR). p300 is an epigenetic regulator that acetylates lysine 27 on histone 3 (H3K27ac) and is activated during fibrosis. Long non-coding RNAs (lncRNAs) are aberrantly expressed in cholangiopathies, but little is known about how they recruit epigenetic complexes and regulate DR. We investigated epigenetic complexes, including transcription factors (TFs) and lncRNAs, contributing to p300-mediated transcription during fibrosis. We evaluated p300 in vivo using tamoxifen-inducible, cholangiocyte-selective, p300 knockout (KO) coupled with bile duct ligation (BDL) and Mdr KO mice treated with SGC-CBP30. Primary cholangiocytes and liver tissue were analyzed for expression of Acta2-as1 lncRNA by qPCR and RNA ISH. In vitro, we performed RNA-sequencing in human cholangiocytes with a p300 inhibitor. Cholangiocytes were exposed to lipopolysaccharide (LPS) as an injury model. We confirmed formation of a p300/ELK1 complex by immunoprecipitation (IP). RNA IP examined interactions between ACTA2-AS1 and p300. Chromatin IP assays evaluated p300/ELK1 occupancy and p300-mediated H3K27ac. Organoids were generated from ACTA2-AS1-depleted cholangiocytes. BDL-induced DR and fibrosis were reduced in Krt19-CreERT/p300fl/fl mice. Similarly, Mdr KO mice were protected from DR and fibrosis after SGC-CBP30 treatment. In vitro, depletion of ACTA2-AS1 reduced expression of proliferative/fibrogenic markers, reduced LPS-induced cholangiocyte proliferation, and impaired organoid formation. ACTA2-AS1 regulated transcription by facilitating p300/ELK1 binding to the PDGFB promoter after LPS. Correspondingly, LPS-induced H3K27ac was mediated by p300/ELK1 and was reduced in ACTA2-AS1-depleted cholangiocytes. Cholangiocyte-selective p300 KO or p300 inhibition attenuate DR/fibrosis in mice. ACTA2-AS1 influences recruitment of p300/ELK1 to specific promoters to drive H3K27ac and epigenetic activation of proliferative/fibrogenic genes. This suggests that cooperation between epigenetic co-activators and lncRNAs facilitates DR/fibrosis in biliary diseases. We identified a three-part complex containing an RNA molecule, a TF, and an epigenetic enzyme. The complex is active in injured bile duct cells and contributes to activation of genes involved in proliferation and fibrosis.
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影响因子：
64.8
作者：
Lasko LM;Jakob CG;Edalji RP;Qiu W;Montgomery D;Digiammarino EL;Hansen TM;Risi RM;Frey R;Manaves V;Shaw B;Algire M;Hessler P;Lam LT;Uziel T;Faivre E;Ferguson D;Buchanan FG;Martin RL;Torrent M;Chiang GG;Karukurichi K;Langston JW;Weinert BT;Choudhary C;de Vries P;Van Drie JH;McElligott D;Kesicki E;Marmorstein R;Sun C;Cole PA;Rosenberg SH;Michaelides MR;Lai A;Bromberg KD
通讯作者：
Bromberg KD
DOI：
10.1056/nejmra1506330
发表时间：
2016-09-22
期刊：
The New England journal of medicine
影响因子：
--
作者：
Lazaridis KN;LaRusso NF
通讯作者：
LaRusso NF
影响因子：
64.5
作者：
Bose DA;Donahue G;Reinberg D;Shiekhattar R;Bonasio R;Berger SL
通讯作者：
Berger SL
影响因子：
29.4
作者：
Aseem SO;Jalan-Sakrikar N;Chi C;Navarro-Corcuera A;De Assuncao TM;Hamdan FH;Chowdhury S;Banales JM;Johnsen SA;Shah VH;Huebert RC
通讯作者：
Huebert RC
DOI：
10.1073/pnas.1115201109
发表时间：
2012-03-06
影响因子：
11.1
作者：
Diril, M. Kasim;Ratnacaram, Chandrahas Koumar;Kaldis, Philipp
通讯作者：
Kaldis, Philipp