A novel transcriptional complex on the VE-cadherin promoter regulated the downregulation of VE-cadherin in the Down Syndrome Candidate Region 1 isoform 1L-mediated angiogenesis.
A novel transcriptional complex on the VE-cadherin promoter regulated the downregulation of VE-cadherin in the Down Syndrome Candidate Region 1 isoform 1L-mediated angiogenesis.
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DOI：
10.1016/j.mvr.2021.104209
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发表时间：
2021-11
影响因子：
3.1
通讯作者：
Zeng, Huiyan
中科院分区：
文献类型：
--
作者：
Hou, Shiqiang;Niu, Gengming;Liu, Xin;Bourbon, Pierre M.;Zhang, Dongmei;Cui, Pengfei;Zhao, Kevin;Zhao, Dezheng;Zeng, Huiyan
血管生成是许多疾病的关键。此前，我们报道了唐氏综合症候选区域1亚型1L(DSCR1-1L)是血管内皮生长因子和组胺诱导内皮细胞表达上调最多的基因之一，并调节小鼠内皮细胞的增殖和Matrigel血管生成。然而，目前尚不清楚DSCR1-1L是否调节体内血管生成，以及其背后的分子机制。本研究通过建立DSCR1-1L基因敲除和过表达模型，研究DSCR1-1L在体内血管生成中的作用。进一步，用分子生物学方法在血管内皮细胞中探索了DSCR1-1L的下游调控靶点。我们发现DSCR11L shRNA显著抑制血管内皮生长因子诱导的小鼠血管生成(p<；0.0001)。在功能增益实验中，EC-FH-DSCR1-1L转基因小鼠内皮细胞中DSCR1-1L基因的过表达足以显著诱导血管生成(p<；0.01)。在早期，DSCR1-1L通过下调VE-钙粘蛋白的表达来调控血管生成，其作用靶向是VE-钙粘蛋白的转录，而不是其mRNA的稳定性。通过启动子报告实验鉴定了VE-cadherin启动子中的三个DSCR1-1L靶向DNA元件，其中发现了一个新的特异转录复合体。通过凝胶迁移率改变分析和DNase I足迹分析，确定VE-钙粘蛋白启动子上的DNA序列(CTTCTG)能够与蛋白质直接相互作用。因此，DSCR1-1L通过下调VE-钙粘蛋白启动子上新的转录复合体的形成，是治疗血管生成疾病的良好靶点。DSCR1-1L shRNAs和cDNAs具有开发临床应用的潜力。我们的结果也对力学研究领域做出了重大贡献。
Angiogenesis is critical for many diseases. Previously, we reported that Down Syndrome Candidate Region 1 isoform 1L (DSCR1–1L) was one of the most up-regulated genes in endothelial cells induced by VEGF and histamine, and regulated endothelial cell proliferation and Matrigel angiogenesis in mice. However, it was not known whether DSCR1–1L regulated angiogenesis in vivo and what was the molecular mechanism underlying it. In this study, gene knockdown and overexpression models were established to study the role of DSCR1–1L in angiogenesis in vivo. Further, the downstream regulatory target of DSCR1–1L was explored with molecular biological methods in vascular endothelial cells. We found that DSCR1–1L shRNAs significantly inhibited angiogenesis induced by VEGF in mice (p < 0.0001). In the gain-of-function assay, overexpression of DSCR1–1L cDNA in mouse endothelium of EC-FH-DSCR1–1L transgenic mice was sufficient to induce angiogenesis significantly (p < 0.01). DSCR1–1L regulated angiogenesis in the early stage by down-regulation of the VE-cadherin expression through targeting its transcription, but not mRNA stability. Three DSCR1–1L-targeted DNA elements in the VE-cadherin promoter were identified by promoter reporter assays, among which, a novel specific transcriptional complex was found. The DNA sequence (CTTCTG) in the VE-cadherin promoter was identified to directly interact with proteins by Electrophoresis Mobility Shift Assays and DNase I footprint assay. Hence, DSCR1–1L is an excellent therapeutic target for angiogenic diseases through down-regulating the formation of a novel transcriptional complex on the VE-cadherin promoter. DSCR1–1L shRNAs and cDNA have the potential to be developed for clinical application. Our results also contribute significantly to the field of mechanistic studies.
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DOI：
10.1084/jem.20091846
发表时间：
2010-03-15
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Helfrich I;Scheffrahn I;Bartling S;Weis J;von Felbert V;Middleton M;Kato M;Ergün S;Augustin HG;Schadendorf D
通讯作者：
Schadendorf D
DOI：
10.1073/pnas.0708148104
发表时间：
2007-10-23
影响因子：
11.1
作者：
Ebos, John M. L.;Lee, Christina R.;Kerbel, Robert S.
通讯作者：
Kerbel, Robert S.
影响因子：
4.8
作者：
Ermak, G;Morgan, TE;Davies, KJA
通讯作者：
Davies, KJA
影响因子：
4.8
作者：
Liu, Xin;Zhao, Dezheng;Zeng, Huiyan
通讯作者：
Zeng, Huiyan
影响因子：
20.3
作者：
Giampietro, Costanza;Taddei, Andrea;Dejana, Elisabetta
通讯作者：
Dejana, Elisabetta