干预非编码转录超保守区uc.261重塑克罗恩病肠黏膜屏障紧密连接的策略研究

It is different to mature views on correlation between genetic susceptibility loci and integral incidence of Crohn's disease that targeted research enrolling genetic factors into the local mechanism of intestinal mucosa barrier damage and restoration is still lacking to date. Furthermore, strategic research to remodel intestinal mucosa barrier in Crohn's disease by interfering the non-coding, transcribed ultraconserved region (T-UCR) with the genetic background is awaiting implementation. This project is based on the preliminary work that the changes of non-coding T-UCR sequence uc.261 may be involved in the intestinal mucosa barrier damage and remodeling in Crohn's disease. The possible axis that "alternation of non-coding T-UCR sequence uc.261 → alternation or modification of PSAT1/TLE4/CNTNAP3 expression → downregulation of CDC42 → suppression of PAR-3-aPKC-PAR-6 complex activity → suppression of transmembrane protein JAMs → tight junction of intestinal mucosa barrier damages" is preliminarily analyzed in Crohn's disease. And then, the basic research strategy to remodel the tight junction of intestinal mucosa barrier will be constructed based on the brand-new perspective by reversing the non-coding T-UCR sequence variation, which can provide novel inspiration exploring potential biomarkers to predict disease recurrence and potential drug targets to achieve mucosal healing in Crohn's disease.

与遗传易感性位点和克罗恩病整体发病率相关的观点已趋于成熟不同，目前尚缺乏将遗传因素纳入肠黏膜屏障破坏和修复交替这一局部机制的针对性研究。而干预具备遗传背景的非编码转录超保守区，继而重塑克罗恩病肠黏膜屏障紧密连接的策略研究更尚待开展。本项目基于前期工作中发现的非编码转录超保守区序列uc.261改变可能参与克罗恩病肠黏膜屏障紧密连接破坏与重塑这一立足点，通过初步分析克罗恩病中可能存在的非编码转录超保守区序列uc.261表达改变→PSAT1/TLE4/CNTNAP3表达或修饰改变→CDC42表达降低→PAR-3-aPKC-PAR-6复合物活性抑制→跨膜蛋白JAMs抑制→肠黏膜屏障紧密连接破坏这一作用轴，继而从逆转非编码转录超保守区序列变异的全新角度，构建重塑肠黏膜屏障紧密连接的基础研究策略，为探究预测克罗恩病疾病复发的潜在生物标记、获取达到黏膜愈合目标的潜在药物靶点提供新的启迪。

与遗传易感性位点和克罗恩病整体发病率相关的观点已趋于成熟不同，目前尚缺乏将遗传因素纳入肠黏膜屏障破坏和修复交替这一局部机制的针对性研究。而干预具备遗传背景的非编码转录超保守区，继而重塑克罗恩病肠黏膜屏障紧密连接的策略研究更尚待开展。本项目基于前期工作中发现的非编码转录超保守区序列uc.261改变可能参与克罗恩病肠黏膜屏障紧密连接破坏与重塑这一立足点，通过初步分析克罗恩病中可能存在的非编码转录超保守区序列uc.261表达改变→PSAT1表达改变→CDC42修饰改变→紧密连接蛋白表达下降→肠黏膜屏障紧密连接破坏这一作用轴，结果发现uc.261通过对下游靶基因PSAT1具有抑制作用继而导致CDC42的活性形式CDC42-GTP降低、PKCζ的活性形式phospho-PKCζ明显升高，最终导致紧密连接蛋白的装配减少；本研究还发现uc.261在克罗恩病中有作为黏膜愈合标志物的可能，逆转uc.261表达，能逆转模型小鼠肠道炎症反应，重塑肠黏膜紧密连接。.本研究首先验证分析了克罗恩病肠黏膜屏障紧密连接破坏过程以及疾病缓解期肠黏膜屏障紧密连接重塑过程中存在的非编码转录超保守区uc.261表达改变；其次探索分析非编码转录超保守区uc.261表达改变对克罗恩病肠黏膜屏障紧密连接破坏和修复的具体作用机制与途径；最后分析了干预非编码转录超保守区uc.261重塑克罗恩病肠黏膜屏障紧密连接策略的可行性。本研究从逆转非编码转录超保守区序列变异的全新角度，构建重塑肠黏膜屏障紧密连接的基础研究策略，为探究预测克罗恩病疾病复发的潜在生物标记、获取达到黏膜愈合目标的潜在药物靶点提供新的启迪。