ciR-0013395/Collagen I 通路参与二氧化硅诱导的肺内皮间质转化的分子机制研究

Silicosis is a untreatable disease characterized by pulmonary fibrosis with occupational exposure to silica dusts..Circular RNAs are ubiquitous in molecular biology, which could modulate the expression of numerous downstream protein via different mechanisms, such as competing endogenous RNA (ceRNA). Our previous studies suggested that SiO2 induced endothelial-mesenchymal transition (EndoMT), associated with the upregulation of collagen I. However, the detailed cellular and molecular mechanisms of involvement of collagen I in EndoMT triggered by silica remain unknown. The bioinformatics analysis showed that ciR-0013395 is homologous circular RNA targeting regulation of collagen I. Therefore, we hypothesize that ciR-0013395/Collagen I pathway will be the potential target of curative treatment for silicosis involving in regulation of EndoMT..The proposed studies will be initiated in human sample followed by study in intact animals, as well as primary cell cultures. Studies using classic pharmacological methods will be combined with molecular biological techniques, as well as immunological methods, all of which are currently well established in our laboratory. Our study will decipher the link between ciR-0013395/Collagen I and EndoMT induced by silica, providing a novel insight into the potential of ciR-0013395/Collagen I in terms of opening up novel therapeutic avenues for silicosis.

矽肺是由于长期吸入大量游离二氧化硅（SiO2）粉尘所引起的以肺纤维化为特征的疾病，目前尚缺乏针对性干预治疗的有效靶点。环状RNA是一类由特殊的选择性剪切产生的首尾相接的非编码RNA，可通过竞争性内源RNA(ceRNA)等机制调控靶蛋白。申请者前期研究发现，SiO2可以诱导内皮细胞的间质转化，引起内皮细胞合成collagen I增加，进而参与肺纤维化进程。通过生物信息学分析结果显示，ciR-0013395是collagen I的母本环状RNA，它可能通过调节collagen I的表达参与内皮细胞的间质转化。本课题拟在前期工作基础上，以SiO2诱导的内皮间质转化为研究对象，以ciR-0013395/Collagen I为切入点，构建矽肺动物模型和细胞模型，重点探讨ciR-0013395/Collagen I 通路在内皮间质转化中的作用及分子机制，为矽肺早期诊断及治疗提供新思路和新靶点。

矽肺是由于长期吸入大量游离二氧化硅（SiO2）粉尘所引起的以肺纤维化为特征的疾病，主要以炎症损伤、组织结构破坏及细胞外基质异常沉积为主要特征，严重影响肺功能。矽肺一旦发生，即使脱离矽尘接触，肺损伤依然进行性发展。目前矽肺尚没有特效治疗措施，因此探寻矽肺发病机制，进而发现矽肺早期诊断分子标志物以及干预肺纤维化进程的靶标已成为迫切需要解决的问题。内皮间质转化参与了肺纤维化过程。本课题在实验室前期研究结果基础上，结合RNA（circRNA，ciR）芯片高通量筛选结果及生物信息学分析，提出“ciR-0013395/Collagen I通路是参与SiO2诱导的内皮间质转化的重要分子机制，通过药理学和分子生物学手段，探讨 ciR-0013395/Collagen I 通路的作用机制，为矽肺早期诊断及治疗提供新的思路和靶点。ciR-0013395 是调控 collagen I 表达的母本 circRNA。研究结果显示，SiO2刺激后内皮细胞发生间质转化，胶原蛋白collagen I明显增加，矽肺小鼠肺中胶原蛋白collagen I明显增加。小鼠肺组织进行脱细胞处理后得到肺细胞外基质，结果显示矽肺小鼠肺细胞基质出现明显的纤维化样改变，胶原蛋白collagen I明显增加。将内皮细胞重新移植到脱细胞处理后的小鼠肺细胞外基质中，内皮细胞功能发生明显改变；考虑到成纤维细胞在肺纤维化中不可忽视的作用，将成纤维细胞重新移植到脱细胞处理后的小鼠肺细胞外基质中，成纤维细胞功能发生明显改变。以上结果提示，纤维化样细胞外基质影响细胞的功能，这可能与胶原蛋白collagen I合成和分泌增加导致在细胞外基质上的异常沉积有关。综上所述，ciR-0013395/Collagen I 通路可能通过改变细胞外基质的结构来影响细胞的正常功能，从而促进肺纤维化进展，为肺纤维化治疗药物的开发提供了新的潜在靶点。