ciR-0001544靶向调控ZC3H4在矽肺炎症和纤维化中的作用研究

The inhalation of silicon dioxide particles causes pneumoconiosis, an untreatable pulmonary disease characterized by alveolar inflammation at the early stage and progressive lung fibrosis at the late stage. Although more and more studies were undertaken to dissect the pathological mechanisms underlying the process of silicosis, the primary cause of this often devastating disease remains elusive. Diagnosis of silicosis could easily be missed, since which needs carefully documented records of occupational exposure and radiological features, with exclusion of other competing diagnoses. As yet, no curative treatment exists, but comprehensive management strategies help to improve quality of life and slow deterioration. Thus, further efforts are needed to find early molecular marker of diagnosis, new target to control the fibrosis induced by silica.. Circular RNAs are ubiquitous in molecular biology. Recent papers have described the presence of circular RNA species from back-spliced exons in mammals (circular exonic RNAs, circRNAs) and have established that they are very abundant and are differentially expressed Circular RNAs may, for example, serve as transcription regulators or as sponges for small RNA regulators. . Our previous studies also suggested that monocyte chemotactic protein 1 (MCP-1) induced protein 1 (MCPIP1, ZC3H12A), an important member of zinc finger protein family, played a critical role in fibroblast proliferation and migration. However, the detailed cellular and molecular mechanisms underlying inflammation and the subsequent fibrosis in response to silica remain unknown. Circular RNA microarray assay suggested that ciR-0001544 level in mouse lung exposure to SiO2 was increased. Interestingly, ciR-0001544 is the regulator of ZC3H4, another important member of zinc finger protein family. Moreover, micro RNA microarray assay found that miR-212 was also decreased, which can inhibit the expression of ZC3H4. . Therefore, we hypothesized that ciR-0001544 targeting regulation of ZC3H4 will be the potential target of curative treatment for silicosis involving in regulation of inflammation and fibrosis. 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. This proposal is both novel and innovative in that the efficacy of regulation of ciR-0001544/ZC3H4 can be of value to prevent or halt progression of silicosis in people. Our study will decipher the link between ciR-0001544/ZC3H4 and inflammation with subsequent fibrosis, induced by silica providing a novel insight into the potential of ciR-0001544/ZC3H4 in terms of opening up novel therapeutic avenues for silicosis.

矽肺是吸入含二氧化硅粉尘颗粒，在人肺内沉积后引发的一种尚无特效治疗措施的肺部疾病，其特征是疾病早期的肺部炎症和后期进行性肺纤维化。矽肺诊治面临早期缺少筛检诊断方法，后期肺纤维化缺乏特效治疗措施。环状RNA（circRNA）是调控生命活动重要的非编码RNA，由于circRNA比线性RNA更稳定，因此是合适的临床诊断标记物与疾病干预靶点。申请者前期研究发现，锌指蛋白家族参与了矽肺发生发展过程；circRNA高通量筛选结果发现，调控锌指蛋白家族成员ZC3H4的ciR-0001544参与矽肺炎性反应和纤维化进程。基于以上结果，申请者提出“ciR-0001544靶向调控ZC3H4是矽肺诊治的有效和关键靶点”假说，拟应用分子生物学手段，结合经典药理学方法，从整体、细胞和分子水平系统揭示此通路在矽肺炎症和肺纤维化中的功能和作用机制，为临床治疗策略的选择和治疗药物靶标的遴选提供重要线索。

本课题针对矽肺临床面临的早期缺少筛检诊断方法，后期肺纤维化缺乏特效治疗措施的问题，研究ciR-0001544靶向调控ZC3H4在矽肺诊治中的应用及其细胞分子机制，阐明从现象→功能→机制的完整证据链，验证以ciR-0001544/ZC3H4为靶点，开发矽肺诊断和治疗新策略，为临床诊治提供新思路。.研究结果表明，在矽肺纤维化发生发展中，单核细胞可能通过转分化为巨噬细胞，失去其保护功能，可能作为矽肺炎性治疗的一个新的靶点，ZC3H4在此过程中发挥重要调控功能，可以成为药物多靶点治疗的候选靶标。进一步研究表示，ZC3H4可以通过ER stress途径参与调节SiO2诱导的巨噬细胞活化，干细胞能够通过下调ZC3H4 的表达从而抑制SiO2诱导的巨噬细胞活化，为干细胞治疗矽肺纤维化提供新的作用靶点。同时，干细胞通过抑制内皮细胞ZC3H4表达及内皮间质转化，进而延缓纤维化进程，这一结果将为临床干细胞治疗矽肺纤维化提供理论依据。同时ZC3H4参与调控成纤维细胞的活化，成纤维细胞在受到SiO2刺激后，ZC3H4表达显著升高，并通过sigmar1引起ER stress，导致成纤维细胞活化、分化和迁移；有趣的是，ER Stress可以进一步通过正反馈，持续升高ZC3H4，导致恶性循环，导致纤维化进程持续进展。.综上所述，本研究通过对矽肺进程中重要的炎症和纤维化病理改变，寻找调控炎症和纤维化进程的关键分子，从表观遗传学调控机制到转录调控机制，分别阐明ZC3H4在矽肺进程中的作用，并探究以其为靶点干细胞治疗的有效性继而可行性，对矽肺的诊断治疗具有重要的意义，随着细胞治疗手段的不断完善，本研究内容为干细胞在矽肺临床中的应用打下了坚实的基础，同时也为药物的开发提供可靠的靶点。