基于自组装细胞芯片大规模筛选研究靶基因 miRNA 调控位点

It has been showed that miRNAs play a critical role in the development of embyro and diseases. As a new class of multitarget regulators, miRNAs have been thought as a type of promising candidates as therapeutical targets. A large body of evidence in animals showed that the addition or inhibition of miRNAs can suppress the growth and metastasis of tumor. However, one central question regarding miRNA, that is, how many targets one miRNAs could directly regulate, remains poorly understood. In this project, we propose to develop a large scale method for the accurate identification of the miRNA targeting sites on a gene based on self-assembled cell microarray. A group of important genes will be selected to identify and analyze their miRNA targeting sites in content of cancer development and two signaling pathwaies, which will help in better understanding the mechanism of miRNAs in cancer development and evaluating their potential in therapy. Our primary data showed that miR-19 can tune the expression of VEGF on three layers, thus regulating tumour angiogenesis on the whole. We believe, it is likely possible that we would make the huge progress in the study of miRNAs if this project would be initiated.

miRNA是一类多靶点调控分子，在个体发育以及疾病发生、发展过程中，发挥着重要的作用，有望成为一类新型的肿瘤治疗靶点。大量的动物实验结果证明，调控miRNA就能明显抑制肿瘤发生和转移。然后，迄今为止，科学家还无法准确知道一条miRNA究竟调控多少条靶基因？针对这一核心问题-miRNA与靶基因作用关系，本课题拟将基于自组装细胞芯片技术，建立一套大规模、准确研究目标靶基因的miRNA调控位点的新方法。通过筛选分析一组重要基因的miRNA调控位点，研究miRNA的作用规律；并从肿瘤发展过程、信号通路等方面，研究miRNA的作用机理，阐释miRNA与靶基因在肿瘤发生、发展过程中的相互作用，全面评估miRNA作为癌症诊断和治疗靶点的可行性。初步结果显示，miRNA-19能够从至少三个层次调控VEGF的表达量，实现对肿瘤血管增生的整体调控。本项目的成功实施，对该领域的发展具有重大的推动作用。

本项目开发一套超高灵敏度筛选研究miRNA靶基因的新方法，详细研究了20余条人鼠保守重要基因与300 条miRNA的相互作用研究，发现人鼠miRNA与靶基因调控的种属差异规律；详细研究了miR-122、miR-19b、miR-190等5条miRNA在肿瘤发生、发展过程中的分子机制，探讨它们作为药靶的可行性。在Nature Communications、ACS Nano、Nucleic Acid Research 等杂志发表高水平SCI论文8篇（4篇影子因子大于10），申报发明专利3项。