RBM47/NLRP3/GSDMD剪接调控网络在肾结石发生发展中的作用及机制研究

选择性剪接可使一个基因产生具有不同功能的蛋白异构体。在正常条件下，选择性剪接的调控是受到严格控制的，选择性剪接的异常可导致多种疾病的发生。我们的前期研究发现NLRP3基因Exon5在肾结石患者肾乳头钙斑组织和非肾结石患者正常肾乳头组织中存在异常的可变剪接，其中NLRP3-exon5+(NLRP3-full)在钙斑组织中高表达，而NLRP3-exon5-(NLRP3-Δ5)在正常肾乳头组织中相对高表达。我们还发现RBM47可调节NLRP3的可变剪接。我们通过一系列体内和体外实验证实NLRP3两个转录本在草酸钙晶体造成的肾小管上皮细胞焦亡和炎症损伤中起着不同的作用从而调控这肾结石的发生发展。在此基础上，本课题拟继续探究RBM47/NLRP3/GSDMD剪接调控网络在肾结石发生发展中的作用机制，并为肾结石的诊断和治疗提供新的理论依据和药物靶点。

Alternative splicing enables one gene to produce multiple protein isoforms with different functions. Under normal conditions, this mechanism is tightly regulated. When deregulated, the aberrant splicing mRNA could cause many kinds of diseases. Here, we identified the upregulation of NLRP3-exon5+ (NLRP3-full) in Randall’s plaque tissues of patients with kidney stones, and relatively upregulation of NLRP3-exon5- (NLRP3-Δ5) in normal renal papillary tissues of controls. We further showed that alternative splicing of NLRP3 pre-mRNA is regulated by RBM47. Through a series of in vivo and in vitro experiments, we confirmed that the two transcripts of NLRP3 played different roles in the pyroptosis and inflammation of renal tubular epithelial cells caused by calcium oxalate crystals. In this project, we plan to systematically study the function and the molecular mechanism of RBM47/NLRP3/GSDMD interaction in kidney stone development and progression, and which will benefit the diagnosis and treatment of kidney stone.