Production of human MUC17 (CRD1-L-CRD2) recombinant protein in rice for the treat

描述(申请人提供)：最近在动物模型中的研究表明，一种名为MUC17的内源性人类肠道粘蛋白片段可以促进肠道细胞恢复并促进实验性结肠炎的愈合，可能为有效治疗溃疡性结肠炎打开一扇新的大门。然而，MUC17在大肠杆菌、酵母和昆虫细胞中的表达受到了极大的挑战，因为它的表达水平低，折叠不当或不能溶解。Ventria的科学家已经开发出一种在大米籽粒中高效的蛋白质表达系统，并使用该系统生产了几种蛋白质，这些蛋白质现在已经进入市场。在目前的提案中，我们想测试使用专有的Ventria水稻表达系统来高水平生产MUC17的可行性。此外，我们还将从稻米中提纯MUC17，并通过体外细胞培养研究其生物学功能。我们推测，从大米中提取的MUC17将促进细胞迁移和抑制细胞凋亡，这在结肠上皮细胞重建中具有重要作用。成功地在水稻籽粒中高水平表达功能性MUC17将极大地加强该项目的研发工作，并为溃疡性结肠炎患者提供一条这种新蛋白的商业化途径。

DESCRIPTION (provided by applicant): Recent studies in animal models demonstrate that an endogenous human intestinal mucin fragment, termed MUC17, augments intestinal cell restitution and enhances healing of experimental colitis, potentially opening a new door to effective treatment of ulcerative colitis. However, expression of MUC17 in E. coli, yeast, and insect cells has met great challenges due to low expression, improper folding or insolubility. Ventria scientists have developed a highly efficient protein expression system in rice grain and have used the system to produce several proteins which are now in the marketplace. In the present proposal, we would like to test the feasibility of employing the proprietary Ventria rice expression system to produce MUC17 at high levels. Furthermore, we will purify MUC17 from rice grain and test its biological function via in vitro cell culture studies. We hypothesize that MUC17, derived from rice grain, will promote cell migration and inhibit apoptosis, important functions in epithelial cell restitution of the colon. Successfully expressing functional MUC17 in rice grain at high levels will greatly enhance the research and development effort of this project and provide a path to commercialization of this novel protein for patients suffering from ulcerative colitis.