Reprogram gastric tissue to functional insulin-secreting cells

 描述(申请人提供)：再生医学的一个主要目标是产生分泌胰岛素的β细胞，用于移植治疗1型糖尿病(T1D)。胚胎干细胞的分化是一项产生功能性胰岛素+细胞的强大技术。然而，畸胎瘤的形成是这种方法的一个严重问题。或者，成人组织可以在一种称为重编程的过程中转化为胰岛素+细胞。对成人细胞重新编程对畸胎瘤的风险很小，但重新编程的效率往往很低，由此产生的胰岛素+细胞的功能有限。对于这种方法，有一个关键的NED来确定最佳的组织来源和重新编程的方法。我的实验室开创了一种基于已定义遗传因子(Ngn3、Pdx1、Mafa，称为NPM因子)鸡尾酒的重新编程方法。在动物模型中，NPM因子足以将胰腺腺泡细胞转化为稳定和功能正常的胰岛素+细胞。在对成人组织的全面筛查中，我们发现驻留在成人胃粘膜中的细胞也对NPM因子快速反应，并产生功能性胰岛素+细胞。值得注意的是，胃胰岛素+细胞比腺泡来源的胰岛素+细胞对葡萄糖的反应更快。因此，胃组织是一个非常有希望的成人组织来源，以产生功能性的胰岛素+细胞。重要的是，人的胃细胞可以从身体来源大量培养和繁殖为类器官。NPM因子的传递导致了C-肽+细胞在人的类器官中的形成，增加了用这种方法产生功能性人胰岛素+细胞的令人兴奋的可能性。该提案的一个主要目的是更深入地了解重新编程过程和由此产生的胃胰岛素+细胞。具体地说，我们将确定诱导的胃胰岛素+细胞与天然胰岛β细胞在分子和功能上的相似性。我们将研究诱导细胞的稳定性，并验证胰岛结构形成将导致传代长期稳定的假设。我们还将确定NPM因素在重新编制程序过程中的单独作用。该提案的第二个主要目标是开发有效地从人胃组织中诱导具有功能的胰岛素+细胞的方法，并结合遗传因素和信号通路调节剂进行治疗。总之，这些研究将为开发一种新的技术来生产用于治疗性移植的功能性人类胰岛素+细胞提供必要的基础。

 DESCRIPTION (provided by applicant): One major goal of regenerative medicine is to produce insulin secreting β-cells for transplantation therapy to treat Type 1 diabetes (T1D). Differentiation of embryonic stem cells is a powerful technology to generate functional insulin+ cells. However, teratoma formation is a serious concern with this approach. Alternatively, adult tissues could be converted into insulin+ cells in a process termed reprogramming. Reprogramming adult cells carries little risk for teratoma, but the reprogramming efficiency tends to be low and the resulting insulin+ cells have limited functionality. There is a critical ned to define the best tissue source and reprogramming method for this approach. My laboratory pioneered a reprogramming method based on a cocktail of defined genetic factors (Ngn3, Pdx1, Mafa, referred to as NPM factors). NPM factors are sufficient to convert pancreatic acinar cells to stable and functional insulin+ cells in animal models. In a comprehensive screen of adult tissues, we discovered that cells residing in the adult gastric mucosa also respond rapidly to NPM factors and generate functional insulin+ cells. Significantly, gastric insulin+ cells become glucose responsive faster than acinar-derived insulin+ cells. Thus, gastric tissue is a highly promising adult tissue source to produce functional insulin+ cells. Importantly, human gastric cells can be cultured and propagated as organoids in large numbers from cadaveric sources. Delivery of NPM factors led to formation of c-peptide+ cells in the human organoids, raising the exciting possibility of generating functional human insulin+ cells with this approach. One major aim of the proposal is to gain deeper understanding of the reprogramming process and the resulting gastric insulin+ cells. Specifically, we will determine the molecular and functional similarity of the induced gastric insulin+ cells with native islet beta-cells. We will investigate stability of the induced cells and test the hypothesis that islet structure formation will lead to heir long-term stability. We will also define the individual function of NPM factors in the reprogramming process. The second major aim of the proposal is to develop methods for efficient induction of functional insulin+ cells from human gastric tissues with combined treatment of genetic factors and signaling pathway modulators. Together, these studies will provide the necessary foundation for developing a novel technology to produce functional human insulin+ cells for therapeutic transplantation.