A High Content Chemical Screen to Identify the Drug Candidates Promoting Human Beta Cell Proliferation

抽象的。 全世界约有3.46亿人患有糖尿病。2004年，估计有340万人死于 高血糖的后果。世界卫生组织预计糖尿病死亡人数将增加一倍 在2005年到2030年之间。1型糖尿病(T1D)和2型糖尿病(T2D)都是由于 人胰腺β细胞团。在诊断为高血糖后，仍有残留的β细胞 在糖尿病患者中检测到。人胰岛β细胞团的再生将提供一种新的途径 用于糖尿病治疗。在健康的成年人中，β细胞团保持在缓慢的增殖率。然而，成人 在某些情况下，如肥胖、怀孕等，胰岛β细胞质量增加。因此，容易和 促进胰岛β细胞增殖的强健方法可作为治疗糖尿病的新方法 病人。 细胞通透性化合物为控制细胞增殖提供了一种方便有效的方法。化学制品 筛查已被应用于确定胰腺β细胞扩增的化学诱导剂。然而，许多人 利用啮齿动物或动物模型鉴定的化合物对原始人几乎没有或缓和了活性 小岛。此外，以前的研究大多使用健康的胰岛样本。目前尚不清楚被确认的 化合物可促进T1D或T2D胰岛的增殖。在我们之前的研究中，我们已经建立了几个 用于鉴定促进人类生成、生存和功能的化合物的化学筛选平台 多能干细胞(HPSC)衍生的β样细胞。在这里，我们建议使用hPSC衍生的类β细胞来 筛选促进人胰腺β细胞复制的化合物，并验证已鉴定的 使用人类健康、T1D、T2D胰岛的化合物。 在初步研究中，我们已经筛选了1200多种化合物，并确定了可能存在的 促进hPSC来源的胰岛β样细胞的增殖。一些受攻击的化合物进一步 已确认使用了主要的人类胰岛。在这里，我们建议将现有的筛查平台扩展到大型 使用后续化验对“化学探针”的清单进行标度和识别。最后，我们将研究其作用机制。 化学探头。所鉴定的化学探针可直接用于抗糖尿病药物的开发。 或被用作发现未来药物发现的新靶点的探针。为了实现这些目标，有三个目标 包括1)进行高含量的筛选，以鉴定促进人类健康的化合物 胰岛β细胞增殖；2)进行后续分析，以最终确定“化学探针”清单；3)研究 化合物的作用机理。

Abstract. Diabetes affects approximately 346 million people worldwide. In 2004, an estimated 3.4 million people died from the consequence of high blood glucose. The World Health Organization projects that diabetes death will double between 2005 and 2030. Both type 1 diabetes (T1D) and type 2 diabetes (T2D) result from the decrease of human pancreatic beta cell mass. After the diagnosis of high blood glucose, there are still residual beta cells detected in diabetes patients. The regeneration of human pancreatic beta cell mass will provide a novel approach for diabetes therapy. In healthy adult beta cell mass is maintained at a slow proliferation rate. However, adult pancreatic beta cell mass increases in certain conditions, such as obesity, pregnancy, etc. Thus, an easy and robust method to enhance pancreatic beta cell proliferation can be used as a new approach to treat diabetes patients. Cell-permeable compounds provide a convenient and efficient approach to control cell proliferation. Chemical screens have been applied to identify chemical inducers of pancreatic beta cell expansion. However, many compounds identified using rodents or animal models show little or moderator activities on primary human islets. In addition, most of the previous studies used healthy islet samples. It is not clear whether the identified compounds improve proliferation of T1D or T2D islets. In our previous studies, we have established several chemical screen platforms to identify the compounds promoting the generation, survival and function of human pluripotent stem cell (hPSC)-derived beta-like cells. Here, we propose to use hPSC-derived beta-like cells to screen for compounds that promote human pancreatic beta cell replication, and to validate the identified compounds using human healthy, T1D, T2D islets. In the preliminary studies, we have screened more than 1,200 compounds and identified hits that potentially increase the proliferation of hPSC-derived pancreatic beta-like cells. Some of hit compounds were further confirmed using primary human islets. Here, we proposed to expand the established screening platform to large scale and identify a list of “chemical probe” using follow-up assays. Finally, we will study the mechanism of action of the chemical probes. The identified chemical probes can be directly used for anti-diabetes drug development or be used as the probes to discover novel targets for future drug discovery. To reach these goals, three aims were proposed, including 1) Carry out a high content screen to identify the compounds promoting human pancreatic beta cell proliferation; 2) Perform follow-up assays to finalize the “chemical probe” list; 3) Study the mechanism of action of the compounds.