A High Throughput Screening to Identify Compounds Rescuing Human Pancreatic Beta Cell Function in Diabetic Conditions

摘要 糖尿病是一种多基因慢性疾病，全世界约有3.46亿人受到影响。在2004年，一个 估计有340万人死于高血糖的后果。世界卫生组织 预计2005年至2030年期间，糖尿病造成的死亡人数将翻一番。失控的糖尿病导致 高血糖，随着时间的推移，这会导致许多器官的严重损害。90%以上的糖尿病患者 人群患有2型糖尿病(T2 DM)。2型糖尿病的经典发病机制与胰岛素抵抗有关 以及胰岛β细胞功能障碍，以葡萄糖刺激的胰岛素分泌受损为标志。只有40%-50%的 胰岛素抵抗的个体进展为T2 DM。进展者的关键缺陷是胰腺 β细胞不能补偿胰岛素抵抗，变得功能失调，最终死亡。因此，胰腺 β细胞功能障碍是从代谢障碍发展到疾病状态的关键一步。 目前大多数抗糖尿病药物的发现都集中在保护胰岛β细胞 细胞死亡或诱导β细胞增殖。只有几种药物被开发出来改善胰岛β细胞。 细胞功能。众所周知，相同的化合物对健康和患病的细胞可能具有相反的作用， 因为细胞生理学与病理生理学的调节可能涉及不同的机制。 因此，抗糖尿病药物发现的一个关键缺口是缺乏拯救人类β细胞的药物 在糖尿病的情况下发挥作用。这一缺陷主要是由于缺乏高通量筛查。 (HTS)平台，用于直接监测人类胰岛β细胞功能。 在我们的初步研究中，我们创建了一种基于发光的HTS分析来识别能够拯救 人的胰岛β细胞在糖尿病状态下的功能。使用库完成引导屏幕后， 超过2,000个小分子，我们确定了可能拯救受损的葡萄糖的命中化合物- 糖脂毒性所致的人胰岛β细胞刺激胰岛素分泌。在此，我们建议 使用我们建立的平台在包含以下内容的多个化学库上执行大规模HTS ~240,000种化合物，通过后续分析验证命中化合物，通过 简单的构效关系分析，并对所鉴定的化学探针的功能进行评价 糖尿病人源化小鼠模型。最后，我们将进行目标识别并研究其机制。 化学探头的作用。鉴定出的化学探针可直接用于抗糖尿病药物 开发或用于发现未来药物发现的新疾病靶点。

Abstract Diabetes is a polygenetic and chronic disease affecting approximately 346 million people worldwide. In 2004, an estimated 3.4 million people died from the consequences of high blood glucose. The World Health Organization projects that deaths caused by diabetes will double between 2005 and 2030. Uncontrolled diabetes results in hyperglycemia, which over time leads to serious damage to many organs. More than 90% of the diabetic population has type 2 diabetes mellitus (T2DM). The classic pathogenesis of T2DM involves insulin resistance and pancreatic beta cell dysfunction, marked by impaired, glucose-stimulated insulin secretion. Only 40-50% of individuals with insulin resistance progress to T2DM. The pivotal defect in those who progress is that pancreatic beta cells fail to compensate for insulin resistance, become dysfunctional, and eventually die. Thus, pancreatic beta cell dysfunction is a key step in the progression from metabolic impairments to a disease state. Most of the current efforts on anti-diabetes drug discovery focus on either protecting pancreatic beta cells from cell death or inducing beta cell proliferation. Only a few drugs have been developed to improve pancreatic beta cell function. It is well known that the same compound may have opposite effects on healthy and diseased cells, since different mechanisms may be involved in the regulation of cellular physiology versus pathophysiology. Therefore, a critical gap in anti-diabetes drug discovery is the lack of drugs rescuing human beta cell function in a diabetic condition. This deficit is mainly due to the lack of a high-throughput screening (HTS) platform to directly monitor human pancreatic beta cell function. In our preliminary studies, we created a luminescence-based HTS assay to identify compounds that rescue human pancreatic beta cell function in a diabetic condition. After completing a pilot screen using a library with more than 2,000 small molecules, we identified hit compounds that potentially rescue the impaired, glucose- stimulated insulin secretion of human pancreatic beta cells exposed to glucolipotoxicity. Here, we propose to perform a large-scale HTS using our established platform on multiple chemical libraries containing ~240,000 compounds, validate the hit compounds using follow-up assays, optimize lead compounds through simple structure activity relationship analyses, and evaluate the function of the identified chemical probes in diabetic-humanized mouse models. Finally, we will perform target identification and study the mechanism of action of the chemical probes. The identified chemical probes can be directly used for anti-diabetes drug development or serve to discover novel disease targets for future drug discovery.