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

高通量筛选以鉴定在糖尿病患者中拯救人类胰腺 β 细胞功能的化合物

• 批准号: 9905514
• 负责人: Shuibing Chen
• 金额: $ 42.38万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9905514
• 关键词: Acetylcysteine; Affect; Antidiabetic Drugs; Antioxidants; Beta Cell; Biochemical; Biological Assay; Blood Glucose; Cause of Death; Cell Death; Cell Line; Cell Proliferation; Cell physiology; Cells; Chemical Actions; Chemicals; Chronic Disease; Defect; Development; Diabetes Mellitus; Disease; Enzyme-Linked Immunosorbent Assay; Exposure to; Fatty Acids; Follow-Up Studies; Functional disorder; Future; Glucose; Goals; High Fat Diet; Human; Hyperglycemia; Impairment; In Vitro; Individual; Insulin Resistance; Libraries; Luciferases; Metabolic; Modeling; Monitor; Non-Insulin-Dependent Diabetes Mellitus; Oleates; Organ; Pathogenesis; Pharmaceutical Preparations; Population; Proinsulin; Rattus; Reporter; Rodent; Signal Transduction; Structure of beta Cell of islet; Structure-Activity Relationship; Study models; Testing; Time; Variant; Work; World Health Organization; base; biophysical techniques; cell growth regulation; cheminformatics; cytotoxicity; diabetic; drug development; drug discovery; follow-up; high throughput screening; humanized mouse; imaging modality; improved; in vivo; insulin secretion; insulinoma; islet; lead optimization; luminescence; mouse model; nano; nanoluciferase; new therapeutic target; novel; screening; small molecule; small molecule libraries

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.

摘要 糖尿病是一种多基因慢性疾病，影响全球约3.46亿人。2004年， 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年间，糖尿病导致的死亡人数将翻一番。不受控制的糖尿病导致 高血糖症，随着时间的推移导致许多器官的严重损害。超过90%的糖尿病患者 2型糖尿病（T2DM）患者。T2DM的经典发病机制涉及胰岛素抵抗 和胰腺β细胞功能障碍，其特征在于受损的葡萄糖刺激的胰岛素分泌。只有40 - 50%的 胰岛素抵抗的个体进展为T2DM。那些进步的人的关键缺陷是胰腺 β细胞不能补偿胰岛素抵抗，变得功能障碍，并最终死亡。因此，胰腺 β细胞功能障碍是从代谢损伤发展到疾病状态的关键步骤。 目前，大多数抗糖尿病药物的研发工作都集中在保护胰腺β细胞免受糖尿病的影响。 细胞死亡或诱导β细胞增殖。只有少数药物被开发出来改善胰腺β 细胞功能众所周知，同一种化合物对健康细胞和病变细胞可能具有相反的作用， 因为不同的机制可能涉及细胞生理学与病理生理学的调节。 因此，抗糖尿病药物发现的一个关键空白是缺乏拯救人类β细胞的药物。 在糖尿病状态下发挥作用。这一缺陷主要是由于缺乏高通量筛选 (HTS)直接监测人类胰腺β细胞功能的平台。 在我们的初步研究中，我们创建了一种基于发光的HTS测定法来鉴定能够拯救 人胰腺β细胞在糖尿病状态下功能。使用库完成试运行屏幕后， 超过2,000个小分子，我们确定了可能拯救受损的化合物，葡萄糖- 刺激暴露于糖脂毒性的人胰腺β细胞的胰岛素分泌。在此，我们建议 使用我们建立的平台在多个化学库上进行大规模HTS， 约240，000种化合物，使用后续分析验证命中化合物，通过以下方法优化先导化合物 简单的构效关系分析，并评价所鉴定的化学探针在 糖尿病人源化小鼠模型。最后，我们将进行目标识别和研究的机制， 化学探针的作用。所鉴定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。