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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的糖尿病是一种多基因慢性疾病，影响全球约 3.46 亿人。 2004年，一个据估计，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，糖尿病造成的死亡人数将翻一番。不受控制的糖尿病会导致高血糖，随着时间的推移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 (T2DM)。 T2DM 的典型发病机制涉及胰岛素抵抗以及胰腺β细胞功能障碍，其特征是葡萄糖刺激的胰岛素分泌受损。仅 40-50% 患有胰岛素抵抗的个体会进展为 T2DM。那些取得进步的人的关键缺陷是胰腺 β细胞无法补偿胰岛素抵抗，变得功能失调，并最终死亡。因此，胰 β细胞功能障碍是从代谢障碍发展到疾病状态的关键步骤。目前抗糖尿病药物发现的大部分努力都集中在保护胰腺β细胞免受细胞死亡或诱导β细胞增殖。仅开发了少数药物来改善胰腺β 细胞功能。众所周知，同一种化合物可能对健康细胞和患病细胞产生相反的作用，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差距是缺乏拯救人类β细胞的药物在糖尿病状态下发挥作用。这种缺陷主要是由于缺乏高通量筛选（HTS）平台直接监测人类胰腺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 测定法来识别可拯救的化合物人类胰腺β细胞在糖尿病状态下的功能。使用库完成试点屏幕后超过 2,000 种小分子，我们鉴定出可能拯救受损葡萄糖的化合物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执行大规模 HTS，其中包含约 240,000 种化合物，使用后续测定验证命中化合物，通过以下方式优化先导化合物简单的结构活性关系分析，并评估已识别化学探针的功能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针的作用。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。