Cytoprotection and the mechanism of action of a natural product Khellin against ER stress

天然产物 Khellin 对抗 ER 应激的细胞保护和作用机制

• 批准号: 9974514
• 负责人: Weidong Wang
• 金额: $ 36.25万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9974514
• 关键词: ATF6 gene; Affect; Ammi visnaga; Animal Model; Animals; Asthma; Atherosclerosis; Attenuated; B-Lymphocytes; Beta Cell; Biochemical; Biological Assay; Cause of Death; Cell Death; Cell model; Cells; Cellular Assay; Cessation of life; Chemicals; Chronic; Coronary heart disease; Cytoprotection; Cytoprotective Agent; Degenerative Disorder; Development; Diabetes Mellitus; Diabetic mouse; Dimerization; Disease; Drug Targeting; Eating; Endoplasmic Reticulum; Event; Exhibits; Foundations; Functional disorder; Grant; Homeostasis; Hyperglycemia; Insulin; Islet Cell; Islets of Langerhans Transplantation; Lead; Mediating; Membrane; Messenger RNA; MicroRNAs; Modeling; Molecular; Molecular Mechanisms of Action; Mus; Natural Products; Neurodegenerative Disorders; Obesity; Organ; Pancreas; Pathogenesis; Pathway interactions; Patients; Peripheral; Pharmaceutical Preparations; Pharmacotherapy; Phosphorylation; Phosphotransferases; Pilot Projects; Plant Extracts; Play; Process; Proteins; Public Health; RNA Decay; RNA Degradation; RNA Splicing; Research; Research Design; Ribonucleases; Role; Severities; Streptozocin; Structure of beta Cell of islet; Technology; Testing; Therapeutic; Time; Transplantation; Weight Gain; Work; analog; base; cell type; clinical development; db/db mouse; diabetic; dimer; endoplasmic reticulum stress; high throughput screening; human disease; improved; in vivo; insight; insulin secretion; insulin sensitivity; misfolded protein; novel; overexpression; response; small molecule; therapeutic target

Endoplasmic reticulum (ER) stress plays an important role in the pathogenesis of a growing list of human diseases, including diabetes, obesity, atherosclerosis, and neurodegenerative diseases. Chronic ER stress leads to cell dysfunction and death through the hyperactivation of the unfolded protein response (UPR), and hence has been proposed as a therapeutic target for the treatment of these diseases. However, no drugs targeting ER stress/UPR-induced cell dysfunction and death have yet been identified. Using high throughput screen technology, we have identified a natural product, a furanochromone derivative, as a molecule of cytoprotection against ER stress. Our preliminary studies revealed that (a) in cell-based assays, the furanochromone derivative protects β cells against ER stress-, glucotoxicity-, and lipotoxicity-induced dysfunction and death, (b) the furanochromone ameliorates hyperglycemia and protects the function and survival of β cells in streptozotocin-induced diabetic animals, and (c) the furanochromone also protects other cell types against ER stress. These studies revealed for the first time that this natural product exhibits cytoprotection against ER stress. Our pilot studies further indicate that the furanochromone derivative selectively inhibits the ER stress-induced activation of one of three unfolded protein response pathways, IRE1α pathway, with no effect on the other two pathways: PERK and ATF6. These findings led to our central hypothesis that this compound inhibits ER stress-induced IRE1α hyperactivation to confer cytoprotection. In this grant, we propose three aims to test this hypothesis. In aim 1, we will determine the mechanism of action of this compound on IRE1α inhibition. We will use biochemical assays to determine the effects of the furanochromone derivative on IRE1a kinase and RNase activities and its phosphorylation and dimeric/oligomeric statuses. In aim 2, we will determine whether the compound confers cytoprotective activity by inhibiting IRE1α. These studies will establish its inhibition on IRE1α activation as the molecular mechanism of the compound’s cytoprotection. Finally, we will determine therapeutic potential of the compound in well-established ER stress-related animal models: two diabetes models of progressive β cell loss (Akita mice and db/db mice). Together, this work will reveal not only a novel cytoprotective activity of the natural product against ER stress but also elucidate its inhibition of IRE1α activity as the molecular mechanism of action underlying its cytoprotection, thus establishing the foundation for the clinical development of the furanochromone derivatives as novel cytoprotective drugs for ER stress-related diseases.

内质网应激在越来越多的人类疾病的发病机制中起着重要作用，包括糖尿病、肥胖、动脉粥样硬化和神经退行性疾病。慢性内质网应激通过未折叠蛋白反应（UPR）的过度激活导致细胞功能障碍和死亡，因此已被提出作为治疗这些疾病的治疗靶点。然而，目前还没有发现针对内质网应激/ upr诱导的细胞功能障碍和死亡的药物。利用高通量筛选技术，我们已经鉴定出一种天然产物，呋喃铬酮衍生物，作为细胞保护分子对抗内质网应激。我们的初步研究显示：(a)在细胞基础实验中，呋喃醌衍生物保护β细胞免受内质网应激、糖毒性和脂毒性诱导的功能障碍和死亡，(b)呋喃醌改善高血糖，保护β细胞的功能和存活，以及(c)呋喃醌还保护其他类型的细胞免受内质网应激。这些研究首次揭示了这种天然产物对内质网应激具有细胞保护作用。我们的前期研究进一步表明，呋喃醌衍生物选择性地抑制内质网应激诱导的三种未折叠蛋白反应途径之一IRE1α途径的激活，而对另外两种途径PERK和ATF6没有影响。这些发现导致了我们的中心假设，即该化合物抑制内质网应激诱导的IRE1α过度激活，从而赋予细胞保护作用。在这项研究中，我们提出了三个目标来检验这一假设。在目的1中，我们将确定该化合物对IRE1α抑制的作用机制。我们将使用生化分析来确定呋喃醌衍生物对IRE1a激酶和RNase活性及其磷酸化和二聚体/低聚体状态的影响。在目的2中，我们将确定该化合物是否通过抑制IRE1α而具有细胞保护活性。这些研究将确定其对IRE1α活化的抑制作用是其细胞保护作用的分子机制。最后，我们将在已建立的内质网应激相关动物模型中确定该化合物的治疗潜力：两种进行性β细胞丢失的糖尿病模型（秋田小鼠和db/db小鼠）。总之，本研究将揭示天然产物对内质网应激的一种新的细胞保护活性，并阐明其抑制IRE1α活性作为其细胞保护作用的分子机制，从而为呋喃醌衍生物作为内质网应激相关疾病的新型细胞保护药物的临床开发奠定基础。