Inhibition of amyloidogenic Islet Amyloid Polypeptide aggregation with designed c

用设计的c抑制淀粉样蛋白形成的胰岛淀粉样蛋白多肽聚集

• 批准号: 8231140
• 负责人: David Aaron Moffet
• 金额: $ 30.05万
• 依托单位: LOYOLA MARYMOUNT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-30 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8231140
• 关键词: Adverse effects; Amino Acids; Amyloid; Atomic Force Microscopy; Bacteria; Binding; Cells; Cessation of life; Chimeric Proteins; Deposition; Development; Disease; Disease Progression; Escherichia coli; Fluorescence; Genes; Human; In Vitro; Islet Cell; Libraries; Life; Link; Mammalian Cell; Non-Insulin-Dependent Diabetes Mellitus; Onset of illness; Pancreas; Patients; Peptide Library; Peptides; Plasmids; Play; Proteins; Reporter; Role; Screening procedure; Synthetic Genes; System; Therapeutic Agents; Thioflavin T; Toxic effect; amyloid formation; combinatorial; design; enhanced green fluorescent protein; extracellular; in vivo; islet; islet amyloid polypeptide; myricetin; novel therapeutics; polypeptide; prevent; protein aggregate; protein aggregation; protein expression; protein folding; research study; self assembly; small molecule

DESCRIPTION (provided by applicant): The aggregation of the 37-amino acid polypeptide Islet Amyloid Polypeptide (IAPP, amylin), as either insoluble amyloid or as small oligomers, appears to play a direct role in the death of pancreatic ¿-islet cells in type 2 diabetes. While IAPP has been known to be the primary component of type 2 diabetes amyloid, the molecular interactions responsible for this aggregation are not fully understood. It is known that IAPP is found as extracellular deposits of amyloid in approximately 95% of patients afflicted with type 2 diabetes. IAPP has also been shown to be a toxic agent in vitro when added to human islet ¿-cells. While it remains unclear how self-assembly of IAPP leads to the development of disease, recent studies have suggested that the formation of lower order protein aggregates (two to ten self-assembled proteins) leads to cellular toxicity and ultimately to the progression of disease. Preventing the formation of these toxic oligomeric species may prevent, or slow, the progression of type 2 diabetes. We propose to use a fluorescent screen to identify peptides and small molecules that inhibit the self-assembly of amyloid-forming IAPP. In this screen, the gene for IAPP is genetically fused to the gene for enhanced green fluorescent protein (EGFP). When the IAPP-EGFP fusion protein is expressed in E. coli the natural propensity of IAPP to aggregate precludes EGFP from folding and fluorescing. However, in the presence of substances that prevent amyloid aggregation, the fused EGFP can fold properly and fluoresce green. We propose to screen designed combinatorial peptide libraries and small molecules to isolate and identify substances that inhibit amyloid formation and, therefore, show potential as therapeutic agents for preventing or slowing the progression of type 2 diabetes. PUBLIC HEALTH RELEVANCE: The aggregation of the small peptide Islet Amyloid Polypeptide (IAPP, amylin) appears to be directly related with loss of ¿-cells and the onset of type 2 diabetes. We propose to screen libraries of short peptides and small molecules to identify substances that inhibit amyloid formation. These experiments have the potential to discover new therapeutic agents for preventing, or slowing, the progression of type 2 diabetes.

描述（由应用提供）：37-氨基酸多肽淀粉样蛋白淀粉样多肽（IAPP，淀粉素）的聚集，作为不溶性淀粉样蛋白或小寡聚物，似乎在2型2型糖尿病中的胰腺细胞死亡中似乎直接作用。尽管已知IAPP是2型糖尿病的主要成分，但尚未完全了解负责该聚集的分子相互作用。众所周知，在大约95％患有2型糖尿病的患者中，IAPP被认为是淀粉样蛋白的细胞外沉积。当将IAPP添加到人类胰岛� -cells中时，IAPP在体外也被证明是一种有毒药物。尽管尚不清楚IAPP的自组装如何导致疾病的发展，但最近的研究表明，低阶蛋白聚集体的形成（2至10个自组装蛋白）会导致细胞毒性，并最终导致疾病的进展。防止这些有毒的寡聚物种的形成可能会阻止或减慢2型糖尿病的进展。我们建议使用荧光筛选来鉴定抑制淀粉样蛋白形成IAPP的自组装的肽和小分子。在此屏幕中，IAPP的基因通常与该基因融合，以增强绿色荧光蛋白（EGFP）。当在大肠杆菌中表达IAPP-EGFP融合蛋白时，IAPP的自然希望将EGFP排除在折叠和荧光中。但是，在有预防淀粉样蛋白聚集的物质的情况下，融合的EGFP可以正确折叠并荧光绿色。我们建议筛选设计的组合胡椒库和小分子，以分离和鉴定抑制淀粉样蛋白形成的物质，因此显示出潜在的治疗剂，以防止或减慢2型糖尿病的发展。 公共卫生相关性：小胡椒胰岛淀粉样多肽（IAPP，淀粉蛋白）的聚集似乎与损失 - 细胞和2型糖尿病的发作直接相关。我们建议筛选短辣椒和小分子的文库，以鉴定抑制淀粉样蛋白形成的物质。这些实验有可能发现新的治疗剂，以预防2型糖尿病的进展。

项目成果

Identification of Plant Extracts that Inhibit the Formation of Diabetes-Linked IAPP Amyloid.

• DOI: 10.1016/j.hermed.2015.11.001
• 发表时间: 2016-03
• 期刊: Journal of herbal medicine
• 影响因子: 2.3
• 作者: Fuentes AL;Hennessy K;Pascual J;Pepe N;Wang I;Santiago A;Chaggan C;Martinez J;Rivera E;Cota P;Cunha C;Nogaj LA;Moffet DA
• 通讯作者: Moffet DA

Myricetin Inhibits Islet Amyloid Polypeptide (IAPP) Aggregation and Rescues Living Mammalian Cells from IAPP Toxicity.

• DOI: 10.2174/1874091x01206010066
• 发表时间: 2012
• 期刊: The open biochemistry journal
• 作者: Zelus C;Fox A;Calciano A;Faridian BS;Nogaj LA;Moffet DA
• 通讯作者: Moffet DA

IAPP aggregation and cellular toxicity are inhibited by 1,2,3,4,6-penta-O-galloyl-β-D-glucose.

• DOI: 10.3109/13506129.2012.762761
• 发表时间: 2013-03
• 期刊: Amyloid : the international journal of experimental and clinical investigation : the official journal of the International Society of Amyloidosis
• 作者: Bruno E;Pereira C;Roman KP;Takiguchi M;Kao PY;Nogaj LA;Moffet DA
• 通讯作者: Moffet DA

Inhibition of Toxic IAPP Amyloid by Extracts of Common Fruits.

• DOI: 10.1016/j.jff.2014.12.013
• 发表时间: 2015-01-01
• 期刊: Journal of functional foods
• 影响因子: 5.6
• 作者: Kao PY;Green E;Pereira C;Ekimura S;Juarez D;Whyte T;Arhar T;Malaspina B;Nogaj LA;Moffet DA
• 通讯作者: Moffet DA