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Project 4

项目4

基本信息

批准号：
8692938
负责人：
FRANKLIN Alan HAYS
金额：
$ 21.08万
依托单位：
UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8692938
关键词：
Affinity Amino Acids Animal Model Apoptosis Apoptotic Applications Grants Attenuated Binding Biological Preservation C-terminal Cell Death Cell Survival Cells Centers of Research Excellence Chemopreventive Agent Clinical Treatment Crystallization Data Diabetes Mellitus Disease Dissociation Dose Drug Targeting Endoplasmic Reticulum Essential Drugs Family Future GRP78 gene Glucose Hepatocyte Heteroarotinoids Homeostasis In Vitro Inbred NOD Mice Instruction Insulin Insulin Receptor Knockout Mice Malignant neoplasm of ovary Mentors Methods Modeling Molecular Molecular Chaperones Molecular Structure Molecular Target Monitor National Institute of General Medical Sciences Non-Insulin-Dependent Diabetes Mellitus Obesity Oklahoma Pancreas Pathology Pathway interactions Peripheral Pharmaceutical Preparations Phase I Clinical Trials Play Positioning Attribute Prevention Production Protein Biosynthesis Proteins Receptor Signaling Regulation Relative (related person)Research Role Solutions Stress Stress Response Signaling Structure Testing Therapeutic Therapeutic Intervention Tissues Translations United States National Institutes of Health Yeasts attenuation base biological adaptation to stress biophysical techniques diabetic drug development endoplasmic reticulum stress expectation flexibility human disease in vivo insight insulin signaling molecular chaperone GRP78 mouse model new therapeutic target novel overexpression polyhistidine protein degradation protein folding protein misfolding response small molecule structural biology tissue culture tool

项目摘要

Diabetes is one of a broad range of human diseases and disorders that are directly associated with endoplasmic reticulum (ER) malfunction. This malfunction is termed "ER stress" (ERS) and results from the accumulation of luminal unfolded/misfolded proteins. Cellular response to ERS is regulated by the Unfolded Protein Response (UPR) and will result in either ERS attenuation or apoptosis. ERS leading to cell death is often observed in type 2 diabetes when pancreatic p-cells are placed under high insulin production loads to maintain euglycemia. This proposal is focused on understanding the function and activity of the soluble 78 kDa Glucose-Regulated protein (GRP78) that serves as a molecular chaperone to facilitate protein folding in the ER lumen. GRP78 is a master regulator of UPR activity and thus plays a significant role in determining cellular response to ERS. To accomplish this objective we will utilize a novel family of small molecules called FlexHets that are now known to target GRP78 function. The focus in AIM 1 is to determine if FlexHet inhibition of GRP78 activity enhances insulin receptor signaling and response. AIM 2 is focused on determining the molecular basis for FlexHet binding to GRP78 as a means to understand GRP78 function and regulation by small molecule therapeutics. AIM 3 uses in vivo mouse models to determine if GRP78 inhibition modulates ERS response and signaling in obese vs. non-obese diabetic mice. Thus, our approach is to combine a range of in vitro and in vivo methods to develop a holistic model of GRP78 function by utilizing FlexHets as a targeted molecular tool to inhibit GRP78 function and activity. GRP78 has been previously shown to play a key role in insulin signaling using heterozygous knockout mice so the current studies are aimed at understanding the molecular basis for this observation and vetting GRP78 as a viable candidate for therapeutic intervention in the treatment of type 2 diabetes.

糖尿病是与多种疾病直接相关的人类疾病和病症之一内质网（ER）功能障碍。这种故障被称为“内质网应激”(ERS)，是由于管腔未折叠/错误折叠蛋白质的积累。细胞对 ERS 的反应受以下因素调节未折叠蛋白反应 (UPR) 将导致 ERS 减弱或细胞凋亡。 ERS 导致当胰腺 p 细胞处于高胰岛素条件下时，2 型糖尿病中经常观察到细胞死亡维持血糖正常的生产负荷。该提案的重点是了解功能和可溶性 78 kDa 葡萄糖调节蛋白 (GRP78) 的活性，该蛋白作为分子伴侣促进内质网腔内的蛋白质折叠。 GRP78 是 UPR 活性的主要调节因子，因此发挥着重要作用在确定细胞对 ERS 的反应中发挥着重要作用。为了实现这一目标，我们将利用一种新颖的称为 FlexHets 的小分子家族目前已知以 GRP78 功能为目标。 AIM 的焦点图1是确定FlexHet对GRP78活性的抑制是否增强胰岛素受体信号传导和反应。 AIM 2 的重点是确定 FlexHet 与 GRP78 结合的分子基础，以此作为了解 GRP78 的功能和小分子疗法的调节。 AIM 3使用活体鼠标确定 GRP78 抑制是否调节肥胖与非肥胖人群的 ERS 反应和信号传导的模型糖尿病小鼠。因此，我们的方法是结合一系列体外和体内方法来开发利用 FlexHets 作为抑制 GRP78 的靶向分子工具，建立 GRP78 功能的整体模型功能和活动。 GRP78 先前已被证明在胰岛素信号传导中发挥关键作用杂合基因敲除小鼠，因此当前的研究旨在了解其分子基础观察和审查 GRP78 作为 2 型治疗干预的可行候选者糖尿病。