Project 4

糖尿病是与糖尿病直接相关的广泛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“ER应力”（ERS）， 管腔未折叠/错误折叠蛋白质的积累。细胞对ERS的反应受 未折叠蛋白应答（UPR），并将导致ERS衰减或凋亡。ERS导致 在2型糖尿病中，当胰腺β细胞处于高胰岛素下时， 生产负荷，以维持稳定。本建议的重点是了解功能， 作为分子伴侣的可溶性78 kDa葡萄糖调节蛋白（GRP 78）的活性， 促进ER腔中的蛋白质折叠。GRP 78是UPR活性的主要调节因子，因此在调节UPR活性中起着重要作用。 在决定细胞对ERS的反应中起重要作用。为了实现这一目标，我们将利用一个新的 称为FlexHets的小分子家族，现在已知其靶向GRP 78功能。AIM的焦点 1是确定FlexHet对GRP 78活性的抑制是否增强胰岛素受体信号传导和应答。 AIM 2专注于确定FlexHet与GRP 78结合的分子基础，作为一种手段， 了解GRP 78的功能和小分子疗法的调节。AIM 3使用体内小鼠 模型，以确定GRP 78抑制是否调节肥胖与非肥胖患者的ERS应答和信号传导。 糖尿病小鼠因此，我们的方法是将一系列体外和体内方法联合收割机来开发一种 利用FlexHets作为靶向分子工具抑制GRP 78功能整体模型 功能和活动。GRP 78先前已被证明在胰岛素信号传导中发挥关键作用， 因此目前的研究旨在了解这种情况的分子基础 观察和审查GRP 78作为治疗2型糖尿病的治疗性干预的可行候选者 糖尿病

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.