GAD65 release in autoimmune diabetes

自身免疫性糖尿病中 GAD65 的释放

• 批准号: 6707177
• 负责人: STEVEN D CHESSLER
• 金额: $ 7.58万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-01 至 2004-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6707177
• 关键词: autoantibody; clinical research; disease /disorder onset; gamma aminobutyrate; glutamate decarboxylase; human tissue; insulin dependent diabetes mellitus; laboratory rat; mass spectrometry; pancreatic islets; serum; technology /technique development; yeast two hybrid system

DESCRIPTION (provided by applicant): This application is submitted in response to an invitation to apply for funds under the Small Grant Program for NIDDK K08/K23 Recipients (PAR-01-066). If awarded, this RO3 grant would provide supplemental funding during the final two years of the applicant's K08 award (DK02944). The additional funding would provide crucial resources, such as technical assistance, that would enhance the applicant's ability to compete for further grant support and would facilitate the transition to full independence. This application proposes two lines of investigation which will build upon and extend ongoing studies of the cell biology of glutamic acid decarboxylase (GAD) in the pancreatic islets. I have found that, surprisingly, the vesicular GABA transporter VIAAT is absent from rat beta cells and human islets. This suggests that there is an as yet unidentified GABA transporter which, I hypothesize, interacts specifically with GAD65. With assistance from the laboratories of Drs. Jay Heinecke and Stanley Fields, I will seek to identify beta-cell proteins that interact with GAD65 and that may therefore influence its trafficking and perhaps its discharge from injured islets. The impetus for the second line of investigation is the importance of detecting GAD65 release in vivo together with my success in developing a sensitive, prototype, blood GAD65 assay. Damage to tissues such as myocardium and liver can be detected and monitored by serum assays for discharged proteins. In contrast, metabolic tests reveal only extensive islet damage after the fact. I hypothesize that a plasma assay for GAD65 will provide a means to detect and monitor ongoing beta-cell damage in individuals with diabetes and asymptomatic insulitis and in animal models of autoimmune diabetes. Such a test would be invaluable for a variety of applications, including investigating the pathogenesis of islet injury and autoimmunity, helping identify candidates for preventative therapies and monitoring transplanted islets for rejection. My goal is to complete the development and validation of a highly sensitive, plasma assay for GAD65 and to conduct a preliminary analysis of plasma GAD65 levels in control and diabetic subjects and in BB rats prior to and after the onset of diabetes. Data generated by these studies will form the basis of further investigations of the intracellular trafficking of GAD65 as well as of in vivo studies of the time-course, triggers and clinical utility of monitoring GAD65 discharge.

描述（由申请人提供）： 本申请是应NIDDK K 08/K23受助人小额赠款计划（PAR-01-066）的邀请提交的。如果授予，此RO 3赠款将在申请人的K 08奖（DK 02944）的最后两年提供补充资金。额外的资金将提供关键资源，如技术援助，这将提高申请者争取更多赠款支持的能力，并将促进向完全独立过渡。本申请提出了两条调查线，这将建立在和扩展正在进行的研究的细胞生物学的谷氨酸脱羧酶（GAD）在胰岛。令人惊讶的是，我发现囊泡GABA转运蛋白VIAAT在大鼠β细胞和人类胰岛中不存在。这表明，有一个尚未确定的GABA转运蛋白，我假设，特别是与GAD 65相互作用。在Jay Heinecke和Stanley Fields博士实验室的协助下，我将试图鉴定与GAD 65相互作用的β细胞蛋白，从而可能影响其运输，并可能影响其从受损胰岛中的排出。第二线研究的动力是检测体内GAD 65释放的重要性，以及我成功开发了一种灵敏的原型血液GAD 65测定法。对组织如心肌和肝脏的损伤可以通过对排出的蛋白质的血清测定来检测和监测。相比之下，代谢测试只显示广泛的胰岛损伤后的事实。我假设GAD 65的血浆测定将提供一种检测和监测糖尿病和无症状胰岛炎个体以及自身免疫性糖尿病动物模型中持续β细胞损伤的方法。这样的测试将是非常宝贵的各种应用，包括研究胰岛损伤和自身免疫的发病机制，帮助确定候选人的预防性治疗和监测移植胰岛排斥反应。我的目标是完成一个高灵敏度的GAD 65血浆检测的开发和验证，并进行对照和糖尿病受试者和BB大鼠糖尿病发作前后的血浆GAD 65水平的初步分析。这些研究产生的数据将形成进一步调查的细胞内贩运的GAD 65以及在体内研究的时间过程，触发器和监测GAD 65放电的临床效用的基础。