Design and use of methods for peptide secretion studies

2型糖尿病的一个特征是朗格汉斯胰岛的β细胞胰岛素分泌受损。 1型糖尿病的特征是对β细胞的自身免疫攻击和随后的胰岛素丢失 分泌物。治疗1型糖尿病的一种有希望的治疗方法是胰岛移植；然而，这种治疗方法 目前由于无法充分评估用于移植的胰岛而受到限制。对小岛有更多的了解 而胰岛素的分泌在确定糖尿病的根本原因和发展中是至关重要的 潜在的治疗方法。对胰岛的研究严重依赖于能够检测到其 生化和生理动力学。 这项工作的总体目标是开发适合于测量化学物质的先进仪器 与朗格汉斯胰岛分泌肽有关的事件。我们最初的两个目标是开发微流体 可以在同时监测激素和代谢物的同时培养单个胰岛的工具 分泌和新陈代谢。为了能够研究细胞与细胞之间的相互作用，微流控芯片将整合一种 微循环系统，允许两种不同类型的细胞在 测量。我们的第三个目标是开发一种代谢组学方法，用于同时测量极点 已知与葡萄糖刺激的胰岛素分泌密切相关的阴离子化合物。该方法将 基于多维分离和质谱学检测。我们的最终目标是使用这些 合作研究中的方法，包括鉴定可用于移植的胰岛 目标是确定测量是否对胰岛评估有用并确定 胰岛生理的关键基因敲除。这些方法将为研究根提供新的机会。 糖尿病的原因和设计新的治疗方法。 版面总结。糖尿病是一个巨大的医疗问题，至少困扰着800万人， 2002年在美国花费了超过1000亿美元。糖尿病的特征是细胞胰岛素分泌受损。 这些星团被称为朗格汉斯岛。我们的研究将产生新的仪器，这将使 对与正常和疾病的胰岛功能相关的化学变化的研究前所未有的详细。 对这种化学变化的评估有望帮助确定新的治疗靶点。

A hallmark of Type 2 diabetes is impaired insulin secretion from beta cells of the islets of Langerhans. Type 1 diabetes is characterized by autoimmune attack on beta cells and subsequent loss of insulin secretion. A promising treatment for type 1 diabetes is transplant of islets; however, this treatment is presently limited by inability to adequately assess islets used in transplant. Greater understanding of islets and insulin secretion is of fundamental importance in determining the root causes of diabetes and developing potential treatments. Studies of islets are critically dependent upon instrumentation that can detect their biochemical and physiological dynamics. The overall objective of this work is to develop advanced instrumentation suitable for measuring chemical events related to peptide secretion from islets of Langerhans. Our first two aims are to develop microfluidic tools that will allow single islets to be cultured while simultaneously monitoring hormone and metabolite secretion and metabolism. To allow cell-cell interactions to be studied, the microfluidic chip will incorporate a microcirculatory system that allows two different cell types to be cultured and interact during the measurements. Our third aim is to develop a metabolomic method for simultaneous measurement of polar anionic compounds known to be critically involved in glucose-stimulated insulin secretion. The method will be based on multi-dimensional separations and mass spectrometry detection. Our final aim is to use these methods in collaborative studies including characterization of islets that may be used for transplant with the goal of determining if the measurements may be useful for islet assessment and determining the effect of key gene knock-outs on islet physiology. These methods will offer new opportunities for studying the root causes of diabetes and devising novel treatments. LAY SUMMARY. Diabetes is an enormous medical problem afflicting at least 8 million people and costing over $100 billion in 2002 in the US. Diabetes is characterized by impaired insulin secretion from cell clusters known as islets of Langerhans. Our research will result in novel instrumentation that will allow studies of the chemical changes associated with normal and diseased islet function in unprecedented detail. Assessment of such chemical changes is expected to help identify novel therapeutic targets.