Multi-Parametric Characterization of Human Islets: Predictor of Islet Quality, Po

描述（由申请人提供）：胰岛移植已成为恢复1型糖尿病患者正常血糖控制的有希望的选择。然而，关键的生物学和体制障碍阻碍了这种治疗策略的广泛应用。其中最主要的是无法恢复足够数量的高质量胰岛，这高度依赖于胰腺的保存。其次，缺乏与移植后功能相关的快速、准确和敏感的移植前测试（即效价测定）限制了我们区分“健康”制剂与那些功能不佳的制剂的能力。此外，高材料和设备成本、对训练有素的人员的需求以及对cGMP法规的强制性遵守，使得胰岛分离技术在每个移植项目中都难以持续复制。由于这些原因，应该开发方法来提高预测胰岛制剂在移植后是否有能力在受体中发挥作用的能力，并且应该开发区域化的胰岛隔离中心网络，以最大限度地减少冷缺血时间，减少体制和财政障碍。一个成功的胰岛移植项目和分离团队已经在威斯康星大学麦迪逊分校建立。本应用程序的主要目标是在区域胰腺恢复和胰岛分布网络中实施一系列胰岛活力和功能效力的定量测量，并验证这些参数预测患者胰岛移植功能的能力。该提案的具体目标是：1)基于质量、效力和功能预测因子的多参数特征，向当地胰岛研究和移植联盟和ICR网络的成员分发优质的人类胰岛用于移植和研究。2)采用三管齐下的方法，依次优化胰腺保存、胰岛分离和分离后培养，以提高胰岛制剂的质量。使用灵敏的评估工具，我们将首先测试一种新的营养因子保存溶液的效果。我们还将开发方法，以减少在分离、培养和运输过程中对胰岛的损害，重点是通过在分离和培养基中添加抗氧化剂以及优化培养pH、氧张力和温度来减少ROS的形成和恢复“正常”代谢状态。有了这个质量改进项目，我们有信心在五年内为ICR研究人员提供超过2000万个高质量的胰岛，并促进其他胰岛移植项目的发展，为糖尿病社区服务。

DESCRIPTION (provided by applicant): Islet transplantation has emerged as a promising option for restoring normal blood sugar control in people with Type 1 diabetes. However, key biological and institutional roadblocks hamper widespread utilization of this therapeutic strategy. Primary among these is the inability to recover a sufficient number of high quality islets, which is highly dependent pancreas preservation. Second, the absence of rapid, accurate, and sensitive pre-transplant test(s) that correlate with post-transplant function (i.e. potency assays) limits our ability to distinguish "healthy" preparations from those that will function poorly. Furthermore, high materials and equipment costs, the need for highly trained personnel and compulsory compliance with cGMP regulations makes islet isolation a difficult technology to consistently reproduce in every transplant program. For these reasons, methods should be developed that improve the ability to predict whether an islet preparation has the ability to function in the recipient after transplantation, and networks of regionalized islet isolation centers should be developed to minimize cold ischemic time and reduce institutional and financial hurdles. A successful Islet Transplant Program and isolation team has been established at the University of Wisconsin-Madison. The main goal of this application is to implement a series of quantitative measures of islet viability and functional potency in a regionalized pancreas recovery and islet distribution network and validate the ability of these parameters to predict islet graft function in patients. The proposal's specific aims are: 1) To distribute superior quality human islets for transplantation and research to members of a local islet research and transplant consortium and the ICR network based on a multi-parametric characterization of predictors of quality, potency and function. 2) To improve the quality of islet preparations by taking a three-pronged approach to sequentially optimize pancreas preservation, islet isolation, and post-isolation culture. Using the sensitive assessment tools, we will first test the effect of a novel trophic factor based preservation solution. We will also develop methods to minimize damage of islets during isolation, culture, and transportation focusing focusing on the reduction of ROS formation and the recovery of a "normal" metabolic state through supplementation of anti-oxidants to the isolation and culture media and optimization of culture pH, oxygen tension, and temperature. With this quality improvement program in place, we are confident we can provide over 20 million high quality islets over a five year period to ICR investigators and facilitate the development of additional islet transplantation programs to serve the diabetes community.