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

描述(申请人提供)：胰岛移植已经成为恢复1型糖尿病患者正常血糖控制的一种有前途的选择。然而，关键的生物学和体制障碍阻碍了这一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(即效价分析)相关的快速、准确和灵敏的移植前检测(S)，限制了我们区分“健康的”制剂和功能较差的制剂的能力。此外，高昂的材料和设备成本，对训练有素的人员的需要，以及强制遵守cGMP法规，使得胰岛分离成为一项难以在每个移植计划中持续复制的技术。出于这些原因，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者身上发挥作用的能力，并且应该发展地区化的胰岛隔离中心网络，以最大限度地减少冷缺血时间，减少制度和经济障碍。威斯康星大学麦迪逊分校已经建立了一个成功的胰岛移植计划和隔离团队。这项应用的主要目标是在区域化的胰腺恢复和胰岛分布网络中实施一系列胰岛活性和功能潜力的定量测量，并验证这些参数预测患者胰岛移植功能的能力。该建议的具体目标是：1)根据质量、效力和功能预测因素的多参数特征，向当地胰岛研究和移植联盟和ICR网络的成员分发用于移植和研究的优质人体胰岛。2)从胰岛保存、胰岛分离、胰岛分离后培养三个方面依次进行优化，提高胰岛制备质量。使用敏感的评估工具，我们将首先测试一种基于营养因子的新型保存液的效果。我们还将开发在分离、培养和运输过程中尽量减少胰岛损害的方法，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值、氧分压和温度来减少ROS的形成和“正常”代谢状态的恢复。有了这一质量改进计划，我们有信心在五年内向ICR研究人员提供超过2000万个高质量的胰岛，并促进开发更多的胰岛移植计划，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.