MULTI-PARAMETRIC CHARACTERIZATION OF HUMAN ISLETS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 研究中心，而研究中心不一定是研究者所在的机构。 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一种有前途的选择。然而，关键的生物学和制度障碍阻碍了这种治疗策略的广泛应用。其中主要的是不能恢复足够数量的高质量胰岛，这是高度依赖于胰腺保存。其次，缺乏与移植后功能相关的快速，准确和灵敏的移植前测试（即效价测定）限制了我们区分“健康”制剂和功能不佳制剂的能力。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成为在每个移植程序中一致地再现的困难技术。由于这些原因，应该开发提高预测胰岛制剂在移植后是否具有在受体中发挥功能的能力的方法，并且应该开发区域化胰岛分离中心的网络以最小化冷缺血时间并减少机构和财政障碍。一个成功的胰岛移植计划和隔离团队已经在威斯康星大学麦迪逊分校成立。本申请的主要目标是在区域化胰腺恢复和胰岛分布网络中实施胰岛活力和功能效力的一系列定量测量，并验证这些参数预测患者胰岛移植物功能的能力。该提案的具体目标是：1）根据质量、效力和功能预测因素的多参数表征，将用于移植和研究的上级质量人类胰岛分配给当地胰岛研究和移植联盟以及ICR网络的成员。2)从胰腺保存、胰岛分离、分离后培养三方面入手，优化胰岛制备工艺，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新的营养因子为基础的保存解决方案的效果。我们还将开发在分离、培养和运输过程中最大限度地减少胰岛损伤的方法，重点关注通过向分离和培养介质中补充抗氧化剂以及优化培养pH值、氧张力和温度来减少ROS形成和恢复“正常”代谢状态。随着这项质量改进计划的实施，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计划的发展，以服务于糖尿病社区。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.