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

人类胰岛的多参数表征：胰岛质量的预测因子 Po

• 批准号: 7500425
• 负责人: LUIS Alberto FERNANDEZ
• 金额: $ 4.73万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-30 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7500425
• 关键词: ABCC1 gene; Activities of Daily Living; Adenine; Antioxidants; Apoptosis; Area Under Curve; Biological; Biological Assay; Biological Preservation; Blood Glucose; Canis familiaris; Cell Extracts; Cells; Characteristics; Chemiluminescence assay; Clinical; Collaborations; Communities; Comparative Study; Complex; Computers; Count; Cryopreservation; Culture Media; Cultured Cells; Cyclic GMP; Data Collection; Development; Diabetes Mellitus; Documentation; Drug Formulations; Ductal; Electron Transport; Elements; Environment; Enzyme-Linked Immunosorbent Assay; Equipment; FADH2; Flow Cytometry; Fluorescein-5-isothiocyanate; Fluorocarbons; Gene Expression; Genes; Genetic Transcription; Glucokinase; Glucose; Glucose Transporter; Goals; High Pressure Liquid Chromatography; Homeodomain Proteins; Hour; Human; Human Resources; Hypoxia; In Vitro; Incubated; Injury; Insulin; Insulin-Dependent Diabetes Mellitus; Ischemia; Islet Cell; Islets of Langerhans; Islets of Langerhans Transplantation; Kidney; Kidney Transplantation; Kinetics; Leukocytes; Luminol; Measurement; Measures; Mediating; Membrane Potentials; Metabolic; Methods; Mitochondria; Mitogen-Activated Protein Kinases; Modeling; Monitor; Necrosis; Network-based; Numbers; Organ; Outcome; Oxidative Stress; Oxygen measurement, partial pressure, arterial; Pancreas; Pancreas Transplantation; Pancreatic Polypeptide; Patients; Phase; Phosphorylation; Ploidies; Preparation; Procedures; Production; Quality Control; Reactive Oxygen Species; Recovery; Regulation; Reporting; Research; Research Personnel; Risk; Rotenone; S pombe hsp9 protein; SCID Mice; Sampling; Score; Series; Signal Pathway; Solutions; Staining method; Stains; Standards of Weights and Measures; Sterility; Streptozocin; Stress; Supplementation; Techniques; Technology; Temperature; Testing; Therapeutic; Time; Training; Translating; Transplantation; Transportation; United States; United States Food and Drug Administration; Universities; University of Wisconsin-lactobionate solution; Vascular Endothelial Growth Factors; Western Blotting; Wisconsin; annexin A5; base; capsule; cost; design; diabetic; glucose metabolism; graft function; improved; in vivo; indexing; inhibitor/antagonist; insulin promoter factor 1; insulin secretion; islet; member; mitochondrial membrane; novel; particle; programs; pyridine nucleotide; quality assurance; response; tool

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.

胰岛移植已成为恢复正常血糖控制的一种有前途的选择 患有1型糖尿病然而，关键的生物和体制障碍阻碍了广泛利用 这种治疗策略。其中主要是无法恢复足够数量的高质量 胰岛，这是高度依赖胰腺保存。第二，缺乏快速、准确、 与移植后功能相关的敏感的移植前测试（即效价测定）限制了我们的 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，需要训练有素的人员和强制遵守cGMP 法规使得胰岛分离成为在每个移植程序中一致再现的困难技术。 由于这些原因，应该开发提高预测胰岛是否受损的能力的方法。 制剂具有在移植后在受体中发挥功能的能力， 应发展隔离中心，以最大限度地减少冷缺血时间， 障碍。牛津大学成功建立了胰岛移植计划和隔离团队 威斯康星州麦迪逊。该应用程序的主要目标是实施一系列定量 区域化胰腺恢复和胰岛移植中胰岛存活力和功能效力的测量 分布网络，并验证这些参数预测胰岛移植功能的能力， 患者该提案的具体目标是：1）分配用于移植的上级质量的人类胰岛 和研究，以当地胰岛研究和移植联盟和ICR网络的成员， 质量、效力和功能预测因子的多参数表征。2)以提高 胰岛制备通过采取三管齐下的方法来依次优化胰腺保存、胰岛 分离和分离后培养。使用敏感的评估工具，我们将首先测试 基于营养因子的新型保存溶液。我们还将开发方法，以尽量减少对胰岛的损害 在分离，培养和运输过程中，重点关注ROS形成的减少和 通过向分离和培养物中补充抗氧化剂恢复"正常"代谢状态 培养基和优化培养物pH、氧张力和温度。随着质量的提高 我们有信心在五年内提供超过2000万个高质量的胰岛 为ICR研究者提供信息，并促进其他胰岛移植计划的发展， 糖尿病社区

项目成果

IL-1beta receptor blockade protects islets against pro-inflammatory cytokine induced necrosis and apoptosis.

• DOI: 10.1002/jcp.21770
• 发表时间: 2009-08
• 期刊: JOURNAL OF CELLULAR PHYSIOLOGY
• 影响因子: 5.6
• 作者: Schwarznau, Alice;Hanson, Matthew S.;Sperger, Jamie M.;Schram, Brian R.;Danobeitia, Juan S.;Greenwood, Krista K.;Vijayan, Ashwanth;Fernandez, Luis A.
• 通讯作者: Fernandez, Luis A.