A Novel Beta Cell Specific Microfluidic Perifusion and Imaging Device for Islet P

一种新型β细胞特异性胰岛P微流控灌注和成像装置

基本信息

批准号：
8234332
负责人：
Jose Oberholzer
金额：
$ 34.01万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-20 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8234332
关键词：
Address Allogenic Beta Cell Biological Biological Assay Biological Products Calcium Calcium Signaling Cell Survival Cell membrane Cell physiology Cells Characteristics Chicago Clinical Clinical Trials Defect Development Foundations Future Glucose Hour Human Illinois Imaging Device Insulin Insulin-Dependent Diabetes Mellitus Investigational New Drug Application Islet Cell Islets of Langerhans Islets of Langerhans Transplantation Kinetics Licensing Licensure Life Liquid substance Measurement Measures Metabolic Methods Microfluidic Microchips Microfluidics Microscopy Mitochondria Modeling Modification Nuclear Envelope Nude Mice Outcome Oxygen Consumption Perfusion Pharmaceutical Preparations Phase Physiological Physiology Population Pre-Clinical Model Preparation Process Ramp Regulation Replacement Therapy Resolution Risk Sample Size Sampling Source Standardization Streptozocin Structure of beta Cell of islet System Technology Testing Time Transplantation Universities base cell type design diabetes mellitus therapy diabetic diabetic patient effective therapy glycemic control graft function improved in vivo indexing innovation insulin secretion islet meetings minimally invasive mouse model novel potency testing pre-clinical prognostic sterility testing success type I diabetic

项目摘要

DESCRIPTION (provided by applicant): Islet transplantation is the only minimally invasive therapy for Type I diabetes that is able to achieve glycemic control without exogenous insulin. However, islet transplantation shows variable success rates, mainly due to the inconsistent quality of human islet preparations. For islet transplantation to become a FDA licensed biologic product, a well- established islet preparation process and product manufacturing consistency will need to be demonstrated. Federal regulations mandate that each biologic product lot be tested for potency before being released for clinical use. At present, there is no reliable potency test available for human pancreatic islets. We hypothesize that an appropriate islet potency test must be beta-cell specific and simultaneously assess key factors associated with islet physiology, including glucose- stimulated changes in mitochondrial potentials, calcium influx and dynamic insulin secretion. To test this hypothesis, an innovative islet perfusions system with functional, live microscopy was developed using microfluidic chip technology to enable simultaneous measurement of glucose-induced changes in mitochondrial potentials, calcium influx and dynamic insulin secretion. Preliminary results indicate that this system can adequately distinguish low potency from high potency human islet preparations. This project will focus on the following aims: (1) To further improve the resolution of the microfluidic system through modification of the chip design. Specifically, our proposal focuses on: a. improving temporal resolution by reducing the volume of the chamber within the microfluidic chip, b. improving flow dynamic control and increasing ease of use by adding a fluid mixer into the chip, c. establishing glucose ramps to evaluate insulin kinetics, d. integrating multiple perfusion chambers into the chip on a motorized platform to increase the sample size of human islets that can be evaluated and provide a better representation of the final islet product. e. developing a rapid insulin secretion measurement. (2) To validate the microfluidic system in a pre-clinical nude mouse model using human islet cell grafts and develop an "Islet Potency Index " predictive of post-transplant islet graft function. Briefly, multivariable regression modeling will determine which islet cell characteristics are significantly associated with in vivo outcome and these will be used to calculate the index. (3) To test the microfluidic system in setting of a clinical human islet transplant trial and investigate the validity of the Islet Potency Index to predict islet graft function. This proposal will test an innovative microfluidic system that provides detailed analysis of pancreatic beta-cell physiology. In the future, it is likely that further developments addressing an unlimited islet cell source and new immunoprotective strategies will make islet transplantation available to a broader proportion of the diabetic population. This microfluidic system could represent a reliable islet potency assay for current and future islet replacement therapies, as well as for the study of new diabetes therapies in general. PUBLIC HEALTH RELEVANCE: An innovative microfluidic system will be optimized and standardized for human islet potency testing. If successful, the microfluidic-based assay could be used for islet product release testing, which represents a yet unmet regulatory requirement for FDA biologic licensure of islet transplants.

描述（申请人提供）：胰岛移植是唯一无需外源胰岛素即可实现血糖控制的 I 型糖尿病微创疗法。然而，胰岛移植的成功率参差不齐，这主要是由于人类胰岛制剂的质量不稳定。为了使胰岛移植成为 FDA 许可的生物产品，需要证明完善的胰岛制备工艺和产品制造的一致性。联邦法规要求每个生物产品批次在投入临床使用之前必须进行效力测试。目前，还没有针对人类胰岛的可靠效价测试。我们假设适当的胰岛效能测试必须是β细胞特异性的，并同时评估与胰岛生理学相关的关键因素，包括葡萄糖刺激的线粒体电位变化、钙流入和动态胰岛素分泌。为了检验这一假设，利用微流控芯片技术开发了一种具有功能性活体显微镜的创新胰岛灌注系统，能够同时测量葡萄糖诱导的线粒体电位、钙流入和动态胰岛素分泌的变化。初步结果表明，该系统可以充分区分低效和高效的人胰岛制剂。该项目将重点实现以下目标：（1）通过芯片设计的修改，进一步提高微流控系统的分辨率。具体来说，我们的建议侧重于：通过减小微流控芯片内的室的体积来提高时间分辨率，b．通过在芯片中添加流体混合器来改善流动动态控制并提高易用性，c．建立葡萄糖斜坡来评估胰岛素动力学，d．将多个灌注室集成到机动平台上的芯片中，以增加可评估的人类胰岛样本量，并更好地代表最终胰岛产品。 e.开发快速胰岛素分泌测量方法。 (2) 使用人胰岛细胞移植物在临床前裸鼠模型中验证微流体系统，并开发预测移植后胰岛移植物功能的“胰岛效力指数”。简而言之，多变量回归模型将确定哪些胰岛细胞特征与体内结果显着相关，并且这些特征将用于计算指数。 (3) 在临床人类胰岛移植试验中测试微流体系统，并研究胰岛效力指数预测胰岛移植物功能的有效性。该提案将测试一种创新的微流体系统，该系统可提供胰腺 β 细胞生理学的详细分析。未来，解决无限胰岛细胞来源和新的免疫保护策略的进一步发展很可能将使胰岛移植适用于更广泛的糖尿病人群。这种微流体系统可以为当前和未来的胰岛替代疗法以及新糖尿病疗法的研究提供可靠的胰岛效力测定。公共健康相关性：创新的微流体系统将针对人类胰岛效力测试进行优化和标准化。如果成功，基于微流体的测定可用于胰岛产品释放测试，这代表了 FDA 胰岛移植生物许可尚未满足的监管要求。