An Integrated Fluorescent Biosensor, Imaging, and Analysis Toolkit for Islet Biologics

用于胰岛生物制品的集成荧光生物传感器、成像和分析工具包

基本信息

批准号：
10379407
负责人：
Huiwang Ai
金额：
$ 40.38万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10379407
关键词：
Affect American Beta Cell Biological Assay Biological Products Biosensor Cell Line Cell Therapy Cell secretion Cells Cellular Phone Chemicals Clinical Collection Color Computer software Custom Development Disease Economics Encapsulated Evaluation Family suidae Funding Future Generations Glucose Glutathione Disulfide Green Fluorescent Proteins Human Hypoxia Image Immune Immune response Immunosuppressive Agents Insulin Insulin-Dependent Diabetes Mellitus Ion Channel Islets of Langerhans Islets of Langerhans Transplantation Lead Metabolic Metabolism Methods Microfluidics Monitor Mus NADP Oranges Organ Transplantation Outcome Oxidation-Reduction Patients Penetration Phenotype Phototoxicity Physiological Physiological Processes Physiology Preparation Proteins Quality Control Research Research Personnel Screening procedure Secretory Vesicles Serotonin Signal Transduction Source Standardization System Tissues Toxic effect Transplantation United States Food and Drug Administration Work Youth allotransplant autocrine base cell replacement therapy diabetes mellitus therapy fluorescence imaging fluorescence microscope genetic element glycemic control human stem cells improved in vitro Assay innovation islet islet stem cells microscopic imaging minimally invasive multiparametric imaging multiplexed imaging next generation novel paracrine public health relevance red fluorescent protein research clinical testing response sensor spatiotemporal stem cells success synergism tool transplant centers

项目摘要

Abstract The objective of this 4-year R01 project is to develop an innovative and comprehensive toolkit, including a collection of fluorescent protein (FP) based biosensors, a microfluidic Islet-On-a-Chip, a smartphone fluorescence microscope, and an imaging analysis application, optimized for the potency evaluation of human islets as cell-based therapy in Type 1 diabetes (T1D), as well as for characterization of the phenotype and physiology of emerging new biologics for future T1D treatment. We propose to use genetically encoded fluorescent biosensors to evaluate key physiological parameters of β- cells and islets, and furthermore, to define the potency, phenotype, and physiology of various promising biologics for T1D cell replacement therapy. Leveraging the complementary expertise of our strong team and our extensive preliminary results, we will pursue the following two Specific Aims: 1. Develop and optimize fluorescent biosensors to monitor key physiological processes in β-cells and islets. 2. Integrate biosensors with microfluidic arrays, a smartphone fluorescence microscope, a customized imaging analysis application for standardized, high-content characterization of islet biologics. The outcome of this project will be an economic and integrated system for convenient and reliable characterization of human islets, xenogeneic islets, encapsulated islets, and stem-cell-derived β-cells. These in vitro assays, which innovatively evaluate several key physiological parameters of β-cells and islets, can not only be used for standardization and quality control, but also be important screening tools to improve the generation, storage, and transport of next-generation biologics. Our proposed work will generate significant impacts on development of current and future cell replacement therapy for diabetes.

抽象的这个为期 4 年的 R01 项目的目标是开发一个创新且全面的工具包，包括基于荧光蛋白 (FP) 的生物传感器、微流控胰岛芯片、智能手机的集合荧光显微镜和成像分析应用程序，针对人类效力评估进行了优化胰岛作为 1 型糖尿病 (T1D) 的细胞疗法，以及用于表征表型和用于未来 T1D 治疗的新兴生物制剂的生理学。我们建议使用基因编码的荧光生物传感器来评估β-的关键生理参数细胞和胰岛，此外，还可以定义各种有前途的生物制剂的效力、表型和生理学利用我们强大的团队和广泛的互补专业知识进行 T1D 细胞替代治疗。根据初步结果，我们将追求以下两个具体目标： 1. 开发和优化荧光生物传感器来监测β细胞和胰岛的关键生理过程。 2. 将生物传感器与微流控阵列、智能手机荧光显微镜、定制的用于对胰岛生物制品进行标准化、高内涵表征的成像分析应用。该项目的成果将是一个经济且集成的系统，方便可靠人类胰岛、异种胰岛、封装胰岛和干细胞衍生的 β 细胞的表征。体外测定创新性地评估了 β 细胞和胰岛的几个关键生理参数，不仅可以可用于标准化和质量控制，也是提高生成率的重要筛选工具，我们提出的工作将对下一代生物制品的储存和运输产生重大影响。当前和未来糖尿病细胞替代疗法的发展。