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

• 批准号: 10200034
• 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
• 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/10200034
• 关键词: Affect; American; Beta Cell; Biological; Biological Assay; 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; 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

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.

摘要 这个为期4年的R01项目的目标是开发一个创新和全面的工具包，包括 收集基于荧光蛋白(FP)的生物传感器、微流控芯片上的胰岛、智能手机 荧光显微镜和一种图像分析应用程序，为评估人类的潜能而优化 胰岛作为1型糖尿病(T1D)的基于细胞的治疗，以及用于表型和 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，并进一步确定各种有前景的生物制剂的效力、表型和生理学 用于T1D细胞替代疗法。利用我们强大团队的互补专业知识和我们广泛的 为了取得初步成果，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，用于监测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 胰岛生物制品标准化、高含量表征的成像分析应用。 该项目的成果将是一个经济、综合的系统，方便可靠 人类胰岛、异种胰岛、被包裹的胰岛和干细胞来源的β细胞的特征。这些输入 体外试验创新性地评估了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，也可作为改进世代的重要筛选工具， 储存和运输下一代生物制品。我们提议的工作将对以下方面产生重大影响 当前和未来糖尿病细胞替代疗法的发展。