Development of an exosome and cargo tracking mouse

外泌体和货物追踪小鼠的开发

• 批准号: 10192854
• 负责人: JANOS ZEMPLENI
• 金额: $ 18.56万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-15 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10192854
• 关键词: Adhesions; Adipose tissue; Alzheimer' s Disease; Animal Model; Area; Attention; Biogenesis; Biology; Biomedical Research; Brain; Breast Cancer Risk Factor; Categories; Cell Communication; Cell Lineage; Cells; Chickens; Code; Colon Carcinoma; Communication; Communities; Cytomegalovirus; Data; Databases; Destinations; Development; Diagnosis; Disease; Distant; Elderly; Embryo; Enhancers; Enterobacteria phage P1 Cre recombinase; Exercise; Funding; Genes; Goals; Grant; Heart Diseases; Human; Impaired cognition; Institutes; Label; Laboratories; Letters; Malignant Neoplasms; Methodology; MicroRNAs; Modeling; Mus; National Cancer Institute; National Institute of Allergy and Infectious Disease; National Institute of Diabetes and Digestive and Kidney Diseases; National Institute of Neurological Disorders and Stroke; National Institute on Aging; Nucleus Accumbens; Obesity; Open Reading Frames; Parkinson Disease; Partner in relationship; Pathologic; Physiological; Plasmids; Play; Prevention; Process; Proteins; RNA; Reporter; Research; Research Personnel; Research Support; Role; Simian virus 40; Site; Sorting - Cell Movement; Streptavidin; Substance abuse problem; Tamoxifen; Terminator Codon; Testing; Time; Tissues; Transgenes; Transmembrane Domain; Traumatic Brain Injury; United States National Institutes of Health; Untranslated RNA; Work; base; beta Actin; design; enhanced green fluorescent protein; exosome; extracellular; heart disease prevention; immunoregulation; innovation; interest; magnetic beads; mouse genome; mouse model; nanoparticle; non-alcoholic fatty liver; non-alcoholic fatty liver disease; promoter; support tools; tool; trafficking; transgene expression; vector; virtual

PROJECT SUMMARY/ABSTRACT Exosomes are nanoparticles that play an essential role in cell-to-cell communication by shuttling a variety of cargos among tissues. Exosome cargos regulate more than 60% of human genes and have been implicated in virtually all physiological and pathological conditions. Supported by compelling preliminary data, this proposal will test the central hypothesis that developing an exosome and cargo tracking (ECT) mouse will provide a tool for identifying the donor and recipient tissues of exosome subsets and assessing the cargos in these exosomes. This project’s long-term goal is to enable the widespread use of this ECT mouse model, which will accelerate the rate of discovery in biomedical and exosome biology research across a variety of fields and illnesses. The main objective of this project will be to develop a tamoxifen-inducible ECT mouse model that allows investigators to exercise spatial and temporal control over the expression of endogenous exosomes in which the exosome marker CD63 is fused to near-infrared protein (iRFP); this will be achieved through one specific aim: to develop and optimize an ECT mouse to assess the origin, destination, and cargo of endogenous exosomes. The ECT vector was designed to allow for sorting of tissue-specific, iRFP-labeled exosomes by streptavidin- coated magnetic beads for subsequent cargo analysis. ECT vector design and iRFP-based exosome capture are methodologically innovative. The proposed work is conceptually innovative because the ECT mouse will be the first model to exercise spatial and temporal control over exosome biogenesis and assess exosome biogenesis, trafficking, and cargo content. One of the strengths of this proposal is that the ECT mouse has already been developed and funding is sought for optimizing the mouse. This project does not overlap with active grants in the NIH RePORTER database (i.e., the project will be unique in the NIH grants portfolio). This project is of great significance and impact because it will break new ground for developing strategies that use exosomes and their cargos in the diagnosis, prevention, and treatment of major diseases such as cancer, cognitive decline in the elderly, and obesity-induced non-alcoholic fatty liver disease. Scholars supported by various NIH institutes will benefit from access to the ECT mouse, and there are many applications in areas relevant to the National Institute of Diabetes and Digestive and Kidney Diseases, the National Institute on Aging, the National Institute of Neurological Disorders and Stroke, the National Cancer Institute, and the National Institute of Allergy and Infectious Diseases. Therefore, this application delivers on the priorities identified in PAR-16-141, which seeks proposals to develop animal models that are applicable to the research interests of two or more categorical NIH institutes.

项目概要/摘要 外泌体是纳米颗粒，通过穿梭多种物质，在细胞间通讯中发挥重要作用 组织之间的货物。外泌体调节超过 60% 的人类基因，并与 几乎所有的生理和病理状况。该提案得到令人信服的初步数据的支持 将测试中心假设，即开发外泌体和货物追踪（ECT）小鼠将提供一种工具 用于识别外泌体子集的供体和受体组织并评估这些外泌体中的货物。 该项目的长期目标是使这种 ECT 小鼠模型得到广泛使用，这将加速 各个领域和疾病的生物医学和外泌体生物学研究的发现率。这 该项目的主要目标是开发一种他莫昔芬诱导的 ECT 小鼠模型，该模型允许 研究人员对内源性外泌体的表达进行空间和时间控制，其中 外泌体标记物 CD63 与近红外蛋白 (iRFP) 融合；这将通过一个具体目标来实现： 开发和优化 ECT 小鼠以评估内源性外泌体的来源、目的地和货物。 ECT 载体的设计目的是通过链霉亲和素对组织特异性、iRFP 标记的外泌体进行分选。 用于后续货物分析的涂层磁珠。 ECT 载体设计和基于 iRFP 的外泌体捕获 在方法上具有创新性。所提出的工作在概念上具有创新性，因为 ECT 小鼠 将是第一个对外泌体生物发生进行空间和时间控制并评估外泌体的模型 生物起源、贩运和货物内容。该提案的优点之一是 ECT 小鼠具有 已经开发出来，并正在寻求资金来优化鼠标。该项目与活动项目不重叠 NIH RePORTER 数据库中的赠款（即该项目在 NIH 赠款组合中将是唯一的）。这个项目 具有重要意义和影响，因为它将为制定使用的战略开辟新天地 外泌体及其货物在癌症等重大疾病的诊断、预防和治疗中的应用 老年人认知能力下降，以及肥胖引起的非酒精性脂肪肝。学者支持 各个 NIH 机构都将受益于 ECT 小鼠的使用，并且在许多领域都有应用 与国家糖尿病、消化和肾脏疾病研究所、国家老龄化研究所相关， 美国国家神经疾病和中风研究所、美国国家癌症研究所和美国国家癌症研究所 过敏和传染病研究所。因此，该应用程序实现了中确定的优先事项 PAR-16-141，寻求开发适用于研究兴趣的动物模型的建议 两个或多个 NIH 分类机构。