Mouse-on-a-chip systems to evaluate pancreas-adipose tissue dynamics in vitro

用于体外评估胰腺脂肪组织动力学的小鼠芯片系统

• 批准号: 9228365
• 负责人: Christopher J Easley
• 金额: $ 36.6万
• 依托单位: AUBURN UNIVERSITY AT AUBURN
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-08 至 2020-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9228365
• 关键词: Acute; Adipocytes; Adipose tissue; Alzheimer' s Disease; Biochemistry; Biological; Biological Assay; Carbohydrates; Chronic; Coculture Techniques; Collaborations; Complex; Devices; Diabetes Mellitus; Diet; Dietary Intervention; Disease; Endocrine; Endocrine Glands; Endocrine system; Fasting; Fatty Acids; Fatty acid glycerol esters; Feedback; Funding; Glucose; Goals; Heart; Hormones; Human; Hyperglycemia; Hyperinsulinism; In Vitro; Incidence; Insulin; Islets of Langerhans; Lead; Link; Lipolysis; Liver; Measurement; Metabolism; Methodology; Methods; Microfluidic Analytical Techniques; Microfluidic Microchips; Microfluidics; Mission; Modeling; Mus; National Institute of Diabetes and Digestive and Kidney Diseases; Nature; Nonesterified Fatty Acids; Nutrient; Obesity; Organ; Output; Overweight; Pancreas; Pancreatic Hormones; Pharmacology; Physiology; Population; Printing; Publications; Recommendation; Regulation; Research; Research Personnel; Role; Sampling; Signal Transduction; Site; Skeletal Muscle; Sucrose; System; Systems Analysis; Techniques; Testing; Tissue Expansion; Tissues; Triglycerides; Work; adipokines; base; body system; detection of nutrient; drug discovery; flexibility; immune function; improved; in vitro Model; innovation; insulin signaling; interest; islet; ketogentic; novel; novel strategies; nutrition; programs; public health relevance; success; tissue culture; uptake

 DESCRIPTION (provided by applicant): Rather than a mere storage site for triglycerides, it is now understood that adipose tissue (fat) is a complex, multicellular endocrine organ that has profound systemic effects, altering the function of nearly all other organ systems. Despite its importance, however, there is a lack of information on the dynamic nature of adipokine secretion and nutrient uptake in adipose tissue, highlighting several unmet needs in methodology. Few techniques exist to interrogate small amounts of adipose tissue, and there is a shortage of methods to explore dynamic function of the organ. Specifically, we have a limited view of the dynamic relationship between glucose, insulin, and adipose function, highlighting an immediate need for better in vitro techniques to study pancreas-adipose tissue crosstalk. As demonstrated in our previous funding period, we propose that our microfluidic systems are ideal to meet these ongoing needs. Our research team developed microfluidic approaches for culture of endocrine tissue, namely pancreatic islets and adipose tissue from C57BL/6J mice, as well as for sampling of hormone secretion. These systems permit dynamic interrogation of the tissues in ways not possible with standard techniques. The long-term goal of this research is to develop in vitro models of the endocrine system for applications in nutrition, metabolism, and drug discovery. In the short term, our objective is to develop a mouse-on-a-chip microfluidic system that permits dynamic and quantitative measurements of both hormone secretion and nutrient uptake from primary tissue. Microfluidic devices will be developed concurrently with small-volume methodology to assay secretion or nutrient uptake from pancreatic islets and adipose tissue, and 3D printing will be used to improve our novel device interfacing and tissue culture methods. Aim 1 of the proposal seeks to develop an automated microfluidic input/output multiplexer (µMUX) for generalizable dynamic control over hormones and nutrients to/from endocrine tissue. Aim 2 will result in targeted small-volume compatible assays for hormones and free fatty acid uptake in adipose tissue. Aims 3 and 4 are biological in nature, using the µMUX system to determine the dynamics of hormone secretion, fatty acid uptake, and lipolysis in endocrine tissues with varied glycemic dynamics (Aim 3), and determining the role of dynamic feedback between the tissues using a co-culture µMUX system (Aim 4). The rationale for this research to provide a flexibly programmable, in vitro micro-model of pancreas-adipose dynamics to test several important biological hypotheses related to gut-pancreas signaling dynamics, insulin/lipolysis/fatty acid uptake dynamics, and regulation of lipolysis at low insulin and glucos (fasting or ketogenic metabolism). The proposed work is significant as a first-of-its-kind in vitro mimic of pancreas-adipose physiology, which we expect will lead to better information on human dietary interventions. The proposal is thus innovative in its technological and its biological approaches. Preliminary evidence strongly supports the feasibility of these proposals, and the research team has a proven track-record of success.

 描述（由申请人提供）：现在人们了解到，脂肪组织（脂肪）不仅仅是甘油三酯的储存场所，而是一种复杂的多细胞内分泌器官，具有深远的系统影响，改变几乎所有其他器官系统的功能。然而，尽管其重要性，但缺乏关于脂肪组织中脂肪因子分泌和营养吸收的动态性质的信息，突出了方法学中的一些未满足的需求。很少有技术可以检测少量脂肪组织，并且缺乏探索器官动态功能的方法。具体来说，我们对葡萄糖、胰岛素和脂肪功能之间的动态关系的看法有限，这突出表明迫切需要更好的体外技术来研究胰腺-脂肪组织的串扰。正如我们之前的资助期所证明的那样，我们建议我们的微流体系统非常适合满足这些持续的需求。我们的研究团队开发了微流体方法，用于培养内分泌组织，即来自 C57BL/6J 小鼠的胰岛和脂肪组织，以及用于激素分泌取样。这些系统允许以标准技术不可能的方式动态询问组织。这项研究的长期目标是开发内分泌系统的体外模型，用于营养、代谢和药物发现。短期内，我们的目标是开发一种芯片上的小鼠微流体系统，该系统可以动态定量测量原代组织的激素分泌和营养吸收。微流体装置将与小体积方法同时开发，以测定胰岛和脂肪组织的分泌或营养吸收，3D 打印将用于改进我们的新型装置接口和组织培养方法。该提案的目标 1 旨在开发一种自动化微流体输入/输出多路复用器 (μMUX)，用于对内分泌组织的激素和营养物质进行通用动态控制。目标 2 将针对脂肪组织中的激素和游离脂肪酸摄取进行有针对性的小体积兼容测定。目标 3 和 4 本质上是生物学的，使用 µMUX 系统确定具有不同血糖动态的内分泌组织中激素分泌、脂肪酸摄取和脂肪分解的动态（目标 3），并使用共培养 µMUX 系统确定组织之间动态反馈的作用（目标 4）。这项研究的基本原理是提供一个灵活可编程的体外胰腺脂肪动力学微观模型，以测试与肠胰腺信号动力学、胰岛素/脂解/脂肪酸摄取动力学以及低胰岛素和葡萄糖（空腹或生酮代谢）下脂解调节相关的几个重要生物学假设。拟议的工作作为首个体外研究具有重要意义 模仿胰腺脂肪生理学，我们期望这将带来有关人类饮食干预的更好信息。因此，该提案在技术和生物学方法方面具有创新性。初步证据有力地支持了这些建议的可行性，并且研究团队拥有良好的成功记录。