Engineered Heterochronic Parabiosis on 3D Microphysiological Systems

3D 微生理系统上的工程异时共生

• 批准号: 10207946
• 负责人: Young Charles Jang
• 金额: $ 46.37万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10207946
• 关键词: 3-Dimensional; Address; Age; Aging; Animals; Biochemical; Biological Models; Biomedical Engineering; Biomimetics; Blood; Blood - brain barrier anatomy; Blood Circulation; Blood Vessels; Brain; Cell physiology; Cells; Characteristics; Clinical; Coculture Techniques; Cues; Cultured Cells; Data; Development; Disease; Endocrine Glands; Engineering; Environment; Exercise; Exhibits; Exposure to; Geroscience; Goals; Human; Image; In Vitro; Investigation; Label; Mechanics; Mediating; Microcirculation; Microfabrication; Microfluidics; Modeling; Muscle; Muscle Contraction; Muscle function; Muscle satellite cell; Muscular Atrophy; Natural regeneration; Nature; Neuromuscular Junction; Operative Surgical Procedures; Optics; Organ; Outcome; Oxidative Stress; Parabiosis; Patients; Physiological; Plasma; Property; Proteins; Proteome; Proteomics; Regenerative capacity; Rejuvenation; Reporter; Rhodopsin; Skeletal Muscle; System; Testing; Transgenes; Translating; Validation; Work; age related; aged; anti aging; base; cell age; cell type; cost effective; genetic approach; human old age (65+); in vitro Model; in vivo; innovation; juvenile animal; microphysiology system; minimally invasive; mortality; mouse model; muscle aging; myogenesis; novel; optogenetics; public health relevance; repaired; satellite cell; stem; stem cell function; time use

PROJECT SUMMARY Heterochronic parabiosis, in which young and aged animals are surgically attached to share circulation, provided evidence of putative ‘anti-geronic’ factors exist but the mechanisms by which circulating factors mediate rejuvenating properties on muscle stem cells and their microenvironment have yet to be elucidated. Due to the complexity of in vivo parabiosis and the dynamic nature of blood-borne factors, reliable identification of these humoral factors remains a major hurdle. To overcome this challenge, Dr. Jang and his team will leverage advanced microengineering approaches to build a 3D microfluidic parabiosis circuit that can control mechanical and biochemical cues in the physiologically relevant 3D microenvironment. In this proposal, his team will further refine and upgrade the in vitro parabiosis platform by integrating a cell-type-specific protein labeling system (MetRSL2774G transgene) to precisely identify muscle secretome, also known as myokines, responsible for rejuvenation effects on muscle stem cells. In addition, the targeted genetics approach will be employed to delineate oxidative stress-induced pro-geronic myokines that negatively impact muscle stem cell function. Finally, the team will also engineer an exercise-induced myokine reporter system within parabiosis-on-a-chip using an optogenetic actuator (Channelrhodopsin 2) co-expressed with protein labeling construct, MetRSL274G. Using this reporter, proposed studies will identify novel contraction-induced myokines with anti-geronic properties and myokines that pass the blood-brain-barrier (BBB) to exert their action on muscle-brain crosstalk. The successful outcomes of this project will have far and broad implications in geroscience. This minimally invasive 3D microphysiological system can be exploited in a variety of studies testing the hallmarks of aging or modeling of age-related diseases in vitro. More importantly, upon validation, the experimental approach used in this proposal can be translated to mimic human parabiosis, which will have significant clinical implications.

项目摘要 杂色抛物比病，其中年轻动物和年龄的动物被外科手术固定在共享循环中，提供了 存在推定的“抗毛”因素的证据，但循环因子介导的机制 肌肉干细胞及其微环境的复兴特性尚未阐明。由于 体内抛物计数的复杂性和血源性因素的动态性质，可靠地识别这些因素 体液因素仍然是一个重大障碍。为了克服这一挑战，张博士和他的团队将利用 高级微型工程方法来构建一个3D微流体抛物性电路，该电路可以控制机械 和物理相关的3D微环境中的生化线索。在这个建议中，他的团队将进一步 通过整合细胞类型的蛋白质标记系统，完善和升级体外抛物线平台 （METRSL2774G转基因）精确地识别肌肉分泌组，也称为肌动物，负责 恢复活力对肌肉干细胞的影响。另外，靶向遗传学方法将被聘为 描绘氧化应激诱导的促晶状肌动物，对肌肉干细胞功能产生负面影响。 最后，该团队还将在芯片上的核恶作症中设计一个运动引起的肌动物记者系统 使用光遗传执行器（通道Rhopopsin 2）与蛋白质标记构建体共表达MetRSL274G。 使用此记者，拟议的研究将鉴定具有抗透性特性的新型收缩诱导的肌动物 通过血脑屏障（BBB）的肌动蛋白（BBB）对肌肉脑串扰发挥作用。这 该项目的成功结果将对Geroscience产生巨大的影响。这种侵入性最低 3D微生物生理系统可以在测试衰老或建模的标志的各种研究中探索 体外与年龄有关的疾病。更重要的是，经过验证，使用的实验方法 提案可以转化为模仿人类核心，这将具有重大的临床意义。