Microphysiological Systems to Study Hypoxic Cardiac Injury

研究缺氧性心脏损伤的微生理系统

• 批准号: 10591258
• 负责人: Megan L. Rexius
• 金额: $ 11.88万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-09 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10591258
• 关键词: Academia; Acute myocardial infarction; Address; Affect; Architecture; Area; Autophagocytosis; Binding; Bioinformatics; Biological Assay; Biomedical Engineering; Blood flow; Calcium; California; Cardiac; Cardiac Myocytes; Cell physiology; Cessation of life; Characteristics; Coenzymes; Communication; Complex; Coronary artery; Coupled; Data; Data Set; Dependence; Development; Engineering; Environment; Fluorescence Microscopy; Functional disorder; Goals; Heart; Heart Injuries; Heterogeneity; Human; Hypoxia; Image; In Vitro; Incubators; Injury; Investigation; Ischemia; Laboratories; Lead; Los Angeles; Measurement; Measures; Mediating; Mediator; Membrane Potentials; Mentors; Mentorship; Metabolic; Metabolism; Methods; MicroRNAs; Microfluidic Microchips; Microfluidics; Modeling; Molecular; Molecular Analysis; Monitor; Motion; Myocardial Infarction; Myocardial Reperfusion Injury; Myocardial Stunning; Myocardial dysfunction; Myocardial tissue; Myocardium; Optics; Oxygen; Paracrine Communication; Pathologic Processes; Pathway interactions; Pediatric Hospitals; Phase; Phenotype; Physiological; Process; Proteomics; Reactive Oxygen Species; Regulation; Reperfusion Injury; Reperfusion Therapy; Research; Research Personnel; Role; Sarcomeres; Site; System; Technology; Therapeutic; Therapeutic Effect; Time; Tissue Viability; Tissues; Traction; Traction Force Microscopy; Training; Universities; Vascular blood supply; blood perfusion; coronary artery occlusion; exosome; experience; extracellular; heart cell; in vitro Model; in vivo Model; induced pluripotent stem cell derived cardiomyocytes; injured; innovation; insight; intercellular communication; medical schools; metabolic imaging; microdevice; microphysiology system; mitochondrial membrane; multi-scale modeling; myocardial damage; novel; optical imaging; organ on a chip; paracrine; pharmacologic; preservation; programs; release of sequestered calcium ion into cytoplasm; repaired; response; restoration; skills; spatiotemporal; tool

Project Summary/Abstract Following the onset of an acute myocardial infarction (MI) with coronary artery occlusion, the restricted blood supply limits oxygenation of the myocardium, resulting in the formation of a steep oxygen (O2) gradient from normoxic, viable tissue to hypoxic, damaged tissue. A site of regional dysfunction exists at the interface between the normoxic and hypoxic tissue, known as the border zone. Reperfusion restores the flow of blood and O2 to the tissue, but also induces ischemia reperfusion injury (IRI), a pathophysiology resulting in further tissue damage. The pathological processes underlying these hypoxic cardiac injuries are not definitively established, in part due to a lack of experimental tools to recapitulate the diverse spatiotemporal O2 gradients characteristic of MI and IRI. The goal of this proposal is to engineer microphysiological systems with tight O2 control to investigate the molecular pathways activated in O2 gradients, and the resulting effects on cardiomyocyte (CM) function, to obtain a comprehensive view of the cardiac response to hypoxic injury. The aims outlined in this proposal will build on the expertise of Dr. Rexius in controlling O2 levels using microfluidics and integrate Heart- on-a-Chip technologies to advance the functional and mechanistic understanding of hypoxic cardiac injury. In the mentored phase, Dr. Rexius will use engineering and pharmacological approaches to control paracrine interactions in an MI border zone microdevice model and determine the role of paracrine-mediated hypoxic- normoxic intercellular communication in defining the spatial metabolic heterogeneity across an O2 gradient (Aim 1). Proteomic and miRNA analysis will be used to identify and validate transfer of exosome cargo as a paracrine mechanism altering CM metabolism. The existing O2 control framework will be utilized to engineer a microphysiological system to model IRI and multiplex measurements of traction force, sarcomere shortening, and calcium transients, and their dependence on O2 tension, to monitor dysfunction with live imaging (Aim 2). In the independent phase, modified versions of these systems will examine the effect of O2 reperfusion rate on CM function and the regulation of autophagy, a process by which cellular material is degraded and recycled (Aim 3). The project and mentorship plan will allow Dr. Rexius to develop skills in (1) non-invasive optical measurements of metabolic parameters, (2) bioinformatics analysis of exosome proteomic and miRNA datasets, (3) traction force microscopy, and (4) communication, mentoring, and laboratory management to prepare to lead an independent research program in academia. Dr. Rexius will be co-mentored by Dr. Megan McCain at the University of Southern California (USC) and Dr. Ching-Ling (Ellen) Lien at the Keck School of Medicine of USC and Children’s Hospital Los Angeles. Dr. Rexius has also enlisted Dr. Keyue Shen (USC) and Dr. Jennifer Van Eyk (Cedars-Sinai) as advisors to support her scientific and professional development. Completion of the aims will reveal novel insights into CM responses in heterogeneous O2 landscapes.

项目摘要/摘要 急性心肌梗塞（MI）伴有冠状动脉闭塞后，受限制 血液供应限制心肌的氧合，导致形成钢氧（O2）梯度 从常规，可行的组织到缺氧，损坏的组织。界面上存在区域功能障碍的位点 在常氧组织和低氧组织之间，称为边界区。再灌注恢复血液的流动和 O2到组织，但也诱导缺血再灌注损伤（IRI），这是一种病理生理学，导致进一步的组织 损害。这些缺氧心脏损伤的基础病理过程并未确定确定， 部分原因是缺乏实验工具来概括潜水员时空o2梯度的特征 Mi和Iri。该建议的目的是设计具有紧密O2控制的微生物生理系统 研究在O2梯度中激活的分子途径，并对心肌细胞（CM）产生影响 功能，以获取对低氧损伤的心脏反应的全面视图。概述的目的 提案将以Rexius博士的专业知识为基础 片上的技术，以提高对低氧心脏损伤的功能和机械理解。在 Rexius博士将使用工程和药物方法来控制旁分泌 MI边框区微电位模型中的相互作用，并确定旁分泌介导的低氧的作用 定义O2梯度的空间代谢异质性时，细胞间通信（AIM） 1）。蛋白质组学和miRNA分析将用于识别和验证外泌体货物作为旁分泌的转移 改变CM代谢的机制。现有的O2控制框架将用于设计 微生物生理系统以建模IRI和牵引力的多个测量，肌节缩短， 和钙瞬变及其对O2张力的依赖性，以通过实时成像监测功能障碍（AIM 2）。在 这些系统的独立阶段修改版本将检查O2再灌注速率对CM的影响 功能和自噬的调节，这是一个降解和回收利用细胞材料的过程（AIM 3）。 该项目和心态计划将使Rexius博士能够在（1）非侵入性光学方面发展技能 代谢参数的测量，（2）外泌体蛋白质组学和miRNA数据集的生物信息学分析， （3）牵引力显微镜以及（4）沟通，心理和实验室管理，以准备领导 学术界的独立研究计划。 Rexius博士将由Megan McCain博士在 南加州大学（USC）和Ching-Ling博士（Ellen）留置权 和儿童医院洛杉矶。 Rexius博士还邀请了Keyue Shen博士（USC）和Jennifer Van博士 EYK（Cedars-Sinai）作为支持她的科学和专业发展的顾问。目标完成 将揭示对异质O2景观中CM反应的新见解。