Mechanical regulation of maturation and pathology of engineered human heart tissues

工程人体心脏组织成熟和病理的机械调节

基本信息

批准号：
10604901
负责人：
Jacob Christopher Scherba
金额：
$ 4.05万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10604901
关键词：
3-Dimensional Acceleration Action Potentials Adjuvant Therapy Adult Affect Bioreactors Birth Cardiac Cardiac Myocytes Cause of Death Cell Respiration Cell Therapy Characteristics Combined Modality Therapy Destinations Development Disease Disease model Electric Stimulation Electron Transport Engineering Environment Enzymes Gene Expression Gene Expression Profile Generations Genetic Goals Growth Growth and Development function Heart Heart Diseases Heart Research Heart Transplantation Heart failure Human Human Engineering Immunohistochemistry In Vitro Maps Measurement Mechanical Stimulation Mechanics Metabolic Methods Mitochondria Modeling Molecular Molecular Profiling Myocardial tissue Myocardium Myosin ATPase Optics Pathologic Pathology Patients Phenotype Physiology Play Process Property Protein Isoforms Protocols documentation Recovery Recovery Support Regulation Reproducibility Research Resistance Rodent Model Role Signal Pathway Slice Stretching Structural Protein Structure Supplementation System Testing Time Tissue Sample Tissues Transmission Electron Microscopy Troponin United States Up-Regulation Ventricular Work cardiac regeneration cardiac tissue engineering cardiogenesis complex IV drug discovery experience fetal fetus cell heart cell human disease implantation in vitro Model induced pluripotent stem cell induced pluripotent stem cell derived cardiomyocytes insight left ventricular assist device mechanical load mechanical stimulus mechanotransduction metabolic abnormality assessment mimetics novel pharmacologic postnatal postnatal development postnatal human protein expression real-time images repository response tissue culture tool transcriptomics

项目摘要

ABSTRACT The advent of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) offers exciting opportunities to study human cardiac disease and development in vitro. However, hiPSC-CMs are structurally and functionally immature and more closely resemble fetal than adult CMs as evident from their reliance on glycolytic instead of oxidative metabolism, weak contractions and excitability, and expression of immature isoforms of structural proteins including myosin and troponin. This makes hiPSC-CMs inadequate for studies of adult-acquired cardiac diseases, such as heart failure (HF), which remains the leading cause of death in the United States and worldwide. The mechanical environment of CMs is widely recognized as a key regulator of cardiac tissue development, physiology, and disease. In particular, dynamic changes in mechanical load experienced by CMs during postnatal development may play a key role in their acquisition of a mature adult phenotype. I hypothesize that presentation of dynamic, time-varying stretch (preload) and resistance to contraction (afterload) to human engineered heart tissues (hEHTs) made of hiPSC-CMs will increase their structural and functional maturity. Furthermore, I predict that mechanical overload of hEHTs will induce pathological features characteristic of HF progression. To test these hypotheses, I have developed a novel bioreactor where mechanical preload and afterload imposed on hEHTs can be independently varied with time of culture via application of stretch and electrical stimulation. Using this platform, I will systematically study how different regimes of progressively increased preload and afterload affect structure, contractile force generation, propagation of action potentials, and transcriptomic and metabolic properties of hEHTs. Additionally, I will employ real-time imaging to identify and characterize the mechanotransduction mechanisms underlying the observed functional changes in the hEHTs. For mechanical overload regimes that induce molecular and functional signatures of an HF phenotype, I will study if different adjuvant therapies combined with applied mechanical unloading akin to use of left ventricular assist devices (LVADs) can reverse-remodel structural and functional deficits in hEHTs. When completed, these studies will identify new mechanobiological drivers of in vitro CM maturation and further the molecular understanding of HF disease and therapy.

抽象的人类诱导多能干细胞来源的心肌细胞 (hiPSC-CM) 的出现提供了令人兴奋的机会有机会在体外研究人类心脏病和发育。然而，hiPSC-CM 在结构上功能上不成熟，比成人 CM 更类似于胎儿，这一点从它们对糖酵解代替氧化代谢，收缩和兴奋性弱，表达不成熟结构蛋白的亚型，包括肌球蛋白和肌钙蛋白。这使得 hiPSC-CM 不足以用于研究成人获得性心脏病，例如心力衰竭（HF），它仍然是成年人死亡的主要原因美国和世界各地。 CM 的机械环境被广泛认为是关键调节因素心脏组织发育、生理学和疾病。特别是机械负载的动态变化 CM 在出生后发育过程中经历的经历可能在他们获得成熟的过程中发挥关键作用表型。我假设动态的、随时间变化的拉伸（预载）和阻力的呈现由 hiPSC-CM 制成的人类工程心脏组织 (hEHT) 的收缩（后负荷）将增加其结构和功能的成熟度。此外，我预测 hEHT 的机械过载会导致心力衰竭进展的病理特征。为了检验这些假设，我写了一本小说生物反应器，其中施加在 hEHT 上的机械预载和后载可以随时间独立变化通过应用拉伸和电刺激进行培养。利用这个平台，我将系统地学习如何逐渐增加的预载和后载的不同机制会影响结构、收缩力的产生、动作电位的传播以及 hEHT 的转录组和代谢特性。此外，我将聘用实时成像来识别和表征观察到的机械传导机制 hEHT 的功能变化。对于诱导分子和功能的机械过载状态 HF 表型的特征，我将研究不同的辅助疗法是否与应用的机械疗法相结合类似于使用左心室辅助装置（LVAD）的卸载可以逆转结构和功能 hEHT 的缺陷。完成后，这些研究将确定体外 CM 的新机械生物学驱动因素成熟并进一步促进心力衰竭疾病和治疗的分子理解。