Institute of Neurophysiology, University of Cologne, Cologne, Germany.
Institute of Neurophysiology, University of Cologne, Cologne, Germany; Department of Pediatric Cardiology, University Hospital Cologne, Cologne, Germany; Department of Clinical and Chemical Pathology, Cairo University, Cairo, Egypt.
Acta Biomater. 2019 Apr 15;89:180-192. doi: 10.1016/j.actbio.2019.03.017. Epub 2019 Mar 9.
Pathophysiological conditions, such as myocardial infarction and mechanical overload affect the mammalian heart integrity, leading to a stiffened fibrotic tissue. With respect to the pathophysiology of cardiac fibrosis but also in the limelight of upcoming approaches of cardiac cell therapy it is of interest to decipher the interaction of cardiomyocytes with fibrotic matrix. Therefore, we designed a hydrogel-based model to engineer fibrotic tissue in vitro as an approach to predict the behavior of cardiomyocytes facing increased matrix rigidity. Here, we generated pure induced pluripotent stem cell-derived cardiomyocytes and cultured them on engineered polyacrylamide hydrogels matching the elasticities of healthy as well as fibrotic cardiac tissue. Only in cardiomyocytes cultured on matrices with fibrotic-like elasticity, transcriptional profiling revealed a substantial up-regulation of a whole panel of cardiac fibrosis-associated transcripts, including collagen I and III, decorin, lumican, and periostin. In addition, matrix metalloproteinases and their inhibitors, known to be essential in cardiac remodeling, were found to be elevated as well as insulin-like growth factor 2. Control experiments with primary cardiac fibroblasts were analyzed and did not show comparable behavior. In conclusion, we do not only present a snapshot on the transcriptomic fingerprint alterations in cardiomyocytes under pathological conditions but also provide a new reproducible approach to study the effects of fibrotic environments to various cell types. STATEMENT OF SIGNIFICANCE: The ageing population in many western countries is faced with an increasing burden of ageing-related diseases such as heart failure which is associated with cardiac fibrosis. A deeper understanding of the interaction of organotypic cells with altered extracellular matrix mechanical properties is of pivotal importance to understand the underlying mechanisms. Here, we present a strategy to combine hydrogel matrices with induced pluripotent stem cell derived cardiomyocytes to study the effect of matrix stiffening on these cells. Our findings suggest an active role of matrix stiffening on cardiomyocyte function and heart failure progression.
病理生理条件，如心肌梗死和机械过载，会影响哺乳动物心脏的完整性，导致纤维化组织变硬。鉴于心脏纤维化的病理生理学，以及即将出现的心脏细胞治疗方法，解析心肌细胞与纤维化基质的相互作用引起了人们的兴趣。因此，我们设计了一种基于水凝胶的模型，以体外构建纤维化组织，从而预测面对基质刚性增加的心肌细胞的行为。在这里，我们生成了纯诱导多能干细胞衍生的心肌细胞，并在匹配健康和纤维化心脏组织弹性的工程化聚丙烯酰胺水凝胶上培养它们。只有在培养在具有纤维化样弹性的基质上的心肌细胞中，转录谱分析显示一整套与心脏纤维化相关的转录物（包括胶原 I 和 III、decorin、lumican 和 periostin）的显著上调。此外，已知在心脏重构中至关重要的基质金属蛋白酶及其抑制剂也被发现升高，以及胰岛素样生长因子 2。对原代心脏成纤维细胞进行的对照实验并未显示出类似的行为。总之，我们不仅呈现了心肌细胞在病理条件下转录组指纹改变的快照，还提供了一种新的可重复方法来研究纤维化环境对各种细胞类型的影响。
许多西方国家的老龄化人口面临着与衰老相关的疾病（如心力衰竭）的负担增加，这些疾病与心脏纤维化有关。深入了解器官样细胞与改变的细胞外基质机械性能的相互作用对于理解潜在机制至关重要。在这里，我们提出了一种将水凝胶基质与诱导多能干细胞衍生的心肌细胞相结合的策略，以研究基质变硬对这些细胞的影响。我们的研究结果表明，基质变硬对心肌细胞功能和心力衰竭进展具有积极作用。
