Mining the stiffness-sensitive transcriptome in human vascular smooth muscle cells identifies long non-coding RNA stiffness regulators. Homo sapiens

Vascular extracellular matrix (ECM) stiffening is a risk factor for aortic and coronary artery disease. How matrix stiffening regulates the transcriptome profile of human aortic (Ao) and coronary (Co) vascular smooth muscle cells (VSMCs) is not well understood. Furthermore, the role of long non-coding RNAs (lncRNAs) in the cellular response to stiffening has never been explored. This study characterizes the stiffness-sensitive transcriptome of human Ao and Co VSMCs and identify potentially key lncRNA regulators of stiffness-dependent VSMC functions. Ao and Co VSMCs were cultured on hydrogel substrates mimicking physiologic and pathologic ECM stiffness. Total RNA-seq was performed to compare the stiffness-sensitive transcriptome profiles of Ao and Co VSMCs. Overall design: 4 unique sample types with 4 replicates (16 total samples). Donor-matched Aortic and Coronary VSMCs were serum starved for 48 hours, then cultured in serum containing media for 24 hours on both soft and stiff fibronectin coated hydrogel matrices.

血管细胞外基质（extracellular matrix, ECM）硬化是主动脉及冠状动脉疾病的高危致病因素。目前学界对细胞外基质硬化如何调控人主动脉（Ao）及冠状动脉（Co）血管平滑肌细胞（vascular smooth muscle cells, VSMCs）的转录组谱尚不明确，且长链非编码RNA（long non-coding RNAs, lncRNAs）在细胞响应硬化刺激中的作用也从未被相关研究探索。
本研究针对人主动脉与冠状动脉VSMCs的刚度敏感型转录组开展系统性特征解析，并鉴定出潜在的刚度依赖性VSMC功能关键lncRNA调控因子。
研究中将供体匹配的主动脉及冠状动脉VSMCs经血清饥饿处理48小时后，接种于模拟生理及病理ECM刚度的水凝胶基质上。
随后通过总RNA测序（Total RNA-seq）比较两类细胞的刚度敏感型转录组谱特征。
本实验整体设计为：共设置4种独特样本类型，每组设置4个生物学重复，总计16份样本。具体处理流程为：供体匹配的主动脉及冠状动脉VSMCs经血清饥饿48小时后，于包被纤连蛋白的软、硬水凝胶基质上，在含血清培养基中培养24小时。