Ji Pengfei, Grande-Allen Kathryn Jane, Balaji Swathi, Birla Ravi K, Keswani Sundeep G
Laboratory for Regenerative Tissue Repair, Texas Children's Hospital, 1102 Bates Ave, Houston, TX, 77030, USA.
Center for Congenital Cardiac Research, Texas Children's Hospital, Houston, TX, USA.
Ann Biomed Eng. 2025 Jun 10. doi: 10.1007/s10439-025-03751-w.
Discrete subaortic stenosis (DSS) is a congenital heart disease characterized by a narrowing of the passage below the aortic valve in the left ventricular outflow tract (LVOT). While endocardial endothelial cells (EECs) are known to play a role in DSS, the response of these cells to shear stress is not known. In this study, we hypothesize that the response of EECs to shear stress in the LVOT is a mediator of DSS.
To test this hypothesis, we conditioned porcine EECs to controlled shear stress regimes using cone-and-plate bioreactors. Subsequently, we quantified the concentration of proinflammatory cytokine in the conditioned media using the Luminex assay. Bulk-RNA sequencing was used to quantify changes in the genotype of the shear stress-conditioned EECs.
The expression of CD31 was knocked down, and subsequently, the changes in release of shear stress-induced proinflammatory cytokines released by EECs were quantified using the Luminex assay. The results of these studies show that the inflammatory cytokines were highly selected in the conditioning medium, and under bioreactor treatment, the cell activated the PI3K-AKT and TNF-a signaling, which also triggered the other immune cell responses through Th1, Th2, and Th17 cell differentiation pathways. Furthermore, CD31 was identified as a potential mediator of the proinflammatory response of shear stress-conditioned EECs.
The studies provide a potential link between shear stress and the subsequent proinflammatory response of EECs as a mediator of DSS.
离散性主动脉瓣下狭窄（DSS）是一种先天性心脏病，其特征是左心室流出道（LVOT）中主动脉瓣下方的通道变窄。虽然已知心内膜内皮细胞（EECs）在DSS中起作用，但这些细胞对剪切应力的反应尚不清楚。在本研究中，我们假设EECs对LVOT中剪切应力的反应是DSS的一个介导因素。
为了验证这一假设，我们使用锥板生物反应器使猪EECs适应可控的剪切应力环境。随后，我们使用Luminex检测法对条件培养基中促炎细胞因子的浓度进行了定量。批量RNA测序用于定量剪切应力处理后的EECs基因型的变化。
CD31的表达被敲低，随后，使用Luminex检测法定量EECs释放的剪切应力诱导的促炎细胞因子的变化。这些研究结果表明，炎症细胞因子在条件培养基中被高度筛选，并且在生物反应器处理下，细胞激活了PI3K-AKT和TNF-α信号通路，这也通过Th1、Th2和Th17细胞分化途径触发了其他免疫细胞反应。此外，CD31被确定为剪切应力处理后的EECs促炎反应的潜在介导因子。
这些研究提供了剪切应力与EECs随后作为DSS介导因子的促炎反应之间的潜在联系。
