Mousso Thomas, Rice Kalina, Tumenbayar Bat-Ider, Pham Khanh, Heo Yuna, Heo Su Chin, Lee Kwonmoo, Lombardo Andrew T, Bae Yongho
Department of Pathology and Anatomical Sciences, Jacobs School of Medicine and Biomedical Sciences, University at Buffalo, NY 14203, USA.
Department of Pharmacology and Toxicology, Jacobs School of Medicine and Biomedical Sciences, University at Buffalo, Buffalo, NY 14203, USA.
bioRxiv. 2024 Dec 12:2024.12.11.628062. doi: 10.1101/2024.12.11.628062.
Arterial stiffness is a key contributor to cardiovascular diseases, including atherosclerosis, restenosis, and coronary artery disease, it has been characterized to be associated with the aberrant migration of vascular smooth muscle cells (VSMCs). However, the underlying molecular mechanisms driving VSMC migration in stiff environments remain incompletely understood. We recently demonstrated that survivin, a member of the inhibitor of apoptosis protein family, is highly expressed in both mouse and human VSMCs cultured on stiff polyacrylamide hydrogels, where it modulates stiffness-mediated cell cycle progression and proliferation. However, its role in stiffness-dependent VSMC migration remains unknown. To assess its impact on migration, we performed time-lapse video microscopy on VSMCs seeded on fibronectin-coated soft and stiff polyacrylamide hydrogels, mimicking the physiological stiffness of normal and diseased arteries, with either survivin inhibition or overexpression. We observed that VSMC motility increased under stiff conditions, while pharmacologic or siRNA-mediated inhibition of survivin reduced stiffness-stimulated migration to rates similar to those observed under soft conditions. Further investigation revealed that cells on stiff hydrogels exhibited greater directional movement and robust lamellipodial protrusion compared to those on soft hydrogels. Interestingly, survivin-inhibited cells on stiff hydrogels showed reduced directional persistence and lamellipodial protrusion compared to control cells. We also examined whether survivin overexpression alone is sufficient to induce cell migration on soft hydrogels, and found that survivin overexpression modestly increased cell motility and partially rescued the lack of directional persistence compared to GFP-expressing control VSMCs on soft hydrogels. In conclusion, our findings demonstrate that survivin plays a key role in regulating stiffness-induced VSMC migration, suggesting that targeting survivin and its signaling pathways could offer therapeutic strategies for addressing arterial stiffness in cardiovascular diseases.
动脉僵硬度是心血管疾病的关键促成因素,包括动脉粥样硬化、再狭窄和冠状动脉疾病,其特征是与血管平滑肌细胞(VSMC)的异常迁移有关。然而,在僵硬环境中驱动VSMC迁移的潜在分子机制仍未完全了解。我们最近证明,凋亡抑制蛋白家族成员survivin在培养于僵硬聚丙烯酰胺水凝胶上的小鼠和人类VSMC中均高度表达,在该环境中它调节硬度介导的细胞周期进程和增殖。然而,其在硬度依赖性VSMC迁移中的作用仍然未知。为了评估其对迁移的影响,我们对接种在纤连蛋白包被的软质和硬质聚丙烯酰胺水凝胶上的VSMC进行了延时视频显微镜观察,模拟正常和患病动脉的生理硬度,分别进行survivin抑制或过表达。我们观察到,在僵硬条件下VSMC的运动性增加,而药物或siRNA介导的survivin抑制将硬度刺激的迁移降低至与在软质条件下观察到的速率相似。进一步研究表明,与软质水凝胶上的细胞相比,硬质水凝胶上的细胞表现出更大的定向运动和强大的片状伪足突出。有趣的是,与对照细胞相比,硬质水凝胶上survivin抑制的细胞表现出定向持续性和片状伪足突出减少。我们还检查了单独的survivin过表达是否足以诱导软质水凝胶上的细胞迁移,发现与软质水凝胶上表达GFP的对照VSMC相比,survivin过表达适度增加了细胞运动性并部分挽救了定向持续性的缺乏。总之,我们的研究结果表明,survivin在调节硬度诱导的VSMC迁移中起关键作用,表明靶向survivin及其信号通路可为解决心血管疾病中的动脉僵硬度提供治疗策略。