Institute of Mechanobiology & Medical Engineering, School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, 800 Dongchuan Road, Minhang, 200240, Shanghai, China.
Department of Pediatric Surgery, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, 1665 Kongjiang Road, Yangpu, Shanghai, 200092, China.
Angiogenesis. 2022 Feb;25(1):71-86. doi: 10.1007/s10456-021-09804-7. Epub 2021 Jun 25.
Aberrant variations in angiogenesis have been observed in tumor tissues with abnormal stiffness of extracellular matrix (ECM). However, it remains largely unclear how ECM stiffness influences tumor angiogenesis. Numerous studies have reported that vascular endothelial growth factor-A (VEGF-A) released from tumor cells plays crucial roles in angiogenesis. Hence, we demonstrated the role of ECM stiffness in VEGF-A release from neuroblastoma (NB) cells and the underlying mechanisms. Based on 17 NB clinical samples, a negative correlation was observed between the length of blood vessels and stiffness of NB tissues. In vitro, an ECM stiffness of 30 kPa repressed the secretion of VEGF from NB cells which subsequently inhibited the tube formation of human umbilical vein endothelial cells (HUVECs). Knocked down VEGF in NB cells or blocked VEGF with neutralizing antibodies both repressed the tube formation of HUVECs. Specifically, 30 kPa ECM stiffness repressed the expression and nuclear accumulation of Yes-associated protein (YAP) to regulate the expression of Serine/Arginine Splicing Factor 1 (SRSF1) via Runt-related transcription factor 2 (RUNX2), which may then subsequently induce the expression and secretion of VEGF in NB tumor cells. Through implantation of 3D col-Tgels with different stiffness into nude mice, the inhibitory effect of 30 kPa on NB angiogenesis was confirmed in vivo. Furthermore, we found that the inhibitory effect of 30 kPa stiffness on NB angiogenesis was reversed by YAP overexpression, suggesting the important role of YAP in NB angiogenesis regulated by ECM stiffness. Overall, our work not only showed a regulatory effect of ECM stiffness on NB angiogenesis, but also revealed a new signaling axis, YAP-RUNX2-SRSF1, that mediates angiogenesis by regulating the expression and secretion of VEGF from NB cells. ECM stiffness and the potential molecules revealed in the present study may be new therapeutic targets for NB angiogenesis.
已经观察到细胞外基质（ECM）异常僵硬的肿瘤组织中存在血管生成的异常变化。然而，ECM 硬度如何影响肿瘤血管生成在很大程度上仍不清楚。许多研究报告称，肿瘤细胞释放的血管内皮生长因子-A（VEGF-A）在血管生成中起着至关重要的作用。因此，我们展示了 ECM 硬度在神经母细胞瘤（NB）细胞中 VEGF-A 释放中的作用及其潜在机制。基于 17 个 NB 临床样本，观察到血管长度与 NB 组织硬度之间存在负相关。在体外，30 kPa 的 ECM 硬度抑制了 NB 细胞中 VEGF 的分泌，进而抑制了人脐静脉内皮细胞（HUVEC）的管形成。NB 细胞中 VEGF 的敲低或中和抗体阻断 VEGF 均抑制了 HUVEC 的管形成。具体而言，30 kPa ECM 硬度通过 Runt 相关转录因子 2（RUNX2）抑制 Yes 相关蛋白（YAP）的表达和核积累来调节丝氨酸/精氨酸剪接因子 1（SRSF1）的表达，从而可能随后诱导 NB 肿瘤细胞中 VEGF 的表达和分泌。通过将不同硬度的 3D col-Tgels 植入裸鼠中，在体内证实了 30 kPa 对 NB 血管生成的抑制作用。此外，我们发现 30 kPa 硬度对 NB 血管生成的抑制作用可被 YAP 过表达逆转，这表明 YAP 在 ECM 硬度调节的 NB 血管生成中起重要作用。总的来说，我们的工作不仅显示了 ECM 硬度对 NB 血管生成的调节作用，而且还揭示了一个新的信号轴，YAP-RUNX2-SRSF1，通过调节 NB 细胞中 VEGF 的表达和分泌来介导血管生成。本研究中揭示的 ECM 硬度和潜在分子可能成为 NB 血管生成的新治疗靶点。
