Avnet Sofia, Mizushima Emi, Severino Beatrice, Lipreri Maria Veronica, Scognamiglio Antonia, Corvino Angela, Baldini Nicola, Cortini Margherita
Department of Biomedical and Neuromotor Sciences, University of Bologna, 40138 Bologna, Italy.
Department of Orthopaedic Surgery, School of Medicine, Sapporo Medical University, Sapporo 060-8543, Hokkaido, Japan.
Metabolites. 2025 Mar 5;15(3):178. doi: 10.3390/metabo15030178.
Angiogenesis, the process of new blood vessel formation, is critically regulated by a balance of pro- and anti-angiogenic factors. This process plays a central role in tumor progression and is modulated by tumor cells. Sphingosine-1-phosphate (S1P), a bioactive lipid signaling molecule acting via G-protein-coupled receptors (S1PR1-5), has emerged as a key mediator of vascular development and pathological angiogenesis in cancer. Consequently, targeting the S1P-S1PRs axis represents a promising strategy for antiangiogenic therapies. This study explores S1PR3 as a potential therapeutic target in osteosarcoma, the most common primary bone malignancy, which we have previously demonstrated to secrete S1P within the acidic tumor microenvironment.
The effects of KRX-725-II and its derivatives, Tic-4-KRX-725-II and [D-Tic]4-KRX-725-II-pepducins acting as S1PR3 antagonists as allosteric modulators of GPCR activity-were tested on metastatic osteosarcoma cells (143B) for proliferation and migration inhibition. Anti-angiogenic activity was assessed using endothelial cells (HUVEC) through proliferation and tubulogenesis assays in 2D, alongside sprouting and migration analyses in a 3D passively perfused microfluidic chip.
S1PR3 inhibition did not alter osteosarcoma cell growth or migration. However, it impaired endothelial cell tubulogenesis up to 75% and sprouting up to 30% in respect to controls. Conventional 2D assays revealed reduced tubule nodes and length, while 3D microfluidic models demonstrated diminished sprouting area and maximum migration distance, indicating S1PR3's role in driving endothelial cell differentiation.
These findings highlight S1PR3 as a critical regulator of angiogenesis and posit its targeting as a novel anti-angiogenic strategy, particularly for aggressive, S1P-secreting tumors with pronounced metastatic potential and an acidic microenvironment.
血管生成是新血管形成的过程，受促血管生成因子和抗血管生成因子平衡的严格调控。这一过程在肿瘤进展中起核心作用，并受肿瘤细胞调节。鞘氨醇-1-磷酸（S1P）是一种通过G蛋白偶联受体（S1PR1 - 5）发挥作用的生物活性脂质信号分子，已成为血管发育和癌症病理血管生成的关键介质。因此，靶向S1P - S1PRs轴是抗血管生成治疗的一个有前景的策略。本研究探讨S1PR3作为骨肉瘤潜在治疗靶点的可能性，骨肉瘤是最常见的原发性骨恶性肿瘤，我们之前已证明其在酸性肿瘤微环境中分泌S1P。
测试KRX - 725 - II及其衍生物Tic - 4 - KRX - 725 - II和[D - Tic]4 - KRX - 725 - II - pepducins作为S1PR3拮抗剂（作为GPCR活性的变构调节剂）对转移性骨肉瘤细胞（143B）增殖和迁移抑制的影响。通过内皮细胞（HUVEC）在二维增殖和管腔形成试验中评估抗血管生成活性，同时在三维被动灌注微流控芯片中进行芽生和迁移分析。
抑制S1PR3并未改变骨肉瘤细胞的生长或迁移。然而，与对照组相比，它使内皮细胞管腔形成减少高达75%，芽生减少高达30%。传统的二维试验显示管腔节点和长度减少，而三维微流控模型显示芽生面积和最大迁移距离减小，表明S1PR3在驱动内皮细胞分化中的作用。
这些发现突出了S1PR3作为血管生成的关键调节因子，并将其靶向作为一种新的抗血管生成策略，特别是对于具有明显转移潜力和酸性微环境的侵袭性、分泌S1P的肿瘤。
