Data Sheet 1_Endothelial GNAQ p.R183Q mutation confers hemoporfin-mediated photodynamic therapy resistance and drives pathological angiogenesis via the angiopoietin-2/TIE2/PI3K/AKT pathway.docx

Hemoporfin-mediated photodynamic therapy (HMME-PDT) has demonstrated significant advantages in the treatment of Port-wine stains (PWSs). However, the therapeutic efficacy of HMME-PDT remains suboptimal in a subset of patients. Somatic mosaic mutations in GNAQ (c.548G>A, p. R183Q) are frequently detected in endothelial cells (ECs) of lesions and represent a common pathogenic mechanism. In this study, we successfully established an in vitro model of PWSs by introducing the GNAQ p. R183Q mutation into HUVECs using lentiviral infection. Our results revealed that GNAQ p. R183Q mutation enhanced ECs proliferation, migration, and angiogenesis. Moreover, the mutation augmented anti-apoptotic mechanisms, thereby conferring heightened resistance to HMME-PDT-induced apoptosis. Residual angiogenic activity persisted following HMME-PDT treatment. These effects are likely mediated by activation of the angiopoietin-2 (ANGPT2)/TIE2/PI3K/AKT signaling axis. Knockdown of ANGPT2 partly reversed these phenotypic alterations and significantly enhanced the efficacy of HMME-PDT. The combination of HMME-PDT with anti-ANGPT2 therapy holds promise for enhancing therapeutic efficacy, suppressing pathological angiogenesis, and ameliorating the clinical manifestations of PWSs.

血卟啉介导的光动力疗法（Hemoporfin-mediated Photodynamic Therapy, HMME-PDT）在鲜红斑痣（Port-wine stains, PWSs）的治疗中已展现出显著优势。然而，HMME-PDT的治疗效果在部分患者中仍未达最优。研究表明，病变内皮细胞（endothelial cells, ECs）中频繁检出GNAQ基因的c.548G>A、p.R183Q体细胞嵌合突变，这是一类常见的致病机制。本研究通过慢病毒感染将GNAQ p.R183Q突变导入人脐静脉内皮细胞（Human Umbilical Vein Endothelial Cells, HUVECs），成功构建了鲜红斑痣的体外模型。实验结果显示，GNAQ p.R183Q突变可增强内皮细胞的增殖、迁移及血管生成能力；同时，该突变强化了抗凋亡机制，使细胞对HMME-PDT诱导的细胞凋亡产生更强抵抗性，且经HMME-PDT处理后仍残留血管生成活性。上述效应可能通过激活血管生成素-2（angiopoietin-2, ANGPT2）/TIE2/PI3K/AKT信号轴介导。敲低ANGPT2可部分逆转上述表型改变，并显著提升HMME-PDT的治疗效能。因此，HMME-PDT联合抗ANGPT2治疗有望增强鲜红斑痣的治疗效果，抑制病理性血管生成，改善患者的临床症状。