García-Caballero M, Blacher S, Paupert J, Quesada A R, Medina M A, Noël A
Laboratory of Tumor and Development Biology, Groupe Interdisciplinaire de Génoprotéomique Appliqué-Cancer (GIGA-Cancer), University of Liège, Liège, Belgium.
Departamento de Biología Molecular y Bioquímica, Facultad de Ciencias, and IBIMA (Biomedical Research Institute of Málaga), Universidad de Málaga, Andalucía Tech, Málaga, Spain.
Br J Pharmacol. 2016 Jun;173(12):1966-87. doi: 10.1111/bph.13488. Epub 2016 May 4.
Lymphangiogenesis is an important biological process associated with the pathogenesis of several diseases, including metastatic dissemination, graft rejection, lymphoedema and other inflammatory disorders. The development of new drugs that block lymphangiogenesis has become a promising therapeutic strategy. In this study, we investigated the ability of toluquinol, a 2-methyl-hydroquinone isolated from the culture broth of the marine fungus Penicillium sp. HL-85-ALS5-R004, to inhibit lymphangiogenesis in vitro, ex vivo and in vivo.
We used human lymphatic endothelial cells (LECs) to analyse the effect of toluquinol in 2D and 3D in vitro cultures and in the ex vivo mouse lymphatic ring assay. For in vivo approaches, the transgenic Fli1:eGFPy1 zebrafish, mouse ear sponges and cornea models were used. Western blotting and apoptosis analyses were carried out to search for drug targets.
Toluquinol inhibited LEC proliferation, migration, tubulogenesis and sprouting of new lymphatic vessels. Furthermore, toluquinol induced apoptosis of LECs after 14 h of treatment in vitro, blocked the development of the thoracic duct in zebrafish and reduced the VEGF-C-induced lymphatic vessel formation and corneal neovascularization in mice. Mechanistically, we demonstrated that this drug attenuates VEGF-C-induced VEGFR-3 phosphorylation in a dose-dependent manner and suppresses the phosphorylation of Akt and ERK1/2.
Based on these findings, we propose toluquinol as a new candidate with pharmacological potential for the treatment of lymphangiogenesis-related pathologies. Notably, its ability to suppress corneal neovascularization paves the way for applications in vascular ocular pathologies.
淋巴管生成是一个重要的生物学过程，与多种疾病的发病机制相关，包括转移扩散、移植排斥、淋巴水肿及其他炎症性疾病。开发阻断淋巴管生成的新药已成为一种有前景的治疗策略。在本研究中，我们研究了从海洋真菌青霉属HL - 85 - ALS5 - R004的培养液中分离出的2 - 甲基对苯二酚——甲苯喹醇，在体外、离体及体内抑制淋巴管生成的能力。
我们使用人淋巴管内皮细胞（LECs）分析甲苯喹醇在二维和三维体外培养以及离体小鼠淋巴环试验中的作用。对于体内实验方法，使用了转基因Fli1:eGFPy1斑马鱼、小鼠耳海绵和角膜模型。进行蛋白质印迹分析和凋亡分析以寻找药物靶点。
甲苯喹醇抑制LECs增殖、迁移、管腔形成及新淋巴管的芽生。此外，甲苯喹醇在体外处理14小时后诱导LECs凋亡，阻断斑马鱼胸导管的发育，并减少小鼠中VEGF - C诱导的淋巴管形成和角膜新生血管形成。从机制上讲，我们证明该药物以剂量依赖的方式减弱VEGF - C诱导的VEGFR - 3磷酸化，并抑制Akt和ERK1/2的磷酸化。
基于这些发现，我们提出甲苯喹醇作为一种具有治疗淋巴管生成相关病症药理学潜力的新候选药物。值得注意的是，其抑制角膜新生血管形成的能力为其在血管性眼部疾病中的应用铺平了道路。
