RESUMEN
Endothelins (EDN) are peptide hormones that activate a GPCR signalling system and contribute to several diseases, including hypertension and cancer. Current knowledge about EDN signalling is fragmentary, and no systems level understanding is available. We investigated phosphoproteomic changes caused by endothelin B receptor (ENDRB) activation in the melanoma cell lines UACC257 and A2058 and built an integrated model of EDNRB signalling from the phosphoproteomics data. More than 5,000 unique phosphopeptides were quantified. EDN induced quantitative changes in more than 800 phosphopeptides, which were all strictly dependent on EDNRB. Activated kinases were identified based on high confidence EDN target sites and validated by Western blot. The data were combined with prior knowledge to construct the first comprehensive logic model of EDN signalling. Among the kinases predicted by the signalling model, AKT, JNK, PKC and AMP could be functionally linked to EDN-induced cell migration. The model contributes to the system-level understanding of the mechanisms underlying the pleiotropic effects of EDN signalling and supports the rational selection of kinase inhibitors for combination treatments with EDN receptor antagonists.
Asunto(s)
Endotelinas/farmacología , Regulación Neoplásica de la Expresión Génica , Melanocitos/metabolismo , Fosfoproteínas/genética , Procesamiento Proteico-Postraduccional , Transducción de Señal , Proteínas Quinasas Activadas por AMP/genética , Proteínas Quinasas Activadas por AMP/metabolismo , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Endotelinas/genética , Endotelinas/metabolismo , Redes Reguladoras de Genes , Humanos , MAP Quinasa Quinasa 4/genética , MAP Quinasa Quinasa 4/metabolismo , Melanocitos/efectos de los fármacos , Melanocitos/patología , Fosfoproteínas/metabolismo , Fosforilación , Proteína Quinasa C/genética , Proteína Quinasa C/metabolismo , Proteómica/métodos , Proteínas Proto-Oncogénicas c-akt/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Receptor de Endotelina B/genética , Receptor de Endotelina B/metabolismoRESUMEN
The World PGX Summit was held in Boston and preceded by a 1-day workshop. The conference aimed at assessing the current 'state-of-the art' in advanced molecular profiling strategies for increased drug development success. The topics varied from regulatory policies, pharmaceutical case examples and vendor presentations on innovative technologies. The subject is obviously closely related to personalized medicine and it was interesting to hear the perspectives from healthcare providers and subscribers. It became clear during the meeting that we are on the verge of important changes in how pharmaceutical research and development operates but also how increased costs and high unmet medical needs require new models in which pharmacogenomics can play an important role.