Mechanisms of endothelial cell protection by blockade of the JAK2 pathway

Resumen

Inhibition of the JAK2/ STAT pathway has been implicated recently in cytoprotective mech- anisms in both vascular smooth muscle cells and astrocytes. The advent of JAK2-specific inhibitors provides a practical tool for the study of this pathway in different cellular types. An interest in finding methods to improve endothelial cell (EC) resistance to injury led us to examine the effect of JAK2/STAT inhibition on EC protection. Furthermore, the signaling pathways involved in JAK2/STAT inhibi- tion-related actions were examined. Our results reveal, for the first time, that blockade of JAK2 with the tyrosine kinase inhibitor AG490 strongly protects cultured EC against cell detachment-dependent death and serum deprivation and increases reseeding efficiency. Con- firmation of the specificity of the effects of JAK2 inhibition was attained by finding protective effects on transfection with a dominant negative JAK2. Furthermore, AG490 blocked serum deprivation- induced phosphorylation of JAK2. In terms of mechanism, treatment with AG490 induces several relevant responses, both in monolayer and detached cells. These mechanisms include the following: 1) Increase and nuclear translocation of the active, dephosphorylated form of -catenin. In functional terms, this translocation is transcrip- tionally active, and its protective effect is further supported by the stimulation of EC cytoprotection by transfectionally induced excess of -catenin. 2) Increase of platelet endothelial cell adhesion molecule (PECAM)/CD31 levels. 3) Increase in total and phosphorylated AKT. 4) Increase in phosphorylated glycogen synthase kinase (GSK)3 / . The present findings imply potential practical applications of JAK2 inhibition on EC. These applications affect not only EC in the monolayer but also circulating detached cells and involve mechanistic interactions not previously described.

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Citación

Mechanisms of endothelial cell protection by blockade of the JAK2 pathway Fernando Neria, Carlos Caramelo, Héctor Peinado, Francisco R. González-Pacheco, Juan JP. Deudero, Alain J. de Solis, Ruth Fernández-Sánchez, Silvia Peñate, Amparo Cano, and Mª Ángeles Castilla American Journal of Physiology-Cell Physiology 2007 292:3, C1123-C1131