NRASQ61R mutation drives elevated angiopoietin‐2 expression in human endothelial cells and a genetic mouse model

Abstract

AbstractBackgroundAngiopoietin‐2 (Ang‐2) is increased in the blood of patients with kaposiform lymphangiomatosis (KLA) and kaposiform hemangioendothelioma (KHE). While the genetic causes of KHE are not clear, a somatic activating NRASQ61R mutation has been found in the lesions of KLA patients.ProcedureOur study tested the hypothesis that the NRASQ61R mutation drives elevated Ang‐2 expression in endothelial cells. Ang‐2 was measured in human endothelial progenitor cells (EPC) expressing NRASQ61R and a genetic mouse model with endothelial targeted NRASQ61R. To determine the signaling pathways driving Ang‐2, NRASQ61R EPC were treated with signaling pathway inhibitors.ResultsAng‐2 levels were increased in EPC expressing NRASQ61R compared to NRASWT by Western blot analysis of cell lysates and ELISA of the cell culture media. Ang‐2 levels were elevated in the blood of NRASQ61R mutant mice. NRASQ61R mutant mice also had reduced platelet counts and splenomegaly with hypervascular lesions, like some KLA patients. mTOR inhibitor rapamycin attenuated Ang‐2 expression by NRASQ61R EPC. However, MEK1/2 inhibitor trametinib was more effective blocking increases in Ang‐2.ConclusionsOur studies show that the NRASQ61R mutation in endothelial cells induces Ang‐2 expression in vitro and in vivo. In cultured human endothelial cells, NRASQ61R drives elevated Ang‐2 through MAP kinase and mTOR‐dependent signaling pathways.