Lyve1-Driven Nras^Q61R Causes Edema, Enlarged Lymphatic Vessels, and Hepatic Vascular Defects in Embryonic Mice

Kaposiform lymphangiomatosis (KLA) is a complex lymphatic anomaly associated with a somatic activating NRAS p.Q61R (NRAS^Q61R) mutation. KLA is characterized by malformed lymphatic vessels that can lead to effusions and coagulopathy. The goal of this study was to generate an in vivo mouse model to determine if prenatal expression of the Nras^Q61R mutation in lymphatic endothelial cells induces disease characteristics found in KLA patients.

A Cre-loxP system was used to conditionally express Nras^Q61R in cells expressing lymphatic vessel endothelial hyaluronan receptor 1 (Lyve1), a marker of lymphatic and other types of endothelial cells that starts being expressed at embryonic day (E) 7.5. Because pups did not survive birth, embryos were collected at E14.5, E15.5, and E18.5 for gross analysis, histology and immunostaining, and organ whole-mounts.

Staining for NRAS^Q61R demonstrated robust recombination in the Nras^Q61R mutant embryos and localization of Nras^Q61R at sites of vascular abnormalities. Nras^Q61R mutant embryos had significant edema and dysmorphic jugular lymph sacs with abnormal Lyve1-positive cellular masses. The lymphatic vessel network in the back skin of the Nras^Q61R mutant embryos had fewer branch points and increased vessel diameter. Nras^Q61R mutant embryos had severe hepatic defects characterized by disordered and enlarged vessels. By E18.5, Nras^Q61R mutant embryos were dead.

Conditional expression of Nras^Q61R in Lyve1-positive cells caused edema, abnormal lymphatic development, and hepatic vascular defects in mouse embryos. These findings further support the role of NRAS^Q61R as a driver of the lymphatic overgrowth, vessel enlargement, and dysfunction in the pathophysiology of KLA.