The potential role of YAP in Axl-mediated resistance to EGFR tyrosine kinase inhibitors.

Yes-associated protein (YAP) is a transcription co-regulator downstream of the Hippo pathway, and plays a critical role in cancer. Although YAP regulation in the canonical Hippo pathway is well established, the Hippo-independent regulation of YAP is not well explored. Here, we showed the possible new mechanism of YAP regulation by the receptor tyrosine kinase Axl. Co-immunoprecipitation and Western blot analysis demonstrated the interaction between YAP and Axl, which was enhanced by Axl ligand Growth Arrest Specific 6 (GAS6) stimulation. Furthermore, we found that YAP is phosphorylated at tyrosine residues by GAS6 stimulation in vivo and Axl directly phosphorylates YAP in vitro. Axl overexpression or GAS6 stimulation increased YAP-mediated transcriptional activity, and YAP-mediated colony forming activity in soft agar was enhanced by co-expression of Axl. In EGFR tyrosine kinase inhibitor (TKI)-sensitive lung cancer cells, YAP protein was downregulated in response to TKI treatment, while overexpression of YAP attenuated TKI sensitivity, suggesting that YAP is a key determinant of TKI response. Moreover Axl overexpression reversed TKI-induced YAP downregulation and induced TKI-resistance, which was reversed by YAP knockdown, further supporting the notion that YAP functions downstream of Axl. Together, these findings suggest a novel role of YAP in Axl-mediated TKI resistance.

Erythrocyte phospholipid and polyunsaturated fatty acid composition in diabetic retinopathy.

BACKGROUND: Long chain polyunsaturated fatty acids (LCPUFAs) including docosahexaenoic acid and arachidonic acid are suspected to play a key role in the pathogenesis of diabetes. LCPUFAs are known to be preferentially concentrated in specific phospholipids termed as plasmalogens. This study was aimed to highlight potential changes in the metabolism of phospholipids, and particularly plasmalogens, and LCPUFAs at various stages of diabetic retinopathy in humans. METHODOLOGY AND PRINCIPAL FINDINGS: We performed lipidomic analyses on red blood cell membranes from controls and mainly type 2 diabetes mellitus patients with or without retinopathy. The fatty acid composition of erythrocytes was determined by gas chromatography and the phospholipid structure was determined by liquid chromatography equipped with an electrospray ionisation source and coupled with a tandem mass spectrometer (LC-ESI-MS/MS). A significant decrease in levels of docosahexaenoic acid and arachidonic acid in erythrocytes of diabetic patients with or without retinopathy was observed. The origin of this decrease was a loss of phosphatidyl-ethanolamine phospholipids esterified with these LCPUFAs. In diabetic patients without retinopathy, this change was balanced by an increase in the levels of several phosphatidyl-choline species. No influence of diabetes nor of diabetic retinopathy was observed on the concentrations of plasmalogen-type phospholipids. CONCLUSIONS AND SIGNIFICANCE: Diabetes and diabetic retinopathy were associated with a reduction of erythrocyte LCPUFAs in phosphatidyl-ethanolamines. The increase of the amounts of phosphatidyl-choline species in erythrocytes of diabetic patients without diabetic retinopathy might be a compensatory mechanism for the loss of LC-PUFA-rich phosphatidyl-ethanolamines.

Plasmalogens in the retina: from occurrence in retinal cell membranes to potential involvement in pathophysiology of retinal diseases.

Plasmalogens (Pls) represent a specific subclass of glycerophospholipids characterized by the presence of a vinyl-ether bond at the sn-1 position of glycerol. Pls are quantitatively important in membranes of neuronal tissues, including the brain and the retina, where they can represent until almost two-third of ethanolamine glycerophospholipids. They are considered as reservoirs of polyunsaturated fatty acids as several studies have shown that arachidonic and docosahexaenoic acids are preferentially esterified on Pls when compared to other glycerophospholipids. Reduced levels of Pls were observed in a number of neurodegenerative disorders such as glaucoma, the second leading cause of blindness worldwide. In a mouse model of Pls deficiency, "glaucoma-like" optic nerve abnormalities were observed as well as developmental defects in the eye. These included microphthalmia, dysgenesis of the anterior segment of the eye, and abnormalities in retinal vessel architecture. Several data from animal and in vitro studies suggest that Pls may be involved in the regulation of retinal vascular development through the release of polyunsaturated fatty acids by a calcium-independent phospholipase A2.

Involvement of plasmalogens in post-natal retinal vascular development.

OBJECTIVE: Proper development of retinal blood vessels is essential to ensure sufficient oxygen and nutrient supplies to the retina. It was shown that polyunsaturated fatty acids (PUFAs) could modulate factors involved in tissue vascularization. A congenital deficiency in ether-phospholipids, also termed "plasmalogens", was shown to lead to abnormal ocular vascularization. Because plasmalogens are considered to be reservoirs of PUFAs, we wished to improve our understanding of the mechanisms by which plasmalogens regulate retinal vascular development and whether the release of PUFAs by calcium-independent phospholipase A2 (iPLA2) could be involved. METHODS AND RESULTS: By characterizing the cellular and molecular steps of retinal vascular development in a mouse model of plasmalogen deficiency, we demonstrated that plasmalogens modulate angiogenic processes during the early phases of retinal vascularization. They influence glial activity and primary astrocyte template formation, endothelial cell proliferation and retinal vessel outgrowth, and impact the expression of the genes involved in angiogenesis in the retina. These early defects led to a disorganized and dysfunctional retinal vascular network at adult age. By comparing these data to those obtained on a mouse model of retinal iPLA2 inhibition, we suggest that these processes may be mediated by PUFAs released from plasmalogens and further signalling through the angiopoietin/tie pathways. CONCLUSIONS: These data suggest that plasmalogens play a crucial role in retinal vascularization processes.