PKCθ-JunB axis via upregulation of VEGFR3 expression mediates hypoxia-induced pathological retinal neovascularization.

Abstract

Pathological retinal neovascularization is the most common cause of vision loss. PKCθ has been shown to play a role in type 2 diabetes, which is linked to retinal neovascularization. Based on these clues, we have studied the role of PKCθ and its downstream target genes JunB and VEGFR3 in retinal neovascularization using global and tissue-specific knockout mouse models along with molecular biological approaches. Here, we show that vascular endothelial growth factor A (VEGFA) induces PKCθ phosphorylation in human retinal microvascular endothelial cells (HRMVECs) and downregulation of its levels attenuates VEGFA-induced HRMVECs migration, sprouting and tube formation. Furthermore, the whole body deletion of PKCθ or EC-specific deletion of its target gene JunB inhibited hypoxia-induced retinal EC proliferation, tip cell formation and neovascularization. VEGFA also induced VEGFR3 expression via JunB downstream to PKCθ in the regulation of HRMVEC migration, sprouting, and tube formation in vitro and OIR-induced retinal EC proliferation, tip cell formation and neovascularization in vivo. In addition, VEGFA-induced VEGFR3 expression requires VEGFR2 activation upstream to PKCθ-JunB axis both in vitro and in vivo. Depletion of VEGFR2 or VEGFR3 levels attenuated VEGFA-induced HRMVEC migration, sprouting and tube formation in vitro and retinal neovascularization in vivo and it appears that these events were dependent on STAT3 activation. Furthermore, the observations using soluble VEGFR3 indicate that VEGFR3 mediates its effects on retinal neovascularization in a ligand dependent and independent manner downstream to VEGFR2. Together, these observations suggest that PKCθ-dependent JunB-mediated VEGFR3 expression targeting STAT3 activation is required for VEGFA/VEGFR2-induced retinal neovascularization.

• Authors: Raj Kumar, Arul M Mani, Nikhlesh K Singh, Gadiparthi N Rao
• Journal: Cell death & disease (Cell Death Dis) Vol. 11 Issue 5 Pg. 325 (05 07 2020) ISSN: 2041-4889 [Electronic] England
• PMID: 32382040 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• JunB protein, human
• Ligands
• Transcription Factors
• Vascular Endothelial Growth Factor A
• Vascular Endothelial Growth Factor Receptor-3
• Protein Kinase C-theta

Topics

• Animals
• Cell Movement (drug effects, genetics)
• Endothelial Cells (drug effects, metabolism)
• Humans
• Hypoxia (complications)
• Ligands
• Mice, Inbred C57BL
• Neovascularization, Physiologic (drug effects, genetics)
• Promoter Regions, Genetic (genetics)
• Protein Kinase C-theta (metabolism)
• Retinal Neovascularization (etiology, genetics, pathology)
• Signal Transduction (drug effects)
• Transcription Factors (metabolism)
• Up-Regulation (drug effects, genetics)
• Vascular Endothelial Growth Factor A (pharmacology)
• Vascular Endothelial Growth Factor Receptor-3 (genetics, metabolism)