Specific fibroblast subpopulations and neuronal structures provide local sources of Vegfc-processing components during zebrafish lymphangiogenesis

Abstract

Proteolytical processing of the growth factor VEGFC through the concerted activity of CCBE1 and ADAMTS3 is required for lymphatic development to occur. How these factors act together in time and space, and which cell types produce these factors is not understood. Here we assess the function of Adamts3 and the related protease Adamts14 during zebrafish lymphangiogenesis and show both proteins to be able to process Vegfc. Only the simultaneous loss of both protein functions results in lymphatic defects identical to vegfc loss-offunction situations. Cell transplantation experiments demonstrate neuronal structures and/or fibroblasts to constitute cellular sources not only for both proteases but also for Ccbe1 and Vegfc. We further show that this locally restricted Vegfc maturation is needed to trigger normal lymphatic sprouting and directional migration. Our data provide a single-cell resolution model for establishing secretion and processing hubs for Vegfc during developmental lymphangiogenesis.

Cellular Origins of the Lymphatic Endothelium: Implications for Cancer Lymphangiogenesis

“…Likewise, loss of the Vegfc-processing components adamts3 and adamts14 in zebrafish also mimicked the vegfc mutant phenotype exhibiting absent trunk, facial and meningeal lining lymphatics (Wang et al, 2020).…”

“…A disintegrin and metalloproteinase with thrombospondin motifs 3 (ADAMTS3) is a VEGF-C protease, whose total deletion (i.e., Adamts3 –/– mice) mirrored the phenotypes observed in Ccbe1 and Vegfc null embryos ( Bui et al, 2016 ). Likewise, loss of the Vegfc-processing components adamts3 and adamts14 in zebrafish also mimicked the vegfc mutant phenotype exhibiting absent trunk, facial and meningeal lining lymphatics ( Wang et al, 2020 ).…”

Endothelial cell-type-specific molecular requirements for angiogenesis drive fenestrated vessel development in the brain

“…Prior studies have shown that Ccbe1, Adamts3, and Adamts14 serve to proteolytically cleave and convert the pro-active form of Vegfc into its mature form during lymphangiogenesis in zebrafish ( Hogan et al, 2009 ; Le Guen et al, 2014 ; Wang et al, 2020 ). Among these three proteins, ccbe1 appears to be highly expressed in the developing brain and meningeal compartments based on its BAC transgenic reporter expression at 48 hpf ( Wang et al, 2020 ), while adamts3 BAC transgenic reporter expression appear to be very low at the same stage. Since 48 hpf is a developmental time point when the DLV is extending toward the mCP, ccbe1 could be a strong candidate that acts to activate Vegfc in this area of the brain.…”

The quest for substrates and binding partners: A critical barrier for understanding the role of ADAMTS proteases in musculoskeletal development and disease

“…Polymorphisms in ADAMTS14 have been associated with delayed onset of Achilles tendon pathology and increased risk for OA 80‐82 . In a recent study in zebrafish, it was demonstrated that both, ADAMTS3 and ADAMTS14 can process VEGF‐C and that inactivation of both proteases was required to elicit a lymphatic phenotype 83 . This suggests that, in contrast to mice and humans, ADAMTS14 can cleave VEGF‐C and compensate for the loss of ADAMTS3 in zebrafish.…”