Endothelin-3 stimulates cell adhesion and cooperates with β1-integrins during enteric nervous system ontogenesis

Abstract

Endothelin-3 (EDN3) and β1-integrins are required for the colonization of the embryonic gut by enteric neural crest cells (ENCCs) to form the enteric nervous system (ENS). β1-integrin-null ENCCs exhibit migratory defects in a region of the gut enriched in EDN3 and in specific extracellular matrix (ECM) proteins. We investigated the putative role of EDN3 on ENCC adhesion properties and its functional interaction with β1-integrins during ENS development. We show that EDN3 stimulates ENCC adhesion to various ECM components in vitro. It induces rapid changes in ENCC shape and protrusion dynamics favouring sustained growth and stabilization of lamellipodia, a process coincident with the increase in the number of focal adhesions and activated β1-integrins. In vivo studies and ex-vivo live imaging revealed that double mutants for Itgb1 and Edn3 displayed a more severe enteric phenotype than either of the single mutants demonstrated by alteration of the ENS network due to severe migratory defects of mutant ENCCs taking place early during the ENS development. Altogether, our results highlight the interplay between the EDN3 and β1-integrin signalling pathways during ENS ontogenesis and the role of EDN3 in ENCC adhesion.

Collagen 18 and agrin are secreted by enteric neural crest cells to remodel their microenvironment and regulate their migration during ENS development

“…ENCDC migration relies on their expression of β1-integrin, an important receptor for ECM adhesion (Nagy et al, 2009). Homozygous deletion of β1-integrin from ENCDCs arrests their migration in the proximal hindgut, leading to distal aganglionosis (Breau et al, 2009;Gazquez et al, 2016). In addition, in both avian and mouse embryos, sonic hedgehog secretion from the gut epithelium promotes mesenchymal expression of chondroitin sulfate proteoglycans (CSPGs), including versican and Col9, which inhibit ENS migration and patterning (Nagy et al, 2016).…”

EDNRBmutations cause Waardenburg syndrome type II in the heterozygous state

“…In particular, the ligand endothelin 3 and its physiological receptor in development, the endothelin receptor type B, are important for the survival, proliferation, migration, and/or differentiation of melanoblasts along the dorsolateral pathway (Lahav, ; Saldana‐Caboverde & Kos, ). The EDNRB/EDN3 (MIM# 131244, MIM# 131242) pathway also regulates cell migration/adhesion and prevents premature differentiation of enteric progenitors (Bondurand, Natarajan, Barlow, Thapar, & Pachnis, ; Gazquez et al., ). Homozygous (or compound heterozygous) mutations in the respective two genes have been described in WS4 (MIM# 277580), that is, WS with HD (Edery et al., ; Hofstra et al., ; Puffenberger et al., ).…”

Biological and translational implications of enteric nerves in colorectal cancer

“…In line with two recent publications by Benesh et al 44 83,84 and that NID1 (a p53-repressed gene) and FBLN2 enhance the proliferation of CRC 85 and lung cancer 86 cells, respectively, in vitro. Besides, with higher levels of NID1 and FBLN2 in mesenchymal-like (CRC) cells and the positive correlation between NID1 expression and mesenchymal-associated genes 85,[87][88][89] , it is likely that Nid1 and Fbln2 also drive the progression of CRC. In fact, similar as in ovarian 83,90 , breast 88,91,92 , endometrial 93 and lung 86,94…”

“…The principle of targeted therapy is that suitable cell surface antigens that are overexpressed, mutated or selectively expressed by tumors in comparison to normal tissue are targeted and through which functional alterations are induced by influencing antigen or receptor functions, modulating the immune system or specific drug delivery 87 . Vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR), have been shown to improve overall survival in metastatic CRC when efficiently targeted 88 .…”

“…First, the activated immune system is able to induce further genetic mutations and epigenetic alterations by the release of reactive oxygen species (ROS) or reactive nitrogen species (RNS) which are known to cause DNA damage 83 . This damage may mutate p53 and mismatch repair genes while cytokines can induce epigenetic alterations or deregulate the expression of miRNAs causing changes in gene silencing [84][85][86][87] . Second, inflammatory signals, often released cytokines, are able to induce CRC development via different mechanisms than in non-CAC.…”