Journal Article

E-cadherin variants associated with oral facial clefts trigger aberrant cell motility in a REG1A-dependent manner

Morais-de-Sá, Eurico, Figueiredo, Ceu, Carneiro, Patrícia, Seruca, Raquel, Figueiredo, Joana, Yang, Vítor, Maia, André F., Machado, José Carlos, Carvalho, João, Melo, Soraia, Pereira, Joana, Ferreira, Rui M.

2024

Overview

Background Germline mutations of E-cadherin contribute to hereditary diffuse gastric cancer (HDGC) and congenital malformations, such as oral facial clefts (OFC). However, the molecular mechanisms through which E-cadherin loss-of-function triggers distinct clinical outcomes remain unknown. We postulate that E-cadherin-mediated disorders result from abnormal interactions with the extracellular matrix and consequent aberrant intracellular signalling, affecting the coordination of cell migration. Methods Herein, we developed in vivo and in vitro models of E-cadherin mutants associated with either OFC or HDGC. Using a Drosophila approach, we addressed the impact of the different variants in cell morphology and migration ability. By combining gap closure migration assays and time-lapse microscopy, we further investigated the migration pattern of cells expressing OFC or HDGC variants. The adhesion profile of the variants was evaluated using high-throughput ECM arrays, whereas RNA sequencing technology was explored for identification of genes involved in aberrant cell motility. Results We have demonstrated that cells expressing OFC variants exhibit an excessive motility performance and irregular leading edges, which prevent the coordinated movement of the epithelial monolayer. Importantly, we found that OFC variants promote cell adhesion to a wider variety of extracellular matrices than HDGC variants, suggesting higher plasticity in response to different microenvironments. We unveiled a distinct transcriptomic profile in the OFC setting and pinpointed REG1A as a putative regulator of this outcome. Consistent with this, specific RNAi-mediated inhibition of REG1A shifted the migration pattern of OFC expressing cells, leading to slower wound closure with coordinated leading edges. Conclusions We provide evidence that E-cadherin variants associated with OFC activate aberrant signalling pathways that support dynamic rearrangements of cells towards improved adaptability to the microenvironment. This proficiency results in abnormal tissue shaping and movement, possibly underlying the development of orofacial malformations. Graphical Abstract

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Adaptability / Adenocarcinoma / Animals / Biomedical and Life Sciences / Birth defects / Cadherins / Cadherins - genetics / Cadherins - metabolism / Cell Adhesion / Cell adhesion & migration / Cell Biology / Cell migration / Cell Movement / Cells / Cleft lip / Congenital defects / Cytokines and Growth Factors / Cytology / Drosophila melanogaster / E-cadherin / Extracellular matrix / Gastric cancer / Gene mutations / Genetic aspects / Genetic variation / Germ-Line Mutation / Health aspects / Hereditary diffuse gastric cancer / Humans / Insects / Intracellular signalling / Life Sciences / Lithostathine - genetics / Medical research / Medicine, Experimental / Microenvironments / Molecular modelling / Motility / Oral facial clefts / Ovaries / Protein-Ligand Interactions / Receptors / REG1A / Risk factors / RNA-mediated interference / Signal transduction / Software / Stomach Neoplasms - metabolism / Transcriptomics / Tumor Microenvironment / Wound healing