Altered regulation of cell migration in IRF6-mutated orofacial cleft patients-derived primary cells reveals a novel role of Rho GTPases in cleft/lip palate development
Loading...
Files
Date
Journal Title
Journal ISSN
Volume Title
Publisher
Elsevier B.V.
Abstract
Orofacial clefts are the most common congenital craniofacial birth defects. They occur from a failure in cell proliferation and fusion of neural crest cells of the lip buds and/or palatal shelves. In this study, we investigate the genetic basis and molecular mechanisms in primary cells derived from a cleft and lip palate patient presenting van der Woude syndrome (VWS). Since mutations in the integrin genes are widely correlated with VWS, Interferon Regulatory Factor 6 (IRF6) screening was conducted in a cohort of 200 participants presenting with orofacial anomalies. Primary fibroblastic cells derived from the upper right gingiva and palatal regions were isolated and two cellular populations from two participants were obtained: a control with no cleft phenotype and a patient with a cleft phenotype typical of van der Woude syndrome (VWS). IRF6 targeted sequencing revealed mutations in two distinct families. Our results showed no alteration in the viability of the CLP/VWS patient cells, suggesting the phenotype associate with the disease is not secondary to a defect in cell proliferation. We did however detect a significant decrease in the migratory ability of the CLP with Van der Woude syndrome (CLP/VWS) patient cells, which could account for the phenotype. When compared to normal cells, patient cells showed a lack of polarization, which would account for their lack of mobility. Patient cells showed protrusions all around the cells and a lack of defined leading edge. This was reflected with actin staining, WAVE2 and Arp2 around the cell, and correlated with an increase in Rac1 activation. Consistently with the increase in Rac1 activation, patient cells showed a loss in the maturation of focal adhesions needed for contractility, which also accounts for the lack in cell migration. Our findings give increased understanding of the molecular mechanisms of VWS and expands the knowledge of van der Woude syndrome (VWS) occurrence by providing a strong molecular evidence that CLP with Van der Woude syndrome (CLP/VWS) phenotype is caused by a defect in normal physiological processes of cells. © 2021 Elsevier B.V.
Description
Keywords
Cell migration, Cleft and lip palate, Irf6, Rac1, Van der woude syndrome, Abnormalities, multiple, Actin-related protein 2, Case-control studies, Cell adhesion, Cell movement, Cell proliferation, Cell survival, Cells, cultured, Cleft lip, Cleft palate, Collagen, Cysts, Female, Genetic predisposition to disease, Humans, Interferon regulatory factors, Lip, Male, Models, biological, Mutation, Pedigree, Phenotype, Rho gtp-binding proteins, Wiskott-aldrich syndrome protein family, Interferon regulatory factor 6, Rac1 protein, Rho guanine nucleotide binding protein, Actin related protein 2, Actr2 protein, human, Interferon regulatory factor, Irf6 protein, human, Wasf2 protein, human, Wiskott aldrich syndrome protein, Adult, Arp2 gene, Article, Cell maturation, Cell population, Cell viability, Child, Cleft face, Cleft lip palate, Cohort analysis, Controlled study, Correlational study, Disease association, Disease course, Fibroblast cell line, Focal adhesion, Gene, Gene expression level, Gene expression regulation, Gene mutation, Gene sequence, Gene targeting, Genetic analysis, Gingiva, Human, Human cell, Human tissue, Immunohistochemistry, Irf6 gene, Major clinical study, Molecular dynamics, Patient mobility, Primary cell, Rac1 gene, Wave2 gene, Biological model, Case control study, Cell culture, Cell motion, Cyst, Genetic predisposition, Genetics, Metabolism, Multiple malformation syndrome, Pathology