title: The Meckel-Gruber syndrome protein TMEM67 controls basal body positioning and epithelial branching morphogenesis in mice via the non-canonical Wnt pathway.
abstract: Ciliopathies are a group of developmental disorders that manifest with multi-organ anomalies. Mutations in TMEM67 (MKS3) cause a range of human ciliopathies, including Meckel-Gruber and Joubert syndromes. In this study we describe multi-organ developmental abnormalities in the Tmem67(tm1Dgen/H1) knockout mouse that closely resemble those seen in Wnt5a and Ror2 knockout mice. These include pulmonary hypoplasia, ventricular septal defects, shortening of the body longitudinal axis, limb abnormalities, and cochlear hair cell stereociliary bundle orientation and basal body/kinocilium positioning defects. The basal body/kinocilium complex was often uncoupled from the hair bundle, suggesting aberrant basal body migration, although planar cell polarity and apical planar asymmetry in the organ of Corti were normal. TMEM67 (meckelin) is essential for phosphorylation of the non-canonical Wnt receptor ROR2 (receptor-tyrosine-kinase-like orphan receptor 2) upon stimulation with Wnt5a-conditioned medium. ROR2 also colocalises and interacts with TMEM67 at the ciliary transition zone. Additionally, the extracellular N-terminal domain of TMEM67 preferentially binds to Wnt5a in an in vitro binding assay. Cultured lungs of Tmem67 mutant mice failed to respond to stimulation of epithelial branching morphogenesis by Wnt5a. Wnt5a also inhibited both the Shh and canonical Wnt/β-catenin signalling pathways in wild-type embryonic lung. Pulmonary hypoplasia phenotypes, including loss of correct epithelial branching morphogenesis and cell polarity, were rescued by stimulating the non-canonical Wnt pathway downstream of the Wnt5a-TMEM67-ROR2 axis by activating RhoA. We propose that TMEM67 is a receptor that has a main role in non-canonical Wnt signalling, mediated by Wnt5a and ROR2, and normally represses Shh signalling. Downstream therapeutic targeting of the Wnt5a-TMEM67-ROR2 axis might, therefore, reduce or prevent pulmonary hypoplasia in ciliopathies and other congenital conditions.
authors.affiliation: Ciliopathy Research Group, Section of Ophthalmology and Neurosciences, Leeds Institute of Molecular Medicine, University of Leeds, Leeds LS9 7TF, UK Department of Anatomy and Embryology, Faculty of Medicine, Al-Azhar University, Cairo 11844, Egypt.
authors.forename: Zakia A
authors.initials: ZA
authors.lastname: Abdelhamed
authors.affiliation: Ciliopathy Research Group, Section of Ophthalmology and Neurosciences, Leeds Institute of Molecular Medicine, University of Leeds, Leeds LS9 7TF, UK.
authors.forename: Subaashini
authors.initials: S
authors.lastname: Natarajan
authors.affiliation: Ciliopathy Research Group, Section of Ophthalmology and Neurosciences, Leeds Institute of Molecular Medicine, University of Leeds, Leeds LS9 7TF, UK.
authors.forename: Gabrielle
authors.initials: G
authors.lastname: Wheway
authors.affiliation: Ciliopathy Research Group, Section of Ophthalmology and Neurosciences, Leeds Institute of Molecular Medicine, University of Leeds, Leeds LS9 7TF, UK.
authors.forename: Christopher F
authors.initials: CF
authors.lastname: Inglehearn
authors.affiliation: Ciliopathy Research Group, Section of Ophthalmology and Neurosciences, Leeds Institute of Molecular Medicine, University of Leeds, Leeds LS9 7TF, UK.
authors.forename: Carmel
authors.initials: C
authors.lastname: Toomes
authors.affiliation: Ciliopathy Research Group, Section of Ophthalmology and Neurosciences, Leeds Institute of Molecular Medicine, University of Leeds, Leeds LS9 7TF, UK c.johnson@leeds.ac.uk d.jagger@ucl.ac.uk.
authors.forename: Colin A
authors.initials: CA
authors.lastname: Johnson
authors.affiliation: UCL Ear Institute, University College London, 332 Gray's Inn Road, London WC1X 8EE, UK c.johnson@leeds.ac.uk d.jagger@ucl.ac.uk.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Jagger
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D019521:Body Patterning; D002454:Cell Differentiation; D016764:Cell Polarity; D002923:Cilia / Q000378:metabolism; D002925:Ciliary Motility Disorders / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000002:abnormalities / Q000378:metabolism; D004677:Encephalocele / Q000378:metabolism*; D004848:Epithelium / Q000196:embryology* / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008168:Lung / Q000196:embryology / Q000378:metabolism; D008565:Membrane Proteins / Q000172:deficiency / Q000378:metabolism*; D051379:Mice; D009024:Morphogenesis; D009154:Mutation / Q000235:genetics; D009925:Organ of Corti / Q000002:abnormalities / Q000196:embryology / Q000473:pathology; D010641:Phenotype; D010766:Phosphorylation; D007690:Polycystic Kidney Diseases / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D057050:Receptor Tyrosine Kinase-like Orphan Receptors / Q000378:metabolism; D012174:Retinitis Pigmentosa; D059547:Stereocilia / Q000378:metabolism; D051153:Wnt Proteins / Q000378:metabolism; D060449:Wnt Signaling Pathway; D000071818:Wnt-5a Protein; D051176:beta Catenin / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; C583064:TMEM67 protein, mouse; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; C438615:Wnt5a protein, mouse; D051176:beta Catenin; D057050:Receptor Tyrosine Kinase-like Orphan Receptors; C477010:Ror2 protein, mouse
keywords: Ciliopathy; Hair bundle; Kinocilia; MKS3; Meckelin; PCP; Planar cell polarity; Primary cilia; Stereocilia; TMEM67; Wnt signalling