title: Transcriptional profiling identifies TNS4 function in epithelial tubulogenesis.
abstract: Hepatocyte growth factor (HGF) plays central roles in tubulogenesis and metastasis [1-4]. HGF treatment of Madin-Darby canine kidney (MDCK) cells grown as cysts in three-dimensional culture induces tubulogenesis [5, 6], which like most tubulogenic processes proceeds through distinct intermediate phases. Identification of genes associated with these phases is central to understanding the molecular mechanisms of tubulogensis; however, because of inefficient, asynchronous tubule formation, isolating such genes has been unfeasible. Here we developed a synchronous, efficient tubulogenesis system and used time-course transcriptional profiling to identify genes temporally regulated in developmental intermediates. Knockdown (KD) of tensin 4 (TNS4), a particularly highly upregulated gene, leads to a decrease in formation of extensions and tubules, two sequential intermediates in tubulogenesis. Exogenous expression of TNS4 marks invasive cells in an epithelial sheet. A mutation in the SH2 domain of TNS4 prevents the transition from extension formation to invasive migration during tubule formation and leads to increased basal activation of STAT3. Exogenous expression of a constitutively active STAT3 mimics the defect by the mutation. Our study highlights the role of the TNS4-STAT3 axis in epithelial sheet invasion and tubulogenesis.
authors.affiliation: Department of Anatomy, University of California, San Francisco, San Francisco, CA 94143-2140, USA.
authors.forename: Sang-Ho
authors.initials: SH
authors.lastname: Kwon
authors.forename: Pavel I
authors.initials: PI
authors.lastname: Nedvetsky
authors.forename: Keith E
authors.initials: KE
authors.lastname: Mostov
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D004285:Dogs; D004848:Epithelium / Q000187:drug effects* / Q000502:physiology; D020869:Gene Expression Profiling; D049452:Green Fluorescent Proteins / Q000378:metabolism; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D007684:Kidney Tubules / Q000187:drug effects / Q000254:growth & development*; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D000070876:Tensins; D014158:Transcription, Genetic / Q000502:physiology*
chemical_list: D008840:Microfilament Proteins; D000070876:Tensins; D049452:Green Fluorescent Proteins; D017228:Hepatocyte Growth Factor