title: TWIST1-Induced miR-424 Reversibly Drives Mesenchymal Programming while Inhibiting Tumor Initiation.
abstract: Epithelial-to-mesenchymal transition (EMT) is a dynamic process that relies on cellular plasticity. Recently, the process of an oncogenic EMT, followed by a reverse mesenchymal-to-epithelial transition (MET), has been implicated as critical in the metastatic colonization of carcinomas. Unlike governance of epithelial programming, regulation of mesenchymal programming is not well understood in EMT. Here, we describe and characterize the first microRNA that enhances exclusively mesenchymal programming. We demonstrate that miR-424 is upregulated early during a TWIST1 or SNAI1-induced EMT, and that it causes cells to express mesenchymal genes without affecting epithelial genes, resulting in a mixed/intermediate EMT. Furthermore, miR-424 increases motility, decreases adhesion, and induces a growth arrest, changes associated with a complete EMT that can be reversed when miR-424 expression is lowered, concomitant with an MET-like process. Breast cancer patient miR-424 levels positively associate with TWIST1/2 and EMT-like gene signatures, and miR-424 is increased in primary tumors versus matched normal breast. However, miR-424 is downregulated in patient metastases versus matched primary tumors. Correspondingly, miR-424 decreases tumor initiation and is posttranscriptionally downregulated in macrometastases in mice, suggesting the need for biphasic expression of miR-424 to transit the EMT-MET axis. Next-generation RNA sequencing revealed miR-424 regulates numerous EMT and cancer stemness-associated genes, including TGFBR3, whose downregulation promotes mesenchymal phenotypes, but not tumor-initiating phenotypes. Instead, we demonstrate that increased MAPK-ERK signaling is critical for miR-424-mediated decreases in tumor-initiating phenotypes. These findings suggest miR-424 plays distinct roles in tumor progression, potentially facilitating earlier, but repressing later, stages of metastasis by regulating an EMT-MET axis.
authors.affiliation: Program in Molecular Biology, University of Colorado Anschutz Medical Campus, Aurora, Colorado.
authors.forename: David J
authors.initials: DJ
authors.lastname: Drasin
authors.affiliation: Department of Pharmacology, University of Colorado Anschutz Medical Campus, Aurora, Colorado.
authors.forename: Anna L
authors.initials: AL
authors.lastname: Guarnieri
authors.affiliation: Program in Molecular Biology, University of Colorado Anschutz Medical Campus, Aurora, Colorado.
authors.forename: Deepika
authors.initials: D
authors.lastname: Neelakantan
authors.affiliation: Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado.
authors.forename: Jihye
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Pharmacology, University of Colorado Anschutz Medical Campus, Aurora, Colorado.
authors.forename: Joshua H
authors.initials: JH
authors.lastname: Cabrera
authors.affiliation: Department of Pharmacology, University of Colorado Anschutz Medical Campus, Aurora, Colorado.
authors.forename: Chu-An
authors.initials: CA
authors.lastname: Wang
authors.affiliation: Department of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, Colorado.
authors.forename: Vadym
authors.initials: V
authors.lastname: Zaberezhnyy
authors.affiliation: Department of Molecular Virology, Immunology and Molecular Genetics, Ohio State University, Columbus, Ohio.
authors.forename: Pierluigi
authors.initials: P
authors.lastname: Gasparini
authors.affiliation: Department of Molecular Virology, Immunology and Molecular Genetics, Ohio State University, Columbus, Ohio.
authors.forename: Luciano
authors.initials: L
authors.lastname: Cascione
authors.affiliation: Department of Molecular Virology, Immunology and Molecular Genetics, Ohio State University, Columbus, Ohio.
authors.forename: Kay
authors.initials: K
authors.lastname: Huebner
authors.affiliation: Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado.
authors.forename: Aik-Choon
authors.initials: AC
authors.lastname: Tan
authors.affiliation: Program in Molecular Biology, University of Colorado Anschutz Medical Campus, Aurora, Colorado. Department of Pharmacology, University of Colorado Anschutz Medical Campus, Aurora, Colorado. Department of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, Colorado. Heide.Ford@ucdenver.edu.
authors.forename: Heide L
authors.initials: HL
authors.lastname: Ford
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D015536:Down-Regulation; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D005260:Female; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics*; D020929:Mitogen-Activated Protein Kinase Kinases / Q000235:genetics; D009362:Neoplasm Metastasis; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D015398:Signal Transduction; D051879:Twist-Related Protein 1 / Q000235:genetics* / Q000378:metabolism; D015854:Up-Regulation
chemical_list: C554214:MIRN424 microrna, human; D035683:MicroRNAs; D009687:Nuclear Proteins; C103173:TWIST1 protein, human; D051879:Twist-Related Protein 1; D020929:Mitogen-Activated Protein Kinase Kinases