title: MicroRNA-199a is induced in dystrophic muscle and affects WNT signaling, cell proliferation, and myogenic differentiation.
abstract: In patients with Duchenne muscular dystrophy (DMD), the absence of a functional dystrophin protein results in sarcolemmal instability, abnormal calcium signaling, cardiomyopathy, and skeletal muscle degeneration. Using the dystrophin-deficient sapje zebrafish model, we have identified microRNAs (miRNAs) that, in comparison to our previous findings in human DMD muscle biopsies, are uniquely dysregulated in dystrophic muscle across vertebrate species. MiR-199a-5p is dysregulated in dystrophin-deficient zebrafish, mdx(5cv) mice, and human muscle biopsies. MiR-199a-5p mature miRNA sequences are transcribed from stem loop precursor miRNAs that are found within the introns of the dynamin-2 and dynamin-3 loci. The miR-199a-2 stem loop precursor transcript that gives rise to the miR-199a-5p mature transcript was found to be elevated in human dystrophic muscle. The levels of expression of miR-199a-5p are regulated in a serum response factor (SRF)-dependent manner along with myocardin-related transcription factors. Inhibition of SRF-signaling reduces miR-199a-5p transcript levels during myogenic differentiation. Manipulation of miR-199a-5p expression in human primary myoblasts and myotubes resulted in dramatic changes in cellular size, proliferation, and differentiation. MiR-199a-5p targets several myogenic cell proliferation and differentiation regulatory factors within the WNT signaling pathway, including FZD4, JAG1, and WNT2. Overexpression of miR-199a-5p in the muscles of transgenic zebrafish resulted in abnormal myofiber disruption and sarcolemmal membrane detachment, pericardial edema, and lethality. Together, these studies identify miR-199a-5p as a potential regulator of myogenesis through suppression of WNT-signaling factors that act to balance myogenic cell proliferation and differentiation.
authors.affiliation: Division of Genetics, Program in Genomics, Boston Children's Hospital, Boston, MA, USA.
authors.forename: M S
authors.initials: MS
authors.lastname: Alexander
authors.forename: G
authors.initials: G
authors.lastname: Kawahara
authors.forename: N
authors.initials: N
authors.lastname: Motohashi
authors.forename: J C
authors.initials: JC
authors.lastname: Casar
authors.forename: I
authors.initials: I
authors.lastname: Eisenberg
authors.forename: J A
authors.initials: JA
authors.lastname: Myers
authors.forename: M J
authors.initials: MJ
authors.lastname: Gasperini
authors.forename: E A
authors.initials: EA
authors.lastname: Estrella
authors.forename: A T
authors.initials: AT
authors.lastname: Kho
authors.forename: S
authors.initials: S
authors.lastname: Mitsuhashi
authors.forename: F
authors.initials: F
authors.lastname: Shapiro
authors.forename: P B
authors.initials: PB
authors.lastname: Kang
authors.forename: L M
authors.initials: LM
authors.lastname: Kunkel
mesh_terms: D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D049109:Cell Proliferation; D034285:Dynamin II / Q000235:genetics; D034286:Dynamin III / Q000235:genetics; D016189:Dystrophin / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D051157:Frizzled Receptors / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D055029:Inverted Repeat Sequences / Q000235:genetics; D000072100:Jagged-1 Protein; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics*; D024510:Muscle Development; D018485:Muscle Fibers, Skeletal / Q000378:metabolism; D018482:Muscle, Skeletal; D009137:Muscular Dystrophy, Animal / Q000235:genetics* / Q000378:metabolism; D032446:Myoblasts / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D000072099:Serrate-Jagged Proteins; D026362:Serum Response Factor / Q000378:metabolism; D015534:Trans-Activators / Q000378:metabolism; D060449:Wnt Signaling Pathway / Q000235:genetics*; D051154:Wnt2 Protein / Q000378:metabolism; D015027:Zebrafish; D029961:Zebrafish Proteins; D000099251:Myocardin
chemical_list: D002135:Calcium-Binding Proteins; D034285:Dynamin II; D034286:Dynamin III; D016189:Dystrophin; D051157:Frizzled Receptors; D036341:Intercellular Signaling Peptides and Proteins; D000072100:Jagged-1 Protein; D008565:Membrane Proteins; D035683:MicroRNAs; D009687:Nuclear Proteins; D000072099:Serrate-Jagged Proteins; D026362:Serum Response Factor; D015534:Trans-Activators; D051154:Wnt2 Protein; D029961:Zebrafish Proteins; D000099251:Myocardin; C402930:FZD4 protein, human; C000606273:JAG1 protein, human; C000606274:Jag1 protein, mouse; C000606276:jag1a protein, zebrafish; C531408:mirn199 microRNA, human