Effects of miR-335-5p in modulating osteogenic differentiation by specifically downregulating Wnt antagonist DKK1

Jin Zhang, Qisheng Tu, Lynda Bonewald, Xi He, Gary Stein, Jane Lian, Jake Chen

Research output: Contribution to journalArticle

153 Citations (Scopus)

Abstract

Dickkopf-related protein 1 (DKK1) is essential to maintain skeletal homeostasis as an inhibitor of Wnt signaling and osteogenic differentiation. The purpose of this study was to investigate the molecular mechanisms underlying the developmental stage-specific regulation of the DKK1 protein level. We performed a series of studies including luciferase reporter assays, micro-RNA microarray, site-specific mutations, and gain- and loss-of-function analyses. We found that the DKK1 protein level was regulated via DKK1 3' UTR by miRNA control, which was restricted to osteoblast-lineage cells. As a result of decreased DKK1 protein level by miR-335-5p, Wnt signaling was enhanced, as indicated by elevated GSK-3β phosphorylation and increased β-catenin transcriptional activity. The effects of miR-335-5p were reversed by anti-miR-335-5p treatment, which downregulated endogenous miR-335-5p. In vivo studies showed high expression levels of miR-335-5p in osteoblasts and hypertrophic chondrocytes of mouse embryos, indicating a pivotal role of miR-335-5p in regulating bone development. In conclusion, miR-335-5p activates Wnt signaling and promotes osteogenic differentiation by downregulating DKK1. This cell- and development-specific regulation is essential and mandatory for the initiation and progression of osteogenic differentiation. miR-335-5p proves to be a potential and useful targeting molecule for promoting bone formation and regeneration.

Original languageEnglish (US)
Pages (from-to)1953-1963
Number of pages11
JournalJournal of Bone and Mineral Research
Volume26
Issue number8
DOIs
StatePublished - Aug 2011
Externally publishedYes

Fingerprint

Down-Regulation
Proteins
Osteoblasts
MicroRNAs
Glycogen Synthase Kinase 3
Catenins
Bone Regeneration
Bone Development
3' Untranslated Regions
Chondrocytes
Luciferases
Osteogenesis
Homeostasis
Embryonic Structures
Phosphorylation
Mutation

Keywords

  • bone formation
  • DKK1
  • MIR-335-5P
  • osteogenic differentiation
  • Wnt

ASJC Scopus subject areas

  • Orthopedics and Sports Medicine
  • Endocrinology, Diabetes and Metabolism

Cite this

Effects of miR-335-5p in modulating osteogenic differentiation by specifically downregulating Wnt antagonist DKK1. / Zhang, Jin; Tu, Qisheng; Bonewald, Lynda; He, Xi; Stein, Gary; Lian, Jane; Chen, Jake.

In: Journal of Bone and Mineral Research, Vol. 26, No. 8, 08.2011, p. 1953-1963.

Research output: Contribution to journalArticle

Zhang, Jin ; Tu, Qisheng ; Bonewald, Lynda ; He, Xi ; Stein, Gary ; Lian, Jane ; Chen, Jake. / Effects of miR-335-5p in modulating osteogenic differentiation by specifically downregulating Wnt antagonist DKK1. In: Journal of Bone and Mineral Research. 2011 ; Vol. 26, No. 8. pp. 1953-1963.
@article{40c3f6482cfb45b2b1e9ef2a397817bd,
title = "Effects of miR-335-5p in modulating osteogenic differentiation by specifically downregulating Wnt antagonist DKK1",
abstract = "Dickkopf-related protein 1 (DKK1) is essential to maintain skeletal homeostasis as an inhibitor of Wnt signaling and osteogenic differentiation. The purpose of this study was to investigate the molecular mechanisms underlying the developmental stage-specific regulation of the DKK1 protein level. We performed a series of studies including luciferase reporter assays, micro-RNA microarray, site-specific mutations, and gain- and loss-of-function analyses. We found that the DKK1 protein level was regulated via DKK1 3' UTR by miRNA control, which was restricted to osteoblast-lineage cells. As a result of decreased DKK1 protein level by miR-335-5p, Wnt signaling was enhanced, as indicated by elevated GSK-3β phosphorylation and increased β-catenin transcriptional activity. The effects of miR-335-5p were reversed by anti-miR-335-5p treatment, which downregulated endogenous miR-335-5p. In vivo studies showed high expression levels of miR-335-5p in osteoblasts and hypertrophic chondrocytes of mouse embryos, indicating a pivotal role of miR-335-5p in regulating bone development. In conclusion, miR-335-5p activates Wnt signaling and promotes osteogenic differentiation by downregulating DKK1. This cell- and development-specific regulation is essential and mandatory for the initiation and progression of osteogenic differentiation. miR-335-5p proves to be a potential and useful targeting molecule for promoting bone formation and regeneration.",
keywords = "bone formation, DKK1, MIR-335-5P, osteogenic differentiation, Wnt",
author = "Jin Zhang and Qisheng Tu and Lynda Bonewald and Xi He and Gary Stein and Jane Lian and Jake Chen",
year = "2011",
month = "8",
doi = "10.1002/jbmr.377",
language = "English (US)",
volume = "26",
pages = "1953--1963",
journal = "Journal of Bone and Mineral Research",
issn = "0884-0431",
publisher = "Wiley-Blackwell",
number = "8",

}

TY - JOUR

T1 - Effects of miR-335-5p in modulating osteogenic differentiation by specifically downregulating Wnt antagonist DKK1

AU - Zhang, Jin

AU - Tu, Qisheng

AU - Bonewald, Lynda

AU - He, Xi

AU - Stein, Gary

AU - Lian, Jane

AU - Chen, Jake

PY - 2011/8

Y1 - 2011/8

N2 - Dickkopf-related protein 1 (DKK1) is essential to maintain skeletal homeostasis as an inhibitor of Wnt signaling and osteogenic differentiation. The purpose of this study was to investigate the molecular mechanisms underlying the developmental stage-specific regulation of the DKK1 protein level. We performed a series of studies including luciferase reporter assays, micro-RNA microarray, site-specific mutations, and gain- and loss-of-function analyses. We found that the DKK1 protein level was regulated via DKK1 3' UTR by miRNA control, which was restricted to osteoblast-lineage cells. As a result of decreased DKK1 protein level by miR-335-5p, Wnt signaling was enhanced, as indicated by elevated GSK-3β phosphorylation and increased β-catenin transcriptional activity. The effects of miR-335-5p were reversed by anti-miR-335-5p treatment, which downregulated endogenous miR-335-5p. In vivo studies showed high expression levels of miR-335-5p in osteoblasts and hypertrophic chondrocytes of mouse embryos, indicating a pivotal role of miR-335-5p in regulating bone development. In conclusion, miR-335-5p activates Wnt signaling and promotes osteogenic differentiation by downregulating DKK1. This cell- and development-specific regulation is essential and mandatory for the initiation and progression of osteogenic differentiation. miR-335-5p proves to be a potential and useful targeting molecule for promoting bone formation and regeneration.

AB - Dickkopf-related protein 1 (DKK1) is essential to maintain skeletal homeostasis as an inhibitor of Wnt signaling and osteogenic differentiation. The purpose of this study was to investigate the molecular mechanisms underlying the developmental stage-specific regulation of the DKK1 protein level. We performed a series of studies including luciferase reporter assays, micro-RNA microarray, site-specific mutations, and gain- and loss-of-function analyses. We found that the DKK1 protein level was regulated via DKK1 3' UTR by miRNA control, which was restricted to osteoblast-lineage cells. As a result of decreased DKK1 protein level by miR-335-5p, Wnt signaling was enhanced, as indicated by elevated GSK-3β phosphorylation and increased β-catenin transcriptional activity. The effects of miR-335-5p were reversed by anti-miR-335-5p treatment, which downregulated endogenous miR-335-5p. In vivo studies showed high expression levels of miR-335-5p in osteoblasts and hypertrophic chondrocytes of mouse embryos, indicating a pivotal role of miR-335-5p in regulating bone development. In conclusion, miR-335-5p activates Wnt signaling and promotes osteogenic differentiation by downregulating DKK1. This cell- and development-specific regulation is essential and mandatory for the initiation and progression of osteogenic differentiation. miR-335-5p proves to be a potential and useful targeting molecule for promoting bone formation and regeneration.

KW - bone formation

KW - DKK1

KW - MIR-335-5P

KW - osteogenic differentiation

KW - Wnt

UR - http://www.scopus.com/inward/record.url?scp=79960633240&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79960633240&partnerID=8YFLogxK

U2 - 10.1002/jbmr.377

DO - 10.1002/jbmr.377

M3 - Article

VL - 26

SP - 1953

EP - 1963

JO - Journal of Bone and Mineral Research

JF - Journal of Bone and Mineral Research

SN - 0884-0431

IS - 8

ER -