Top

Journal of Bone and Mineral Metabolism

Published in:

01-01-2022 | Osteoporosis | Original Article

LINC02381, a sponge of miR-21, weakens osteogenic differentiation of hUC-MSCs through KLF12-mediated Wnt4 transcriptional repression

Authors: Gang Zhao, Wen-Dong Luo, Yong Yuan, Feng Lin, Li-Min Guo, Jing-Jing Ma, Han-Bo Chen, Huang Tang, Jun Shu

Published in: Journal of Bone and Mineral Metabolism | Issue 1/2022

Abstract

Introduction

Human umbilical cord blood-derived MSCs (hUC-MSCs) have the potential to differentiate into osteoblasts. This study investigated the function and potential mechanisms of a novel lncRNA LINC02381 in hUC-MSC osteogenic differentiation.

Materials and methods

hUC-MSCs were maintained in osteogenic differentiation medium. RT-qPCR assay was performed to assess LINC02381 expression. Alizarin Red S (ARS) and alkaline phosphatase (ALP) staining were performed to evaluate osteogenic differentiation. The interaction between miR-21 and LINC0238/KLF12 was determined by luciferase reporter and RNA immunoprecipitation (RIP) assays. Chromatin immunoprecipitation (ChIP) assay was used to confirm the transcriptional regulation of KLF12 on Wnt4 promoter. The nuclear translocation of β-catenin was evaluated using immunofluorescence. hUC-MSCs seeded on Bio-Oss Collagen scaffolds were transplanted into nude mice to assess in vivo osteogenesis. Bone formation was observed by H&E and Masson's trichrome staining. OSX and OPN levels were assessed by immunohistochemistry.

Results

LINC02381 was up-regulated in the clinical samples of osteoporotic patients. However, LINC02381 expression was reduced during osteogenic differentiation of hUC-MSCs. Enforced expression of LINC02381 suppressed the osteogenic differentiation of hUC-MSCs. Mechanistically, LINC02381 sponged miR-21 to enhance KLF12 expression, which led to the inactivation of Wnt/β-catenin signaling pathway. Furthermore, miR-21 mimics or KLF12 silencing counteracted LINC02381-induced inhibition of osteogenic differentiation, whereas IWP-4 (an inhibitor of Wnt pathway) abolished this effect.

Conclusion

In summary, LINC02381 repressed osteogenic differentiation of hUS-MSCs through sponging miR-21 to enhance KLF12-mediated inactivation of Wnt/β-catenin pathway, indicating that LINC02381 might be a therapeutic target for osteoporosis.

Available only for authorised users

Yang L, Li Y, Gong R, Gao M, Feng C et al (2019) The long non-coding RNA-ORLNC1 regulates bone mass by directing mesenchymal stem cell fate. Mol Ther 27:394–410. https://doi.org/10.1016/j.ymthe.2018.11.019CrossRefPubMed

Tang P, Xiong Q, Ge W, Zhang L (2014) The role of microRNAs in osteoclasts and osteoporosis. RNA Biol 11:1355–1363. https://doi.org/10.1080/15476286.2014.996462CrossRefPubMed

Ko NY, Chen LR, Chen KH (2020) The role of micro RNA and long-non-coding RNA in osteoporosis. Int J Mol Sci. https://doi.org/10.3390/ijms21144886CrossRefPubMedPubMedCentral

Mora-Raimundo P, Lozano D, Manzano M, Vallet-Regi M (2019) Nanoparticles to knockdown osteoporosis-related gene and promote osteogenic marker expression for osteoporosis treatment. ACS Nano 13:5451–5464. https://doi.org/10.1021/acsnano.9b00241CrossRefPubMedPubMedCentral

Griffin M, Iqbal SA, Bayat A (2011) Exploring the application of mesenchymal stem cells in bone repair and regeneration. J Bone Joint Surg Br 93:427–434. https://doi.org/10.1302/0301-620X.93B4.25249CrossRefPubMed

Bougioukli S, Saitta B, Sugiyama O, Tang AH, Elphingstone J, Evseenko D, Lieberman JR (2019) Lentiviral gene therapy for bone repair using human umbilical cord blood-derived mesenchymal stem cells. Hum Gene Ther 30:906–917. https://doi.org/10.1089/hum.2018.054CrossRefPubMedPubMedCentral

Wang Y, You C, Wei R, Zu J, Song C, Li J, Yan J (2017) Modification of human umbilical cord blood stem cells using polyethylenimine combined with modified TAT peptide to enhance BMP-2 production. Biomed Res Int 2017:2971413. https://doi.org/10.1155/2017/2971413CrossRefPubMedPubMedCentral

Hong B, Lee S, Shin N, Ko Y, Kim D, Lee J, Lee W (2018) Bone regeneration with umbilical cord blood mesenchymal stem cells in femoral defects of ovariectomized rats. Osteoporos Sarcopenia 4:95–101. https://doi.org/10.1016/j.afos.2018.08.003CrossRefPubMedPubMedCentral

Antebi B, Pelled G, Gazit D (2014) Stem cell therapy for osteoporosis. Curr Osteoporos Rep 12:41–47. https://doi.org/10.1007/s11914-013-0184-xCrossRefPubMed

10.

Xiao T, Liu L, Li H, Sun Y, Luo H, Li T, Wang S, Dalton S, Zhao RC, Chen R (2021) Long noncoding RNA ADINR regulates adipogenesis by transcriptionally activating C/EBPalpha. Stem Cell Rep 16:1006–1008. https://doi.org/10.1016/j.stemcr.2021.03.024CrossRef

11.

Zhang J, Jiang T, Liang X, Shu S, Xiang X, Zhang W, Guo T, Xie W, Deng W, Tang X (2019) lncRNA MALAT1 mediated high glucose-induced HK-2 cell epithelial-to-mesenchymal transition and injury. J Physiol Biochem 75:443–452. https://doi.org/10.1007/s13105-019-00688-2CrossRefPubMed

12.

Xu Q, Cheng D, Liu Y, Pan H, Li G, Li P, Li Y, Sun W, Ma D, Ni C (2021) LncRNA-ATB regulates epithelial-mesenchymal transition progression in pulmonary fibrosis via sponging miR-29b-2-5p and miR-34c-3p. J Cell Mol Med 25:7294–7306. https://doi.org/10.1111/jcmm.16758CrossRefPubMedPubMedCentral

13.

Han Y, Kang C, Kang M, Quan W, Gao H, Zhong Z (2019) Long non-coding RNA Mirt2 prevents TNF-alpha-triggered inflammation via the repression of microRNA-101. Int Immunopharmacol 76: https://doi.org/10.1016/j.intimp.2019.105878CrossRefPubMed

14.

Liu J, Wu M, Feng G, Li R, Wang Y, Jiao J (2020) Downregulation of LINC00707 promotes osteogenic differentiation of human bone marrowderived mesenchymal stem cells by regulating DKK1 via targeting miR103a3p. Int J Mol Med 46:1029–1038. https://doi.org/10.3892/ijmm.2020.4672CrossRefPubMedPubMedCentral

15.

Jin C, Jia L, Tang Z, Zheng Y (2020) Long non-coding RNA MIR22HG promotes osteogenic differentiation of bone marrow mesenchymal stem cells via PTEN/ AKT pathway. Cell Death Dis 11:601. https://doi.org/10.1038/s41419-020-02813-2CrossRefPubMedPubMedCentral

16.

Jafarzadeh M, Soltani BM, Soleimani M, Hosseinkhani S (2020) Epigenetically silenced LINC02381 functions as a tumor suppressor by regulating PI3K-Akt signaling pathway. Biochimie 171–172:63–71. https://doi.org/10.1016/j.biochi.2020.02.009CrossRefPubMed

17.

Chen X, Zhang Z, Ma Y, Su H, Xie P, Ran J (2020) LINC02381 promoted cell viability and migration via targeting miR-133b in cervical cancer cells. Cancer Manag Res 12:3971–3979. https://doi.org/10.2147/CMAR.S237285CrossRefPubMedPubMedCentral

18.

Yang SE, Ha CW, Jung M, Jin HJ, Lee M, Song H, Choi S, Oh W, Yang YS (2004) Mesenchymal stem/progenitor cells developed in cultures from UC blood. Cytotherapy 6:476–486. https://doi.org/10.1080/14653240410005041CrossRefPubMed

19.

Cao L, Liu W, Zhong Y, Zhang Y, Gao D, He T, Liu Y, Zou Z, Mo Y, Peng S, Shuai C (2020) Linc02349 promotes osteogenesis of human umbilical cord-derived stem cells by acting as a competing endogenous RNA for miR-25-3p and miR-33b-5p. Cell Prolif 53: https://doi.org/10.1111/cpr.12814CrossRefPubMedPubMedCentral

20.

Chen B, Dodge ME, Tang W, Lu J, Ma Z, Fan CW, Wei S, Hao W, Kilgore J, Williams NS, Roth MG, Amatruda JF, Chen C, Lum L (2009) Small molecule-mediated disruption of Wnt-dependent signaling in tissue regeneration and cancer. Nat Chem Biol 5:100–107. https://doi.org/10.1038/nchembio.137CrossRefPubMedPubMedCentral

21.

Xiaoling G, Shuaibin L, Kailu L (2020) MicroRNA-19b-3p promotes cell proliferation and osteogenic differentiation of BMSCs by interacting with lncRNA H19. BMC Med Genet 21:11. https://doi.org/10.1186/s12881-020-0948-yCrossRefPubMedPubMedCentral

22.

Vescini F, Attanasio R, Balestrieri A, Bandeira F, Bonadonna S et al (2016) Italian association of clinical endocrinologists (AME) position statement: drug therapy of osteoporosis. J Endocrinol Investig 39:807–834. https://doi.org/10.1007/s40618-016-0434-8CrossRef

23.

Mi B, Yan C, Xue H, Chen L, Panayi AC, Hu L, Hu Y, Cao F, Sun Y, Zhou W, Xiong Y, Liu G (2020) Inhibition of circulating miR-194-5p reverses osteoporosis through Wnt5a/beta-catenin-dependent induction of osteogenic differentiation. Mol Ther Nucleic Acids 21:814–823. https://doi.org/10.1016/j.omtn.2020.07.023CrossRefPubMedPubMedCentral

24.

Feng L, Zhang JF, Shi L, Yang ZM, Wu TY, Wang HX, Lin WP, Lu YF, Lo JHT, Zhu DH, Li G (2020) MicroRNA-378 suppressed osteogenesis of MSCs and impaired bone formation via inactivating Wnt/beta-catenin signaling. Mol Ther Nucleic Acids 21:1017–1028. https://doi.org/10.1016/j.omtn.2020.07.018CrossRefPubMedPubMedCentral

25.

Wei F, Liu D, Feng C, Zhang F, Yang S, Hu Y, Ding G, Wang S (2015) microRNA-21 mediates stretch-induced osteogenic differentiation in human periodontal ligament stem cells. Stem Cells Dev 24:312–319. https://doi.org/10.1089/scd.2014.0191CrossRefPubMed

26.

Meng YB, Li X, Li ZY, Zhao J, Yuan XB, Ren Y, Cui ZD, Liu YD, Yang XJ (2015) microRNA-21 promotes osteogenic differentiation of mesenchymal stem cells by the PI3K/beta-catenin pathway. J Orthop Res 33:957–964. https://doi.org/10.1002/jor.22884CrossRefPubMed

27.

Sun X, Li X, Qi H, Hou X, Zhao J, Yuan X, Ma X (2020) MiR-21 nanocapsules promote early bone repair of osteoporotic fractures by stimulating the osteogenic differentiation of bone marrow mesenchymal stem cells. J Orthop Transl 24:76–87. https://doi.org/10.1016/j.jot.2020.04.007CrossRef

28.

Wu T, Wang S, Wang L, Zhang W, Chen W, Lv X, Li Y, Hussain Z, Sun W (2020) Long noncoding RNA (lncRNA) CTTN-IT1 elevates skeletal muscle satellite cell proliferation and differentiation by acting as ceRNA for YAP1 through absorbing miR-29a in Hu sheep. Front Genet 11:843. https://doi.org/10.3389/fgene.2020.00843CrossRefPubMedPubMedCentral

29.

Zhang L, Xie H, Li S (2020) LncRNA LOXL1-AS1 controls osteogenic and adipocytic differentiation of bone marrow mesenchymal stem cells in postmenopausal osteoporosis through regulating the miR-196a-5p/Hmga2 axis. J Bone Miner Metab. https://doi.org/10.1007/s00774-020-01123-zCrossRefPubMed

30.

Wu D, Yin L, Sun D, Wang F, Wu Q, Xu Q, Xin B (2020) Long noncoding RNA TUG1 promotes osteogenic differentiation of human periodontal ligament stem cell through sponging microRNA-222-3p to negatively regulate Smad2/7. Arch Oral Biol 117: https://doi.org/10.1016/j.archoralbio.2020.104814CrossRefPubMed

31.

Yan Q, Yan G, Zhang C, Wang Z, Huang C, Wang J, Zhou J, Liu Y, Ding L, Zhang Q, Zhen X, Jiang Y, Sun H (2019) miR-21 reverses impaired decidualization through modulation of KLF12 and NR4A1 expression in human endometrial stromal cellsdagger. Biol Reprod 100:1395–1405. https://doi.org/10.1093/biolre/ioz026CrossRefPubMed

32.

Takai H, van Wijnen AJ, Ogata Y (2019) Induction of chondrogenic or mesenchymal stem cells from human periodontal ligament cells through inhibition of Twist2 or Klf12. J Oral Sci 61:313–320. https://doi.org/10.2334/josnusd.18-0224CrossRefPubMed

33.

Yang C, Wang C, Zhou J, Liang Q, He F, Li F, Li Y, Chen J, Zhang F, Han C, Liu J, Li K, Tang Y (2020) Fibronectin 1 activates WNT/beta-catenin signaling to induce osteogenic differentiation via integrin beta1 interaction. Lab Investig. https://doi.org/10.1038/s41374-020-0451-2CrossRefPubMed

34.

Shen J, Sun Y, Liu X, Zhu Y, Bao B, Gao T, Chai Y, Xu J, Zheng X (2021) EGFL6 regulates angiogenesis and osteogenesis in distraction osteogenesis via Wnt/beta-catenin signaling. Stem Cell Res Ther 12:415. https://doi.org/10.1186/s13287-021-02487-3CrossRefPubMedPubMedCentral

35.

Huang Y, Xiao D, Huang S, Zhuang J, Zheng X, Chang Y, Yin D (2020) Circular RNA YAP1 attenuates osteoporosis through up-regulation of YAP1 and activation of Wnt/beta-catenin pathway. Biomed Pharmacother 129: https://doi.org/10.1016/j.biopha.2020.110365CrossRefPubMed

Title: LINC02381, a sponge of miR-21, weakens osteogenic differentiation of hUC-MSCs through KLF12-mediated Wnt4 transcriptional repression
Authors: Gang Zhao
Wen-Dong Luo
Yong Yuan
Feng Lin
Li-Min Guo
Jing-Jing Ma
Han-Bo Chen
Huang Tang
Jun Shu
Publication date: 01-01-2022
Publisher: Springer Singapore
Keywords: Osteoporosis
Osteoporosis
Published in: Journal of Bone and Mineral Metabolism / Issue 1/2022
Print ISSN: 0914-8779
Electronic ISSN: 1435-5604
DOI: https://doi.org/10.1007/s00774-021-01277-4

Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Reserve your place

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin

Prof. Florian Limbourg

Prof. Anoop Chauhan

Developed by: Springer Medicine

Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Pediatric Cardiology Video

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

Cardiopulmonary Comorbidity Video

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Valvular Heart Disease Video

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.

Watch now

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

16-04-2024 Type 2 Diabetes News

Incentivised to exercise

11-04-2024 Lifestyle Modifications News

New intervention for STEMI-related cardiogenic shock

10-04-2024 Myocardial Infarction News

» View more highlights on the ACC 2024 congress hub page

Image Credits

Illustration of figure on mountain summit/© Orapun / Stock.adobe.com, STEP AAG HeroTeaserCollection image/© Tarek / Generated with AI / Stock.adobe.com, ACC 2024 Pediatric and Congenital Heart Disease: Year in Review/© 2024 American College of Cardiology, ACC 2024 Pulmonary Vascular Disease: Year in Review/© 2024 American College of Cardiology, ACC 2024 Valvular Heart Disease: Year in Review/© 2024 American College of Cardiology, ACC 2024 HF and Cardiomyopathies: Year in Review/© 2024 American College of Cardiology, Woman out walking/© Seventyfour / stock.adobe.com (symbolic image with model), Woman checking smartwatch /© saltdium / stock.adobe.com (symbolic image with model), Cardiac catheterization/© Damian / stock.adobe.com

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Introduction

Materials and methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2022

PGC-1: a key regulator in bone homeostasis

Imaging findings and treatment outcomes of a rare subtype of medication-related osteonecrosis of the jaw

Disruption of the mouse Bmal1 locus promotes heterotopic ossification with aging via TGF-beta/BMP signaling

The relatıonshıp between serum hydroxyvıtamın D levels and postural stabılıty ın young adults

Performance evaluation of the new chemiluminescent intact FGF23 assay relative to the existing assay system

Polypharmacy, chronic kidney disease, and incident fragility fracture: a prospective cohort study

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

Highlights from the ACC 2024 Congress

ACC 2024 Congress Coverage