Hearing loss is considered the most common sensory disorder across the world. Nowadays, a cochlear implant can be an effective treatment for patients. Moreover, it is often believed that sensorineural hearing loss in humans is caused by loss or disruption of the function of hair cells in the cochlea. In this respect, mesenchymal cells can be a good candidate for cell-based therapeutic approaches. To this end, the potential of human bone marrow-derived mesenchymal stem cells to differentiate into hair cells with the help of transfection of microRNA in vitro was investigated. MicroRNA mimics (miRNA-96, 182, and 183) were transfected to human bone marrow-derived mesenchymal stem cells using Lipofec-tamine as a common transfection reagent following the manufacturer’s instructions at 50 nM for microRNA mimics and 50 nM for the scramble. The changes in cell morphology were also observed under an inverted microscope. Then, the relative expression levels of SOX2, POU4F3, MYO7A, and calretinin were assayed using real-time polymerase chain reaction according to the ΔΔCt method. The ATOH1 level was similarly measured via real-time polymerase chain reaction and Western blotting. The results showed that increased expression of miRNA-182, but neither miRNA-96 nor miRNA-183, could lead to higher expression levels in some hair cell markers. The morphology of the cells also did not change in this respect, but the evaluation of gene expression at the levels of mRNA could promote the expression of the ATOH1, SOX2, and POU4F3 markers. Furthermore, miRNA-182 could enhance the expression of ATOH1 at the protein level. According to the results of this study, it was concluded that miRNA-182 could serve as a crucial function in hair cell differentiation by the upregulation of SOX2, POU4F3, and ATOH1 to promote a hair cell’s fate.

Keywords:
Hearing loss, MicroRNA-183 family, Hair cell, Human bone marrow-derived mesenchymal stem cells
1.
Ahmadi H, Sani HM, Farnoosh G, Sani MRM: Comparative study of speech and language development in children with normal hearing and cochlear implant in Iran. Indian J Otol 2017; 23: 135.
2.
Bermingham NA, Hassan BA, Price SD, Vollrath MA, Ben-Arie N, Eatock RA, Bellen HJ, Lysakowski A, Zoghbi HY: Math1: an essential gene for the generation of inner ear hair cells. Science 1999; 284: 1837–1841.
3.
Cafaro J, Lee GS, Stone JS: Atoh1 expression defines activated progenitors and differentiating hair cells during avian hair cell regeneration. Dev Dyn 2007; 236: 156–170.
4.
Dechesne CJ, Rabejac D, Desmadryl G: Development of calretinin immunoreactivity in the mouse inner ear. J Comp Neurol 1994; 346: 517–529.
5.
Fettiplace R, Ricci AJ: Mechanoelectrical transduction in auditory hair cells; in Eatock RA, Fay RR, Popper AN (eds): Vertebrate Hair Cells. New York, Springer, 2006, pp 154–203.
6.
Friedman RC, Farh KK, Burge CB, Bartel DP: Most mammalian mRNAs are conserved targets of microRNAs. Genome Res 2009; 19: 92–105.
7.
Gao FB: Context-dependent functions of specific microRNAs in neuronal development. Neural Dev 2010; 5: 25.
8.
Hasson T, Heintzelman MB, Santos-Sacchi J, Corey DP, Mooseker MS: Expression in cochlea and retina of myosin VIIa, the gene product defective in Usher syndrome type 1B. Proc Natl Acad Sci USA 1995; 92: 9815–9819.
9.
Hu X, Huang J, Feng L, Fukudome S, Hamajima Y, Lin J: Sonic hedgehog (SHH) promotes the differentiation of mouse cochlear neural progenitors via the Math1-Brn3.1 signaling pathway in vitro. J Neurosci Res 2010; 88: 927–935.
10.
Jeon SJ, Oshima K, Heller S, Edge AS: Bone marrow mesenchymal stem cells are progenitors in vitro for inner ear hair cells. Mol Cell Neurosci 2007; 34: 59–68.
11.
Kamiya K, Fujinami Y, Hoya N, Okamoto Y, Kouike H, Komatsuzaki R, Kusano R, Nakagawa S, Satoh H, Fujii M, Matsunaga T: Mesenchymal stem cell transplantation accelerates hearing recovery through the repair of injured cochlear fibrocytes. Am J Pathol 2007; 171: 214–226.
12.
Kros C, Marcotti W, Van Netten S, Self T, Libby R, Brown S, Richardson G, Steel K: Reduced climbing and increased slipping adaptation in cochlear hair cells of mice with Myo7a mutations. Nat Neurosci 2002; 5: 41.
13.
Kuss AW, Chen W: MicroRNAs in brain function and disease. Curr Neurol Neurosci Rep 2008; 8: 190–197.
14.
Li Y, Li A, Wu J, He Y, Yu H, Chai R, Li H: MiR-182-5p protects inner ear hair cells from cisplatin-induced apoptosis by inhibiting FOXO3a. Cell Death Dis 2016; 7:e2362.
15.
Mahmoodian-Sani MR, Hashemzadeh-Chaleshtori M, Saidijam M, Jami MS, Ghasemi-Dehkordi P: MicroRNA-183 family in inner ear: hair cell development and deafness. J Audiol Otol 2016; 20: 131–138.
16.
Mahmoodian-Sani MR, Mehri-Ghahfarrokhi A: The potential of miR-183 family expression in inner ear for regeneration, treatment, diagnosis and prognosis of hearing loss. J Otol 2017; 12: 55–61.
17.
Mahmoudian-Sani MR, Hashemzadeh-Chaleshtori M, Jami MS, Saidijam M: In vitro differentiation of human bone marrow mesenchymal stem cells to hair cells using growth factors. Int Tinnitus J 2017a; 21: 179–184.
18.
Mahmoudian-Sani MR, Mehri-Ghahfarrokhi A, Ahmadinejad F, Hashemzadeh-Chaleshtori M, Saidijam M, Jami MS: MicroRNAs: effective elements in ear-related diseases and hearing loss. Eur Arch Otorhinolaryngol 2017b; 274: 2373–2380.
19.
Mahmoudian-Sani MR, Mehri-Ghahfarrokhi A, Hashemzadeh-Chaleshtori M, Saidijam M, Jami MS: Comparison of three types of mesenchymal stem cells (bone marrow, adipose tissue, and umbilical cord-derived) as potential sources for inner ear regeneration. Int Tinnitus J 2017c; 21: 122–127.
20.
Mangiardi DA, McLaughlin-Williamson K, May KE, Messana EP, Mountain DC, Cotanche DA: Progression of hair cell ejection and molecular markers of apoptosis in the avian cochlea following gentamicin treatment. J Comp Neurol 2004; 475: 1–18.
21.
Naito Y, Nakamura T, Nakagawa T, Iguchi F, Endo T, Fujino K, Kim TS, Hiratsuka Y, Tamura T, Kanemaru S, Shimizu Y, Ito J: Transplantation of bone marrow stromal cells into the cochlea of chinchillas. Neuroreport 2004; 15: 1–4.
22.
Olex-Zarychta D: Successful treatment of sudden sensorineural hearing loss by means of pharmacotherapy combined with early hyperbaric oxygen therapy: Case report. Medicine 2017; 96:e9397.
23.
Oshima K, Grimm CM, Corrales CE, Senn P, Martinez Monedero R, Geleoc GS, Edge A, Holt JR, Heller S: Differential distribution of stem cells in the auditory and vestibular organs of the inner ear. J Assoc Res Otolaryngol 2007; 8: 18–31.
24.
Self T, Mahony M, Fleming J, Walsh J, Brown S, Steel KP: Shaker-1 mutations reveal roles for myosin VIIA in both development and function of cochlear hair cells. Development 1998; 125: 557–566.
25.
Sharif S, Nakagawa T, Ohno T, Matsumoto M, Kita T, Riazuddin S, Ito J: The potential use of bone marrow stromal cells for cochlear cell therapy. Neuroreport 2007; 18: 351–354.
26.
Stone JS, Cotanche DA: Identification of the timing of S phase and the patterns of cell proliferation during hair cell regeneration in the chick cochlea. J Comp Neurol 1994; 341: 50–67.
27.
Ushakov K, Rudnicki A, Avraham KB: MicroRNAs in sensorineural diseases of the ear. Front Mol Neurosci 2013; 6: 52.
28.
Walters BJ, Coak E, Dearman J, Bailey G, Yamashita T, Kuo B, Zuo J: In vivo interplay between p27(Kip1), GATA3, ATOH1, and POU4F3 converts non-sensory cells to hair cells in adult mice. Cell Rep 2017; 19: 307–320.
29.
Wang XR, Zhang XM, Du J, Jiang H: MicroRNA-182 regulates otocyst-derived cell differentiation and targets T-box1 gene. Hear Res 2012; 286: 55–63.
30.
Xiang M, Gan L, Li D, Chen ZY, Zhou L, O’Malley BW Jr, Klein W, Nathans J: Essential role of POU-domain factor Brn-3c in auditory and vestibular hair cell development. Proc Natl Acad Sci USA 1997; 94: 9445–9450.
31.
Zheng JL, Gao WQ: Analysis of rat vestibular hair cell development and regeneration using calretinin as an early marker. J Neurosci 1997; 17: 8270–8282.
© 2018 S. Karger AG, Basel
2018
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.