Autophagy induction in the skeletal myogenic differentiation of human tonsil-derived mesenchymal stem cells

Park,Saeyoung; Choi,Yoonyoung; Jung,Namhee; Kim,Jieun; Oh,Seiyoon; Yu,Yeonsil; Ahn,Jung-Hyuck; Jo,Inho; Choi,Byung-Ok; Jung,Sung-Chul

doi:10.3892/ijmm.2017.2898

Autophagy induction in the skeletal myogenic differentiation of human tonsil-derived mesenchymal stem cells

Authors:
- Saeyoung Park
- Yoonyoung Choi
- Namhee Jung
- Jieun Kim
- Seiyoon Oh
- Yeonsil Yu
- Jung-Hyuck Ahn
- Inho Jo
- Byung-Ok Choi
- Sung-Chul Jung
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Affiliations: Department of Biochemistry, School of Medicine, Ewha Womans University, Seoul 07985, Republic of Korea, Department of Human Biology, College of Human Ecology, Cornell University, Ithaca, NY 14850, USA, Department of Molecular Medicine, School of Medicine, Ewha Womans University, Seoul 07985, Republic of Korea, Department of Neurology, Samsung Medical Center, Sungkyunkwan University, Seoul 06351, Republic of Korea
Published online on: February 20, 2017 https://doi.org/10.3892/ijmm.2017.2898
Pages: 831-840
Copyright: © Park et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Mesenchymal stem cells (MSCs) are capable of self-renewal and differentiation and are thus a valuable source for the replacement of diseased or damaged organs. Previously, we reported that the tonsils can be an excellent reservoir of MSCs for the regeneration of skeletal muscle (SKM) damage. However, the mechanisms involved in the differentiation from tonsil-derived MSCs (T-MSCs) to myocytes via myoblasts remain unclear. To clarify these mechanisms, we analyzed gene expression profiles of T-MSCs during differentiation into myocytes compared with human skeletal muscle cells (hSKMCs). Total RNA was extracted from T-MSCs, T-MSC-derived myoblasts and myocytes, and hSKMCs and was subjected to analysis using a microarray. Microarray analysis of the three phases of myogenic differentiation identified candidate genes associated with myogenic differentiation. The expression pattern of undifferentiated T-MSCs was distinguishable from the myogenic differentiated T-MSCs and hSKMCs. In particular, we selected FNBP1L, which among the upregulated genes is essential for antibacterial autophagy, since autophagy is related to SKM metabolism and myogenesis. T-MSCs differentiated toward myoblasts and skeletal myocytes sequentially, as evidenced by increased expression of autophagy-related markers (including Beclin-1, LC3B and Atg5) and decreased expression of Bcl-2. Furthermore, we reconfirmed that autophagy has an effect on the mechanism of skeletal myogenic differentiation derived from T-MSCs by treatment with 5-azacytidine and bafilomycin A1. These data suggest that the transcriptome of the T-MSC-derived myocytes is similar to that of hSKMCs, and that autophagy has an important role in the mechanism of myogenic differentiation of T-MSCs.

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1	Ryu KH, Cho KA, Park HS, Kim JY, Woo SY, Jo I, Choi YH, Park YM, Jung SC, Chung SM, et al: Tonsil-derived mesenchymal stromal cells: Evaluation of biologic, immunologic and genetic factors for successful banking. Cytotherapy. 14:1193–1202. 2012. View Article : Google Scholar : PubMed/NCBI
2	Park S, Choi Y, Jung N, Yu Y, Ryu KH, Kim HS, Jo I, Choi BO and Jung SC: Myogenic differentiation potential of human tonsil-derived mesenchymal stem cells and their potential for use to promote skeletal muscle regeneration. Int J Mol Med. 37:1209–1220. 2016.PubMed/NCBI
3	Dezawa M, Ishikawa H, Itokazu Y, Yoshihara T, Hoshino M, Takeda S, Ide C and Nabeshima Y: Bone marrow stromal cells generate muscle cells and repair muscle degeneration. Science. 309:314–317. 2005. View Article : Google Scholar : PubMed/NCBI
4	Di Rocco G, Iachininoto MG, Tritarelli A, Straino S, Zacheo A, Germani A, Crea F and Capogrossi MC: Myogenic potential of adipose-tissue-derived cells. J Cell Sci. 119:2945–2952. 2006. View Article : Google Scholar : PubMed/NCBI
5	Kim JA, Shon YH, Lim JO, Yoo JJ, Shin HI and Park EK: MYOD mediates skeletal myogenic differentiation of human amniotic fluid stem cells and regeneration of muscle injury. Stem Cell Res Ther. 4:1472013. View Article : Google Scholar : PubMed/NCBI
6	Nunes VA, Cavaçana N, Canovas M, Strauss BE and Zatz M: Stem cells from umbilical cord blood differentiate into myotubes and express dystrophin in vitro only after exposure to in vivo muscle environment. Biol Cell. 99:185–196. 2007. View Article : Google Scholar
7	Park S, Kim E, Koh SE, Maeng S, Lee WD, Lim J, Shim I and Lee YJ: Dopaminergic differentiation of neural progenitors derived from placental mesenchymal stem cells in the brains of Parkinson's disease model rats and alleviation of asymmetric rotational behavior. Brain Res. 1466:158–166. 2012. View Article : Google Scholar : PubMed/NCBI
8	Kerkis I, Kerkis A, Dozortsev D, Stukart-Parsons GC, Gomes Massironi SM, Pereira LV, Caplan AI and Cerruti HF: Isolation and characterization of a population of immature dental pulp stem cells expressing OCT-4 and other embryonic stem cell markers. Cells Tissues Organs. 184:105–116. 2006. View Article : Google Scholar
9	Husmann I, Soulet L, Gautron J, Martelly I and Barritault D: Growth factors in skeletal muscle regeneration. Cytokine Growth Factor Rev. 7:249–258. 1996. View Article : Google Scholar : PubMed/NCBI
10	Wang K, Wang C, Xiao F, Wang H and Wu Z: JAK2/STAT2/STAT3 are required for myogenic differentiation. J Biol Chem. 283:34029–34036. 2008. View Article : Google Scholar : PubMed/NCBI
11	Conboy IM and Rando TA: The regulation of Notch signaling controls satellite cell activation and cell fate determination in postnatal myogenesis. Dev Cell. 3:397–409. 2002. View Article : Google Scholar : PubMed/NCBI
12	Fortini P, Ferretti C, Iorio E, Cagnin M, Garribba L, Pietraforte D, Falchi M, Pascucci B, Baccarini S, Morani F, et al: The fine tuning of metabolism, autophagy and differentiation during in vitro myogenesis. Cell Death Dis. 7:e21682016. View Article : Google Scholar : PubMed/NCBI
13	Maley MA, Fan Y, Beilharz MW and Grounds MD: Intrinsic differences in MyoD and myogenin expression between primary cultures of SJL/J and BALB/C skeletal muscle. Exp Cell Res. 211:99–107. 1994. View Article : Google Scholar : PubMed/NCBI
14	Miller KJ, Thaloor D, Matteson S and Pavlath GK: Hepatocyte growth factor affects satellite cell activation and differentiation in regenerating skeletal muscle. Am J Physiol Cell Physiol. 278:C174–C181. 2000.PubMed/NCBI
15	Heszele MF and Price SR: Insulin-like growth factor I: The yin and yang of muscle atrophy. Endocrinology. 145:4803–4805. 2004. View Article : Google Scholar : PubMed/NCBI
16	Nakanishi K, Dohmae N and Morishima N: Endoplasmic reticulum stress increases myofiber formation in vitro. FASEB J. 21:2994–3003. 2007. View Article : Google Scholar : PubMed/NCBI
17	Levine B and Klionsky DJ: Development by self-digestion: Molecular mechanisms and biological functions of autophagy. Dev Cell. 6:463–477. 2004. View Article : Google Scholar : PubMed/NCBI
18	Nuschke A, Rodrigues M, Stolz DB, Chu CT, Griffith L and Wells A: Human mesenchymal stem cells/multipotent stromal cells consume accumulated autophagosomes early in differentiation. Stem Cell Res Ther. 5:1402014. View Article : Google Scholar : PubMed/NCBI
19	Sin J, Andres AM, Taylor DJ, Weston T, Hiraumi Y, Stotland A, Kim BJ, Huang C, Doran KS and Gottlieb RA: Mitophagy is required for mitochondrial biogenesis and myogenic differentiation of C2C12 myoblasts. Autophagy. 12:369–380. 2016. View Article : Google Scholar :
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
21	Yu Y, Park YS, Kim HS, Kim HY, Jin YM, Jung SC, Ryu KH and Jo I: Characterization of long-term in vitro culture-related alterations of human tonsil-derived mesenchymal stem cells: Role for CCN1 in replicative senescence-associated increase in osteogenic differentiation. J Anat. 225:510–518. 2014. View Article : Google Scholar : PubMed/NCBI
22	Pestonjamasp KN, Pope RK, Wulfkuhle JD and Luna EJ: Supervillin (p205): A novel membrane-associated, F-actin-binding protein in the villin/gelsolin superfamily. J Cell Biol. 139:1255–1269. 1997. View Article : Google Scholar
23	Morris NJ, Ross SA, Lane WS, Moestrup SK, Petersen CM, Keller SR and Lienhard GE: Sortilin is the major 110-kDa protein in GLUT4 vesicles from adipocytes. J Biol Chem. 273:3582–3587. 1998. View Article : Google Scholar : PubMed/NCBI
24	Hu H, Gao X, Sun Y, Zhou J, Yang M and Xu Z: Alpha-actinin-2, a cytoskeletal protein, binds to angiogenin. Biochem Biophys Res Commun. 329:661–667. 2005. View Article : Google Scholar : PubMed/NCBI
25	Huett A, Ng A, Cao Z, Kuballa P, Komatsu M, Daly MJ, Podolsky DK and Xavier RJ: A novel hybrid yeast-human network analysis reveals an essential role for FNBP1L in antibacterial autophagy. J Immunol. 182:4917–4930. 2009. View Article : Google Scholar : PubMed/NCBI
26	Marquez RT and Xu L: Bcl-2:Beclin 1 complex: Multiple, mechanisms regulating autophagy/apoptosis toggle switch. Am J Cancer Res. 2:214–221. 2012.PubMed/NCBI
27	Nakatogawa H, Suzuki K, Kamada Y and Ohsumi Y: Dynamics and diversity in autophagy mechanisms: Lessons from yeast. Nat Rev Mol Cell Biol. 10:458–467. 2009. View Article : Google Scholar : PubMed/NCBI
28	Moran JL, Li Y, Hill AA, Mounts WM and Miller CP: Gene expression changes during mouse skeletal myoblast differentiation revealed by transcriptional profiling. Physiol Genomics. 10:103–111. 2002. View Article : Google Scholar : PubMed/NCBI
29	Osses N and Brandan E: ECM is required for skeletal muscle differentiation independently of muscle regulatory factor expression. Am J Physiol Cell Physiol. 282:C383–C394. 2002. View Article : Google Scholar : PubMed/NCBI
30	Hinz B, Phan SH, Thannickal VJ, Galli A, Bochaton-Piallat ML and Gabbiani G: The myofibroblast: One function, multiple origins. Am J Pathol. 170:1807–1816. 2007. View Article : Google Scholar : PubMed/NCBI
31	Oh SW, Pope RK, Smith KP, Crowley JL, Nebl T, Lawrence JB and Luna EJ: Archvillin, a muscle-specific isoform of supervillin, is an early expressed component of the costameric membrane skeleton. J Cell Sci. 116:2261–2275. 2003. View Article : Google Scholar : PubMed/NCBI
32	Ariga M, Nedachi T, Katagiri H and Kanzaki M: Functional role of sortilin in myogenesis and development of insulin-responsive glucose transport system in C2C12 myocytes. J Biol Chem. 283:10208–10220. 2008. View Article : Google Scholar : PubMed/NCBI
33	Kostin S, Hein S, Arnon E, Scholz D and Schaper J: The cytoskeleton and related proteins in the human failing heart. Heart Fail Rev. 5:271–280. 2000. View Article : Google Scholar
34	Sakoh-Nakatogawa M, Matoba K, Asai E, Kirisako H, Ishii J, Noda NN, Inagaki F, Nakatogawa H and Ohsumi Y: Atg12-Atg5 conjugate enhances E2 activity of Atg3 by rearranging its catalytic site. Nat Struct Mol Biol. 20:433–439. 2013. View Article : Google Scholar : PubMed/NCBI
35	Neel BA, Lin Y and Pessin JE: Skeletal muscle autophagy: A new metabolic regulator. Trends Endocrinol Metab. 24:635–643. 2013. View Article : Google Scholar : PubMed/NCBI
36	Wakitani S, Saito T and Caplan AI: Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-azacytidine. Muscle Nerve. 18:1417–1426. 1995. View Article : Google Scholar : PubMed/NCBI
37	Zheng JK, Wang Y, Karandikar A, Wang Q, Gai H, Liu AL, Peng C and Sheng HZ: Skeletal myogenesis by human embryonic stem cells. Cell Res. 16:713–722. 2006. View Article : Google Scholar : PubMed/NCBI
38	Mizushima N, Ohsumi Y and Yoshimori T: Autophagosome formation in mammalian cells. Cell Struct Funct. 27:421–429. 2002. View Article : Google Scholar
39	Jackson MJ: Reactive oxygen species and redox-regulation of skeletal muscle adaptations to exercise. Philos Trans R Soc Lond B Biol Sci. 360:2285–2291. 2005. View Article : Google Scholar : PubMed/NCBI
40	Malicdan MC, Noguchi S, Nonaka I, Saftig P and Nishino I: Lysosomal myopathies: An excessive build-up in autophagosomes is too much to handle. Neuromuscul Disord. 18:521–529. 2008. View Article : Google Scholar : PubMed/NCBI