open access
Effect of miR-195 inhibition on human skeletal muscle-derived stem/progenitor cells


- Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
open access
Abstract
BACKGROUND: Application of a circulating miR-195 inhibitor could be a helping factor in in vitro model of human skeletal muscle- derived stem/progenitor cells (SkMDS/PCs). Previously, miR-195 expression has been reported to be a negative factor for myogenesis.
AIMS: The aim of the study was to obtain anti-apoptotic and anti-aging effects in in vitro cultured myoblasts and to improve their ability to form myotubes by suppressing miR-195 expression.
METHODS: Human wild-type (WT) SkMDS/PC cells incubated with control (nonspecific) miRNA inhibitor and miR-195-inhibited SkMDS/PCs were studied. Functional assays (myotube formation and cell ageing), antioxidant, and myogenic gene expression analyses were performed at two time points, at the 7th and 11th cell passages.
RESULTS: Myotube formation was found to be almost 2-fold higher in the miR-195-inhibited SkMDS/PCs population (p<0.05) compared to WT cells. miR-195 inhibition did not appear to affect cell ageing or rejuvenate human SkMDS/PCs. Antioxidant (SOD3 and FOXO) gene expression was augmented in the miR-195-inhibited SkMDS/PCs population, but no positive effect on the remaining antioxidant genes (SOD1, SOD2, and catalase) was observed. A significant increase in MyoD gene expression with a concomitant decrease in MyoG (p<0.05) was further documented in miR-195-inhibited SkMDS/PCs compared to WT cells (11th cell passage).
CONCLUSIONS: The performed studies may lead to the preconditioning of myogenic stem cells to extend their potential for pro-regenerative activity. miR-195 inhibitor may serve as conditioning factor augmenting selective antioxidant genes expression and proliferative potential of SkMDS/PCs, but not having an impact on cell aging and/ or apoptosis.
Abstract
BACKGROUND: Application of a circulating miR-195 inhibitor could be a helping factor in in vitro model of human skeletal muscle- derived stem/progenitor cells (SkMDS/PCs). Previously, miR-195 expression has been reported to be a negative factor for myogenesis.
AIMS: The aim of the study was to obtain anti-apoptotic and anti-aging effects in in vitro cultured myoblasts and to improve their ability to form myotubes by suppressing miR-195 expression.
METHODS: Human wild-type (WT) SkMDS/PC cells incubated with control (nonspecific) miRNA inhibitor and miR-195-inhibited SkMDS/PCs were studied. Functional assays (myotube formation and cell ageing), antioxidant, and myogenic gene expression analyses were performed at two time points, at the 7th and 11th cell passages.
RESULTS: Myotube formation was found to be almost 2-fold higher in the miR-195-inhibited SkMDS/PCs population (p<0.05) compared to WT cells. miR-195 inhibition did not appear to affect cell ageing or rejuvenate human SkMDS/PCs. Antioxidant (SOD3 and FOXO) gene expression was augmented in the miR-195-inhibited SkMDS/PCs population, but no positive effect on the remaining antioxidant genes (SOD1, SOD2, and catalase) was observed. A significant increase in MyoD gene expression with a concomitant decrease in MyoG (p<0.05) was further documented in miR-195-inhibited SkMDS/PCs compared to WT cells (11th cell passage).
CONCLUSIONS: The performed studies may lead to the preconditioning of myogenic stem cells to extend their potential for pro-regenerative activity. miR-195 inhibitor may serve as conditioning factor augmenting selective antioxidant genes expression and proliferative potential of SkMDS/PCs, but not having an impact on cell aging and/ or apoptosis.
Keywords
miR-195 inhibitor, human skeletal muscle-derived stem/progenitor cells (SkMDS/PCs), apoptosis, oxidative stress, regenerative medicine




Title
Effect of miR-195 inhibition on human skeletal muscle-derived stem/progenitor cells
Journal
Kardiologia Polska (Polish Heart Journal)
Issue
Article type
Original article
Published online
2022-05-13
Page views
9
Article views/downloads
6
DOI
10.33963/KP.a2022.0127
Pubmed
Keywords
miR-195 inhibitor
human skeletal muscle-derived stem/progenitor cells (SkMDS/PCs)
apoptosis
oxidative stress
regenerative medicine
Authors
Magdalena Nowaczyk
Agnieszka Malcher
Agnieszka Zimna
Natalia Rozwadowska
Maciej Kurpisz


- Eschenhagen T, Bolli R, Braun T, et al. Cardiomyocyte Regeneration: A Consensus Statement. Circulation. 2017; 136(7): 680–686.
- Bertoli G, Cava C, Castiglioni I. MicroRNAs: new biomarkers for diagnosis, prognosis, therapy prediction and therapeutic tools for breast cancer. Theranostics. 2015; 5(10): 1122–1143.
- Femminella GD, Ferrara N, Rengo G. The emerging role of microRNAs in Alzheimer's disease. Front Physiol. 2015; 6: 40.
- Kondkar AA, Abu-Amero KK. Utility of circulating microRNAs as clinical biomarkers for cardiovascular diseases. Biomed Res Int. 2015; 2015: 821823.
- He JF, Luo YM, Wan XH, et al. Biogenesis of MiRNA-195 and its role in biogenesis, the cell cycle, and apoptosis. J Biochem Mol Toxicol. 2011; 25(6): 404–408.
- Vriend LEM, De Witt Hamer PC, Van Noorden CJF, et al. WEE1 inhibition and genomic instability in cancer. Biochim Biophys Acta. 2013; 1836(2): 227–235.
- Wang J, Martin JF. Macro advances in microRNAs and myocardial regeneration. Curr Opin Cardiol. 2014; 29(3): 207–213.
- Singh R, Yadav V, Kumar S, et al. MicroRNA-195 inhibits proliferation, invasion and metastasis in breast cancer cells by targeting FASN, HMGCR, ACACA and CYP27B1. Sci Rep. 2015; 5: 17454.
- Porrello ER, Johnson BA, Aurora AB, et al. MiR-15 family regulates postnatal mitotic arrest of cardiomyocytes. Circ Res. 2011; 109(6): 670–679.
- Hodgkinson CP, Kang MH, Dal-Pra S, et al. MicroRNAs and cardiac regeneration. Circ Res. 2015; 116(10): 1700–1711.
- Okada M, Kim HaW, Matsu-ura K, et al. Abrogation of age-induced microrna-195 rejuvenates the senescent mesenchymal stem cells by reactivating telomerase. Stem Cells. 2016; 34(1): 148–159.
- Wang H, Zhang Q, Wang B, et al. miR-22 regulates C2C12 myoblast proliferation and differentiation by targeting TGFBR1. Eur J Cell Biol. 2018; 97(4): 257–268.
- Long G, Wang F, Duan Q, et al. Circulating miR-30a, miR-195 and let-7b associated with acute myocardial infarction. PLoS One. 2012; 7(12): e50926.
- Kolanowski TJ, Rozwadowska N, Malcher A, et al. In vitro and in vivo characteristics of connexin 43-modified human skeletal myoblasts as candidates for prospective stem cell therapy for the failing heart. Int J Cardiol. 2014; 173(1): 55–64.
- Nowaczyk M, Malcher A, Zimna A, et al. Transient and stable overexpression of extracellular superoxide dismutase is positively associated with the myogenic function of human skeletal muscle-derived stem/progenitor cells. Antioxidants (Basel). 2020; 9(9).
- Nowaczyk M, Malcher A, Zimna A, et al. Addition of popular exogenous antioxidant agent, PBN, to culture media may be an important step to optimization of myogenic stem/progenitor cell preparation protocol. Antioxidants (Basel). 2021; 10(6).
- Sugihara H, Teramoto N, Yamanouchi K, et al. Oxidative stress-mediated senescence in mesenchymal progenitor cells causes the loss of their fibro/adipogenic potential and abrogates myoblast fusion. Aging (Albany NY). 2018; 10(4): 747–763.
- Zha JP, Wang XQ, Di J. MiR-920 promotes osteogenic differentiation of human bone mesenchymal stem cells by targeting HOXA7. J Orthop Surg Res. 2020; 15(1): 254.
- Braga L, Ali H, Secco I, et al. Non-coding RNA therapeutics for cardiac regeneration. Cardiovasc Res. 2021; 117(3): 674–693.
- Kondo H, Kim HaW, Wang L, et al. Blockade of senescence-associated microRNA-195 in aged skeletal muscle cells facilitates reprogramming to produce induced pluripotent stem cells. Aging Cell. 2016; 15(1): 56–66.
- Pajcini KV, Pomerantz JH, Alkan O, et al. Myoblasts and macrophages share molecular components that contribute to cell-cell fusion. J Cell Biol. 2008; 180(5): 1005–1019.
- Dueñas A, Expósito A, Muñoz MD, et al. MiR-195 enhances cardiomyogenic differentiation of the proepicardium/septum transversum by Smurf1 and Foxp1 modulation. Sci Rep. 2020; 10(1): 9334.
- Naylor RM, Baker DJ, van Deursen JM. Senescent cells: a novel therapeutic target for aging and age-related diseases. Clin Pharmacol Ther. 2013; 93(1): 105–116.
- Zhao DL, Wu QL. Effect of inhibition to Yes-related proteins-mediated Wnt/β-catenin signaling pathway through miR-195-5p on apoptosis of gastric cancer cells. Eur Rev Med Pharmacol Sci. 2019; 23(15): 6486–6496.
- Yuan T, Zhang Li, Yao S, et al. miR‑195 promotes LPS‑mediated intestinal epithelial cell apoptosis via targeting SIRT1/eIF2a. Int J Mol Med. 2020; 45(2): 510–518.
- Gopinath SD, Webb AE, Brunet A, et al. FOXO3 promotes quiescence in adult muscle stem cells during the process of self-renewal. Stem Cell Reports. 2014; 2(4): 414–426.
- Xiong S, Salazar G, Patrushev N, et al. FoxO1 mediates an autofeedback loop regulating SIRT1 expression. J Biol Chem. 2011; 286(7): 5289–5299.
- Zheng D, Ma J, Yu Y, et al. Silencing of miR-195 reduces diabetic cardiomyopathy in C57BL/6 mice. Diabetologia. 2015; 58(8): 1949–1958.
- Zheng D, Yu Y, Li M, et al. Inhibition of microRNA 195 prevents apoptosis and multiple-organ injury in mouse models of sepsis. J Infect Dis. 2016; 213(10): 1661–1670.
- Seale P, Rudnicki MA. A new look at the origin, function, and "stem-cell" status of muscle satellite cells. Dev Biol. 2000; 218(2): 115–124.
- Milewska M, Grabiec K, Grzelkowska-Kowalczyk K. Interactions of proliferation and differentiation signaling pathways in myogenesis [in Polish]. Postepy Hig Med Dosw (Online). 2014; 68: 516–526.
- Sato T, Yamamoto T, Sehara-Fujisawa A. miR-195/497 induce postnatal quiescence of skeletal muscle stem cells. Nat Commun. 2014; 5: 4597.
- Le Moal E, Pialoux V, Juban G, et al. Redox control of skeletal muscle regeneration. Antioxid Redox Signal. 2017; 27(5): 276–310.
- Blais A, Tsikitis M, Acosta-Alvear D, et al. An initial blueprint for myogenic differentiation. Genes Dev. 2005; 19(5): 553–569.
- Qiu H, Zhong J, Luo L, et al. Regulatory axis of miR-195/497 and HMGA1-Id3 governs muscle cell proliferation and differentiation. Int J Biol Sci. 2017; 13(2): 157–166.
- Fiedorowicz K, Rozwadowska N, Zimna A, et al. Tissue-specific promoter-based reporter system for monitoring cell differentiation from iPSCs to cardiomyocytes. Sci Rep. 2020; 10(1): 1895.
- Szklarczyk D, Gable AL, Nastou KC, et al. The STRING database in 2021: customizable protein-protein networks, and functional characterization of user-uploaded gene/measurement sets. Nucleic Acids Res. 2021; 49(D1): D605–D612.