Co‑culturing with hypoxia pre‑conditioned mesenchymal stem cells as a new strategy for the prevention of irradiation‑induced fibroblast‑to‑myofibroblast transition

Zhuang,Lei; Xia,Wenzheng; Hou,Meng

doi:10.3892/or.2019.7293

Co‑culturing with hypoxia pre‑conditioned mesenchymal stem cells as a new strategy for the prevention of irradiation‑induced fibroblast‑to‑myofibroblast transition

Authors:
- Lei Zhuang
- Wenzheng Xia
- Meng Hou
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Affiliations: Department of Hepatobiliary Surgery, First Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China, Department of Neurosurgery, First Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China, Department of Radiation Oncology, First Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China
Published online on: August 23, 2019 https://doi.org/10.3892/or.2019.7293
Pages: 1781-1792
Copyright: © Zhuang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cardiac fibrosis is a pathological consequence of radiation‑induced fibroblast proliferation and fibroblast‑to‑myofibroblast transition (FMT). Mesenchymal stem cell (MSC) transplantation has been revealed to be an effective treatment strategy to inhibit cardiac fibrosis. We identified a novel MSC‑driven mechanism that inhibited cardiac fibrosis, via the regulation of multiple fibrogenic pathways. Hypoxia pre‑conditioned MSCs (MSCsHypoxia) were co‑cultured with fibroblasts using a Transwell system. Radiation‑induced fibroblast proliferation was assessed using an MTT assay, and FMT was confirmed by assessing the mRNA levels of various markers of fibrosis, including type I collagen (Col1) and alpha smooth muscle actin (α‑SMA). α‑SMA expression was also confirmed via immunocytochemistry. The expression levels of Smad7 and Smad3 were detected by western blotting, and Smad7 was silenced using small interfering RNAs. The levels of oxidative stress following radiation were assessed by the detection of reactive oxygen species (ROS) and the activity of superoxide dismutase (SOD), malondialdehyde (MDA), and 4‑hydroxynonenal (HNE). It was revealed that co‑culturing with MSCsHypoxia could inhibit fibroblast proliferation and FMT. In addition, the present results indicated that MSCs are necessary and sufficient for the inhibition of fibroblast proliferation and FMT by functionally targeting TGF‑β1/Smad7/Smad3 signaling via the release of hepatocyte growth factor (HGF). Furthermore, it was observed that MSCs inhibited fibrosis by modulating oxidative stress. Co‑culturing with MSCsHypoxia alleviated fibroblast proliferation and FMT via the TGF‑β1/Smad7/Smad3 pathway. MSCs may represent a novel therapeutic approach for the treatment of radiation‑related cardiac fibrosis.

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1	Swerdlow AJ, Higgins CD, Smith P, Cunningham D, Hancock BW, Horwich A, Hoskin PJ, Lister A, Radford JA, Rohatiner AZ and Linch DC: Myocardial infarction mortality risk after treatment for Hodgkin disease: A collaborative British cohort study. J Natl Cancer Inst. 99:206–214. 2007. View Article : Google Scholar : PubMed/NCBI
2	Darby SC, Ewertz M, McGale P, Bennet AM, Blom-Goldman U, Brønnum D, Correa C, Cutter D, Gagliardi G, Gigante B, et al: Risk of ischemic heart disease in women after radiotherapy for breast cancer. N Engl J Med. 368:987–998. 2013. View Article : Google Scholar : PubMed/NCBI
3	Darby S, McGale P, Peto R, Granath F, Hall P and Ekbom A: Mortality from cardiovascular disease more than 10 years after radiotherapy for breast cancer: Nationwide cohort study of 90000 Swedish women. BMJ. 326:256–257. 2003. View Article : Google Scholar : PubMed/NCBI
4	Tukenova M, Guibout C, Oberlin O, Doyon F, Mousannif A, Haddy N, Guérin S, Pacquement H, Aouba A, Hawkins M, et al: Role of cancer treatment in long-term overall and cardiovascular mortality after childhood cancer. J Clin Oncol. 28:1308–1315. 2010. View Article : Google Scholar : PubMed/NCBI
5	Tapio S: Pathology and biology of radiation-induced cardiac disease. J Radiat Res. 57:439–448. 2016. View Article : Google Scholar : PubMed/NCBI
6	Subramanian V, Borchard S, Azimzadeh O, Sievert W, Merl-Pham J, Mancuso M, Pasquali E, Multhoff G, Popper B, Zischka H, et al: PPARα is necessary for radiation-induced activation of noncanonical TGFβ signaling in the heart. J Proteome Res. 17:1677–1689. 2018. View Article : Google Scholar : PubMed/NCBI
7	Curigliano G, Cardinale D, Dent S, Criscitiello C, Aseyev O, Lenihan D and Cipolla CM: Cardiotoxicity of anticancer treatments: Epidemiology, detection, and management. CA Cancer J Clin. 66:309–325. 2016. View Article : Google Scholar : PubMed/NCBI
8	Bagno L, Hatzistergos KE, Balkan W and Hare JM: Mesenchymal stem cell-based therapy for cardiovascular disease: Progress and challenges. Mol Ther. 26:1610–1623. 2018. View Article : Google Scholar : PubMed/NCBI
9	Golpanian S, Wolf A, Hatzistergos KE and Hare JM: Rebuilding the damaged heart: Mesenchymal stem cells, cell-based therapy, and engineered heart tissue. Physiol Rev. 96:1127–1168. 2016. View Article : Google Scholar : PubMed/NCBI
10	Ma S, Xie N, Li W, Yuan B, Shi Y and Wang Y: Immunobiology of mesenchymal stem cells. Cell Death Differ. 21:216–225. 2014. View Article : Google Scholar : PubMed/NCBI
11	Le Blanc K and Mougiakakos D: Multipotent mesenchymal stromal cells and the innate immune system. Nat Rev Immunol. 12:383–396. 2012. View Article : Google Scholar : PubMed/NCBI
12	Wu T, Xie Y, Huang J, Li P, Wang X, Yan Y, Xia T, Li L, Zhu F, Li H and Wu R: The optimal intervention time of bone marrow mesenchymal stem cells in ameliorating cardiac fibrosis induced by viral myocarditis: A randomized controlled trial in mice. Stem Cells Int. 2017:32580352017. View Article : Google Scholar : PubMed/NCBI
13	Huang L, Ma W, Ma Y, Feng D, Chen H and Cai B: Exosomes in mesenchymal stem cells, a new therapeutic strategy for cardiovascular diseases? Int J Biol Sci. 11:238–245. 2015. View Article : Google Scholar : PubMed/NCBI
14	Meng SS, Xu XP, Chang W, Lu ZH, Huang LL, Xu JY, Liu L, Qiu HB, Yang Y and Guo FM: LincRNA-p21 promotes mesenchymal stem cell migration capacity and survival through hypoxic preconditioning. Stem Cell Res Ther. 9:2802018. View Article : Google Scholar : PubMed/NCBI
15	Dobaczewski M, Chen W and Frangogiannis NG: Transforming growth factor (TGF)-β signaling in cardiac remodeling. J Mol Cell Cardiol. 51:600–606. 2011. View Article : Google Scholar : PubMed/NCBI
16	Yamamura Y, Hua X, Bergelson S and Lodish HF: Critical role of Smads and AP-1 complex in transforming growth factor-beta-dependent apoptosis. J Biol Chem. 275:36295–36302. 2000. View Article : Google Scholar : PubMed/NCBI
17	Hayashi H, Abdollah S, Qiu Y, Cai J, Xu YY, Grinnell BW, Richardson MA, Topper JN, Gimbrone MA Jr, Wrana JL and Falb D: The MAD-related protein Smad7 associates with the TGFβ receptor and functions as an antagonist of TGFbeta signaling. Cell. 89:1165–1173. 1997. View Article : Google Scholar : PubMed/NCBI
18	Wei LH, Huang XR, Zhang Y, Li YQ, Chen HY, Yan BP, Yu CM and Lan HY: Smad7 inhibits angiotensin II-induced hypertensive cardiac remodelling. Cardiovasc Res. 99:665–673. 2013. View Article : Google Scholar : PubMed/NCBI
19	Berk BC, Fujiwara K and Lehoux S: ECM remodeling in hypertensive heart disease. J Clin Invest. 117:568–575. 2007. View Article : Google Scholar : PubMed/NCBI
20	Fredj S, Bescond J, Louault C and Potreau D: Interactions between cardiac cells enhance cardiomyocyte hypertrophy and increase fibroblast proliferation. J Cell Physiol. 202:891–899. 2005. View Article : Google Scholar : PubMed/NCBI
21	Lucas JA, Zhang Y, Li P, Gong K, Miller AP, Hassan E, Hage F, Xing D, Wells B, Oparil S and Chen YF: Inhibition of transforming growth factor-beta signaling induces left ventricular dilation and dysfunction in the pressure-overloaded heart. Am J Physiol Heart Circ Physiol. 298:H424–H432. 2010. View Article : Google Scholar : PubMed/NCBI
22	Cheema AK, Pathak R, Zandkarimi F, Kaur P, Alkhalil L, Singh R, Zhong X, Ghosh S, Aykin-Burns N and Hauer-Jensen M: Liver metabolomics reveals increased oxidative stress and fibrogenic potential in gfrp transgenic mice in response to ionizing radiation. J Proteome Res. 13:3065–3074. 2014. View Article : Google Scholar : PubMed/NCBI
23	Yin TC, Wu RW, Sheu JJ, Sung PH, Chen KH, Chiang JY, Hsueh SK, Chung WJ, Lin PY, Hsu SL, et al: Combined therapy with extracorporeal shock wave and adipose-derived mesenchymal stem cells remarkably improved acute ischemia-reperfusion injury of quadriceps muscle. Oxid Med Cell Longev. 2018:60126362018. View Article : Google Scholar : PubMed/NCBI
24	Moon J, Schwarz SC, Lee HS, Kang JM, Lee YE, Kim B, Sung MY, Höglinger G, Wegner F, Kim JS, et al: Preclinical analysis of fetal human mesencephalic neural progenitor cell lines: Characterization and safety in vitro and in vivo. Stem Cells Transl Med. 6:576–588. 2017. View Article : Google Scholar : PubMed/NCBI
25	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 : PubMed/NCBI
26	Teekakirikul P, Eminaga S, Toka O, Alcalai R, Wang L, Wakimoto H, Nayor M, Konno T, Gorham JM, Wolf CM, et al: Cardiac fibrosis in mice with hypertrophic cardiomyopathy is mediated by non-myocyte proliferation and requires Tgf-β. J Clin Invest. 120:3520–3529. 2010. View Article : Google Scholar : PubMed/NCBI
27	Zhang Q, Ye H, Xiang F, Song LJ, Zhou LL, Cai PC, Zhang JC, Yu F, Shi HZ, Su Y, et al: miR-18a-5p inhibits sub-pleural pulmonary fibrosis by targeting TGF-β receptor II. Mol Ther. 25:728–738. 2017. View Article : Google Scholar : PubMed/NCBI
28	Ruart M, Chavarria L, Campreciós G, Suárez-Herrera N, Montironi C, Guixé-Muntet S, Bosch J, Friedman SL, Garcia-Pagán JC and Hernández-Gea V: Impaired endothelial autophagy promotes liver fibrosis by aggravating the oxidative stress response during acute liver injury. J Hepatol. 70:458–469. 2019. View Article : Google Scholar : PubMed/NCBI
29	Serrano NA, Mikkelsen R, Canada J, Mezzaroma E, Weiss E and Abbate A: Biomarkers of cardiac injury in patients undergoing thoracic radiation therapy. Int J Cardiol. 223:507–509. 2016. View Article : Google Scholar : PubMed/NCBI
30	Hancock SL, Donaldson SS and Hoppe RT: Cardiac disease following treatment of Hodgkin's disease in children and adolescents. J Clin Oncol. 11:1208–1215. 1993. View Article : Google Scholar : PubMed/NCBI
31	McGale P and Darby SC: Low doses of ionizing radiation and circulatory diseases: A systematic review of the published epidemiological evidence. Radiat Res. 163:247–257. 2005. View Article : Google Scholar : PubMed/NCBI
32	Demirci S, Nam J, Hubbs JL, Nguyen T and Marks LB: Radiation-induced cardiac toxicity after therapy for breast cancer: Interaction between treatment era and follow-up duration. Int J Radiat Oncol Biol Phys. 73:980–987. 2009. View Article : Google Scholar : PubMed/NCBI
33	Heidenreich PA, Hancock SL, Lee BK, Mariscal CS and Schnittger I: Asymptomatic cardiac disease following mediastinal irradiation. J Am Coll Cardiol. 42:743–749. 2003. View Article : Google Scholar : PubMed/NCBI
34	Gabbiani G: The myofibroblast in wound healing and fibrocontractive diseases. J Pathol. 200:500–503. 2003. View Article : Google Scholar : PubMed/NCBI
35	Hare JM, Traverse JH, Henry TD, Dib N, Strumpf RK, Schulman SP, Gerstenblith G, DeMaria AN, Denktas AE, Gammon RS, et al: A randomized, double-blind, placebo-controlled, dose-escalation study of intravenous adult human mesenchymal stem cells (prochymal) after acute myocardial infarction. J Am Coll Cardiol. 54:2277–2286. 2009. View Article : Google Scholar : PubMed/NCBI
36	Ranganath S, Levy O, Inamdar M and Karp JM: Harnessing the mesenchymal stem cell secretome for the treatment of cardiovascular disease. Cell Stem Cell. 10:244–258. 2012. View Article : Google Scholar : PubMed/NCBI
37	Molina EJ, Palma J, Gupta D, Torres D, Gaughan JP, Houser S and Macha M: Reverse remodeling is associated with changes in extracellular matrix proteases and tissue inhibitors after mesenchymal stem cell (MSC) treatment of pressure overload hypertrophy. J Tissue Eng Regen Med. 3:85–91. 2009. View Article : Google Scholar : PubMed/NCBI
38	Shao L, Zhang Y, Lan B, Wang J, Zhang Z, Zhang L, Xiao P, Meng Q, Geng YJ, Yu XY and Li Y: MiRNA-sequence indicates that mesenchymal stem cells and exosomes have similar mechanism to enhance cardiac repair. Biomed Res Int. 2017:41507052017. View Article : Google Scholar : PubMed/NCBI
39	Hu X, Xu Y, Zhong Z, Wu Y, Zhao J, Wang Y, Cheng H, Kong M, Zhang F, Chen Q, et al: A large-scale investigation of hypoxia-preconditioned allogeneic mesenchymal stem cells for myocardial repair in nonhuman primates: Paracrine activity without remuscularization. Circ Res. 118:970–983. 2016. View Article : Google Scholar : PubMed/NCBI
40	Shin HS, Lee S, Kim YM and Lim JY: Hypoxia-activated adipose mesenchymal stem cells prevents irradiation-induced salivary hypofunction by enhanced paracrine effect through fibroblast growth factor 10. Stem Cells. 36:1020–1032. 2018. View Article : Google Scholar : PubMed/NCBI
41	Seemann I, Gabriels K, Visser NL, Hoving S, te Poele JA, Pol JF, Gijbels MJ, Janssen BJ, van Leeuwen FW, Daemen MJ, et al: Irradiation induced modest changes in murine cardiac function despite progressive structural damage to the myocardium and microvasculature. Radiother Oncol. 103:143–150. 2012. View Article : Google Scholar : PubMed/NCBI
42	Derynck R and Zhang YE: Smad-dependent and Smad-independent pathways in TGF-beta family signalling. Nature. 425:577–584. 2003. View Article : Google Scholar : PubMed/NCBI
43	Taylor C, Mcgale P, Brønnum D, Correa C, Cutter D, Duane FK, Gigante B, Jensen MB, Lorenzen E, Rahimi K, et al: Cardiac structure injury after radiotherapy for breast cancer: Cross-sectional study with individual patient data. J Clin Oncol. 36:2288–2296. 2018. View Article : Google Scholar : PubMed/NCBI
44	Hirshfeld JW Jr, Fiorilli PN and Silvestry FE: Important strategies to reduce occupational radiation exposure in the cardiac catheterization laboratory: No lower limit. J Am Coll Cardiol. 71:1255–1258. 2018. View Article : Google Scholar : PubMed/NCBI
45	Viczenczova C, Kura B, Egan Benova T, Yin C, Kukreja RC, Slezak J, Tribulova N and Szeiffova Bacova B: Irradiation-induced cardiac connexin-43 and miR-21 responses are hampered by treatment with atorvastatin and aspirin. Int J Mol Sci. 19:2018. View Article : Google Scholar : PubMed/NCBI
46	Seawright JW, Samman Y, Sridharan V, Mao XW, Cao M, Singh P, Melnyk S, Koturbash I, Nelson GA, Hauer-Jensen M and Boerma M: Effects of low-dose rate γ-irradiation combined with simulated microgravity on markers of oxidative stress, DNA methylation potential, and remodeling in the mouse heart. PLoS One. 12:e01805942017. View Article : Google Scholar : PubMed/NCBI