WNT11-Conditioned Medium Promotes Angiogenesis through the Activation of Non-Canonical WNT-PKC-JNK Signaling Pathway
Abstract
:1. Introduction
2. Material and Methods
2.1. Animals
2.2. Cell Culture and Transduction with Wnt11 Plasmid
2.3. Quantitative Real-Time PCR
2.4. Western Blotting
2.5. Immunocytochemistry
2.6. Collection of Conditioned Medium (CdM)
2.7. CLT Formation and Endothelial Cell Migration
2.8. Rat Myocardial Infarction (MI) Model
2.9. Cardiac Function Assessment
2.10. Myocardial Blood Flow Measurement and Myocardial Blood Vessel Staining
2.11. Statistical Analysis
3. Results
3.1. Wnt11 Is Upregulated in MSCWnt11
3.2. CdMWnt11 Promoted Capillary-Like Tube (CLT) Formation and Sprouting of HUVEC Spheroids
3.3. Wnt11 Plays an Important Role in CdMWnt11-Mediated Angiogenesis
3.4. Wnt11 Activates Non-Canonical PKC/JNK Signaling Pathway
3.5. CdMWnt11 Improved Cardiac Function via Increasing Regional Blood Flow and Blood Vessel Density
4. Discussion
Author Contributions
Funding
Conflicts of Interest
References
- Mathiasen, A.B.; Qayyum, A.A.; Jorgensen, E.; Helqvist, S.; Kofoed, K.F.; Haack-Sørensen, M.; Ekblond, A.; Kastrup, J. Bone marrow-derived mesenchymal stromal cell treatment in patients with ischaemic heart failure: Final 4-year follow-up of the MSC-HF trial. Eur. J. Heart Fail. 2020, 22, 884–892. [Google Scholar] [CrossRef] [PubMed]
- Chan, J.L.; Miller, J.G.; Zhou, Y.; Robey, P.G.; Stroncek, D.F.; Arai, A.E.; Sachdev, V.; Horvath, K.A. Intramyocardial Bone Marrow Stem Cells in Patients Undergoing Cardiac Surgical Revascularization. Ann. Thorac. Surg. 2020, 109, 1142–1149. [Google Scholar] [CrossRef] [PubMed]
- Tang, Y.L.; Zhao, Q.; Qin, X.; Shen, L.; Cheng, L.; Ge, J.; Ian Phillips, M. Paracrine action enhances the effects of autologous mesenchymal stem cell transplantation on vascular regeneration in rat model of myocardial infarction. Ann. Thorac. Surg. 2005, 80, 229–236, discussion 236–237. [Google Scholar] [CrossRef] [PubMed]
- Uemura, R.; Xu, M.; Ahmad, N.; Ashraf, M. Bone marrow stem cells prevent left ventricular remodeling of ischemic heart through paracrine signaling. Circ. Res. 2006, 98, 1414–1421. [Google Scholar] [CrossRef] [Green Version]
- Hnatiuk, A.P.; Ong, S.G.; Olea, F.D.; Locatelli, P.; Riegler, J.; Lee, W.H.; Jen, C.H.; De Lorenzi, A.; Giménez, C.S.; Laguens, R.; et al. Allogeneic Mesenchymal Stromal Cells Overexpressing Mutant Human Hypoxia-Inducible Factor 1-α (HIF1-α) in an Ovine Model of Acute Myocardial Infarction. J. Am. Heart Assoc. 2016, 5, e003714. [Google Scholar] [CrossRef] [Green Version]
- Rabbani, S.; Soleimani, M.; Sahebjam, M.; Imani, M.; Nassiri, S.M.; Atashi, A.; Joupari, M.D.; Ghiaseddin, A.; Latifpour, M.; Tafti, S.H.A. Effects of Endothelial and Mesenchymal Stem Cells on Improving Myocardial Function in a Sheep Animal Model. J. Tehran Heart Cent. 2017, 12, 65–71. [Google Scholar] [PubMed]
- Rahbarghazi, R.; Nassiri, S.M.; Ahmadi, S.H.; Mohammadi, E.; Rabbani, S.; Araghi, A.; Hosseinkhani, H. Dynamic induction of pro-angiogenic milieu after transplantation of marrow-derived mesenchymal stem cells in experimental myocardial infarction. Int. J. Cardiol. 2014, 173, 453–466. [Google Scholar] [CrossRef] [PubMed]
- Santos Nascimento, D.; Mosqueira, D.; Sousa, L.M.; Teixeira, M.; Filipe, M.; Resende, T.P.; Araújo, A.F.; Valente, M.; Almeida, J.; Martins, J.P. Human umbilical cord tissue-derived mesenchymal stromal cells attenuate remodeling after myocardial infarction by proangiogenic, antiapoptotic, and endogenous cell-activation mechanisms. Stem Cell Res. Ther. 2014, 5, 5. [Google Scholar] [CrossRef] [Green Version]
- Balbi, C.; Lodder, K.; Costa, A.; Moimas, S.; Moccia, F.; van Herwaarden, T.; Rosti, R.; Campagnoli, F.; Palmeri, A.; De Biasio, P.; et al. Reactivating endogenous mechanisms of cardiac regeneration via paracrine boosting using the human amniotic fluid stem cell secretome. Int. J. Cardiol. 2019, 287, 87–95. [Google Scholar] [CrossRef]
- Tilokee, E.L.; Latham, N.; Jackson, R.; Mayfield, A.E.; Ye, B.; Mount, S.; Lam, B.-K.; Suuronen, E.J.; Ruel, M.; Stewart, D.J.; et al. Paracrine Engineering of Human Explant-Derived Cardiac Stem Cells to Over-Express Stromal-Cell Derived Factor 1alpha Enhances Myocardial Repair. Stem Cells 2016, 34, 1826–1835. [Google Scholar] [CrossRef]
- Moon, R.T.; Bowerman, B.; Boutros, M.; Perrimon, N. The promise and perils of Wnt signaling through β-catenin. Science 2002, 296, 1644–1666. [Google Scholar] [CrossRef] [PubMed]
- Logan, C.Y.; Nusse, R. The Wnt signaling pathway in development and disease. Annu. Rev. Cell. Dev. Biol. 2004, 20, 781–810. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Nakatsu, M.N.; Ding, Z.; Ng, M.Y.; Truong, T.T.; Yu, F.; Deng, S.X. Wnt/β-catenin signaling regulates proliferation of human cornea epithelial stem/progenitor cells. Investig. Ophthalmol. Vis. Sci. 2011, 52, 4734–4741. [Google Scholar] [CrossRef]
- DiRenzo, D.M.; Chaudhary, M.A.; Shi, X.; Franco, S.R.; Zent, J.; Wang, K.; Guo, L.-W.; Kent, K.C. A crosstalk between TGF-β/Smad3 and Wnt/β-catenin pathways promotes vascular smooth muscle cell proliferation. Cell Signal. 2016, 28, 498–505. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Tenjin, Y.; Kudoh, S.; Kubota, S.; Yamada, T.; Matsuo, A.; Sato, Y.; Ichimura, T.; Kohrogi, H.; Sashida, G.; Sakagami, T. Ascl1-induced Wnt11 regulates neuroendocrine differentiation, cell proliferation, and E-cadherin expression in small-cell lung cancer and Wnt11 regulates small-cell lung cancer biology. Lab. Investig. 2019, 99, 1622–1635. [Google Scholar] [CrossRef]
- Mori, H.; Yao, Y.; Learman, B.S.; Kurozumi, K.; Ishida, J.; Ramakrishnan, S.K.; Overmyer, K.A.; Xue, X.; Cawthorn, W.P.; Reid, M.A.; et al. Induction of WNT11 by hypoxia and hypoxia-inducible factor-1alpha regulates cell proliferation, migration and invasion. Sci. Rep. 2016, 6, 21520. [Google Scholar] [CrossRef] [Green Version]
- Kuhl, M.; Sheldahl, L.C.; Park, M.; Miller, J.R.; Moon, R.T. The Wnt/Ca2+ pathway: A new vertebrate Wnt signaling pathway takes shape. Trends Genet. 2000, 16, 279–283. [Google Scholar] [CrossRef]
- Sheldahl, L.C.; Slusarski, D.C.; Pandur, P.; Miller, J.R.; Kuhl, M.; Moon, R.T. Dishevelled activates Ca2+ flux, PKC, and CamKII in vertebrate embryos. J. Cell Biol. 2003, 161, 769–777. [Google Scholar] [CrossRef]
- Kuhl, M.; Sheldahl, L.C.; Malbon, C.C.; Moon, R.T. Ca(2+)/calmodulin-dependent protein kinase II is stimulated by Wnt and Frizzled homologs and promotes ventral cell fates in Xenopus. J. Biol. Chem. 2000, 275, 12701–12711. [Google Scholar] [CrossRef] [Green Version]
- Morishita, Y.; Kobayashi, K.; Klyachko, E.; Jujo, K.; Maeda, K.; Losordo, D.W.; Murohara, T. Wnt11 Gene Therapy with Adeno-associated Virus 9 Improves Recovery from Myocardial Infarction by Modulating the Inflammatory Response. Sci. Rep. 2016, 6, 21705. [Google Scholar] [CrossRef] [Green Version]
- Chen, B.; Chen, X.; Liu, C.; Li, J.; Liu, F.; Huang, Y. Co-expression of Akt1 and Wnt11 promotes the proliferation and cardiac differentiation of mesenchymal stem cells and attenuates hypoxia/reoxygenation-induced cardiomyocyte apoptosis. Biomed. Pharmacother. 2018, 108, 508–514. [Google Scholar] [CrossRef]
- Sasaki, T.; Hwang, H.; Nguyen, C.; Kloner, R.A.; Kahn, M. The small molecule Wnt signaling modulator ICG-001 improves contractile function in chronically infarcted rat myocardium. PLoS ONE 2013, 8, e75010. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Fan, Y.; Ho, B.X.; Pang, J.K.S.; Pek, N.M.Q.; Hor, J.H.; Ng, S.Y.; Soh, B.-S. Wnt/β-catenin-mediated signaling re-activates proliferation of matured cardiomyocytes. Stem Cell Res. Ther. 2018, 9, 338. [Google Scholar] [CrossRef] [Green Version]
- Mills, K.M.; Szczerkowski, J.L.A.; Habib, S.J. Wnt ligand presentation and reception: From the stem cell niche to tissue engineering. Open Biol. 2017, 7, 170140. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Alfaro, M.P.; Vincent, A.; Saraswati, S.; Thorne, C.A.; Hong, C.C.; Lee, E.; Young, P.P. sFRP2 suppression of bone morphogenic protein (BMP) and Wnt signaling mediates mesenchymal stem cell (MSC) self-renewal promoting engraftment and myocardial repair. J. Biol. Chem. 2010, 285, 35645–35653. [Google Scholar] [CrossRef] [Green Version]
- Xiang, G.; Yang, Q.; Wang, B.; Sekiya, N.; Mu, X.; Tang, Y.; Chen, C.W.; Okada, M.; Cummins, J.; Gharaibeh, B.; et al. Lentivirus-mediated Wnt11 gene transfer enhances Cardiomyogenic differentiation of skeletal muscle-derived stem cells. Mol. Ther. 2011, 19, 790–796. [Google Scholar] [CrossRef] [PubMed]
- Chen, H.T.; Huang, A.B.; He, Y.L.; Bian, J.; Li, H.J. Wnt11 overexpression promote adipose-derived stem cells differentiating to the nucleus pulposus-like phenotype. Eur. Rev. Med. Pharmacol. Sci. 2017, 21, 1462–1470. [Google Scholar]
- He, Z.; Li, H.; Zuo, S.; Pasha, Z.; Wang, Y.W.; Yang, Y.; Jiang, W.; Ashraf, M.; Xu, M. Transduction of Wnt11 promotes mesenchymal stem cell transdifferentiation into cardiac phenotypes. Stem Cells Dev. 2011, 20, 1771–1778. [Google Scholar] [CrossRef]
- Koyanagi, M.; Haendeler, J.; Badorff, C.; Brandes, R.P.; Hoffmann, J.; Pandur, P.; Zeiher, A.M.; Kühl, M.; Dimmeler, S. Non-canonical Wnt signaling enhances differentiation of human circulating progenitor cells to cardiomyogenic cells. J. Biol. Chem. 2005, 280, 16838–16842. [Google Scholar] [CrossRef] [Green Version]
- Flaherty, M.P.; Abdel-Latif, A.; Li, Q.; Hunt, G.; Ranjan, S.; Ou, Q.; Tang, X.-L.; Johnson, R.K.; Bolli, R.; Dawn, B. Noncanonical Wnt11 signaling is sufficient to induce cardiomyogenic differentiation in unfractionated bone marrow mononuclear cells. Circulation 2008, 117, 2241–2252. [Google Scholar] [CrossRef] [Green Version]
- Zuo, S.; Jones, W.K.; Li, H.; He, Z.; Pasha, Z.; Yang, Y.; Wang, Y.; Fan, G.-G.; Ashraf, M.; Xu, M. Paracrine effect of Wnt11-overexpressing mesenchymal stem cells on ischemic injury. Stem Cells Dev. 2012, 21, 598–608. [Google Scholar] [CrossRef] [PubMed]
- Kinnaird, T.; Stabile, E.; Burnett, M.S.; Lee, C.W.; Barr, S.; Fuchs, S.; Epstein, S.E. Marrow-derived stromal cells express genes encoding a broad spectrum of arteriogenic cytokines and promote in vitro and in vivo arteriogenesis through paracrine mechanisms. Circ. Res. 2004, 94, 678–685. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Hou, L.; Kim, J.J.; Woo, Y.J.; Huang, N.F. Stem cell-based therapies to promote angiogenesis in ischemic cardiovascular disease. Am. J. Physiol. Heart Circ. Physiol. 2016, 310, H455–H465. [Google Scholar] [CrossRef]
- Ertl, J.; Pichlsberger, M.; Tuca, A.C.; Wurzer, P.; Fuchs, J.; Geyer, S.H.; Maurer-Gesek, B.; Weninger, W.J.; Pfeiffer, D.; Bubalo, V. Comparative study of regenerative effects of mesenchymal stem cells derived from placental amnion, chorion and umbilical cord on dermal wounds. Placenta 2018, 65, 37–46. [Google Scholar] [CrossRef]
- Li, H.; Zuo, S.; He, Z.; Yang, Y.; Pasha, Z.; Wang, Y.; Xu, M. Paracrine factors released by GATA-4 overexpressed mesenchymal stem cells increase angiogenesis and cell survival. Am. J. Physiol. Heart Circ. Physiol. 2010, 299, H1772–H1781. [Google Scholar] [CrossRef] [Green Version]
- Timmers, L.; Lim, S.K.; Hoefer, I.E.; Arslan, F.; Lai, R.C.; van Oorschot, A.A.M.; Goumans, M.J.; Strijder, C.; Sze, S.K.; Choo, A.; et al. Human mesenchymal stem cell-conditioned medium improves cardiac function following myocardial infarction. Stem Cell Res. 2011, 6, 206–214. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Abreu, S.C.; Weiss, D.J.; Rocco, P.R. Extracellular vesicles derived from mesenchymal stromal cells: A therapeutic option in respiratory diseases? Stem Cell Res. Ther. 2016, 7, 53. [Google Scholar] [CrossRef] [Green Version]
- Ibrahim, A.G.; Cheng, K.; Marban, E. Exosomes as critical agents of cardiac regeneration triggered by cell therapy. Stem Cell Rep. 2014, 2, 606–619. [Google Scholar] [CrossRef] [Green Version]
- Wang, N.; Chen, C.; Yang, D.; Liao, Q.; Luo, H.; Wang, X.; Zhou, F.; Yang, X.; Yang, J.; Zeng, C.; et al. Mesenchymal stem cells-derived extracellular vesicles, via miR-210, improve infarcted cardiac function by promotion of angiogenesis. Biochim. Biophys. Acta 2017, 1863, 2085–2092. [Google Scholar] [CrossRef]
- Venetsanakos, E.; Mirza, A.; Fanton, C.; Romanov, S.R.; Tlsty, T.; McMahon, M. Induction of tubulogenesis in telomerase-immortalized human microvascular endothelial cells by glioblastoma cells. Exp. Cell Res. 2002, 273, 21–33. [Google Scholar] [CrossRef]
- Kern, J.; Steurer, M.; Gastl, G.; Gunsilius, E.; Untergasser, G. Vasohibin inhibits angiogenic sprouting in vitro and supports vascular maturation processes in vivo. BMC Cancer 2009, 9, 284. [Google Scholar] [CrossRef] [Green Version]
- Boodhwani, M.; Sodha, N.R.; Laham, R.J.; Sellke, F.W. The future of therapeutic myocardial angiogenesis. Shock 2006, 26, 332–341. [Google Scholar] [CrossRef] [PubMed]
- Bastakoty, D.; Saraswati, S.; Joshi, P.; Atkinson, J.; Feoktistov, I.; Liu, J.; Harris, J.L.; Young, P.P. Temporary, Systemic Inhibition of the WNT/β-Catenin Pathway promotes Regenerative Cardiac Repair following Myocardial Infarct. Cell Stem Cells Regen. Med. 2016, 2. [Google Scholar] [CrossRef] [PubMed]
- Paik, D.T.; Rai, M.; Ryzhov, S.; Sanders, L.N.; Aisagbonhi, O.; Funke, M.J.; Feoktistov, I.; Hatzopoulos, A.K. Wnt10b Gain-of-Function Improves Cardiac Repair by Arteriole Formation and Attenuation of Fibrosis. Circ. Res. 2015, 117, 804–816. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- McBride, J.D.; Rodriguez-Menocal, L.; Guzman, W.; Candanedo, A.; Garcia-Contreras, M.; Badiavas, E.V. Bone Marrow Mesenchymal Stem Cell-Derived CD63(+) Exosomes Transport Wnt3a Exteriorly and Enhance Dermal Fibroblast Proliferation, Migration, and Angiogenesis In Vitro. Stem Cells Dev. 2017, 26, 1384–1398. [Google Scholar] [CrossRef] [PubMed]
- Guo, X.; Gu, X.; Hareshwaree, S.; Rong, X.; Li, L.; Chu, M. Induced pluripotent stem cell-conditional medium inhibits H9C2 cardiomyocytes apoptosis via autophagy flux and Wnt/β-catenin pathway. J. Cell Mol. Med. 2019, 23, 4358–4374. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Cheng, C.W.; Yeh, J.C.; Fan, T.P.; Smith, S.K.; Charnock-Jones, D.S. Wnt5a-mediated non-canonical Wnt signalling regulates human endothelial cell proliferation and migration. Biochem. Biophys. Res. Commun. 2008, 365, 285–290. [Google Scholar] [CrossRef]
- Masckauchan, T.N.; Agalliu, D.; Vorontchikhina, M.; Ahn, A.; Parmalee, N.L.; Li, C.-M.; Khoo, A.; Tycko, B.; Brown, A.M.C.; Kitajewski, J. Wnt5a signaling induces proliferation and survival of endothelial cells in vitro and expression of MMP-1 and Tie-2. Mol. Biol. Cell. 2006, 17, 5163–5172. [Google Scholar] [CrossRef]
- Buga, A.M.; Margaritescu, C.; Scholz, C.J.; Radu, E.; Zelenak, C.; Popa-Wagner, A. Transcriptomics of post-stroke angiogenesis in the aged brain. Front. Aging Neurosci. 2014, 6, 44. [Google Scholar] [CrossRef]
- Seifert, J.R.; Mlodzik, M. Frizzled/PCP signalling: A conserved mechanism regulating cell polarity and directed motility. Nat. Rev. Genet. 2007, 8, 126–138. [Google Scholar] [CrossRef]
- Reis, M.; Liebner, S. Wnt signaling in the vasculature. Exp. Cell Res. 2013, 319, 1317–1323. [Google Scholar] [CrossRef] [PubMed]
- Pandur, P.; Lasche, M.; Eisenberg, L.M.; Kuhl, M. Wnt-11 activation of a non-canonical Wnt signalling pathway is required for cardiogenesis. Nature 2002, 418, 636–641. [Google Scholar] [CrossRef]
- Xu, H.; Czerwinski, P.; Hortmann, M.; Sohn, H.Y.; Forstermann, U.; Li, H. Protein kinase C α promotes angiogenic activity of human endothelial cells via induction of vascular endothelial growth factor. Cardiovasc. Res. 2008, 78, 349–355. [Google Scholar] [CrossRef] [Green Version]
- Abdul-Ghani, M.; Dufort, D.; Stiles, R.; De Repentigny, Y.; Kothary, R.; Megeney, L.A. Wnt11 promotes cardiomyocyte development by caspase-mediated suppression of canonical Wnt signals. Mol. Cell Biol. 2011, 31, 163–178. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Min, J.K.; Park, H.; Choi, H.J.; Kim, Y.; Pyun, B.-J.; Agrawal, V.; Song, B.-W.; Jeon, J.; Maeng, Y.-S.; Rho, S.-S.; et al. The WNT antagonist Dickkopf2 promotes angiogenesis in rodent and human endothelial cells. J. Clin. Investig. 2011, 121, 1882–1893. [Google Scholar] [CrossRef] [PubMed] [Green Version]
Primers | Forward | Reverse |
---|---|---|
Wnt11 | 5′-CAGGATCCCAAGCCAATAAA | 5′-GACAGGTAGCGGGTCTTGAG |
GFP | 5′-AAGTTCATCTGCACCACCG | 5′-TCCTTGAAGAAGATGGTGCG |
GAPDH | 5′-GTATGACAACGAATTTGGCTACAG | 5′-TGAGGGTCTCTCTTCCTCTTGT |
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Wang, J.; Gong, M.; Zuo, S.; Xu, J.; Paul, C.; Li, H.; Liu, M.; Wang, Y.-G.; Ashraf, M.; Xu, M. WNT11-Conditioned Medium Promotes Angiogenesis through the Activation of Non-Canonical WNT-PKC-JNK Signaling Pathway. Genes 2020, 11, 1277. https://0-doi-org.brum.beds.ac.uk/10.3390/genes11111277
Wang J, Gong M, Zuo S, Xu J, Paul C, Li H, Liu M, Wang Y-G, Ashraf M, Xu M. WNT11-Conditioned Medium Promotes Angiogenesis through the Activation of Non-Canonical WNT-PKC-JNK Signaling Pathway. Genes. 2020; 11(11):1277. https://0-doi-org.brum.beds.ac.uk/10.3390/genes11111277
Chicago/Turabian StyleWang, Jingcai, Min Gong, Shi Zuo, Jie Xu, Chris Paul, Hongxia Li, Min Liu, Yi-Gang Wang, Muhammad Ashraf, and Meifeng Xu. 2020. "WNT11-Conditioned Medium Promotes Angiogenesis through the Activation of Non-Canonical WNT-PKC-JNK Signaling Pathway" Genes 11, no. 11: 1277. https://0-doi-org.brum.beds.ac.uk/10.3390/genes11111277