miR-126 regulates platelet-derived growth factor receptor-α expression and migration of primary human osteoblasts
-
Yvonne Schmidt
, Filip Simunovic, Sandra Strassburg
, Dietmar Pfeifer , G. Björn Stark und Günter Finkenzeller
Abstract
Adequate vascularization is an essential requirement for bone development, fracture healing and bone tissue engineering. We have previously described the coculture of primary human osteoblasts (hOBs) and human endothelial cells (HUVECs), designed to investigate the interactions between these cells. In this system, we showed that cocultivation of these two cell types leads to a downregulation of platelet-derived growth factor receptor-α (PDGFR-α) in hOBs, which was a consequence of reduced mRNA stability. In the current study we investigated the possible involvement of microRNAs in this process. Firstly, we performed a microarray analysis of osteoblastic miRNAs following cocultivation with HUVECs, revealing an upregulation of miR-126. This result was confirmed by RT-qPCR, and we observed that the increase is dependent on direct cell-to-cell contacts. Gain-of-function and loss-of-function experiments showed that miR-126 is a negative regulator of PDGFR-α mRNA. Additionally, migration of hOBs was inhibited by miR-126 overexpression and stimulated by miR-126 inhibition. Addition of PDGFR-α blocking antibody to hOB culture also inhibited hOB migration. There was no effect of miR-126 modulation on osteoblast proliferation, apoptosis rate or differentiation. In conclusion, we report that the miR-126/PDGFR-α system regulates the migratory behavior of human osteoblasts, without exerting effects on cell survival and differentiation.
Acknowledgments
This work was supported by Deutsche Forschungsgemeinschaft (DFG), grant number: FI 790/4-1 to GF and by a research grant from the Medical School of Freiburg University (SIM916/13) to FS.
References
Bartel, D.P. (2004). MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116, 281–297.10.1016/S0092-8674(04)00045-5Suche in Google Scholar
Bolstad, B.M., Irizarry, R.A., Astrand, M., and Speed, T.P. (2003). A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics 19, 185–193.10.1093/bioinformatics/19.2.185Suche in Google Scholar PubMed
Caplan, A.I., and Correa, D. (2011). PDGF in Bone Formation and Regeneration: New Insights into a Novel Mechanism Involving MSCs. J. Orthop. Res. 29, 1795–1803.10.1002/jor.21462Suche in Google Scholar PubMed
Chaudhary, L.R., Hofmeister, A.M., and Hruska, K.A. (2004). Differential growth factor control of bone formation through osteoprogenitor differentiation. Bone 34, 402–411.10.1016/j.bone.2003.11.014Suche in Google Scholar PubMed
Dong, S., Yang, B., Guo, H., and Kang, F. (2012). MicroRNAs regulate osteogenesis and chondrogenesis. Biochem. Biophys. Res. Commun. 418, 587–591.10.1016/j.bbrc.2012.01.075Suche in Google Scholar PubMed
Donovan, J., Abraham, D., and Norman, J. (2013). Platelet-derived growth factor signaling in mesenchymal cells. Front. Biosci. (Landmark Ed) 18, 106–119.10.2741/4090Suche in Google Scholar PubMed
Finkenzeller, G., Arabatzis, G., Geyer, M., Wenger, A., Bannasch, H., and Stark, G.B. (2006). Gene expression profiling reveals platelet-derived growth factor receptor α as a target of cell contact-dependent gene regulation in an endothelial cell-osteoblast co-culture model. Tissue Eng. 12, 2889–2903.10.1089/ten.2006.12.2889Suche in Google Scholar PubMed
Finkenzeller, G., Mehlhorn, A.T., Schmal, H., and Stark, G.B. (2010). Post-transcriptional regulation of osteoblastic platelet-derived growth factor receptor-α expression by co-cultured primary endothelial cells. Cells Tissues Organs 192, 28–38.10.1159/000276590Suche in Google Scholar PubMed
Fish, J.E., Santoro, M.M., Morton, S.U., Yu, S., Yeh, R.F., Wythe, J.D., Ivey, K.N., Bruneau, B.G., Stainier, D.Y., and Srivastava, D. (2008). miR-126 regulates angiogenic signaling and vascular integrity. Dev. Cell 15, 272–284.10.1016/j.devcel.2008.07.008Suche in Google Scholar PubMed PubMed Central
Fitter, S., Dewar, A.L., Kostakis, P., To, L.B., Hughes, T.P., Roberts, M.M., Lynch, K., Vernon-Roberts, B., and Zannettino, A.C. (2008). Long-term imatinib therapy promotes bone formation in CML patients. Blood 111, 2538–2547.10.1182/blood-2007-07-104281Suche in Google Scholar PubMed
Fuchs, S., Hofmann, A., and Kirkpatrick, C. (2007). Microvessel-like structures from outgrowth endothelial cells from human peripheral blood in 2-dimensional and 3-dimensional co-cultures with osteoblastic lineage cells. Tissue Eng. 13, 2577–2588.10.1089/ten.2007.0022Suche in Google Scholar PubMed
Grabher, C., Payne, E.M., Johnston, A.B., Bolli, N., Lechman, E., Dick, J.E., Kanki, J.P., and Look, A.T. (2011). Zebrafish microRNA-126 determines hematopoietic cell fate through c-Myb. Leukemia 25, 506–514.10.1038/leu.2010.280Suche in Google Scholar PubMed PubMed Central
Grellier, M., Bordenave, L., and Amedee, J. (2009). Cell-to-cell communication between osteogenic and endothelial lineages: implications for tissue engineering. Trends Biotechnol. 27, 562–571.10.1016/j.tibtech.2009.07.001Suche in Google Scholar PubMed
Guo, C., Sah, J.F., Beard, L., Willson, J.K., Markowitz, S.D., and Guda, K. (2008). The noncoding RNA, miR-126, suppresses the growth of neoplastic cells by targeting phosphatidylinositol 3-kinase signaling and is frequently lost in colon cancers. Genes Chromosomes Cancer 47, 939–946.10.1002/gcc.20596Suche in Google Scholar PubMed PubMed Central
Hengartner, N.E., Fiedler, J., Ignatius, A., and Brenner, R.E. (2013). IL-1beta inhibits human osteoblast migration. Mol. Med. 19, 36–42.10.2119/molmed.2012.00058Suche in Google Scholar PubMed PubMed Central
Hock, J.M. and Canalis, E. (1994). Platelet-derived growth factor enhances bone cell replication, but not differentiated function of osteoblasts. Endocrinology 134, 1423–1428.10.1210/endo.134.3.8119182Suche in Google Scholar PubMed
Hollinger, J.O., Hart, C.E., Hirsch, S.N., Lynch, S., and Friedlaender, G.E. (2008). Recombinant human platelet-derived growth factor: biology and clinical applications. J. Bone Joint. Surg. Am. 90 (Suppl 1), 48–54.10.2106/JBJS.G.01231Suche in Google Scholar PubMed
Irizarry, R.A., Bolstad, B.M., Collin, F., Cope, L.M., Hobbs, B., and Speed, T.P. (2003a). Summaries of Affymetrix GeneChip probe level data. Nucleic Acids Res. 31, e15.10.1093/nar/gng015Suche in Google Scholar PubMed PubMed Central
Irizarry, R.A., Hobbs, B., Collin, F., Beazer-Barclay, Y.D., Antonellis, K.J., Scherf, U., and Speed, T.P. (2003b). Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics 4, 249–264.10.1093/biostatistics/4.2.249Suche in Google Scholar PubMed
Kaipel, M., Schutzenberger, S., Schultz, A., Ferguson, J., Slezak, P., Morton, T.J., Van Griensven, M., and Redl, H. (2012). BMP-2 but not VEGF or PDGF in fibrin matrix supports bone healing in a delayed-union rat model. J. Orthop. Res. 30, 1563–1569.10.1002/jor.22132Suche in Google Scholar PubMed
Kaully, T., Kaufman-Francis, K., Lesman, A., and Levenberg, S. (2009). Vascularization–the conduit to viable engineered tissues. Tissue Eng. Part B Rev. 15, 159–169.10.1089/ten.teb.2008.0193Suche in Google Scholar
Kratchmarova, I., Blagoev, B., Haack-Sorensen, M., Kassem, M., and Mann, M. (2005). Mechanism of divergent growth factor effects in mesenchymal stem cell differentiation. Science 308, 1472–1477.10.1126/science.1107627Suche in Google Scholar
Kubota, K., Sakikawa, C., Katsumata, M., Nakamura, T., and Wakabayashi, K. (2002a). PDGF BB purified from osteoclasts acts as osteoblastogenesis inhibitory factor (OBIF). J. Biomol. Tech. 13, 62–71.Suche in Google Scholar
Kubota, K., Sakikawa, C., Katsumata, M., Nakamura, T., and Wakabayashi, K. (2002b). Platelet-derived growth factor BB secreted from osteoclasts acts as an osteoblastogenesis inhibitory factor. J. Bone Miner. Res. 17, 257–265.10.1359/jbmr.2002.17.2.257Suche in Google Scholar
Kumar, A., Salimath, B.P., Stark, G.B., and Finkenzeller, G. (2010). Platelet-derived growth factor receptor signaling is not involved in osteogenic differentiation of human mesenchymal stem cells. Tissue Eng. Part A 16, 983–993.10.1089/ten.tea.2009.0230Suche in Google Scholar
Li, Z., Li, N., Wu, M., Li, X., Luo, Z., and Wang, X. (2013). Expression of miR-126 suppresses migration and invasion of colon cancer cells by targeting CXCR4. Mol. Cell Biochem. 381, 233–242.10.1007/s11010-013-1707-6Suche in Google Scholar
Lian, J.B., Stein, G.S., van Wijnen, A.J., Stein, J.L., Hassan, M.Q., Gaur, T., and Zhang, Y. (2012). MicroRNA control of bone formation and homeostasis. Nat. Rev. Endocrinol. 8, 212–227.10.1038/nrendo.2011.234Suche in Google Scholar
Pechak, D.G., Kujawa, M.J., and Caplan, A.I. (1986). Morphological and histochemical events during first bone formation in embryonic chick limbs. Bone 7, 441–458.10.1016/8756-3282(86)90004-9Suche in Google Scholar
Raines, E.W., Ross R. (1992). Compartmentalization of PDGF on extracellular binding sites dependent on exon-6-encoded sequences. J. Cell Biol. 116, 533–543.10.1083/jcb.116.2.533Suche in Google Scholar PubMed PubMed Central
Rouwkema, J., de Boer, J., and Van Blitterswijk, C.A. (2006). Endothelial cells assemble into a 3-dimensional prevascular network in a bone tissue engineering construct. Tissue Eng. 12, 2685–2693.10.1089/ten.2006.12.2685Suche in Google Scholar PubMed
Sato, N., Beitz, J.G., Kato, J., Yamamoto, M., Clark, J.W., Calabresi, P., Raymond, A., and Frackelton, A.R., Jr. (1993). Platelet-derived growth factor indirectly stimulates angiogenesis in vitro. Am. J. Pathol. 142, 1119–1130.Suche in Google Scholar
Simunovic, F., Steiner, D., Pfeifer, D., Stark, G.B., Finkenzeller, G., and Lampert, F. (2013). Increased extracellular matrix and proangiogenic factor transcription in endothelial cells after cocultivation with primary human osteoblasts. J. Cell. Biochem. 114, 1584–1594.10.1002/jcb.24500Suche in Google Scholar PubMed
Sonntag, K.C., Woo, T.U., and Krichevsky, A.M. (2012). Converging miRNA functions in diverse brain disorders: a case for miR-124 and miR-126. Exp. Neurol. 235, 427–435.10.1016/j.expneurol.2011.11.035Suche in Google Scholar PubMed PubMed Central
Stahl, A., Wenger, A., Weber, H., Stark, G.B., Augustin, H.G., and Finkenzeller, G. (2004). Bi-directional cell contact-dependent regulation of gene expression between endothelial cells and osteoblasts in a three-dimensional spheroidal coculture model. Biochem. Biophys. Res. Commun. 322, 684–692.10.1016/j.bbrc.2004.07.175Suche in Google Scholar PubMed
Steffens, L., Wenger, A., Stark, G.B., and Finkenzeller, G. (2009). In vivo engineering of a human vasculature for bone tissue engineering applications. J. Cell. Mol. Med. 13, 3380–3386.10.1111/j.1582-4934.2008.00418.xSuche in Google Scholar PubMed PubMed Central
Steiner, D., Lampert, F., Stark, G.B., and Finkenzeller, G. (2012). Effects of endothelial cells on proliferation and survival of human mesenchymal stem cells and primary osteoblasts. J. Orthop. Res. 30, 1682–1689.10.1002/jor.22130Suche in Google Scholar PubMed
Sun, H., Qu, Z., Guo, Y., Zang, G., and Yang, B. (2007). In vitro and in vivo effects of rat kidney vascular endothelial cells on osteogenesis of rat bone marrow mesenchymal stem cells growing on polylactide-glycoli acid (PLGA) scaffolds. Biomed. Eng. Online 6, 41.Suche in Google Scholar
Taipaleenmaki, H., Bjerre Hokland, L., Chen, L., Kauppinen, S., and Kassem, M. (2012). Mechanisms in endocrinology: micro-RNAs: targets for enhancing osteoblast differentiation and bone formation. Eur. J. Endocrinol. 166, 359–371.10.1530/EJE-11-0646Suche in Google Scholar PubMed
Villars, F., Guillotin, B., Amedee, T., Dutoya, S., Bordenave, L., Bareille, R., and Amedee, J. (2002). Effect of HUVEC on human osteoprogenitor cell differentiation needs heterotypic gap junction communication. Am. J. Physiol. Cell Physiol. 282, C775–785.10.1152/ajpcell.00310.2001Suche in Google Scholar PubMed
Wang, S., Aurora, A.B., Johnson, B.A., Qi, X., McAnally, J., Hill, J.A., Richardson, J.A., Bassel-Duby, R., and Olson, E.N. (2008). The endothelial-specific microRNA miR-126 governs vascular integrity and angiogenesis. Dev. Cell. 15, 261–271.10.1016/j.devcel.2008.07.002Suche in Google Scholar PubMed PubMed Central
Wenger, A., Stahl, A., Weber, H., Finkenzeller, G., Augustin, H.G., Stark, G.B., and Kneser, U. (2004). Modulation of in vitro angiogenesis in a three-dimensional spheroidal coculture model for bone tissue engineering. Tissue Eng. 10, 1536–1547.10.1089/ten.2004.10.1536Suche in Google Scholar PubMed
Wu, Z., Irizarry, R.A., Gentleman, R., Murillo, F.M., and Spencer, F. (2004). A Model-Based Background Adjustment for Oligonucleotide Expression Arrays. J. Am. Statist. Assoc. 99, 909–917.10.1198/016214504000000683Suche in Google Scholar
Yoshida, S., Iwasaki, R., Kawana, H., Miyauchi, Y., Hoshi, H., Miyamoto, H., Mori, T., Kanagawa, H., Katsuyama, E., Fujie, A., et al. (2012). PDGFBB promotes PDGFRα-positive cell migration into artificial bone in vivo. Biochem. Biophys. Res. Commun. 421, 785–789.10.1016/j.bbrc.2012.04.084Suche in Google Scholar PubMed
Yu, H., Vandevord, P.J., Gong, W., Wu, B., Song, Z., Matthew, H.W., Wooley, P.H., and Yang, S.Y. (2008). Promotion of osteogenesis in tissue-engineered bone by pre-seeding endothelial progenitor cells-derived endothelial cells. J. Orthop. Res. 26, 1147–1152.10.1002/jor.20609Suche in Google Scholar PubMed
©2014 by De Gruyter
Artikel in diesem Heft
- Frontmatter
- Reviews
- Cholesterol lowering: role in cancer prevention and treatment
- The role of the Mpv17 protein mutations of which cause mitochondrial DNA depletion syndrome (MDDS): lessons from homologs in different species
- Research Articles/Short Communications
- Genes and Nucleic Acids
- Identification of a novel PHEX mutation in a Chinese family with X-linked hypophosphatemic rickets using exome sequencing
- Protein Structure and Function
- Complement activation by salivary agglutinin is secretor status dependent
- Synthesis, biological evaluation, and docking studies of PAR2-AP-derived pseudopeptides as inhibitors of kallikrein 5 and 6
- Membranes, Lipids, Glycobiology
- Recombinant HDL (Milano) protects endotoxin-challenged rats from multiple organ injury and dysfunction
- Cell Biology and Signaling
- miR-126 regulates platelet-derived growth factor receptor-α expression and migration of primary human osteoblasts
- Legumain expression, activity and secretion are increased during monocyte-to-macrophage differentiation and inhibited by atorvastatin
- Proteolysis
- Cell surface serine protease matriptase-2 suppresses fetuin-A/AHSG-mediated induction of hepcidin
Artikel in diesem Heft
- Frontmatter
- Reviews
- Cholesterol lowering: role in cancer prevention and treatment
- The role of the Mpv17 protein mutations of which cause mitochondrial DNA depletion syndrome (MDDS): lessons from homologs in different species
- Research Articles/Short Communications
- Genes and Nucleic Acids
- Identification of a novel PHEX mutation in a Chinese family with X-linked hypophosphatemic rickets using exome sequencing
- Protein Structure and Function
- Complement activation by salivary agglutinin is secretor status dependent
- Synthesis, biological evaluation, and docking studies of PAR2-AP-derived pseudopeptides as inhibitors of kallikrein 5 and 6
- Membranes, Lipids, Glycobiology
- Recombinant HDL (Milano) protects endotoxin-challenged rats from multiple organ injury and dysfunction
- Cell Biology and Signaling
- miR-126 regulates platelet-derived growth factor receptor-α expression and migration of primary human osteoblasts
- Legumain expression, activity and secretion are increased during monocyte-to-macrophage differentiation and inhibited by atorvastatin
- Proteolysis
- Cell surface serine protease matriptase-2 suppresses fetuin-A/AHSG-mediated induction of hepcidin