بررسی اثر آگونیست -آدرنرژیکی ایزوپروترنول بر بیان miR-886-3p و miR-23a در سلولهای بنیادی مزانشیمی مغز استخوان انسان

Background and Objective: Mobilization of Hematopoietic Stem Cells (HSCs) for transplantation and the importance of -adrenergic signals in induction of this process have been well investigated. However, little is known about the role of -adrenergic signals in mobilization of HSCs and factors influenced by these signals. The Chemokine Stromal Derived Factor -1 (SDF-1) which is expressed by human bone marrow-derived mesenchymal stem cells (hMSCs), has a key role in mobilization of HSCs. In addition, miR-886-3p and miR-23a can regulate the expression of SDF-1 in hMSCs. In this study, to investigate the role of miR-886-3p and miR-23a in mobilization process, expression of both miRNAs was evaluated in hMSCs treated by Isoproterenol (a -adrenergic agonist). Materials and Methods: hMSCs were isolated from human bone marrow and cultured. Following flowcytometric analysis, the cells were treated with 100 M isoproterenol. Total RNA was extracted at 12 and 48 hours post treatment, and also from untreated hMSCs as control. Then, miR-886-3p and miR-23a expression levels were quantified by quantitative Reverse Transcriptase PCR. Results: The expression level of miR-886-3p increased significantly at 12 and 48 hours post treatment (P<0.05). In addition, the expression level of miR-23a decreased at 12 hours post treatment and increased significantly at 48 hours post treatment (P<0.05). Conclusion: Isoproterenol induces miR-886-3p in hMSCs. MiR-23a is primarily decreased, and then increased due to treating with isoproterenol. So both miRNAs can contribute to mobilization process. References 1- Horowitz MM, Gale RP, Sondel PM, et al. Graft-versus-leukemia reactions after bone marrow transplantation. Blood. 1990 75: 555-62. 2- Sykes M, Nikolic B. Treatment of severe autoimmune disease by stem-cell transplantation. Nature. 2005 435: 620-7. 3- Copelan EA. Hematopoietic stem-cell transplantation. N Engl J Med. 2006 354: 1813-26. 4- Wright DE, Wagers AJ, Gulati AP, Johnson FL, Weissman IL. Physiological migration of hematopoietic stem and progenitor cells. Science. 2001 294: 1933-6. 5- Anderlini P, Körbling M. The use of mobilized peripheral blood stem cells from normal donors for allografting. Stem cells. 1997 15: 9-17. 6- Orkin SH, Zon LI. Hematopoiesis: an evolving paradigm for stem cell biology. Cell. 2008 132: 631-44. 7- Levesque J-P, Winkler IG. Mobilization of hematopoietic stem cells: state of the art. Curr Opin Organ Transplant. 2008 13: 53-8. 8- Motabi IH, DiPersio JF. Advances in stem cell mobilization. Blood Rev. 2012 26: 267-78. 9- Siena S, Bregni M, Brando B, Ravagnani F, Bonadonna G, Gianni AM. Circulation of CD34+ hematopoietic stem cells in the peripheral blood of high-dose cyclophosphamide-treated patients: enhancement by intravenous recombinant human granulocyte-macrophage colony-stimulating factor. Blood. 1989 74: 1905-14. 10- Dar A, Kalinkovich A, Netzer N, et al. AMD3100 signals via the nervous system, inducing release to the circulation of bone marrow SDF-1, which is crucial for progenitor cell mobilization. Blood. (ASH Annual Meeting Abstracts). 2006 108: Abstract, 1315. 11- Mendez-Ferrer S, Battista M, Frenette PS. Cooperation of 2-and 3-adrenergic receptors in hematopoietic progenitor cell mobilization. Ann N Y Acad Sci. 2010 1192: 139-44. 12- Spiegel A, Shivtiel S, Kalinkovich A, et al. Catecholaminergic neurotransmitters regulate migration and repopulation of immature human CD34+ cells through Wnt signaling. Nat Immunol. 2007 8: 1123-31. 13- Katayama Y, Battista M, Kao W-M, et al. Signals from the sympathetic nervous system regulate hematopoietic stem cell egress from bone marrow. Cell. 2006 124: 407-21. 14- Dar A, Schajnovitz A, Lapid K, et al. Rapid mobilization of hematopoietic progenitors by AMD3100 and catecholamines is mediated by CXCR4-dependent SDF-1 release from bone marrow stromal cells. Leukemia. 2011 25: 1286-96. 15- Muthu K, Iyer S, He L-K, et al. Murine hematopoietic stem cells and progenitors express adrenergic receptors. J Neuroimmunol. 2007 186: 27-36. 16- Méndez-Ferrer S, Lucas D, Battista M, Frenette PS. Haematopoietic stem cell release is regulated by circadian oscillations. Nature. 2008 452: 442-7. 17- Almquist A, Goldenberg IF, Milstein S, et al. Provocation of bradycardia and hypotension by isoproterenol and upright posture in patients with unexplained syncope. N Engl J Med. 1989 320: 346-51. 18- Sorrentino A, Ferracin M, Castelli G, et al. Isolation and characterization of CD146 multipotent mesenchymal stromal cells. Exp Hematol. 2008 36: 1035-46. 19- Ponomaryov T, Peled A, Petit I, et al. Induction of the chemokine stromal-derived factor-1 following DNA damage improves human stem cell function. J Clin Invest. 2000 106: 1331-9. 20- Ceradini DJ, Kulkarni AR, Callaghan MJ, et al. Progenitor cell trafficking is regulated by hypoxic gradients through HIF-1 induction of SDF-1. Nat Med. 2004 10: 858-64. 21- Sugiyama T, Kohara H, Noda M, Nagasawa T. Maintenance of the hematopoietic stem cell pool by CXCL12-CXCR4 chemokine signaling in bone marrow stromal cell niches. Immunity. 2006 25: 977-88. 22- Petit I, Szyper-Kravitz M, Nagler A, et al. G-CSF induces stem cell mobilization by decreasing bone marrow SDF-1 and up-regulating CXCR4. Nat Immunol. 2002 3: 687-94. 23- Heissig B, Hattori K, Dias S, et al. Recruitment of stem and progenitor cells from the bone marrow niche requires MMP-9 mediated release of kit-ligand. Cell. 2002 109: 625-37. 24- Fedyk ER, Jones D, Critchley HOD, Phipps RP, Blieden TM, Springer TA. Expression of stromal-derived factor-1 is decreased by IL-1 and TNF and in dermal wound healing. J Immunol. 2001 166: 5749-54. 25- Nakayama T, Mutsuga N, Tosato G. Effect of fibroblast growth factor 2 on stromal cell-derived factor 1 production by bone marrow stromal cells and hematopoiesis. J Natl Cancer Inst. 2007 99: 223-35. 26- Lu MH, Li CZ, Hu CJ, et al. microRNA-27b suppresses mouse MSC migration to the liver by targeting SDF-1 alpha in vitro. Biochem Biophys Res Commun. 2012 421: 389-95. 27- Solingen C, Boer HC, Bijkerk R, et al. MicroRNA-126 modulates endothelial SDF-1 expression and mobilization of Sca-1+/Lin− progenitor cells in ischaemia. Cardiovasc Res. 2011 92: 449-55. 28- Morenos L, Saffery R, Mechinaud F, et al. Evaluation of microRNA expression in patient bone marrow aspirate slides. PloS one. 2012 7: e42951. 29- Wu L, Zhou H, Zhang Q, et al. DNA methylation mediated by a microRNA pathway. Mol Cell. 2010 38: 465-75. 30- Davis-Dusenbery BN, Hata A. Mechanisms of control of microRNA biogenesis. J Biochem. 2010 148: 381-92. 31- Budhu A, Ji J, Wang X. The clinical potential of microRNAs. J Hematol Oncol. 2010 3: 37. 32- Minayi N, Alizadeh S, Dargahi H, et al. The Effect of miR-210 Up-regulation on proliferation and survival of mouse bone marrow derived mesenchymal stem cell. Int J Hematol Oncol Stem Cell Res. 2014 8: 15. 33- Pillai MM, Yang X, Balakrishnan I, Bemis L, Torok-Storb B. MiR-886-3p down regulates CXCL12 (SDF1) expression in human marrow stromal cells. PloS one. 2010 5: e14304. 34- Fierro F, Poitz D, Nolta JA, et al. MicroRNA mir-23a regulates SDF-1alpha expression in human bone marrow-derived mesenchymal stem cells. Blood. 2008: 112: 841. 35- Levesque J-P, Liu F, Simmons PJ, et al. Characterization of hematopoietic progenitor mobilization in protease-deficient mice. Blood. 2004 104: 65-72.