The Effect of Erythropoietin Treatment on Gene Expression Profile of Mesenchymal Stem Cells

Introduction: Mesenchymal Stem Cells are one of the most important elements of bone marrow microenvironment, which has a role in stromal support and stem cell differentiation. Exosomes are small vesicles that responsible from various cellular roles such as cell-cell communication and cell signaling, which may affect nearby and distant cells/tissues. Mesenchymal Stem Cells have therapeutic importance because of their multipotency and immune modulation potentials also with their exosomes. Erythropoietin, produced by liver hepatocytes, is responsible for erythroid differenti-ation (erythropoiesis) in bone marrow. In addition, Erythropoietin treatment of sev-eral cell types including Mesenchymal Stem Cells, showed therapeutic effects in var-ious diseases. Objective: The aim of this study is to examine the effect of erythropoietin on bone marrow Mesenchymal Stem Cells transcriptome and exosome derived miRNA profile. Materials and Methods: Effect of 3 different doses of Erythropoietin (1 IU/ml, 10 IU/ml and 100 IU/ml) for 48 hours on Mesenchymal Stem Cells transcriptome profile was analyzed. The results illustrated that 10 IU/ml Erythropoietin treatment has the most effective concentration in terms of gene expression profile. Therefore, small RNA libraries targeting miRNA was analyzed with 10 IU/ml Erythropoietin treated versus non treated groups with next generation sequencing. Results: We found that Erythropoietin treatment slightly changed global gene ex-pression profile. On the other hand, it was observed that Erythropoietin treated Mesenchymal Stem Cells have different exosomal miRNA profile. Conclusion: Differentially expressed exosomal miRNAs may have therapeutic effects in different conditions. It will be important to perform further studies with in vitro models, mimicking different physiological conditions and diseases for Mesenchymal Stem Cells and exosome biology.


Kobolak J, Dinnyes A, Memic A, et al. Mesenchymal stem cells: Identification, phenotypic characterization, biolog-ical properties and potential for regenerative medicine through biomaterial micro-engineering of their niche. Methods 2016; 99: 62-8.

Spees JL, Lee RH, Gregory CA. Mechanisms of mesenchy-mal stem/stromal cell function. Stem Cell Res Ther 2016; 7(1): 12 5.

Vizoso FJ, Eiro N, Cid S, et al. Mesenchymal Stem Cell Secretome: Toward Cell-Free Therapeutic Strategies in Regenerative Medicine. Int J Mol Sci. 2017; 18(9).

Thery C, Ostrowski M, Segura E. Membrane vesicles as conveyors of immune responses. Nat Rev Immunol 2009; 9(8): 581-93.

Zhang H-G, Grizzle WE. Exosomes: A Novel Pathway of Local and Distant Intercellular Communication that Facilitates the Growth and Metastasis of Neoplastic Lesions. The American Journal of Pathology 2014; 184(1): 28 - 41.

Ophelders DR, Wolfs TG, Jellema RK, et al. Mesenchymal Stromal Cell-Derived Extracellular Vesicles Protect the Fetal Brain After Hypoxia-Ischemia. Stem Cells Transl Med 2016; 5(6): 754-63.

Collino F, Bruno S, Incarnato D, et al. AKI Recovery Induced by Mesenchymal Stromal Cell-Derived Extracellular Vesicles Carrying MicroRNAs. J Am Soc Nephrol 2015; 26(10): 2349-60.

Jarmalaviciute A, Pivoriunas A. Exosomes as a poten-tial novel therapeutic tools against neurodegenerative diseases. Pharmacol Res 2016; 113(Pt B): 816-22

Tomasoni S, Longaretti L, Rota C, et al. Transfer of growth factor receptor mRNA via exosomes unravels the regen-erative effect of mesenchymal stem cells. Stem Cells Dev 2013; 22(5): 772-80.

Phinney DG, Pittenger MF. Concise Review: MSC-Derived Exosomes for Cell-Free Therapy. Stem Cells 2017; 35(4): 851- 8.

Miyake T, Kung CK, Goldwasser E. Purification of human erythropoietin. J Biol Chem 1977; 252(15): 5558-64.

Onal EM, Sag AA, Sal O, et al. Erythropoietin mediates brain-vascular-kidney crosstalk and may be a treatment target for pulmonary and resistant essential hyperten-sion. Clin Exp Hypertens 2017; 39(3): 197-209.

Powell JS, Berkner KL, Lebo RV, et al. Human erythropoie-tin gene: high level expression in stably transfected mam-malian cells and chromosome localization. Proc Natl Acad Sci USA 1986; 83(17): 6465-9.

Noguchi CT, Wang L, Rogers HM, el at. Survival and prolif-erative roles of erythropoietin beyond the erythroid lin-eage. Expert Rev Mol Med 2008; 10: e36.

Maiese K, Li F, Chong ZZ. New avenues of exploration for erythropoietin. JAMA 2005; 293(1): 90-5.

Li J, Guo W, Xiong M, et al. Erythropoietin facilitates the recruitment of bone marrow mesenchymal stem cells to sites of spinal cord injury. Exp Ther Med 2017; 13(5): 18 0 6 -12 .

Wang Y, Lu X, He J, et al. Influence of erythropoietin on microvesicles derived from mesenchymal stem cells pro-tecting renal function of chronic kidney disease. Stem Cell Res Ther 2015; 6: 100.

Tari K, Atashi A, Kaviani S, et al. Erythropoietin induces production of hepatocyte growth factor from bone mar-row mesenchymal stem cells in vitro. Biologicals 2017; 45: 15 - 9.

Ercan E, Bagla AG, Aksoy A, et al. In vitro protection of ad-ipose tissue-derived mesenchymal stem cells by erythro-poietin. Acta Histochem 2014; 116(1): 117-25.

Koh SH, Noh MY, Cho GW, et al. Erythropoietin increases the motility of human bone marrow-multipotent stromal cells (hBM-MSCs) and enhances the production of neu-rotrophic factors from hBM-MSCs. Stem Cells Dev 2009; 18(3): 411-21.

Liao Y-C, Wang Y-S, Guo Y-C, et al. Let-7g Improves Multiple Endothelial Functions Through Targeting Transforming Growth Factor-Beta and SIRT-1 Signaling. Journal of the American College of Cardiology 2014; 63(16): 1685-94.

Zhang Y, Fan M, Zhang X, et al. Cellular microRNAs up-reg-ulate transcription via interaction with promoter TATA- box motifs. RNA 2014; 20(12): 1878-89.

Zhang J, Ma J, Long K, et al. Overexpression of Exosomal Cardioprotective miRNAs Mitigates Hypoxia-Induced H9c2 Cells Apoptosis. Int J Mol Sci 2017; 18(4).

Beltrami C, Besnier M, Shantikumar S, et al. Human Pericardial Fluid Contains Exosomes Enriched with Cardiovascular-Expressed MicroRNAs and Promotes Therapeutic Angiogenesis. Molecular therapy: the jour-nal of the American Society of Gene Therapy 2017; 25(3): 679 -93.

Yang H, Fang F, Chang R, et al. MicroRNA-140-5p suppress-es tumor growth and metastasis by targeting transform-ing growth factor beta receptor 1 and fibroblast growth factor 9 in hepatocellular carcinoma. Hepatology 2013; 5 8 (1) : 2 0 5 -17.

Si X, Zhang X, Hao X, et al. Upregulation of miR-99a is associated with poor prognosis of acute myeloid leu-kemia and promotes myeloid leukemia cell expansion. Oncotarget 2016; 7(47): 78095-109.

Chen C-F, He X, Arslan AD, et al. Novel regulation of NF-YB by miR-485-3p affects expression of DNA topoisomer-ase IIα and drug responsiveness. Molecular Pharmacology 2011; 79(4): 735-41.

Li Y, Chen D, Li Y, et al. Oncogenic cAMP responsive ele-ment binding protein 1 is overexpressed upon loss of tu-mor suppressive miR-10b-5p and miR-363-3p in renal can-cer. Oncol Rep 2016; 35(4): 1967- 78.

Liu Z-R, Song Y, Wan L-H, et al. Over-expression of miR- 451a can enhance the sensitivity of breast cancer cells to tamoxifen by regulating 14-3-3ζ, estrogen receptor α, and autophagy. Life Sciences 2016; 149: 104-13.

Rasheed Z, Al-Shobaili HA, Rasheed N, et al. MicroRNA- 26a-5p regulates the expression of inducible nitric oxide synthase via activation of NF-κB pathway in human os-teoarthritis chondrocytes. Archives of Biochemistry and Biophysics 2016; 594: 61-7.

Wang X, Liu S, Cao L, et al. miR-29a-3p suppresses cell pro-liferation and migration by downregulating IGF1R in hepa-tocellular carcinoma. Oncotarget 2017; 8(49): 86592-603.

Wang T, Ren Y, Liu R, et al. miR-195-5p Suppresses the Proliferation, Migration, and Invasion of Oral Squamous Cell Carcinoma by Targeting TRIM14. Biomed Res Int 2017; 2 017: 7 37814 8 .

Dong W, Yao C, Teng X, et al. MiR-140-3p suppressed cell growth and invasion by downregulating the expression of ATP8A1 in non- small cell lung cancer. Tumor Biology 2016; 37(3): 2973-85.

Yang XW, Zhang LJ, Huang XH, et al. miR-145 suppress-es cell invasion in hepatocellular carcinoma cells: miR-145 targets ADAM17. Hepatol Res 2014; 44(5): 551-9.[

Hsu KW, Fang WL, Huang KH, et al. Notch1 pathway-medi-ated microRNA-151-5p promotes gastric cancer progres-sion. Oncotarget 2016; 7(25): 38036-51.

Geng L, Sun B, Gao B, et al. MicroRNA-103 promotes col-orectal cancer by targeting tumor suppressor DICER and PTEN. International journal of molecular sciences 2014; 15(5): 8458-72.

Yang J, Zhang Z, Chen C, et al. MicroRNA-19a-3p inhib-its breast cancer progression and metastasis by induc-ing macrophage polarization through downregulated ex-pression of Fra-1 proto-oncogene. Oncogene 2013; 33: 3014.[

Busch S, Auth E, Scholl F, et al. 5- lipoxygenase is a direct target of miR-19a-3p and miR-125b-5p. J Immunol 2015; 19 4(4): 16 4 6 -53.

Wang J, He Q, Han C, et al. p53-facilitated miR-199a-3p regulates somatic cell reprogramming. Stem Cells 2012; 30 (7 ): 14 05 -13.[

Liu J, Wang Y, Cui J, et al. miR199a-3p regulates P53 by tar-geting CABLES1 in mouse cardiac c-kit(+) cells to promote proliferation and inhibit apoptosis through a negative feedback loop. Stem Cell Res Ther 2017; 8(1): 127.

Kaynak Göster