Expression of autocrine macrophage migration inhibitory factor and its receptors of human embryonic stem cells

doi:10.12307/2022.985

Abstract

Abstract: BACKGROUND: Embryonic stem cells have a great potential in helping to understand cell development, regenerative medicine, tissue engineering, and cell therapy due to their self-renewal and multidirectional differentiation characteristics. To have a deeper understanding and effective use of human embryonic stem cells, it is necessary to explore and discover the potential molecules that affect the proliferation and survival of human embryonic stem cells. Macrophage migration inhibitory factor is expressed in a variety of human cells. At first, it was thought to be mainly involved in inflammation, but it was later found to play an important role in cell proliferation, differentiation, angiogenesis, and tumorigenesis. However, whether human embryonic stem cells express this important factor and their functions in human embryonic stem cells remain unclear.
OBJECTIVE: To determine whether human embryonic stem cells express macrophage migration inhibitory factor and its related receptors, determine which receptor interacts with macrophage migration inhibitory factor, and initially explore the effect of macrophage migration inhibitory factor on the proliferation and survival of human embryonic stem cells.
METHODS: Human embryonic stem cells were cultured. The level of macrophage migration inhibitory factor in the cell culture medium was detected by enzyme-linked immunosorbent assay. The distribution of macrophage migration inhibitory factor in the cell was observed by immunofluorescence confocal microscope. Cell immunofluorescence and western blot assay were utilized to detect the expression of embryonic stem cell related factors. Immunofluorescence confocal microscopy and immunoprecipitation were applied to detect the binding of macrophage migration inhibitory factor and its receptor. Group intervention: Human embryonic stem cells were intervened with ISO-1 (0, 12, 24, 48, 96, 192 μmol/L) for 24 hours to determine the best inhibitory concentration. Different mass concentrations of macrophage migration inhibitory factor (30, 100, 300 μg/L) were incubated with the optimal inhibitory concentration of ISO-1 for 24 hours. CCK-8 assay was used to detect cell proliferation activity. TUNEL staining was used to detect cell apoptosis level.
RESULTS AND CONCLUSION: (1) Macrophage migration inhibitory factor was expressed in the cytoplasm, membrane and culture medium of human embryonic stem cells. (2) Human embryonic stem cells expressed macrophage migration inhibitory factor, mainly expressing its receptors CXCR2 and CXCR7. (3) Macrophage migration inhibitory factor mainly bound to the receptor CXCR7. (4) The effect of macrophage migration inhibitory factor inhibitor ISO-1 on cell proliferation and survival: Compared with the blank group, as the ISO-1 concentration increased, the cell gap increased significantly and the cell viability gradually decreased (P < 0.0001). TUNEL assay showed that cell apoptosis rate was significantly increased (P < 0.000 1). (5) After adding different mass concentrations of migration inhibitory factor (30, 100, 300 μg/L), it could weaken ISO-1 (192 μmol/L) induced negative effects on cells; cell viability was significantly increased (P < 0.05); and apoptosis rate was significantly reduced (P < 0.01). (6) The results have shown that human embryonic stem cells express and secrete macrophage migration inhibitory factor, which is an important factor and can be detected in the cytoplasm, cell membrane, and outside the cell. Human embryonic stem cells mainly express the receptors CXCR2 and CXCR7 of macrophage migration inhibitors, instead of the classic receptor CD74. The protein receptor that interacts with macrophage migration inhibitory factor on human embryonic stem cells is CXCR7. No evidence of interaction with CXCR2 has been found, and its role needs further study. In addition, studies have found that macrophage migration inhibitory factors play an important role in maintaining the proliferation and survival of human embryonic stem cells.

Key words: embryonic stem cell, stem cell, factor, macrophage migration inhibitory factor, cell proliferation, proliferation

CLC Number:

Wei Yanzhao, Zheng Xiaohan, Gao Shijun, Huang Ting, Wei Xufang, Chen Xinxu, Zhao Zhenqiang. Expression of autocrine macrophage migration inhibitory factor and its receptors of human embryonic stem cells[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(1): 34-41.

Figures/Tables 12

References

[1] CHEN J, WANG Y, WANG C, et al. LncRNA Functions as a New Emerging Epigenetic Factor in Determining the Fate of Stem Cells. Front Genet. 2020;11:277.
[2] THOMSON JA, ITSKOVITZ-ELDOR J, SHAPIRO SS, et al. Embryonic stem cell lines derived from human blastocysts. Science.1998;282(5391):1145-1147.
[3] ZAKRZEWSKI W, DOBRZYNSKI M, SZYMONOWICZ M, et al. Stem cells: past, present, and future. Stem Cell Res Ther. 2019;10(1):68.
[4] NERI S. Genetic Stability of Mesenchymal Stromal Cells for Regenerative Medicine Applications: A Fundamental Biosafety Aspect. Int J Mol Sci. 2019;20(10):2406.
[5] TILLMANN S, BERNHAGEN J, NOELS H. Arrest Functions of the MIF Ligand/Receptor Axes in Atherogenesis. Front Immunol. 2013;4:115.
[6] GUNTHER S, FAGONE P, JALCE G, et al. Role of MIF and D-DT in immune-inflammatory, autoimmune, and chronic respiratory diseases: from pathogenic factors to therapeutic targets. Drug Discov Today. 2019;24(2):428-439.
[7] KLEMKE L, DE OLIVEIRA T, WITT D, et al. Hsp90-stabilized MIF supports tumor progression via macrophage recruitment and angiogenesis in colorectal cancer. Cell Death Dis. 2021;12(2):155.
[8] NAGARAJAN P, TOBER KL, RIGGENBACH JA, et al. MIF antagonist (CPSI-1306) protects against UVB-induced squamous cell carcinoma. Mol Cancer Res. 2014; 12(9):1292-1302.
[9] GORDON-WEEKS AN, LIM SY, YUZHALIN AE, et al. Macrophage migration inhibitory factor: a key cytokine and therapeutic target in colon cancer. Cytokine Growth Factor Rev. 2015;26(4):451-461.
[10] OLIVEIRA CS, DE BOCK CE, MOLLOY TJ, et al. Macrophage migration inhibitory factor engages PI3K/Akt signalling and is a prognostic factor in metastatic melanoma. BMC Cancer. 2014;14: 630.
[11] LIAO B, ZHONG BL, LI Z, et al. Macrophage migration inhibitory factor contributes angiogenesis by up-regulating IL-8 and correlates with poor prognosis of patients with primary nasopharyngeal carcinoma. J Surg Oncol. 2010;102(7):844-851.
[12] WADGAONKAR R, SOMNAY K, GARCIA JG. Thrombin induced secretion of macrophage migration inhibitory factor (MIF) and its effect on nuclear signaling in endothelium. J Cell Biochem. 2008;105(5):1279-1288.
[13] AMIN MA, HAAS CS, ZHU K, et al. Migration inhibitory factor up-regulates vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 via Src, PI3 kinase, and NFkappaB. Blood. 2006;107(6):2252-2261.
[14] XIE J, YANG L, TIAN L, et al. Macrophage Migration Inhibitor Factor Upregulates MCP-1 Expression in an Autocrine Manner in Hepatocytes during Acute Mouse Liver Injury. Sci Rep. 2016;6:27665.
[15] MA H, WANG J, THOMAS DP, et al. Impaired macrophage migration inhibitory factor-AMP-activated protein kinase activation and ischemic recovery in the senescent heart. Circulation. 2010;122(3):282-292.
[16] LUE H, THIELE M, FRANZ J, et al. Macrophage migration inhibitory factor (MIF) promotes cell survival by activation of the Akt pathway and role for CSN5/JAB1 in the control of autocrine MIF activity. Oncogene. 2007;26(35):5046-5059.
[17] LANG T, FOOTE A, LEE J P, et al. MIF: Implications in the Pathoetiology of Systemic Lupus Erythematosus. Front Immunol. 2015;6:577.
[18] SINITSKI D, KONTOS C, KRAMMER C, et al. Macrophage Migration Inhibitory Factor (MIF)-Based Therapeutic Concepts in Atherosclerosis and Inflammation. Thromb Haemost. 2019;119(4):553-566.
[19] KANG I, BUCALA R. The immunobiology of MIF:function, genetics and prospects for precision medicine. Nat Rev Rheumatol. 2019;15(7):427-437.
[20] ALAMPOUR-RAJABI S, EL BO, ROT A, et al. MIF interacts with CXCR7 to promote receptor internalization, ERK1/2 and ZAP-70 signaling, and lymphocyte chemotaxis. FASEB J. 2015;29(11):4497-4511.
[21] KLASEN C, OHL K, STERNKOPF M, et al. MIF promotes B cell chemotaxis through the receptors CXCR4 and CD74 and ZAP-70 signaling. J Immunol. 2014; 192(11):5273-5284.
[22] JANKAUSKAS SS, WONG D, BUCALA R, et al. Evolving complexity of MIF signaling. Cell Signal. 2019;57:76-88.
[23] XU X, PANG J, CHEN Y, et al. Macrophage Migration Inhibitory Factor (MIF) Deficiency Exacerbates Aging-Induced Cardiac Remodeling and Dysfunction Despite Improved Inflammation: Role of Autophagy Regulation. Sci Rep. 2016;6: 22488.
[24] LIU Y, ZHAO L, JU Y, et al. A novel androstenedione derivative induces ROS-mediated autophagy and attenuates drug resistance in osteosarcoma by inhibiting macrophage migration inhibitory factor (MIF). Cell Death Dis. 2014;5:e1361.
[25] YAO Y, DENG Q, SONG W, et al. MIF Plays a Key Role in Regulating Tissue-Specific Chondro-Osteogenic Differentiation Fate of Human Cartilage Endplate Stem Cells under Hypoxia. Stem Cell Reports. 2016;7(2):249-262.
[26] CUI J, ZHANG F, WANG Y, et al. Macrophage migration inhibitory factor promotes cardiac stem cell proliferation and endothelial differentiation through the activation of the PI3K/Akt/mTOR and AMPK pathways. Int J Mol Med. 2016;37(5):1299-1309.
[27] ZHANG X, CHEN L, WANG Y, et al. Macrophage migration inhibitory factor promotes proliferation and neuronal differentiation of neural stem/precursor cells through Wnt/beta-catenin signal pathway. Int J Biol Sci. 2013;9(10):1108-1120.
[28] GILFILLAN M, DAS P, SHAH D, et al. Inhibition of microRNA-451 is associated with increased expression of Macrophage Migration Inhibitory Factor and mitgation of the cardio-pulmonary phenotype in a murine model of Bronchopulmonary Dysplasia. Respir Res. 2020;21(1):92.
[29] BAYRAKTAR S, TANYERI BB, KILIC U. Umbilical cord levels of macrophage migration inhibitory factor in neonatal respiratory distress syndrome. Turk J Med Sci. 2021; 51(2):722-726.
[30] SUMAIYA K, LANGFORD D, NATARAJASEENIVASAN K, et al. Macrophage migration inhibitory factor (MIF): A multifaceted cytokine regulated by genetic and physiological strategies. Pharmacol Ther. 2021;108024. doi: 10.1016/j.pharmthera.2021.108024.
[31] VERJANS E, NOETZEL E, BEKTAS N, et al. Dual role of macrophage migration inhibitory factor (MIF) in human breast cancer. BMC Cancer. 2009;9:230.
[32] FUJIHARA Y, HIKITA A, TAKATO T, et al. Roles of macrophage migration inhibitory factor in cartilage tissue engineering. J Cell Physiol. 2018;233(2):1490-1499.
[33] YAO J, LENG L, SAULER M, et al. Transcription factor ICBP90 regulates the MIF promoter and immune susceptibility locus. J Clin Invest. 2016;126(2):732-744.
[34] KIM J, GEE HY, LEE MG. Unconventional protein secretion - new insights into the pathogenesis and therapeutic targets of human diseases. J Cell Sci. 2018; 131(12):jcs213686.
[35] EICKHOFF R, WILHELM B, RENNEBERG H, et al. Purification and characterization of macrophage migration inhibitory factor as a secretory protein from rat epididymis: evidences for alternative release and transfer to spermatozoa. Mol Med. 2001;7(1):27-35.
[36] BILSBORROW JB, DOHERTY E, TILSTAM P V, et al. Macrophage migration inhibitory factor (MIF) as a therapeutic target for rheumatoid arthritis and systemic lupus erythematosus. Expert Opin Ther Targets. 2019.23(9):733-744.
[37] TILSTAM PV, PANTOURIS G, CORMAN M, et al. A selective small-molecule inhibitor of macrophage migration inhibitory factor-2 (MIF-2), a MIF cytokine superfamily member, inhibits MIF-2 biological activity. J Biol Chem. 2019;294(49):18522-18531.
[38] STADTMANN A, ZARBOCK A. CXCR2: From Bench to Bedside. Front Immunol. 2012;3:263.
[39] PUCHERT M, ENGELE J. The peculiarities of the SDF-1/CXCL12 system: in some cells, CXCR4 and CXCR7 sing solos, in others, they sing duets. Cell Tissue Res. 2014;355(2):239-253.
[40] CHATTERJEE M, BORST O, WALKER B, et al. Macrophage migration inhibitory factor limits activation-induced apoptosis of platelets via CXCR7-dependent Akt signaling. Circ Res. 2014;115(11):939-949.
[41] STARLETS D, GORE Y, BINSKY I, et al. Cell-surface CD74 initiates a signaling cascade leading to cell proliferation and survival. Blood. 2006;107(12):4807-4816.
[42] GORE Y, STARLETS D, MAHARSHAK N, et al. Macrophage migration inhibitory factor induces B cell survival by activation of a CD74-CD44 receptor complex. J Biol Chem. 2008;283(5):2784-2792.
[43] COURNIA Z, LENG L, GANDAVADI S, et al. Discovery of human macrophage migration inhibitory factor (MIF)-CD74 antagonists via virtual screening. J Med Chem. 2009;52(2):416-424.
[44] SHAUL YD, SEGER R. The MEK/ERK cascade: from signaling specificity to diverse functions. Biochim Biophys Acta. 2007;1773(8):1213-1226.
[45] STAMATIADES GA, KAISER UB. Gonadotropin regulation by pulsatile GnRH: Signaling and gene expression. Mol Cell Endocrinol. 2018;463:131-141.
[46] VOSKAS D, LING LS, WOODGETT JR. Signals controlling un-differentiated states in embryonic stem and cancer cells: role of the phosphatidylinositol 3’ kinase pathway. J Cell Physiol. 2014;229(10):1312-1322.
[47] SATO N, SANJUAN IM, HEKE M, et al. Molecular signature of human embryonic stem cells and its comparison with the mouse. Dev Biol. 2003;260(2):404-413.
[48] PALING NR, WHEADON H, BONE HK, et al. Regulation of embryonic stem cell self-renewal by phosphoinositide 3-kinase-dependent signaling. J Biol Chem. 2004;279(46):48063-48070.
[49] SHPARBERG RA, GLOVER HJ, MORRIS MB. Modeling Mammalian Commitment to the Neural Lineage Using Embryos and Embryonic Stem Cells. Front Physiol. 2019;10:705.
[50] BRAZIL DP, YANG ZZ, HEMMINGS BA. Advances in protein kinase B signalling: AKTion on multiple fronts. Trends Biochem Sci. 2004;29(5):233-242.
[51] TAHIR SA, GAO J, MIURA Y, et al. Autoimmune antibodies correlate with immune checkpoint therapy-induced toxicities. Proc Natl Acad Sci U S A. 2019;116(44): 22246-22251.
[52] RAJASEKARAN D, GRONING S, SCHMITZ C, et al. Macrophage Migration Inhibitory Factor-CXCR4 Receptor Interactions: EVIDENCE FOR PARTIAL ALLOSTERIC AGONISM IN COMPARISON WITH CXCL12 CHEMOKINE. J Biol Chem. 2016;291(30):15881-15895.