Exosomes from miR-149-3p-transfected menstrual blood-derived mesenchymal stem cells ameliorate inflammation and migration of endometriosis cells

Document Type : Original Article

Authors

1 Department of Biology, Faculty of Sciences and Converging Technologies, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Department of Biology, Faculty of Sciences, Islamic Azad University, Roudehen Branch, Roudehen, Iran

3 Department of Cell Biology and Regenerative Medicine, Academic Center for Education, Culture and Research (ACECR), Qom branch, Qom, Iran

10.22038/ijbms.2025.86443.18677

Abstract

Objective(s): Endometriosis carries remarkable social, public health, and financial consequences. Based on two theories of retrograde menstruation and stem cells, menstrual blood-derived stem cells (MenSCs) play a significant role in endometriosis since key genes of critical cellular processes are differentially expressed in the MenSCs of endometriosis and non-endometriosis women (E- and NE-MenSCs, respectively). In this study, E-MenSCs were isolated from the menstrual blood of women with various endometriosis subtypes. We tried to find the proper microRNA (miRNA) and assayed the effects of exosome-encapsulated miRNA on modulating the gene expression profile and functional pattern of E-MenSCs.
Materials and Methods: After in silico selection of miR-149-3p using publicly accessible algorithm-based databases, E- and NE-MenSCs were cultured as controls, and the other experimental groups were as follows: E-MenSCs transfected with empty and miRNA vectors (E-MenSC+BB and E-MenSC+miR), and E-MenSCs treated with exosomes derived from non-transfected and miRNA-transfected NE-MenSCs (E-MenSC+Exo and E-MenSC+T-Exo). Then, the expression level of selected genes, the level of interleukins (ILs) and oxygen reactive species (ROS), the protein level of β-catenin and Ki-67, and the migratory ability were assessed through real-time PCR, ELISA, western blot, and scratching tests, respectively.
Results: Although both E-MenSCs+T-Exo and E-MenSC+miR showed down-regulation of IL-6, -8, and -10, neither had decreased IL-1β, vascular endothelial growth factor, IDO1, and KRAS levels. Furthermore, only the IL-6 protein level was significantly decreased in the E-MenSC+miR group, but the levels of IL-6, IL-8, ROS, β-catenin, and Ki67 were significantly lower in the E-MenSCs+T-Exo group compared to the E-MenSCs.
Conclusion: The potential of exosomes as miRNA carriers could be considered in developing novel endometriosis therapies.

Keywords

Main Subjects

References

1.    Taylor HS, Kotlyar AM, Flores VA. Endometriosis is a chronic systemic disease: Clinical challenges and novel innovations. Lancet 2021;397:839-852.
2.    Koninckx PR, Fernandes R, Ussia A, Schindler L, Wattiez A, Al-Suwaidi S, et al. Pathogenesis based diagnosis and treatment of endometriosis. Front Endocrinol 2021;12:745548-745560.
3.    Horne AW, Missmer SA. Pathophysiology, diagnosis, and management of endometriosis. Bmj 2022;379:e070750.
4.    Zondervan KT, Becker CM, Missmer SA. Endometriosis. N Engl J Med 2020;382:1244-1256.
5.    Sampson JA. Peritoneal endometriosis due to the menstrual dissemination of endometrial tissue into the peritoneal cavity. Am J Obstet Gynecol 1927;14:422-469.
6.    Brichant G, Laraki I, Henry L, Munaut C, Nisolle M. New therapeutics in endometriosis: A review of hormonal, non-Hormonal, and non-Coding RNA Treatments. Int J Mol Sci 2021;22:10498-10515.
7.    Barra F, Ferrero S. Adhesion proteins: Suitable therapeutic targets or biomarkers of therapy response for endometriosis? Acta Obstet Gynecol Scand 2019;98:810-811.
8.    Meng X, Ichim TE, Zhong J, Rogers A, Yin Z, Jackson J, et al. Endometrial regenerative cells: A novel stem cell population. J Transl Med 2007;5:57-66.
9.    Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 2006;8:315-317.
10.    Bjørge IM, Kim SY, Mano JF, Kalionis B, Chrzanowski W. Extracellular vesicles, exosomes and shedding vesicles in regenerative medicine - a new paradigm for tissue repair. Biomater Sci 2017;6:60-78.
11. Maas SLN, Breakefield XO, Weaver AM. Extracellular vesicles: Unique intercellular delivery vehicles. Trends Cell Biol 2017;27:172-188.
12.    Lai RC, Arslan F, Lee MM, Sze NS, Choo A, Chen TS, et al. Exosome secreted by MSC reduces myocardial ischemia/reperfusion injury. Stem Cell Res 2010;4:214-222.
13.    Colombo M, Raposo G, Théry C. Biogenesis, secretion, and intercellular interactions of exosomes and other extracellular vesicles. Annu Rev Cell Dev Biol 2014;30:255-289.
14.    Lo Cicero A, Stahl PD, Raposo G. Extracellular vesicles shuffling intercellular messages: for good or for bad. Curr Opin Cell Biol 2015;35:69-77.
15.    Ma ZJ, Yang JJ, Lu YB, Liu ZY, Wang XX. Mesenchymal stem cell-derived exosomes: Toward cell-free therapeutic strategies in regenerative medicine. World J Stem Cells 2020;12:814-840.
16.    Sahraei SS, Davoodi Asl F, Kalhor N, Sheykhhasan M, Fazaeli H, Moud SS, et al. A comparative study of gene expression in menstrual blood-derived stromal cells between endometriosis and healthy women. Biomed Res Int 2022;2022:7053521.
17.    Davoodi Asl F, Sahraei SS, Kalhor N, Fazaeli H, Sheykhhasan M, Soleimani Moud S, et al. Promising effects of exosomes from menstrual blood-derived mesenchymal stem cells on endometriosis. Reprod Biol 2023;23:100788.
18.    Adams BD, Parsons C, Walker L, Zhang WC, Slack FJ. Targeting noncoding RNAs in disease. J Clin Invest 2017;127:761-771.
19.    Santamaria X, Taylor H. MicroRNA and gynecological reproductive diseases. Fertil Steril 2014;101:1545-1551.
20.    Filigheddu N, Gregnanin I, Porporato PE, Surico D, Perego B, Galli L, et al. Differential expression of microRNAs between eutopic and ectopic endometrium in ovarian endometriosis. J Biomed Biotechnol 2010;2010:369549.
21.    Sheikholeslami A, Nabiuni M, Arefian E. Suppressing the molecular signaling pathways involved in inflammation and cancer in breast cancer cell lines MDA-MB-231 and MCF-7 by miR-590. Tumour Biol 2017;39:1010428317697570.
22.    Nikoo S, Ebtekar M, Jeddi-Tehrani M, Shervin A, Bozorgmehr M, Vafaei S, et al. Menstrual blood-derived stromal stem cells from women with and without endometriosis reveal different phenotypic and functional characteristics. Mol Hum Reprod 2014;20:905-918.
23.    Sahin C, Mamillapalli R, Yi KW, Taylor HS. microRNA Let-7b: A Novel treatment for endometriosis. J Cell Mol Med 2018;22:5346-5353.
24.    Sahraei SS, Davoodi Asl F, Kalhor N, Sheykhhasan M, Fazaeli H, Moud SS, et al. A comparative study of gene expression in menstrual blood-derived stromal cells between endometriosis and healthy women. Biomed Res Int 2022;2022:7053521.
25.    Fazaeli H, Kalhor N, Naserpour L, Davoodi F, Sheykhhasan M, Hosseini SKE, et al. A comparative study on the effect of exosomes secreted by mesenchymal stem cells derived from adipose and bone marrow tissues in the treatment of osteoarthritis-induced mouse model. Biomed Res Int 2021;2021:9688138-9688150.
26. Siavashi V, Nassiri SM, Rahbarghazi R, Vafaei R, Sariri R. ECM-dependence of endothelial progenitor cell features. J Cell Biochem 2016;117:1934-1946.
27. Jiang R, Zhang H, Zhou J, Wang J, Xu Y, Zhang H, et al. Inhibition of long non-coding RNA XIST upregulates microRNA-149-3p to repress ovarian cancer cell progression. Cell Death Dis 2021;12:145-156.
28. Cao D, Jia Z, You L, Wu Y, Hou Z, Suo Y, et al. 18β-glycyrrhetinic acid suppresses gastric cancer by activation of miR-149-3p-Wnt-1 signaling. Oncotarget 2016;7:71960-71973.
29. Si L, Xu L, Yin L, Qi Y, Han X, Xu Y, et al. Potent effects of dioscin against pancreatic cancer via miR-149-3P-mediated inhibition of the Akt1 signalling pathway. Br J Pharmacol 2017;174:553-568.
30. Yuxue J, Ran S, Minghui F, Minjia S. Applications of nanomaterials in endometriosis treatment. Front Bioeng Biotechnol 2023;11:1184155-1184165.
31. Wu P, Zhang B, Ocansey DKW, Xu W, Qian H. Extracellular vesicles: A bright star of nanomedicine. Biomaterials 2021;269:120467.
32. Wu D, Lu P, Mi X, Miao J. Exosomal miR-214 from endometrial stromal cells inhibits endometriosis fibrosis. Mol Hum Reprod 2018;24:357-365.
33. Ohlsson Teague EM, Van der Hoek KH, Van der Hoek MB, Perry N, Wagaarachchi P, Robertson SA, et al. MicroRNA-regulated pathways associated with endometriosis. Mol Endocrinol 2009;23:265-275.
34. Papari E, Noruzinia M, Kashani L, Foster WG. Identification of candidate microRNA markers of endometriosis with the use of next-generation sequencing and quantitative real-time polymerase chain reaction. Fertil Steril 2020;113:1232-1241.
35. Riccio LdGC, Santulli P, Marcellin L, Abrão MS, Batteux F, Chapron C. Immunology of endometriosis. Best Pract Res Clin Obstet Gynaecol 2018;50:39-49.
36. Kang YJ, Jeung IC, Park A, Park YJ, Jung H, Kim TD, et al. An increased level of IL-6 suppresses NK cell activity in peritoneal fluid of patients with endometriosis via regulation of SHP-2 expression. Hum Reprod 2014;29:2176-2189.
37. Sikora J, Smycz-Kubańska M, Mielczarek-Palacz A, Kondera-Anasz Z. Abnormal peritoneal regulation of chemokine activation-The role of IL-8 in pathogenesis of endometriosis. Am J Reprod Immunol 2017;77: doi: 10.1111/aji.12622.
38. Matsuzaki S, Pouly JL, Canis M. IL-10 is not anti-fibrotic but pro-fibrotic in endometriosis: IL-10 treatment of endometriotic stromal cells in vitro promotes myofibroblast proliferation and collagen type I protein expression. Hum Reprod 2023;38:14-29.
39. Munn DH, Mellor AL, Rossi M, Young JW. Dendritic cells have the option to express IDO-mediated suppression or not. Blood 2005;105:2618.
40.    Jeddi-Tehrani M, Abbasi N, Dokouhaki P, Ghasemi J, Rezania S, Ostadkarampour M, et al. Indoleamine 2,3-dioxygenase is expressed in the endometrium of cycling mice throughout the oestrous cycle. J Reprod Immunol 2009;80:41-48.
41.    Mei J, Jin LP, Ding D, Li MQ, Li DJ, Zhu XY. Inhibition of IDO1 suppresses cyclooxygenase-2 and matrix metalloproteinase-9 expression and decreases proliferation, adhesion and invasion of endometrial stromal cells. Mol Hum Reprod 2012;18:467-476.
42. Schubbert S, Shannon K, Bollag G. Hyperactive Ras in developmental disorders and cancer. Nat Rev Cancer 2007;7:295-308.
43.    Grechukhina O, Petracco R, Popkhadze S, Massasa E, Paranjape T, Chan E, et al. A polymorphism in a let-7 microRNA binding site of KRAS in women with endometriosis. EMBO Mol Med 2012;4:206-217.
44.    Calcagno A, Grassi T, Mariuzzi L, Marzinotto S, Londero AP, Orsaria M, et al. Expression patterns of Aurora A and B kinases, Ki-67 and the estrogen and progesterone receptors determined using an endometriosis tissue microarray model. Hum reprod 2011;26:2731-2741.
45. Shin CH, Lee H, Kim HR, Choi KH, Joung JG, Kim HH. Regulation of PLK1 through competition between hnRNPK, miR-149-3p and miR-193b-5p. Cell Death Differ 2017;24:1861-1871.
46. Lin RJ, Lin YC, Yu AL. miR-149* induces apoptosis by inhibiting Akt1 and E2F1 in human cancer cells. Mol Carcinog 2010;49:719-727.
47. Ru Q, Tian X, Pi MS, Chen L, Yue K, Xiong Q, et al. Voltage‑gated K+ channel blocker quinidine inhibits proliferation and induces apoptosis by regulating expression of microRNAs in human glioma U87‑MG cells. Int J Oncol 2015;46:833-840.
48. Tosti C, Pinzauti S, Santulli P, Chapron C, Petraglia F. Pathogenetic mechanisms of deep infiltrating endometriosis. Reprod Sci 2015;22:1053-1059.
49. Ngô C, Chéreau C, Nicco C, Weill B, Chapron C, Batteux F. Reactive oxygen species controls endometriosis progression. Am J Pathol 2009;175:225-234.
50. Reactive oxygen species-induced activation of the MAP kinase signaling pathways. Antioxid Redox Signal 2006;8:1775-1789.
51. Matsuoka T, Takanashi K, Dan K, Yamamoto K, Tomobe K, Shinozuka T. Effects of mesenchymal stem cell-derived exosomes on oxidative stress responses in skin cells. Mol Biol Rep 2021;48:4527-4535.
52.    Matsuzaki S, Darcha C. Involvement of the Wnt/β-catenin signaling pathway in the cellular and molecular mechanisms of fibrosis in endometriosis. PLoS One 2013;8:e76808.
53.    Yang D, Du G, Xu A, Xi X, Li D. Expression of miR-149-3p inhibits proliferation, migration, and invasion of bladder cancer by targeting S100A4. Am J Cancer Res 2017;7:2209-2219.
54.    Ebralidze A, Tulchinsky E, Grigorian M, Afanasyeva A, Senin V, Revazova E, et al. Isolation and characterization of a gene specifically expressed in different metastatic cells and whose deduced gene product has a high degree of homology to a Ca2+-binding protein family. Genes Dev 1989;3:1086-1093.
55.    Varma R, Rollason T, Gupta JK, Maher ER. Endometriosis and the neoplastic process. Reproduction 2004;127:293-304.
56.    Jiang WS, Huang CL, Zhang J, Xu F, Dai XH. MicroRNA-149 inhibits the progression of lung adenocarcinoma through targeting RAP1B and inactivating Wnt/β-catenin pathway. Eur Rev Med Pharmacol Sci 2020;24:4846-4854.
57.    Valentijn AJ, Palial K, Al-Lamee H, Tempest N, Drury J, Von Zglinicki T, et al. SSEA-1 isolates human endometrial basal glandular epithelial cells: Phenotypic and functional characterization and implications in the pathogenesis of endometriosis. Hum Reprod 2013;28:2695-2708.
58.    Ma B, Hottiger MO. Crosstalk between Wnt/β-Catenin and NF-κB signaling pathway during inflammation. Front Immunol 2016;7:378-391.
59.    Koh HB, Kim HJ, Kang SW, Yoo TH. Exosome-based drug delivery: Translation from bench to clinic. Pharmaceutics 2023;15:2042-2070.
60.    Palakurthi SS, Shah B, Kapre S, Charbe N, Immanuel S, Pasham S, et al. A comprehensive review of challenges and advances in exosome-based drug delivery systems. Nanoscale Adv 2024;6:5803-5826.
Iranian Journal of Basic Medical Sciences
Volume 28, Issue 11
November 2025
Pages 1575-1588

Files

Share

How to cite

Statistics