Unleashing the Impact of Exosomes Derived from Human Placental Mesenchymal Stem Cells (hPMSCs) on U-266 Myeloma Cell Line
,
Abstract
Background: Multiple myeloma (MM) is a malignancy of plasma cells, terminally differentiated B cells, with complications like hypercalcemia, renal failure, anemia, and bone disease, which are also known as CRAB criteria. MM develops from monoclonal gammopathy of unknown significance (MGUS), a pre-malignant plasma cell dyscrasia. Over some time, MGUS has the potential to progress into smoldering multiple myeloma (SMM), which can evolve into MM. MM rarely progresses into plasma cell leukemia (PCL), a condition in which malignant plasma cells no longer stay in the bone marrow niche and circulate in the peripheral blood. In MM, various soluble factors play important roles, and interleukin-6 has different vital roles.
Interleukin-6, an inflammatory cytokine, has significant roles in the growth, survival, angiogenesis, metastasis, and apoptosis resistance in MM. Interleukin-6 is produced and secreted by both autocrine from myeloma cells and paracrine from bone marrow stromal cells. To tackle MM, various therapeutic approaches were applied over many years, and according to the results, most patients with MM can respond well to first-line treatment. However, the majority of patients may relapse as conventional treatment may not be curative. So, there is an urgent need for novel cell-based and cell-free therapeutic strategies, such as mesenchymal stem cell-based therapies and their products to offer new therapeutic strategies for MM.
Materials and Methods: In the present study, we investigated the impacts of exosomes derived from human placental mesenchymal stem cells (hPMSCs) on apoptosis and interleukin-6 expression in a myeloma cell line, U-266, for the first time. hPMSCs were isolated from the human placenta and cultured in a DMEM medium. After characterizing the cells and acknowledging their identity, they underwent several passages and their supernatant was collected to harvest exosomes. The exosomes were isolated by ultracentrifugation and characterized by DLS and TEM, and their concentration was measured by BCA protein assay. U266 cells were treated with different concentrations of exosomes and then MTT and annexin/propidium iodide flow cytometry tests were performed to evaluate cell viability. Afterward, a real-time PCR test was performed to evaluate interleukin-6 gene expression.
Results: According to our findings, treatment of U-266 cells with hPMSCS-derived exosomes led to the preservation of myeloma cells without changes in their cell cycle. Surprisingly, treatments did not hinder the expression of interleukin-6 in the myeloma cells.
Conclusion: In MM patients, interleukin-6 plays different roles, and it is a desirable target to design new therapeutic strategies. To evaluate the effects of new therapeutic strategies, we designed and performed our study to estimate the effects of cell-free therapeutic strategy. In the present study, the impacts of hPMSCS-derived exosomes on the viability of MM cells and interleukin-6 gene expression were evaluated. The results showed that hPMSCS-derived exosomes resulted in the perseverance of myeloma cells without changes in the cell cycle. Furthermore, the interleukin-6 gene expression level showed no significant change.
2. Bianchi G, Anderson KC. Understanding biology to tackle the disease: Multiple myeloma from bench to bedside, and back. 2014;64(6):422-44.
3. Börger V, Bremer M, Ferrer-Tur R, et al. Mesenchymal Stem/Stromal Cell-Derived Extracellular Vesicles and Their Potential as Novel Immunomodulatory Therapeutic Agents. Int J Mol Sci. 2017;18(7):1450.
4. Cargnoni A, Piccinelli EC, Ressel L, et al. Conditioned medium from amniotic membrane-derived cells prevents lung fibrosis and preserves blood gas exchanges in bleomycin-injured mice-specificity of the effects and insights into possible mechanisms. Cytotherapy. 2014;16(1):17-32.
5. Catlett-Falcone R, Landowski TH, Oshiro MM, et al. Constitutive activation of Stat3 signaling confers resistance to apoptosis in human U266 myeloma cells. Immunity. 1999;10(1):105-15.
6. Cejalvo MJ, de la Rubia J. Which therapies will move to the front line for multiple myeloma?. Expert Rev Hematol. 2017;10(5):383-392.
7. Chen CP, Chen YY, Huang JP, et al. The effect of conditioned medium derived from human placental multipotent mesenchymal stromal cells on neutrophils: possible implications for placental infection. Mol Hum Reprod. 2014;20(11):1117-25.
8. Deng ZB, Liu Y, Liu C, et al. Immature myeloid cells induced by a high-fat diet contribute to liver inflammation. Hepatology. 2009; 50(4): 1412-20.
9. El Andaloussi S, Mäger I, Breakefield XO, et al. Extracellular vesicles: biology and emerging therapeutic opportunities. Nat Rev Drug Discov. 2013; 12(5):347-57.
10. Fairfield H, Falank C, Avery L, et al. Multiple myeloma in the marrow: pathogenesis and treatments. Ann N Y Acad Sci. 2016; 1364(1): 32-51.
11. Fu Q, Fu TM, Cruz AC, et al. Structural basis and functional role of intramembrane trimerization of the Fas/CD95 death receptor. Mol Cell. 2016; 61(4):602-613.
12. Galipeau J, Sensébé L. Mesenchymal stromal cells: clinical challenges and therapeutic opportunities. Cell stem cell. 2018; 22(6): 824-33.
13. Gorodetsky R, Aicher WK. Allogenic Use of Human Placenta-Derived Stromal Cells as a Highly Active Subtype of Mesenchymal Stromal Cells for Cell-Based Therapies. Int J Mol Sci. 2021;22(10):5302.
14. Gu YZ, Xue Q, Chen YJ, et al. Different roles of PD-L1 and FasL in immunomodulation mediated by human placenta-derived mesenchymal stem cells. Hum Immunol. 2013;74(3): 267-76.
15. Gurung S, Perocheau D, Touramanidou L, et al. The exosome journey: from biogenesis to uptake and intracellular signalling. Cell Commun Signal. 2021;19(1):47.
16. Harrell CR, Jovicic N, Djonov V, et al. Therapeutic Use of Mesenchymal Stem Cell-Derived Exosomes: From Basic Science to Clinics. Pharmaceutics. 2020; 12(5):474.
17. Hashemian SMR, Aliannejad R, Zarrabi M, et al. Mesenchymal stem cells derived from perinatal tissues for treatment of critically ill COVID-19-induced ARDS patients: a case series. Stem Cell Res Ther. 2021; 12(1):91.
18. Johnstone RM, Adam M, Hammond JR, et al. Vesicle formation during reticulocyte maturation. Association of plasma membrane activities with released vesicles (exosomes). J Biol Chem.1987; 262(19): 9412-20.
19. Michio K, Hirano T, Matsuda T, et al. Autocrine generation and requirement of BSF-2/IL-6 for human multiple myelomas. Nature.1988; 332(6159): 83-5.
20. Soo-Hwan K, Jung J, Jin Cho K, et al. Immunomodulatory Effects of Placenta-derived Mesenchymal Stem Cells on T Cells by Regulation of FoxP3 Expression. Int J Stem Cells. 2018; 11(2): 196–204.
21. Kishimoto T. IL-6: from its discovery to clinical applications. Int Immunol. 2010; 22(5):347-52.
22. Klein B, Zhang XG, Jourdan M, et al. Paracrine rather than autocrine regulation of myeloma-cell growth and differentiation by interleukin-6'. Blood. 1989;73(2):517-26.
23. Kumar S K, Rajkumar V, Kyle RA, et al. 2017. Multiple myeloma. Nat Rev Dis Primers. 2017:3:17046.
24. Kuriyan AE, Albini TA, Townsend JH, et al. Vision Loss after Intravitreal Injection of Autologous "Stem Cells" for AMD. N Engl J Med. 2017; 376(11):1047-1053.
25. Kyle R A, Larson DR, Therneau TM, et al. Long-Term Follow-up of Monoclonal Gammopathy of Undetermined Significance. N Engl J Med. 2018;378(3): 241-49.
26. Landgren O, Kyle R A, Pfeiffer RM, et al. Monoclonal gammopathy of undetermined significance (MGUS) consistently precedes multiple myeloma: a prospective study. Blood. 2009;113(22): 5412-7.
27. Landowski TH, Qu N, Buyuksal I, et al. Mutations in the Fas antigen in patients with multiple myeloma. Blood.1997; 90(11): 4266-70.
28. Changdong Li, Zhang W, Jiang X, et al. Human-placenta-derived mesenchymal stem cells inhibit proliferation and function of allogeneic immune cells. Cell Tissue Res. 2007; 330(3): 437-46.
29. Lipinska N, Romaniuk A, Paszel-Jaworska A, et al. Telomerase and drug resistance in cancer. Cell Mol Life Sci. 2017;74(22): 4121-32.
30. Men Y, Yelick J, Jin S, et al. Exosome reporter mice reveal the involvement of exosomes in mediating neuron to astroglia communication in the CNS. Nat Commun. 2019; 10(1): 4136.
31. Moodley Y, Vaghjiani V, Chan J, et al. Anti-inflammatory effects of adult stem cells in sustained lung injury: a comparative study. PLoS One. 2013; 8(8): e69299.
32. Osaki M, Okada F. 2019. Exosomes and Their Role in Cancer Progression. Yonago Acta Med. 2019; 62(2): 182-90.
33. Pearce A, Haas M, Viney R, et al. Incidence and severity of self-reported chemotherapy side effects in routine care: A prospective cohort study. PLoS One. 2017; 12(10): e0184360.
34. Puthier D, Derenne S, Barillé S, et al. Mcl‐1 and Bcl‐xL are co‐regulated by IL‐6 in human myeloma cells. Br J Haematol. 1999;107(2):392-5.
35. Rajkumar S V. Multiple myeloma: Every year a new standard?, Hematol Oncol.2019; 37 Suppl 1(Suppl 1):62-65.
36. Rajkumar S V, Landgren O, Mateos MV. Smoldering multiple myeloma. Blood. 2015; 125(20): 3069-75.
37. Ratajczak J, Miekus K, Kucia M, et al. Embryonic stem cell-derived microvesicles reprogram hematopoietic progenitors: evidence for horizontal transfer of mRNA and protein delivery. Leukemia. 2006; 20(5): 847-56.
38. Stenqvist AC, Nagaeva O, Baranov V, et al. Exosomes secreted by human placenta carry functional Fas ligand and TRAIL molecules and convey apoptosis in activated immune cells, suggesting exosome-mediated immune privilege of the fetus. J Immunol. 2013; 191(11): 5515-23.
39. Tanabe O, Kawano M, Tanaka H, et al. BSF‐2/IL‐6 does not augment lg secretion but stimulates proliferation in myeloma cells. Am J Hematol. 1989; 31(4): 258-62.
40. Weiss BM, Abadie J, Verma P, et al. A monoclonal gammopathy precedes multiple myeloma in most patients. Blood.2009;113(22): 5418-22.
41. Mingjun Wu, Zhang R, Zou Q, et al. Comparison of the biological characteristics of mesenchymal stem cells derived from the human placenta and umbilical cord. Sci Rep. 2018; 8(1): 5014.
42. Wu Q, Yang Z, Nie Y, et al. Multi-drug resistance in cancer chemotherapeutics: mechanisms and lab approaches. Cancer Lett. 2014; 347(2): 159-66.
43. Zhang B, Wang M, Gong A, et al. HucMSC-Exosome Mediated-Wnt4 Signaling Is Required for Cutaneous Wound Healing. Stem Cells. 2015; 33(7): 2158-68.
44. Zhang L, Yu J, Park BH, et al. Role of BAX in the apoptotic response to anticancer agents. Science.2000; 290(5493): 989-92.
| Files | ||
| Issue | Vol 18 No 3 (2024) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijhoscr.v18i3.16109 | |
| Keywords | ||
| Mesenchymal stem cells Exosomes Multiple myeloma Interleukin-6 | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
