Biological and Immunological Properties of Mesenchymal Stromal Cells Derived From Bone Marrow in Childhood Acute Myeloid Leukemia

Abstract

Objective: Pediatric acute myeloid leukemia (AML) is a common form of pediatric leukemia and is characterized by the accumulation of abnormal white blood cells, called blasts, in the bone marrow (BM). The aim of this study was to understand the BM microenvironment by studying the biological and immunological properties of BM-derived mesenchymal stromal cells (MSCs) and mononuclear cells (MNCs) to elucidate the potential role of phytohaemag glutinin in cell viability. Materials and Methods: BM and peripheral blood samples were obtained from seven pediatric AML patients and seven donors. BM-MSCs and MNCs were isolated and characterized. Population doubling (PD) values, adipogenic and osteogenic differentiation capacity, cell viability, phytohemagglutinin (PHA) assay, and flow cytometry were performed. Results: Mononucleated cells from peripheral blood of AML patients and donors and T-cell activation markers (CD3+CD69+, CD4+C25+, CD3+HLA-DR+) were measured by flow cytometry (𝜒2=2.184; p=0.823). BM-MSCs were co-cultured with MNCs, and PD values for AML patients were similar to those of donors (z=1.074; p=0.394). It was statistically significant when healthy MNC and healthy MNC PHA(+) groups were compared (p=0.015). When healthy MNC PHA(+) and healthy MSC+AML MNC PHA(+) groups were compared, it was found to be statistically significant (p=0.014). Conclusion: This interaction is also not unidirectional. This interaction serves as a marker for understanding the immunological effects of AML.

Keywords

• Pediatric AML
• BM-MSCs
• PHA
• Cell viability
• MNCs

References

1. 1. Miari KE, Williams MTS. Stromal bone marrow fibroblasts and mesenchYmal stem cells support acute mYeloid leukaemia cells and promote therapY resistance. Br J Pharmacol 2024; 181(2): 216-37. google scholar
2. 2. Umeda M, Ma J, Westover T, Ni Y, Song G, Maciaszek JL, et al. A new genomic framework to categorize pediatric acute mYeloid leukemia. Nat Genet 2024; 56(2): 281-93. google scholar
3. 3. Elgarten CW, Aplenc R. Pediatric acute mYeloid leukemia: Updates on biologY, risk stratification, and therapY. Curr Opin Pediatr 2020; 32(1): 57-66. google scholar
4. 4. Baker N, BoYette LB, Tuan RS. Characterization of bone marrow-derived mesenchYmal stem cells in aging. Bone 2015; 70(1): 37-47. google scholar
5. 5. Koung Ngeun S, Shimizu M, Kaneda M. Characterization of rabbit mesenchYmal stem/stromal cells after crYopreservation. BiologY (Basel) 2023; 12(10): 1312. google scholar
6. 6. Sabbah R, Saadi S, Shahar-GabaY T, GerassY S, Yehudai-Resheff S, Zuckerman T. Abnormal adipogenic signaling in the bone marrow mesenchYmal stem cells contributes to supportive microenvironment for leukemia development. Cell Commun Signal 2023; 10; 21(1): 277 google scholar
7. 7. Lin H, Sohn J, Shen H, Langhans MT, Tuan RS. Bone marrow mesenchYmal stem cells: Aging and tissue engineering applications to enhance bone healing. Biomaterials 2019; 203: 96-110. google scholar
8. 8. Gulden G, Sert B, TeYmur T, AY Y, TirYaki NN, Mishra AK, et al. CAR-T Cells with phYtohemagglutinin (PHA) provide anti-cancer capacitY with better proliferation, rejuvenated effector memorY, and reduced exhausted T cell frequencies. Vaccines (Basel) 2023; 31; 11(2): 313. google scholar

9. 9. Ok BozkaYa I, Azik F, Tavil B, Koksal Y, Ozguner M, Tunc B, et al. The Effect of granulocYte colonY-stimulating factor on immune-modulatorY cYtokines in the bone marrow microenvironment and mesenchYmal stem cells of healthY donors Biol. Blood Marrow Transplant 2015; 21(11): 1888-94. google scholar
10. 10. Bennett JM, CatovskY D, Daniel MT, Flandrin G, Galton DA, Gralnick HR, et al. Proposals for the classification of the acute leukaemias. French-American-British (FAB) co-operative group. Br J Haematol 1976; 33(4): 451-8. google scholar
11. 11. Vardiman JW, Thiele J, Arber DA, Brunning RD, Borowitz MJ, Porwit A, et al. The 2008 revision of the World Health Organization (WHO) classification of mYeloid neoplasms and acute leukemia: Rationale and important changes. Blood 2009; 114(5): 937-51. google scholar
12. 12. Conforti A, Biagini S, Del Bufalo F, Sirleto P, Angioni A, Starc N, et al. Biological, functional and genetic characterization of bone marrow-derived mesenchYmal stromal cells from pediatric patients affected bY acute lYmphoblastic leukemia. PLoS One 2013; 7; 8(11): e76989. google scholar
13. 13. Barachini S, Trombi L, Danti S, D'Alessandro D, Battolla B, Legitimo A, et al. Morpho-functional characterization of human mesenchYmal stem cells from umbilical cord blood for potential uses in regenerative medicine. Stem Cells Dev 2009; 18(2): 293-305. google scholar
14. 14. de Rooij JDE, Michel Zwaan C, van den Heuvel-Eibrink M. Pediatric AML: From biologY to clinical management. J Clin Med 2015; 4(1): 127-49. google scholar
15. 15. KfourY Y, Scadden DT. MesenchYmal cell contributions to the stem cell niche. Cell Stem Cell 2015; 16(3): 239-53. google scholar
16. 16. Caplan AI. MesenchYmal stem cells. J Orthop Res 1991; 9(5): 641-50. google scholar
17. 17. Haddad R, Saldanha-Araujo F. Mechanisms of T-cell immunosuppression bY mesenchYmal stromal cells: What do we know so far? Biomed Res Int 2014; 2014: 216806. google scholar
18. 18. Ribeiro A, Laranjeira P, Mendes S, Velada I, Leite C, Andrade P, et al. MesenchYmal stem cells from umbilical cord matrix, adipose tissue and bone marrow exhibit different capabilitY to suppress peripheral blood B, natural killer and T cells. Stem Cell Res Ther 2013; 4(5): 125. google scholar
19. 19. Stein PH, Singer A. Similar co-stimulation requirements of CD4+ and CD8+ primarY T helper cells: Role of IL-1 and IL-6 in inducing IL-2 secretion and subsequent proliferation. Int Immunol 1992; 4(3): 327-35. google scholar
20. 20. Li CY, Wu XY, Tong JB, Yang XX, Zhao JL, Zheng QF, et al. Comparative analYsis of human mesenchYmal stem cells from bone marrow and adipose tissue under xeno-free conditions for cell therapY. Stem Cell Res Ther 2015; 13; 6(1): 55. google scholar
21. 21. Zhang W, Ge W, Li C, You S, Liao L, Han Q, et al. Effects of mesenchYmal stem cells on differentiation, maturation, and function of human monocYte-derived dendritic cells. Stem Cells Dev 2004; 13(3): 263-71. google scholar
22. 22. Simms PE, Ellis TM. UtilitY of flow cYtometric detection of CD69 expression as a rapid method for determining polY- and oligoclonal lYmphocYte activation. Clin Diagn Lab Immunol 1996; 3(3): 301-4. google scholar
23. 23. Kuhweide R, Van Damme J, Lorre K, Baroja ML, Tsudo M, Ceuppens JL. AccessorY cell-derived helper signals in human T-cell activation with phYtohemagglutinin: Induction of interleukin 2, responsiveness of interleukin 6, and production of interleukin 2 bY interleukin 1. CYtokine 1990; 2(1): 45-54. google scholar
24. 24. Rea IM, McNerlan SE, Alexander HD. CD69, CD25, and HLA-DR activation antigen expression on CD3lYmphocYtes and relationship to serum TNF-a, IFN-y, and sIL-2R levels in aging. Exp Gerontol 1999; 34(1): 79-93. google scholar
25. 25. Damle RN, Ghiotto F, Valetto A, Albesiano E, Fais F, Yan XJ, et al. B-cell chronic lYmphocYtic leukemia cells express a surface membrane phenotYpe of activated, antigen-experienced B lYmphocYtes. Blood 2002; 99(11): 4087-93. google scholar
26. 26. Del Poeta G, Del Principe MI, Zucchetto A, Luciano F, Buccisano F, Rossi FM, et al. CD69 is independentlY prognostic in chronic lYmphocYtic leukemia: A comprehensive clinical and biological profiling studY. Haematologica 2012; 97(2): 279-87. google scholar
27. 27. Dimitriou H, Linardakis E, Martimianaki G, Stiakaki E, Perdikogianni CH, Charbord P, et al. Properties and potential of bone marrow mesenchYmal stromal cells from children with hematologic diseases. CYtotherapY 2008; 10(2): 125-33. google scholar
28. 28. Stenderup K, Justesen J, Clausen C, Kassem M. Aging is associated with decreased maximal life span and accelerated senescence of bone marrow stromal cells. Bone 2003; 33(6): 919-26. google scholar 29. Machado Cde V, Telles PD, Nascimento IL. Immunological characteristics of mesenchYmal stem cells. Rev Bras Hematol Hemoter 2013; 35(1): 62-7. google scholar
29. 30. Peng Y, Wang Y, Wang M, Lan J, Chen Y. Therapeutic applications of toll-like receptors (TLRs) agonists in AML. Clin Transl Oncol 2022; 24(12): 2319-29. google scholar
30. 31. Zeng AGX, Bansal S, Jin L, Mitchell A, Chen WC, Abbas HA, et al. A cellular hierarchY framework for understanding heterogeneitY and predicting drug response in acute mYeloid leukemia. Nat Med 2022; 28(6): 1212-23. google scholar
31. 32. Bochev I, Elmadjian G, KYurkchiev D, Tzvetanov L, Altankova I, Tivchev P, et al. MesenchYmal stem cells from human bone marrow or adipose tissue differentlY modulate mitogen-stimulated B-cell immunoglobulin production in vitro. Cell Biol Int 2008; 32(4): 384-93. google scholar
32. 33. Tabera S, Perez-Simon JA, Dıez-Campelo M, Sânchez-Abarca Ll, Blanco B, Lopez A, et al. The effect of mesenchymal stem cells on the viability, proliferation and differentiation of B-lymphocytes. Haematologica 2008; 93(9): 1301-09. google scholar