Cytotoxic and Cytostatic Effects of Targeting mTOR and Hedgehog Pathways in Acute Myeloid Leukemia

Year 2022, Volume: 12 Issue: 3, 202 - 208, 31.12.2022

Enes Çiçek Fulya Mina Küçüktaş Münevver Yenigül Emel Gencer Akcok

https://doi.org/10.26650/experimed.1193721

Abstract

Objectives: Acute myeloid leukemia (AML) is a highly aggressive heterogeneous hematopoietic malignancy characterized by a rapid and abnormal proliferation of immature myeloid leukemia cells in the bone marrow and peripheral blood. Aberrant alterations in signal transduction pathways are strongly associated with the progression of AML. This study aimed to investigate cell viability and the cell cycle in AML cells by targeting the Hedgehog and mTOR signaling pathways with rapamycin and GANT61.

Materials and Method: The antiproliferative effect of rapamycin and GANT61 was assessed by the MTT cell viability assay in two AML cell lines: CMK and MOLM-13. The effect of the inhibitors on cell-cycle distribution was determined using propidium iodide staining and measured with flow cytometry.

Results: Rapamycin, an mTOR inhibitor, and GANT61, a Gli-1 inhibitor, decreased the cell proliferation of CMK and MOLM-13 cells. The IC20 values, which is the drug concentration that inhibits cell growth by 20%, were combined and administered to the cells. The results show the drugs to have a combinatorial inhibitory effect on CMK cells but not on MOLM-13 cells. In addition, the combination of drugs arrested the cells during the G0/G1 phase.

Conclusion: This study suggests a novel combination therapy approach for AML via mTOR and Hedgehog signaling pathway inhibition using rapamycin and GANT61, respectively. It also suggest further studies be performed to reveal the mechanism of action.

Keywords

Hedgehog, mTOR, Leukemia, Combination therapy, Cell cycle

References

• 1. Darici S, Alkhaldi H, Horne G, J0rgensen HG, Marmiroli S, Huang X. Targeting PI3K/Akt/mTOR in AML: Rationale and clinical evidence. J Clin Med 2020; 9(9): 2934. [CrossRef] google scholar
• 2. Nepstad I, Hatfield KJ, Gronnings^ter IS, Reikvam H. The PI3K-Akt-mTOR signaling pathway in human acute myeloid leukemia (AML) cells. Int J Mol Sci 2020; 21(8): 2907. [CrossRef] google scholar
• 3. Saultz JN, Garzon R. Acute myeloid leukemia: A concise review. J Clin Med 2016; 5(3): 33. [CrossRef] google scholar
• 4. Scholl C, Gilliland DG, Frohling S. Deregulation of signaling pathways in acute myeloid leukemia. Semin Oncol 2008; 35(4): 336-45. [CrossRef] google scholar
• 5. Terao T, Minami Y. Targeting Hedgehog (Hh) Pathway for the acute myeloid leukemia treatment. Cells 2019; 8(4): 312. [CrossRef] google scholar
• 6. Niyaz M, Khan MS, Mudassar S. Hedgehog signaling: An achilles' heel in cancer. Transl Oncol 2019; 12(10): 1334-44. [CrossRef] google scholar
• 7. Sari IN, Phi LTH, Jun,N, Wijaya YT, Lee S, Kwon HY. Hedgehog signaling in cancer: A prospective therapeutic target for eradicating cancer stem cells. Cells 2018; 7(11): 208. [CrossRef] google scholar
• 8. Shallis RM, Bewersdorf JP, Boddu PC, Zeidan AM. Hedgehog pathway inhibition as a therapeutic target in acute myeloid leukemia. Expert Rev Anticancer Ther 2019; 19: 717-29. [CrossRef] google scholar
• 9. Long B, Wang LX, Zheng FM, Lai SP, Xu DR, Hu Y, et al. Targeting GLI1 suppresses cell growth and enhances chemosensitivity in CD34+ enriched acute myeloid leukemia progenitor cells. Cell Physiol Biochem 2016; 38(4): 1288-302. [CrossRef] google scholar
• 10. Ruch JM, Kim EJ. Hedgehog signaling pathway and cancer therapeutics: Progress to date. Drugs. 2013; 73, 613-23. [CrossRef] google scholar
• 11. Harada K, Ohashi R, Naito K, Kanki K. Hedgehog signal inhibitor GANT61 inhibits the malignant behavior of undifferentiated hepatocellular carcinoma cells by targeting non-canonical GLI signaling. Int J Mol Sci 2020; 21(9): 3126. [CrossRef] google scholar
• 12. Saxton RA, Sabatini DM. mTOR signaling in growth, metabolism, and disease. Cell 2017; 168(6): 960-76. [CrossRef] google scholar
• 13. Zou Z, Tao T, Li H, Zhu X. mTOR signaling pathway and mTOR inhibitors in cancer: Progress and challenges. Cell Biosci 2020; 10(1): 31. [CrossRef] google scholar
• 14. Park S, Chapuis N, Tamburini J, Bardet V, Cornillet-Lefebvre P, Willems L, et al. Role of the PI3K/AKT and mTOR signaling pathways in acute myeloid leukemia. Haematologica 2010; 95(5): 819-28. [CrossRef] google scholar
• 15. Feng Y, Chen X, Cassady K, Zou Z, Yang S, Wang Z and Zhang X. The Role of mTOR inhibitors in hematologic disease: From bench to bedside. Front Oncol 2021; 10: 611690. [CrossRef] google scholar
• 16. Yenigül M, Ak^ok i, Gencer Ak^ok EB. Ethacrynic acid and cinnamic acid combination exhibits selective anticancer effects on K562 chronic myeloid leukemia cells. Mol Biol Rep 2022; 49(8): 7521-30. [CrossRef] google scholar
• 17. Chou TC. Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Res 2010; 70: 440-6. [CrossRef] google scholar
• 18. PDQ Adult Treatment Editorial Board. Adult Acute Myeloid Leukemia Treatment (PDQ®): Patient Version. In PDQ Cancer Information Summaries. National Cancer Institute (US); 2002. Available from: http://www.ncbi.nlm.nih.gov/books/NBK65939/ google scholar
• 19. Aberger F, Hutterer E, Sternberg C, del Burgo PJ, Hartmann TN. Acute myeloid leukemia -strategies and challenges for targeting oncogenic Hedgehog/GLI signaling. Cell Commun Signal 2017; 15(1): 8. [CrossRef] google scholar
• 20. Li X, Chen F, Zhu Q, Ding B, Zhong Q, Huang K, et al. Gli-1/PI3K/ AKT/NF-kB pathway mediates resistance to radiation and is a target for reversion of responses in refractory acute myeloid leukemia cells. Oncotarget 2016; 7: 33004-15. [CrossRef] google scholar
• 21. Queiroz KC, Ruela-de-Sousa RR, Fuhler GM, Aberson HL, Ferreira CV, Peppelenbosch MP, et al. Hedgehog signaling maintains chemoresistance in myeloid leukemic cells. Oncogene 2010; 29(48): 6314-22. [CrossRef] google scholar
• 22. Jamieson C, Martinelli G, Papayannidis C, Cortes JE. Hedgehog pathway inhibitors: A new therapeutic class for the treatment of acute myeloid leukemia. Blood Cancer Discov 2020; 1(2): 134-45. [CrossRef] google scholar
• 23. Laplante M, Sabatini DM. mTOR signaling at a glance. J Cell Sci 2009; 122(20): 3589-94. [CrossRef] google scholar
• 24. Arriola Apelo SI, Lamming DW. Rapamycin: An inhibitor of aging emerges from the soil of Easter Island. J Gerontol A Biol Sci Med Sci 2016; 71(7): 841-9. [CrossRef] google scholar
• 25. Ballou LM, Lin RZ. Rapamycin and mTOR kinase inhibitors. J Chem Biol 2008; 1(1-4): 27-36. [CrossRef] google scholar
• 26. Récher C, Beyne-Rauzy O, Demur C, Chicanne G, Dos Santos C, Mansat-De Mas V, et al. Antileukemic activity of rapamycin in acute myeloid leukemia. Blood 2005; 105 (6): 2527-34. [CrossRef] google scholar
• 27. Wang Y, Ding Q, Yen CJ, Xia W, Izzo JG, Lang JY, et al. The Crosstalk of mTOR/S6K1 and Hedgehog pathways. Cancer Cell 2012; 21(3): 374-87. [CrossRef] google scholar
• 28. Bacelar Sacramento de Araujo T, de Oliveira Siquara da Rocha L, Torres Andion Vidal M, Cerqueira Coelho PL, Galvao dos Reis M, Solano de Freitas Souza B, et al. GANT61 Reduces Hedgehog Molecule (GLI1) Expression and Promotes Apoptosis in Metastatic Oral Squamous Cell Carcinoma Cells. Int J Mol Sci 2020; 21(17): 6076. [CrossRef] google scholar
• 29. Miyazaki Y, Matsubara S, Ding Q, Tsukasa K, Yoshimitsu M, Kosai K, et al. Efficient elimination of pancreatic cancer stem cells by hedgehog/GLI inhibitor GANT61 in combination with mTOR inhibition. Mol Cancer 2016; 15(1): 49. [CrossRef] google scholar
• 30. Pan D, Li Y, Li Z, Wang Y, Wang P, Liang Y. Gli inhibitor GANT61 causes apoptosis in myeloid leukemia cells and acts in synergy with rapamycin. Leuk Res 2012; 36(6): 742-8. [CrossRef] google scholar
• 31. Zhou C, Du J, Zhao L, Liu W, Zhao T, Liang H, Fang P, et al. GLI1 reduces drug sensitivity by regulating cell cycle through PI3K/ AKT/GSK3/CDK pathway in acute myeloid leukemia. Cell Death Dis 12(3): 231. [CrossRef] google scholar
• 32. Latuske E, Stamm H, Klokow M, Vohwinkel G, Muschhammer J, Bokemeyer C, Jücker M, Kebenko M, Fiedler W, Wellbrock J. Combined inhibition of GLI and FLT3 signaling leads to effective anti-leukemic effects in human acute myeloid leukemia. Oncotarget 2017; 8: 29187-201. [CrossRef] google scholar
• 33. SiY,ChuH,ZhuW,XiaoT,ShenX,FuY,etal.Concentration-dependent effects of rapamycin on proliferation, migration and apoptosis of endothelial cells in human venous malformation. Exp Ther Med 2018; 16: 4595-601. [CrossRef] google scholar
• 34. Cheng KY, Hao M. Mammalian target of rapamycin (mTOR) regulates transforming growth factor-ß1 (TGF-ßl)-induced epithelial-mesenchymal transition via decreased pyruvate kinase M2 (PKM2) expression in cervical cancer cells. Med Sci Monit 2017; 23: 2017-28. [CrossRef] google scholar