Cytotoxicity Assessment and Apoptosis-related Gene Profiling of Antibody Treated Acute Myeloid Leukemia (AML) and Acute Lymphocytic Leukemia (ALL) Cancerous Cell Lines
-
Mahdi Habibi-Anbouhi
,
Zahra Kafi
,
Leila Ghazizadeh
,
Shabnam Kharazi
,
Mahdi Behdani
,
Fatemeh Faraji
,
Mohammadi Ali Shokrgozar
Abstract
Acute myeloid leukemia (AML) and acute lymphocytic leukemia (ALL) are common acute leukemia in adults and children, respectively. In these malignancies, chemotherapy is the main treatment strategy that fails in many cases and is usually associated with adverse effects on healthy cells. In this regard, the development of new therapies is essential. Monoclonal antibodies directed to the cell surface markers of leukemic blasts may have promising consequences with minimal toxic effects on normal cells. Since cluster of differentiation 45Ra (CD45Ra) and CD123 antigens, two considered surface markers of leukemic blasts in AML and ALL respectively, are overexpressed on AML and ALL blasts, CD34+ leukemic progenitors, and AML-LSCs in comparison with normal hematopoietic stem cells (HSCs), they were selected to be targeted; using specific monoclonal antibodies. In this project, CD45Ra+ cells and CD123+ cells were targeted by anti-CD45Ra and/or anti-CD123 monoclonal antibodies. Cytotoxicity effect and cell death induction was determined by 3-(4,5-dimethylthiazol-2-yl)-2–5-diphenyltetrazolium bromide (MTT) assay and flow cytometry. Changes in the expression profile of MCL1, cMyc, Survivin, Id1, and PIM1 genes were assessed by real-time PCR. Statistical analysis of the results showed effective antibody-mediated cytotoxicity and induction of apoptosis in KG1α (CD45Ra+) and Nalm6 (CD123+) cell lines. Also, a significant change in the expression level of some of the apoptosis-related genes was observed. According to the results of this study, it can be concluded that an effective targeting of AML and ALL cancerous cell lines can be performed by anti-CD45Ra and anti-CD123 monoclonal antibodies through their effector functions and apoptosis induction.
1. Society AC. Cancer Facts & Figures 2014. 2014.
2. Kuchinskaya E, Heyman M, Grander D, Linderholm M, Soderhall S, Zaritskey A, et al. Children and adults with acute lymphoblastic leukaemia have similar gene expression profiles. European journal of haematology. 2005;74(6):466-80.
3. Meshinchi S, Arceci RJ. Prognostic factors and risk-based therapy in pediatric acute myeloid leukemia. The oncologist. 2007;12(3):341-55.
4. Leszczyniecka M, Roberts T, Dent P, Grant S, Fisher PB. Differentiation therapy of human cancer: basic science and clinical applications. Pharmacology & therapeutics. 2001;90(2-3):105-56.
5. Nahar R, Muschen M. Pre-B cell receptor signaling in acute lymphoblastic leukemia. Cell cycle. 2009;8(23):3874-7.
6. Zafir-Lavie I, Michaeli Y, Reiter Y. Novel antibodies as anticancer agents. Oncogene. 2007;26(25):3714-33.
7. Uckun FM, Gesner TG, Song CW, Myers DE, Mufson A. Leukemic B-cell precursors express functional receptors for human interleukin-3. Blood. 1989;73(2):533-42.
8. Djokic M, Bjorklund E, Blennow E, Mazur J, Soderhall S, Porwit A. Overexpression of CD123 correlates with the hyperdiploid genotype in acute lymphoblastic leukemia. Haematologica. 2009;94(7):1016-9.
9. Goardon N, Marchi E, Atzberger A, Quek L, Schuh A, Soneji S, et al. Coexistence of LMPP-like and GMP-like leukemia stem cells in acute myeloid leukemia. Cancer cell. 2011;19(1):138-52.
10. Ruella M, Barrett DM, Kenderian SS, Shestova O, Hofmann TJ, Perazzelli J, et al. Dual CD19 and CD123 targeting prevents antigen-loss relapses after CD19-directed immunotherapies. The Journal of Clinical Investigation. 2016;126(10):3814-26.
11. Saunders AE, Johnson P. Modulation of immune cell signalling by the leukocyte common tyrosine phosphatase, CD45. Cellular signalling. 2010;22(3):339-48.
12. Kersten B, Valkering M, Wouters R, Amerongen R, Hanekamp D, Kwidama Z, et al. CD45RA, a specific marker for leukaemia stem cell sub‐populations in acute myeloid leukaemia. British journal of haematology. 2016.
13. Moradi-Kalbolandi S, Davani D, Golkar M, Habibi-Anbouhi M, Abolhassani M, Shokrgozar MA. Soluble Expression and Characterization of a New scFv Directed to Human CD123. Applied biochemistry and biotechnology. 2016;178(7):1390-406.
14. Aguirre E, Renner O, Narlik-Grassow M, Blanco-Aparicio C. Genetic Modeling of PIM Proteins in Cancer: Proviral Tagging and Cooperation with Oncogenes, Tumor Suppressor Genes, and Carcinogens. Front Oncol. 2014;4:109.
15. Belmar J, Fesik SW. Small molecule Mcl-1 inhibitors for the treatment of cancer. Pharmacology & therapeutics. 2015;145:76-84.
16. Ling F, Kang B, Sun XH. Id proteins: small molecules, mighty regulators. Curr Top Dev Biol. 2014;110:189-216.
17. Krishna A, Singh S, Kumar V, Pal US. Molecular concept in human oral cancer. Natl J Maxillofac Surg. 2015;6(1):9-15.
18. Soleimanpour E, Babaei E. Survivin as a Potential Target for Cancer Therapy. Asian Pac J Cancer Prev. 2015;16(15):6187-91.
19. Dang CV. c-Myc target genes involved in cell growth, apoptosis, and metabolism. Molecular and cellular biology. 1999;19(1):1-11.
20. Luo H, Li Q, O'Neal J, Kreisel F, Le Beau MM, Tomasson MH. c-Myc rapidly induces acute myeloid leukemia in mice without evidence of lymphoma-associated antiapoptotic mutations. Blood. 2005;106(7):2452-61.
21. Kawagoe H, Kandilci A, Kranenburg TA, Grosveld GC. Overexpression of N-Myc rapidly causes acute myeloid leukemia in mice. Cancer research. 2007;67(22):10677-85.
22. Miller DM, Thomas SD, Islam A, Muench D, Sedoris K. c-Myc and cancer metabolism. Clinical cancer research : an official journal of the American Association for Cancer Research. 2012;18(20):5546-53.
23. Kaufmann SH, Karp JE, Svingen PA, Krajewski S, Burke PJ, Gore SD, et al. Elevated expression of the apoptotic regulator Mcl-1 at the time of leukemic relapse. Blood. 1998;91(3):991-1000.
24. Malin S, McManus S, Busslinger M. STAT5 in B cell development and leukemia. Current opinion in immunology. 2010;22(2):168-76.
25. Barille-Nion S, Bah N, Vequaud E, Juin P. Regulation of cancer cell survival by BCL2 family members upon prolonged mitotic arrest: opportunities for anticancer therapy. Anticancer research. 2012;32(10):4225-33.
26. Tam WF, Gu TL, Chen J, Lee BH, Bullinger L, Frohling S, et al. Id1 is a common downstream target of oncogenic tyrosine kinases in leukemic cells. Blood. 2008;112(5):1981-92.
27. Ma J, Shi M, Li G, Wang N, Wei J, Wang T, et al. Regulation of Id1 expression by epigallocatechin3gallate and its effect on the proliferation and apoptosis of poorly differentiated AGS gastric cancer cells. International journal of oncology. 2013;43(4):1052-8.
28. Carter BZ, Milella M, Altieri DC, Andreeff M. Cytokine-regulated expression of survivin in myeloid leukemia. Blood. 2001;97(9):2784-90.
29. McKenzie JA, Grossman D. Role of the apoptotic and mitotic regulator survivin in melanoma. Anticancer research. 2012;32(2):397-404.
30. Tyner JW, Jemal AM, Thayer M, Druker BJ, Chang BH. Targeting survivin and p53 in pediatric acute lymphoblastic leukemia. Leukemia. 2012;26(4):623-32.
31. Kantarjian H, Thomas D, Wayne AS, O'Brien S. Monoclonal antibody-based therapies: a new dawn in the treatment of acute lymphoblastic leukemia. Journal of Clinical Oncology. 2012;30(31):3876-83.
32. Jabbour E, O’Brien S, Ravandi F, Kantarjian H. Monoclonal antibodies in acute lymphoblastic leukemia. Blood. 2015;125(26):4010-6.
33. Qu Z, Goldenberg DM, Cardillo TM, Shi V, Hansen HJ, Chang C-H. Bispecific anti-CD20/22 antibodies inhibit B-cell lymphoma proliferation by a unique mechanism of action. Blood. 2008;111(4):2211-9.
34. Busfield S, Biondo M, Wong M, Ramshaw H, Lee E, Ghosh S, et al. Targeting of acute myeloid leukemia in vitro and in vivo with an anti-CD123 mAb engineered for optimal ADCC. Leukemia. 2014;28(11):2213-21.
35. Jin L, Lee EM, Ramshaw HS, Busfield SJ, Peoppl AG, Wilkinson L, et al. Monoclonal antibody-mediated targeting of CD123, IL-3 receptor α chain, eliminates human acute myeloid leukemic stem cells. Cell stem cell. 2009;5(1):31-42.
36. Smith BD, Roboz GJ, Walter RB, Altman JK, Ferguson A, Curcio TJ, et al. First-in man, phase 1 study of CSL362 (anti-IL3Rα/anti-CD123 monoclonal antibody) in patients with CD123+ acute myeloid leukemia (AML) in CR at high risk for early relapse. Blood. 2014;124(21):120-.
37. Sun Q, Woodcock JM, Rapoport A, Stomski FC, Korpelainen EI, Bagley CJ, et al. Monoclonal antibody 7G3 recognizes the N-terminal domain of the human interleukin-3 (IL-3) receptor alpha-chain and functions as a specific IL-3 receptor antagonist. Blood. 1996;87(1):83-92.
38. He SZ, Busfield S, Ritchie DS, Hertzberg MS, Durrant S, Lewis ID, et al. A Phase 1 study of the safety, pharmacokinetics and anti-leukemic activity of the anti-CD123 monoclonal antibody CSL360 in relapsed, refractory or high-risk acute myeloid leukemia. Leukemia & lymphoma. 2015;56(5):1406-15.
39. Orozco JJ, Balkin ER, Gooley TA, Kenoyer A, Hamlin DK, Wilbur DS, et al. Anti-CD45 Radioimmunotherapy with 90 Y but Not 177 Lu Is Effective Treatment in a Syngeneic Murine Leukemia Model. PloS one. 2014;9(12):e113601.
40. Kim J-E, Son JE, Jeong H, Kim DJ, Seo SG, Lee E, et al. A novel cinnamon-related natural product with Pim-1 inhibitory activity inhibits leukemia and skin cancer. Cancer research. 2015;75(13):2716-28.
41. Keeton EK, McEachern K, Dillman KS, Palakurthi S, Cao Y, Grondine MR, et al. AZD1208, a potent and selective pan-Pim kinase inhibitor, demonstrates efficacy in preclinical models of acute myeloid leukemia. Blood. 2014;123(6):905-13.
42. Feng W, Yoshida A, Ueda T. YM155 induces caspase-8 dependent apoptosis through downregulation of survivin and Mcl-1 in human leukemia cells. Biochemical and biophysical research communications. 2013;435(1):52-7.
43. Chandele A, Prasad V, Jagtap JC, Shukla R, Shastry PR. Upregulation of survivin in G2/M cells and inhibition of caspase 9 activity enhances resistance in staurosporine-induced apoptosis. Neoplasia. 2004;6(1):29-40.
44. Tirrò E, Consoli ML, Massimino M, Manzella L, Frasca F, Sciacca L, et al. Altered expression of c-IAP1, survivin, and Smac contributes to chemotherapy resistance in thyroid cancer cells. Cancer research. 2006;66(8):4263-72.
45. Belmar J, Fesik SW. Small molecule Mcl-1 inhibitors for the treatment of cancer. Pharmacology & therapeutics. 2015;145:76-84.
46. Varadarajan S, Vogler M, Butterworth M, Dinsdale D, Walensky L, Cohen G. Evaluation and critical assessment of putative MCL-1 inhibitors. Cell Death & Differentiation. 2013;20(11):1475-84.
47. Mistry H, Hsieh G, Buhrlage SJ, Huang M, Park E, Cuny GD, et al. Small-molecule inhibitors of USP1 target ID1 degradation in leukemic cells. Molecular cancer therapeutics. 2013;12(12):2651-62.
48. Pan SY, Morrison H. Physical activity and hematologic cancer prevention. Physical Activity and Cancer: Springer; 2010. p. 135-58.
| Files | ||
| Issue | Vol 18, No 6 (2019) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijaai.v18i6.2181 | |
| PMID | 32245312 | |
| Keywords | ||
| Cytotoxicity assessment Apoptotic genes Acute leukemia cell lines Immunotherapy | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
