Transcriptome analysis of ABCB1, ABCG2 and the BCL2/BAX ratio in refractory and relapsed canine lymphomas under treatment and rescue protocol

Open access

Abstract

The main problems that cause unresponsiveness to an anti-neoplastic drug are the overexpression of drug resistant and anti-apoptotic proteins in tumor cells. In a rescue protocol we evaluated the ability of toceranib phosphate concurrent with lomustine (CCNU) or L-asparaginase and vincristine to decrease drug resistant and apoptotic proteins in relapsed and refractory canine lymphomas. The peripheral blood samples were collected before and after the rescue treatment from fourteen dogs that were refractory to cyclophosphamide-vincristine-prednisolone (COP) or COP-doxorubicin (CHOP) treatment and had recurrent multicentric lymphoma. The mRNA expression level of ABCB1, ABCG2, Bcl2 and Bax were determined by quantitative real-time PCR. The fold-change in ABCB1, ABCG2, Bcl2 and Bax mRNA levels were analyzed in correlation with the progression-free survival (PFS). After the rescue treatment, the ABCB1 and ABCG2 mRNA expression levels were 1.57- and 1.85-fold lower (p = 0.4 and p = 0.87), respectively, compared to pre-treatment. Bcl2/Bax ratio was numerically but not significantly decreased 1.02-fold (p = 0.74). The overall response rate of this protocol was 50% with a median PFS of 79 days (range 14-207 days). The low medians of relative expression levels of ABCB1, ABCG2 and Bcl2/Bax ratio group did not correlate with the clinical outcomes when compared to the high medians of relative expression levels, and likewise with the clinical stage, immunophenotype, histological grade and sub-stage. Therefore, the administration of a rescue drug with toceranib phosphate might be beneficial in refractory and relapsed canine lymphoma.

1. Uozurmi K, Nakaichi M, Yamamoto Y, Une S, Taura Y: Development of multidrug resistance in a canine lymphoma cell line. Res Vet Sci 2005,78:217-224.

2. Moore AS, Leveille CR, Reimann KA, Shu H, Arias IM: The expression of P-glycoprotein in canine lymphoma and its association with multidrug resistance. Cancer Invest 1995,13:475-479.

3. Bergman PJ, Ogilvie GK, Powers BE: Monoclonal antibody C219 immunohistochemistry against P-glycoprotein: sequential analysis and predictive ability in dogs with lymphoma. J Vet Intern Med 1996,10:354-359.

4. Lee JJ, Hughes CS, Fine RL, Page RL: P-glycoprotein expression in canine lymphoma: a relevant, intermediate model of multidrug resistance. Cancer 1996,77:1892-1898.

5. Culmsee K, Gruber AD, von Samson-Himmelstjerna G, Nolte I: Quantification of MDR-1 gene expression in canine tissues by real-time reverse transcription quantitative polymerase chain reaction. Res Vet Sci 2004,77:223-229.

6. Tomiyasu H, Goto-Koshino Y, Takahashi M, Fujino Y, Ohno K, Tsujimoto H: Quantitative analysis of mRNA for 10 different drug resistance factors in dogs with lymphoma. J Vet Med Sci 2010,72:1165-1172.

7. Zandvliet M, Teske E, Schrickx JA, Mol JA: A longitudinal study of ABC transporter expression in canine multicentric lymphoma. Vet J 2015,205:263-271.

8. Lemmon MA, Schlessinger J: Cell signaling by receptor tyrosine kinases. Cell 2010,141:1117-1134.

9. London CA, Malpas PB, Wood-Follis SL, Boucher JF, Rusk AW, Rosenberg MP, Henry CJ, Mitchener KL, Klein MK, Hintermeister JG, Bergman PJ, Couto GC, Mauldin GN, Michels GM: Multi-center, placebo-controlled, double-blind, randomized study of oral toceranib phosphate (SU11654), a receptor tyrosine kinase inhibitor, for the treatment of dogs with recurrent (either local or distant) mast cell tumor following surgical excision. Clin Cancer Res 2009,15:3856-3865.

10. London C, Mathie T, Stingle N, Clifford C, Haney S, Klein MK, Beaver L, Vickery K, Vail DM, Hershey B, Ettinger S, Vaughan A, Alvarez F, Hillman L, Kiselow M, Thamm D, Higginbotham ML, Gauthier M, Krick E, Phillips B, Ladue T, Jones P, Bryan J, Gill V, Novasad A, Fulton L, Carreras J, McNeill C, Henry C, Gillings S: Preliminary evidence for biologic activity of toceranib phosphate (Palladia((R))) in solid tumours. Vet Comp Oncol 2012,10:194-205.

11. Pan X, Tsimbas K, Kurzman ID, Vail DM: Safety evaluation of combination CCNU and continuous toceranib phosphate (Palladia((R))) in tumour-bearing dogs: a phase I dose-finding study. Vet Comp Oncol 2016,14:202-209.

12. Zandvliet M, Teske E, Chapuis T, Fink-Gremmels J, Schrickx JA: Masitinib reverses doxorubicin resistance in canine lymphoid cells by inhibiting the function of P-glycoprotein. J Vet Pharmacol Ther 2013,36:583-587.

13. Llambi F, Moldoveanu T, Tait SW, Bouchier-Hayes L, Temirov J, McCormick LL, Dillon CP, Green DR: A unified model of mammalian BCL-2 protein family interactions at the mitochondria. Mol Cell 2011,44:517-531.

14. Dettwiler M, Croci M, Vaughan L, Guscetti F: Immunohistochemical expression study of proapoptotic BH3-only protein bad in canine nonneoplastic tissues and canine lymphomas. Vet Pathol 2013,50:789-796.

15. Jung JT, Kim DH, Kwak EK, Kim JG, Park TI, Sohn SK, Do YR, Kwon KY, Song HS, Park EH, Lee KB: Clinical role of Bcl-2, Bax, or p53 overexpression in peripheral T-cell lymphomas. Ann Hematol 2006,85:575-581.

16. Koshino A, Goto-Koshino Y, Setoguchi A, Ohno K, Tsujimoto H: Mutation of p53 Gene and Its Correlation with the Clinical Outcome in Dogs with Lymphoma. J Vet Intern Med 2016,30:223-229.

17. Meichner K, Fogle JE, English L, Suter SE: Expression of Apoptosis-regulating Proteins Bcl-2 and Bax in Lymph Node Aspirates from Dogs with Lymphoma. J Vet Intern Med 2016,30:819-826.

18. Sohn SK, Jung JT, Kim DH, Kim JG, Kwak EK, Park T, Shin DG, Sohn KR, Lee KB: Prognostic significance of bcl-2, bax, and p53 expression in diffuse large B-cell lymphoma. Am J Hematol 2003,73:101-107.

19. Sirivisoot S, Techangamsuwan S, Tangkawattana S, Rungsipipat A: Pax5 as a potential candidate marker for canine B-cell lymphoma. Vet Med (Praha) 2017,62:74-80.

20. Peters IR, Peeters D, Helps CR, Day MJ: Development and application of multiple internal reference (housekeeper) gene assays for accurate normalisation of canine gene expression studies. Vet Immunol Immunopathol 2007,117:55-66.

21. Del Puerto HL, Martins AS, Moro L, Milsted A, Alves F, Braz GF, Vasconcelos AC: Caspase-3/-8/-9, Bax and Bcl-2 expression in the cerebellum, lymph nodes and leukocytes of dogs naturally infected with canine distemper virus. Genet Mol Res 2010,9:151-161.

22. Silver N, Best S, Jiang J, Thein SL: Selection of housekeeping genes for gene expression studies in human reticulocytes using real-time PCR. BMC Mol Biol 2006,7:33.

23. Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F: Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 2002,3:RESEARCH0034.

24. Andersen CL, Jensen JL, Orntoft TF: Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 2004,64:5245-5250.

25. Pfaffl MW, Tichopad A, Prgomet C, Neuvians TP: Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: BestKeeper--Excel-based tool using pair-wise correlations. Biotechnol Lett 2004,26:509-515.

26. Xie F, Xiao P, Chen D, Xu L, Zhang B: miRDeepFinder: a miRNA analysis tool for deep sequencing of plant small RNAs. Plant Mol Biol 2012.

27. Saba CF, Thamm DH, Vail DM: Combination chemotherapy with L-asparaginase, lomustine, and prednisone for relapsed or refractory canine lymphoma. J Vet Intern Med 2007,21:127-132.

28. London CA, Hannah AL, Zadovoskaya R, Chien MB, Kollias-Baker C, Rosenberg M, Downing S, Post G, Boucher J, Shenoy N, Mendel DB, McMahon G, Cherrington JM: Phase I dose-escalating study of SU11654, a small molecule receptor tyrosine kinase inhibitor, in dogs with spontaneous malignancies. Clin Cancer Res 2003,9:2755-2768.

29. Houghton PJ, Germain GS, Harwood FC, Schuetz JD, Stewart CF, Buchdunger E, Traxler P: Imatinib mesylate is a potent inhibitor of the ABCG2 (BCRP) transporter and reverses resistance to topotecan and SN-38 in vitro. Cancer Res 2004,64:2333-2337.

30. Tiwari AK, Sodani K, Wang SR, Kuang YH, Ashby CR, Jr., Chen X, Chen ZS: Nilotinib (AMN107, Tasigna) reverses multidrug resistance by inhibiting the activity of the ABCB1/Pgp and ABCG2/BCRP/MXR transporters. Biochem Pharmacol 2009,78:153-161.

31. Sen R, Natarajan K, Bhullar J, Shukla S, Fang HB, Cai L, Chen ZS, Ambudkar SV, Baer MR: The novel BCR-ABL and FLT3 inhibitor ponatinib is a potent inhibitor of the MDR-associated ATP-binding cassette transporter ABCG2. Mol Cancer Ther 2012,11:2033-2044.

32. Chimura N, Shibata S, Kimura T, Kondo N, Mori T, Hoshino Y, Kamishina H, Maeda S: Suitable reference genes for quantitative real-time rt-pcr in total RNA extracted from canine whole blood using the PAXgene system. J Vet Med Sci 2011,73:1101-1104.

33. Gulmann C, Espina V, Petricoin E, 3rd, Longo DL, Santi M, Knutsen T, Raffeld M, Jaffe ES, Liotta LA, Feldman AL: Proteomic analysis of apoptotic pathways reveals prognostic factors in follicular lymphoma. Clin Cancer Res 2005,11:5847-5855.

34. Frantz AM, Sarver AL, Ito D, Phang TL, Karimpour-Fard A, Scott MC, Valli VE, Lindblad-Toh K, Burgess KE, Husbands BD, Henson MS, Borgatti A, Kisseberth WC, Hunter LE, Breen M, O'Brien TD, Modiano JF: Molecular profiling reveals prognostically significant subtypes of canine lymphoma. Vet Pathol 2013,50:693-703.

35. Valli VE, Kass PH, San Myint M, Scott F: Canine lymphomas: association of classification type, disease stage, tumor subtype, mitotic rate, and treatment with survival. Vet Pathol 2013,50:738-748.

36. Tainton KM, Smyth MJ, Jackson JT, Tanner JE, Cerruti L, Jane SM, Darcy PK, Johnstone RW: Mutational analysis of P-glycoprotein: suppression of caspase activation in the absence of ATP-dependent drug efflux. Cell Death Differ 2004,11:1028-1037.

37. Gollapud S, Gupta S: Anti-P-glycoprotein antibody-induced apoptosis of activated peripheral blood lymphocytes: a possible role of P-glycoprotein in lymphocyte survival. J Clin Immunol 2001,21:420-430.

38. Luciano F, Ricci JE, Herrant M, Bertolotto C, Mari B, Cousin JL, Auberger P: T and B leukemic cell lines exhibit different requirements for cell death: correlation between caspase activation, DFF40/DFF45 expression, DNA fragmentation and apoptosis in T cell lines but not in Burkitt's lymphoma. Leukemia 2002,16:700-707.

39. Bram EE, Stark M, Raz S, Assaraf YG: Chemotherapeutic drug-induced ABCG2 promoter demethylation as a novel mechanism of acquired multidrug resistance. Neoplasia 2009,11:1359-1370.

40. Bai M, Skyrlas A, Agnantis NJ, Kamina S, Kitsoulis P, Kanavaros P: Cluster analysis of apoptosis-associated bcl2 family proteins in diffuse large B-cell lymphomas. Relations with the apoptotic index, the proliferation profile and the B-cell differentiation immunophenotypes. Anticancer Res 2004,24:3081-3088.

41. Cayrol F, Praditsuktavorn P, Fernando TM, Kwiatkowski N, Marullo R, Calvo-Vidal MN, Phillip J, Pera B, Yang SN, Takpradit K, Roman L, Gaudiano M, Crescenzo R, Ruan J, Inghirami G, Zhang T, Cremaschi G, Gray NS, Cerchietti L: THZ1 targeting CDK7 suppresses STAT transcriptional activity and sensitizes T-cell lymphomas to BCL2 inhibitors. Nat Commun 2017,8:14290.

Acta Veterinaria

The Journal of University of Belgrade

Journal Information

IMPACT FACTOR 2017: 0.604
5-year IMPACT FACTOR: 0.439

CiteScore 2018: 0.71

SCImago Journal Rank (SJR) 2018: 0.281
Source Normalized Impact per Paper (SNIP) 2018: 0.547

Metrics

All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 388 308 24
PDF Downloads 147 133 10