Molecular screening and the clinical impacts of BCR‑ABL KD mutations in patients with imatinib‑resistant chronic myeloid leukemia

Koçkan,Betül; Toptaş,Tayfur; Atagündüz,Işik; Tuğlular,Ayşe  Tülin; Özer,Ayşe; Akkiprik,Mustafa

doi:10.3892/ol.2017.7606

Molecular screening and the clinical impacts of BCR‑ABL KD mutations in patients with imatinib‑resistant chronic myeloid leukemia

Authors:
- Betül Koçkan
- Tayfur Toptaş
- Işik Atagündüz
- Ayşe Tülin Tuğlular
- Ayşe Özer
- Mustafa Akkiprik
View Affiliations

Affiliations: Department of Medical Services and Techniques, Vocational School, Beykent University, Buyukcekmece, 34550 Istanbul, Turkey, Department of Hematology, School of Medicine, Marmara University, Maltepe, 34854 Istanbul, Turkey, Department of Medical Biology, School of Medicine, Marmara University, Maltepe, 34854 Istanbul, Turkey
Published online on: December 13, 2017 https://doi.org/10.3892/ol.2017.7606
Pages: 2419-2424

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The present study aimed to detect the frequency of kinase domain (KD) mutations in order to evaluate their clinical significance and functional importance in 45 patients with chronic myeloid leukemia (CML) who were resistant to imatinib therapy. Sanger sequencing was used (45 patients), along with allele‑specific oligonucleotide polymerase chain reaction (ASO‑PCR; 3 patients), for the screening of mutations. BCR/ABL KD was amplified by nested PCR and sequencing was performed. Secondly, ASO‑PCR was performed to confirm the results of the sequence analysis for E255K mutations. Mutations were detected in 11/45 patients (24.44%) via Sanger sequencing. D241G (4.4%), C369C (4.4%), K285N (2.2%), A380T (2.2%) and A366V (2.2%) mutations were detected. E255K (8.8%) was detected by ASO‑PCR and Sanger sequencing. Mutations are a primary reason for suboptimal responses, loss of response and resistance to imatinib. In particular, the E255K mutation, which is characterized by resistance to imatinib and nilotinib, was detected in four patients. Analyzing the mutations and monitoring patients with CML may improve their prognosis and survival rate. ASO‑PCR assays will be beneficial for the routine monitoring of mutations.

View Figures

View References

1	Deininger MW, Goldman JM and Melo JV: The molecular biology of chronic myeloid leukemia. Blood. 96:3343–3356. 2000.PubMed/NCBI
2	Corbin AS, La Rosée P, Stoffregen EP, Druker BJ and Deininger MW: Several Bcr-Abl kinase domain mutants associated with imatinib mesylate resistance remain sensitive to imatinib. Blood. 101:4611–4614. 2003. View Article : Google Scholar : PubMed/NCBI
3	Litzow MR: Imatinib resistance, obstacles and opportunities. Arch Pathol Lab Med. 130:669–679. 2006.PubMed/NCBI
4	Wongboonma W, Thongnoppakhun W and Auewarakul CU: BCR-ABL kinase domain mutations in tyrosine kinase inhibitors-naïve and -exposed Southeast Asian chronic myeloid leukemia patients. Exp Mol Pathol. 92:259–265. 2012. View Article : Google Scholar : PubMed/NCBI
5	Sawyers CL: Chronic myeloid leukemia. N Engl J Med. 340:1330–1340. 1999. View Article : Google Scholar : PubMed/NCBI
6	O'Brien S, Berman E, Borghaei H, DeAngelo DJ, Devetten MP, Devine S, Erba HP, Gotlib J, Jagasia M, Moore JO, et al: NCCN clinical practice guidelines in oncology: Chronic myelogenous leukemia. J Natl Compr Canc Netw. 7:984–1023. 2009. View Article : Google Scholar : PubMed/NCBI
7	Iqbal N and Iqbal N: Imatinib: A breakthrough of targeted therapy in cancer. Chemother Res Pract. 2014:3570272014.PubMed/NCBI
8	Soverini S, Hochhaus A, Nicolini FE, Gruber F, Lange T, Saglio G, Pane F, Müller MC, Ernst T, Rosti G, et al: BCR-ABL kinase domain mutation analysis in chronic myeloid leukemia patients treated with tyrosine kinase inhibitors: Recommendations from an expert panel on behalf of European Leukemia net. Blood. 118:1208–1215. 2011. View Article : Google Scholar : PubMed/NCBI
9	Ursan ID, Jiang R, Pickard EM, Lee TA, Ng D and Pickard AS: Emergence of BCR-ABL kinase domain mutations associated with newly diagnosed chronic myeloid leukemia: A meta-analysis of clinical trials of tyrosine kinase inhibitors. J Manag Care Spec Pharm. 21:114–122. 2015. View Article : Google Scholar : PubMed/NCBI
10	La Rosée P and Deininger MW: Resistance to imatinib: Mutations and beyond. Semin Hematol. 47:335–343. 2010. View Article : Google Scholar : PubMed/NCBI
11	Vaidya S, Vundinti BR, Shanmukhaiah C, Chakrabarti P and Ghosh K: Evolution of BCR/ABL gene mutation in CML is time dependent and dependent on the pressure exerted by tyrosine kinase inhibitor. PLoS One. 10:e01148282015. View Article : Google Scholar : PubMed/NCBI
12	Wang Z, Liu Z, Wu X, Chu S, Wang J, Yuan H, Roth M, Yuan YC, Bhatia R and Chen WY: Correction: ATRA-induced cellular differentiation and CD38 expression inhibits acquisition of BCR-ABL mutations for CML acquired resistance. PLoS Genet. 10:e10044142014. View Article : Google Scholar : PubMed/NCBI
13	Cortes J, Jabbour E, Kantarjian H, Yin CC, Shan J, O'Brien S, Garcia-Manero G, Giles F, Breeden M, Reeves N, et al: Dynamics of BCR-ABL kinase domain mutations in chronic myeloid leukemia after sequential treatment with multiple tyrosine kinase inhibitors. Blood. 110:4005–4011. 2007. View Article : Google Scholar : PubMed/NCBI
14	Apperley JF: Part I: Mechanisms of resistance to imatinib in chronic myeloid leukaemia. Lancet Oncol. 8:1018–1029. 2007. View Article : Google Scholar : PubMed/NCBI
15	Jabbour E, Hochhaus A, Cortes J, La Rosée P and Kantarjian HM: Choosing the best treatment strategy for chronic myeloid leukemia patients resistant to imatinib: Weighing the efficacy and safety of individual drugs with BCR-ABL mutations and patient history. Leukemia. 24:6–12. 2010. View Article : Google Scholar : PubMed/NCBI
16	Baccarani M, Castagnetti F, Gugliotta G and Rosti G: A review of the european leukemianet recommendations for the management of CML. Ann Hematol. 94 Suppl 2:S141–S147. 2015. View Article : Google Scholar : PubMed/NCBI
17	Ai J and Tiu RV: Practical management of patients with chronic myeloid leukemia who develop tyrosine kinase inhibitor-resistant BCR-ABL1 mutations. Ther Adv Hematol. 5:107–120. 2014. View Article : Google Scholar : PubMed/NCBI
18	Sorel N, Roy L, Martineau G, Guilhot F, Turhan AG and Chomel JC: Sequential emergence of ABL-kinase mutations with loss of unmutated BCR-ABL allele during targeted therapies of CML. Blood. 108:1782–1783. 2006. View Article : Google Scholar : PubMed/NCBI
19	Ernst T, Erben P, Muller MC, Paschka P, Schenk T, Hoffmann J, Kreil S, La Rosée P, Hehlmann R and Hochhaus A: Dynamics of BCR-ABL mutated clones prior to hematologic or cytogenetic resistance to imatinib. Haematologica. 93:186–192. 2008. View Article : Google Scholar : PubMed/NCBI
20	Deininger MW, McGreevey L, Willis S, Bainbridge TM, Druker BJ and Heinrich MC: Detection of ABL kinase domain mutations with denaturing high performance liquid chromatography. Leukemia. 18:864–871. 2004. View Article : Google Scholar : PubMed/NCBI
21	Iqbal Z, Aleem A, Iqbal M, Naqvi MI, Gill A, Taj AS, Qayyum A, ur-Rehman N, Khalid AM, Shah IH, et al: Sensitive detection of pre-existing BCR-ABL kinase domain mutations in CD34+ cells of newly diagnosed chronic-phase chronic myeloid leukemia patients is associated with imatinib resistance: Implications in the post-imatinib era. PLoS One. 8:e557172013. View Article : Google Scholar : PubMed/NCBI
22	Erbilgin Y, Çatal S, Eşkazan AE, Hatirnaz Ö, Soysal T and Özbek U: ABL gene kinase domain mutation scanning by denaturing high performance liquid chromatography sequencing method. Turk J Hematol. 28:97–102. 2011. View Article : Google Scholar
23	Kang HY, Hwang JY, Kim SH, Goh HG, Kim M and Kim DW: Comparison of allele specific oligonucleotide-polymerase chain reaction and direct sequencing for high throughput screening of ABL kinase domain mutations in chronic myeloid leukemia resistant to imatinib. Haematologica. 91:659–662. 2006.PubMed/NCBI
24	Srivastava S and Dutt S: Imatinib mesylate resistance and mutations: An Indian experience. Indian J Med Paediatr Oncol. 34:213–220. 2013. View Article : Google Scholar : PubMed/NCBI
25	Sherbenou DW, Hantschel O, Kaupe I, Willis S, Bumm T, Turaga LP, Lange T, Dao K, Press RD, Druker BJ, et al: BCR-ABL SH3-SH2 domain mutations in chronic myeloid leukemia patients on imatinib. Blood. 116:3278–3285. 2010. View Article : Google Scholar : PubMed/NCBI
26	Gruber F, Hansen H, Olsen M, Eggen L, Skogen B, Gedde-Dahl T, Lundan T, Porkka K, Simonsson B, Markevärn B, et al: Quantitative analyses of BCR-ABL mutations associated with imatinib treatment in CML. Blood. 106:20002005.
27	Khorashad JS, Kelley TW, Szankasi P, Mason CC, Soverini S, AdriaL T, Eide CA, Zabriskie MS, Lange T, Estrada JC, et al: BCR-ABL1 compound mutations in tyrosine kinase inhibitor-resistant CML: Frequency and clonal relationships. Blood. 121:489–498. 2013. View Article : Google Scholar : PubMed/NCBI
28	Rajasekaran R, Doss PDC, Prasad GA and Sethumadhavan R: In silico identification and analysis of drug resistant mutants of ABL tyrosine kinase based on detrimental missense mutations. Curr Signal Trans Ther. 6:396–404. 2011. View Article : Google Scholar
29	von Bubnoff N, Veach DR, van der Kuip H, Aulitzky WE, Sänger J, Seipel P, Bornmann WG, Peschel C, Clarkson B and Duyster J: A cell-based screen for resistance of Bcr-Abl-positive leukemia identifies the mutation pattern for PD166326, an alternative Abl kinase inhibitor. Blood. 105:1652–1659. 2005. View Article : Google Scholar : PubMed/NCBI
30	Soverini S, Baccarani M, Iacobucci I and Martinelli G: Resistance to tyrosine kinase inhibitors in Philadelphia chromosome-positive leukemias: Which mutations matter? Clinical Leukemia. 1:223–228. 2007. View Article : Google Scholar
31	Chomel JC, Sorel N and Turhan AG: ABL-Kinase mutations in progenitors and stem cells from chronic myeloid leukemia patients. Stem Cells Cancer Stem Cells. 8:305–315. 2012. View Article : Google Scholar