Phenotypic and functional characterization of subpopulation of Imatinib resistant chronic myeloid leukemia cell line

Yalda Hekmatshoar; Aynur Karadag Gurel; Tulin Ozkan; Yalda Rahbar Saadat; Asli Koc; Arzu Zeynep Karabay; Sureyya Bozkurt; Asuman Sunguroglu

doi:10.1016/j.advms.2023.06.002

Phenotypic and functional characterization of subpopulation of Imatinib resistant chronic myeloid leukemia cell line

Adv Med Sci. 2023 Sep;68(2):238-248. doi: 10.1016/j.advms.2023.06.002. Epub 2023 Jul 6.

Authors

Yalda Hekmatshoar¹, Aynur Karadag Gurel², Tulin Ozkan³, Yalda Rahbar Saadat⁴, Asli Koc⁵, Arzu Zeynep Karabay⁵, Sureyya Bozkurt⁶, Asuman Sunguroglu³

Affiliations

¹ Department of Medical Biology, School of Medicine, Altinbas University, Istanbul, Turkey; Department of Medical Biology, School of Medicine, Ankara University, Ankara, Turkey. Electronic address: yalda.hekmatshoar@altinbas.edu.tr.
² Department of Medical Biology, School of Medicine, Usak University, Usak, Turkey. Electronic address: aynur.karadag@usak.edu.tr.
³ Department of Medical Biology, School of Medicine, Ankara University, Ankara, Turkey.
⁴ Kidney Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
⁵ Faculty of Pharmacy, Department of Biochemistry, Ankara University, Ankara, Turkey.
⁶ Department of Medical Biology, School of Medicine, Istinye University, Istanbul, Turkey.

PMID: 37421850
DOI: 10.1016/j.advms.2023.06.002

Abstract

Purpose: Chronic myeloid leukemia (CML) is a hematological malignancy characterized by the presence of BCR-ABL protein. Imatinib (IMA) is considered as the first line therapy in management of CML which particularly targets the BCR-ABL tyrosine kinase protein. However, emergence of resistance to IMA hinders its clinical efficiency. Hence, identifying novel targets for therapeutic approaches in CML treatment is of great importance. Here, we characterize a new subpopulation of highly adherent IMA-resistant CML cells that express stemness and adhesion markers compared to naive counterparts.

Materials and methods: We performed several experimental assays including FISH, flow cytometry, and gene expression assays. Additionally, bioinformatics analysis was performed by normalized web-available microarray data (GSE120932) to revalidate and introduce probable biomarkers. Protein-protein interactions (PPI) network was analyzed by the STRING database employing Cytoscape v3.8.2.

Results: Our findings demonstrated that constant exposure to 5 μM IMA led to development of the adherent phenotype (K562R-adh). FISH and BCR-ABL expression analysis indicated that K562R-adh cells were derived from the original cells (K562R). In order to determine the role of various genes involved in epithelial-mesenchymal transition (EMT) and stem cell characterization, up/down-regulation of various genes including cancer stem cell (CSC), adhesion and cell surface markers and integrins were observed which was similar to the findings of the GSE120932 dataset.

Conclusion: Treating CML patients with tyrosine kinase inhibitors (TKIs) as well as targeting adhesion molecules deemed to be effective approaches in prevention of IMA resistance emergence which in turn may provide promising effects in the clinical management of CML patients.

Keywords: Chronic myeloid leukemia; Drug resistance; Epithelial–mesenchymal transition; Phenotype switching; Tyrosine kinase inhibitors.

MeSH terms

Apoptosis
Drug Resistance, Neoplasm / genetics
Fusion Proteins, bcr-abl / genetics
Fusion Proteins, bcr-abl / metabolism
Fusion Proteins, bcr-abl / pharmacology
Humans
Imatinib Mesylate / pharmacology
Imatinib Mesylate / therapeutic use
K562 Cells
Leukemia, Myelogenous, Chronic, BCR-ABL Positive* / drug therapy
Leukemia, Myelogenous, Chronic, BCR-ABL Positive* / genetics
Leukemia, Myelogenous, Chronic, BCR-ABL Positive* / pathology
Phenotype
Protein Kinase Inhibitors* / pharmacology
Protein Kinase Inhibitors* / therapeutic use

Substances

Imatinib Mesylate
Protein Kinase Inhibitors
Fusion Proteins, bcr-abl