Exosomes released by imatinib‑resistant K562 cells contain specific membrane markers, IFITM3, CD146 and CD36 and increase the survival of imatinib‑sensitive cells in the presence of imatinib

Hrdinova,Tereza; Toman,Ondrej; Dresler,Jiri; Klimentova,Jana; Salovska,Barbora; Pajer,Petr; Bartos,Oldrich; Polivkova,Vaclava; Linhartova,Jana; Machova Polakova,Katerina; Kabickova,Hana; Brodska,Barbora; Krijt,Matyas; Zivny,Jan; Vyoral,Daniel; Petrak,Jiri

doi:10.3892/ijo.2020.5163

Exosomes released by imatinib‑resistant K562 cells contain specific membrane markers, IFITM3, CD146 and CD36 and increase the survival of imatinib‑sensitive cells in the presence of imatinib

Authors:
- Tereza Hrdinova
- Ondrej Toman
- Jiri Dresler
- Jana Klimentova
- Barbora Salovska
- Petr Pajer
- Oldrich Bartos
- Vaclava Polivkova
- Jana Linhartova
- Katerina Machova Polakova
- Hana Kabickova
- Barbora Brodska
- Matyas Krijt
- Jan Zivny
- Daniel Vyoral
- Jiri Petrak
View Affiliations

Affiliations: Institute of Hematology and Blood Transfusion, 128 20 Prague 2, Czech Republic, Institute of Hematology and Blood Transfusion, 128 20 Prague 2, Czech Republic, Military Health Institute, Military Medical Agency, 160 01 Prague 6, Czech Republic, Faculty of Military Health Sciences, University of Defense in Brno, 500 02 Hradec Kralove, Czech Republic, Department of Genome Integrity, Institute of Molecular Genetics of The Czech Academy of Sciences, 142 20 Prague 4, Czech Republic, Department of Infectious Diseases, First Faculty of Medicine, Charles University and Military University Hospital Prague, 169 02 Prague 6, Czech Republic, Institute of Pathological Physiology, First Faculty of Medicine, Charles University, 128 20 Prague 2, Czech Republic
Published online on: December 23, 2020 https://doi.org/10.3892/ijo.2020.5163
Pages: 238-250

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Abstract

Chronic myeloid leukemia (CML) is a malignant hematopoietic disorder distinguished by the presence of a BCR‑ABL1 fused oncogene with constitutive kinase activity. Targeted CML therapy by specific tyrosine kinase inhibitors (TKIs) leads to a marked improvement in the survival of the patients and their quality of life. However, the development of resistance to TKIs remains a critical issue for a subset of patients. The most common cause of resistance are numerous point mutations in the BCR‑ABL1 gene, followed by less common mutations and multiple mutation-independent mechanisms. Recently, exosomes, which are extracellular vesicles excreted from normal and tumor cells, have been associated with drug resistance and cancer progression. The aim of the present study was to characterize the exosomes released by imatinib‑resistant K562 (K562IR) cells. The K562IR‑derived exosomes were internalized by imatinib‑sensitive K562 cells, which thereby increased their survival in the presence of 2 µM imatinib. The exosomal cargo was subsequently analyzed to identify resistance‑associated markers using a deep label‑free quantification proteomic analysis. There were >3,000 exosomal proteins identified of which, 35 were found to be differentially expressed. From this, a total of 3, namely the membrane proteins, interferon‑induced transmembrane protein 3, CD146 and CD36, were markedly upregulated in the exosomes derived from the K562IR cells, and exhibited surface localization. The upregulation of these proteins was verified in the K562IR exosomes, and also in the K562IR cells. Using flow cytometric analysis, it was possible to further demonstrate the potential of CD146 as a cell surface marker associated with imatinib resistance in K562 cells. Taken together, these results suggested that exosomes and their respective candidate surface proteins could be potential diagnostic markers of TKI drug resistance in CML therapy.

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