Temesfői, Viktória and Molnár, Kinga and Kaltenecker, Péter and Réger, Barbara and Szomor, Árpád and Horváth-Szalai, Zoltán and Alizadeh, Hussain and Kajtár, Béla and Kőszegi, Tamás and Miseta, Attila János and Nagy, Tamás and Faust, Zsuzsanna (2021) O-GlcNAcylation in early stages of chronic lymphocytic leukemia; protocol development for flow cytometry. CANCER BIOMARKERS, 32 (3). pp. 353-362. ISSN 1574-0153
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Abstract
Recent studies proved that metabolic changes in malignant disorders have an impact on protein glycosylation, however, only a few attempts have been made so far to use O-GlcNAc analysis as a prognostic tool. Glucose metabolism is reported to be altered in hematological malignancies thus, we hypothesized that monitoring intracellular O-GlcNAc levels in Rai stage 0-I (Binet A) CLL patients could give deeper insights regarding subtle metabolic changes of progression which are not completely detected by the routine follow-up procedures.In this proof of concept study we established a flow cytometric detection method for the assessment of O-GlcNAcylation as a possible prognostic marker in CLL malignancy which was supported by fluorescence microscopy.Healthy volunteers and CLL patients were recruited for this study. Lymphocytes were isolated, fixed and permeabilised by various methods to find the optimal experimental condition for O-GlcNAc detection by flow cytometry. O-GlcNAc levels were measured and compared to lymphocyte count and various blood parameters including plasma glucose level.The protocol we developed includes red blood cell lysis, formalin fixation, 0.1% Tween 20 permeabilisation and employs standardized cell number per sample and unstained controls. We have found significant correlation between O-GlcNAc levels and WBC (R2= 0.8535, p< 0.0029) and lymphocyte count (R2= 0.9225, p< 0.0006) in CLL patients. Interestingly, there was no such correlation in healthy individuals (R2= 0.05664 for O-GlcNAc vs WBC and R2= 0.04379 for O-GlcNAc vs lymphocytes).Analyzing O-GlcNAc changes in malignant disorders, specifically in malignant hematologic diseases such as CLL, could be a useful tool to monitor the progression of the disease.
| Item Type: | Article |
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| Additional Information: | Funding Agency and Grant Number: University of Pecs, Medical School [KA-2018-17, KA-2018-21, KA-2019-36, KA-2019-28, EFOP 3.6.1-16-201600004, GINOP-2.3.2-15-2016-00021]; Higher Education Institutional Excellence Program of the Ministry for Innovation and Technology in Hungary [FIKP II.2] Funding text: This work was supported by grants from University of Pecs, Medical School, KA-2018-17; KA-2018-21; KA-2019-36; KA-2019-28 and EFOP 3.6.1-16-201600004 (Comprehensive Development for Implementing Smart Specialization Strategies at the University of Pecs); GINOP-2.3.2-15-2016-00021; and Higher Education Institutional Excellence Program of the Ministry for Innovation and Technology in Hungary, within the framework of the second thematic program of the University of Pecs (FIKP II.2). |
| Uncontrolled Keywords: | Flow Cytometry; Chronic lymphocytic leukemia; Immunometabolism; O-GlcNAcylation; RL2; |
| Subjects: | R Medicine / orvostudomány > RC Internal medicine / belgyógyászat > RC0254 Neoplasms. Tumors. Oncology (including Cancer) / daganatok, tumorok, onkológia |
| SWORD Depositor: | MTMT SWORD |
| Depositing User: | MTMT SWORD |
| Date Deposited: | 08 Feb 2022 14:50 |
| Last Modified: | 08 Feb 2022 14:50 |
| URI: | http://real.mtak.hu/id/eprint/137496 |
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