O-GlcNAcylation in early stages of chronic lymphocytic leukemia: Protocol development for flow cytometry

Autor:	Kinga Molnár, Attila Miseta, Árpád Szomor, Péter Kaltenecker, Hussain Alizadeh, Viktória Temesfői, Tamás Nagy, Béla Kajtár, Barbara Réger, Zsuzsanna Faust, Tamás Kőszegi, Zoltán Horváth-Szalai
Rok vydání:	2021
Předmět:	Male Cancer Research Glycosylation Lymphocyte Chronic lymphocytic leukemia Disease Carbohydrate metabolism Malignancy Proof of Concept Study Flow cytometry 03 medical and health sciences 0302 clinical medicine Genetics medicine Humans Stage (cooking) 030304 developmental biology Neoplasm Staging 0303 health sciences medicine.diagnostic_test business.industry General Medicine medicine.disease Flow Cytometry Prognosis Leukemia Lymphocytic Chronic B-Cell Red blood cell medicine.anatomical_structure Oncology 030220 oncology & carcinogenesis Case-Control Studies Immunology Female business
Zdroj:	Cancer biomarkers : section A of Disease markers. 32(3)
ISSN:	1875-8592
Popis:	BACKGROUND: 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. OBJECTIVE: 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. METHODS: 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. RESULTS: 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). CONCLUSION: 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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::256a4ae26d0ffe30163e685205c237b2 https://pubmed.ncbi.nlm.nih.gov/34151834 Zobrazit plný text záznamu Plný text ve formátu PDF Plný text ve formátu HTML
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