BCR::ABL1 expression in chronic myeloid leukemia cells in low oxygen is regulated by glutamine via CD36-mediated fatty acid uptake

Abstract Background Chronic myeloid leukemia (CML) is influenced by microenvironmental nutrients, glucose (Glc), and glutamine (Gln) which regulate cell proliferation, viability, and the expression of the driver oncoprotein (BCR::ABL1). Results Our study revealed that Glc, while partially supporting...

Full description

Saved in:
Bibliographic Details
Main Authors: Caterina Mancini, Giulio Menegazzi, Silvia Peppicelli, Giampaolo Versienti, Daniele Guasti, Giuseppe Pieraccini, Elisabetta Rovida, Matteo Lulli, Laura Papucci, Persio Dello Sbarba, Alessio Biagioni
Format: Article
Language:English
Published: BMC 2025-05-01
Series:Cancer Cell International
Subjects:
Online Access:https://doi.org/10.1186/s12935-025-03805-y
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850154763320033280
author Caterina Mancini
Giulio Menegazzi
Silvia Peppicelli
Giampaolo Versienti
Daniele Guasti
Giuseppe Pieraccini
Elisabetta Rovida
Matteo Lulli
Laura Papucci
Persio Dello Sbarba
Alessio Biagioni
author_facet Caterina Mancini
Giulio Menegazzi
Silvia Peppicelli
Giampaolo Versienti
Daniele Guasti
Giuseppe Pieraccini
Elisabetta Rovida
Matteo Lulli
Laura Papucci
Persio Dello Sbarba
Alessio Biagioni
author_sort Caterina Mancini
collection DOAJ
description Abstract Background Chronic myeloid leukemia (CML) is influenced by microenvironmental nutrients, glucose (Glc), and glutamine (Gln) which regulate cell proliferation, viability, and the expression of the driver oncoprotein (BCR::ABL1). Results Our study revealed that Glc, while partially supporting alone cell growth in normoxia, is essential in low oxygen conditions, whereas Gln is ineffective. Under low oxygen, Gln reduced oxidative respiratory activity while enhancing glycolysis. In these conditions, fatty acid (FA) metabolism becomes crucial, as evidenced by increased lipid droplets (LD) accumulation when Glc was absent. Gln, in particular, drives CD36-mediated FA uptake, suppressing the BCR::ABL1 oncoprotein and facilitating cell survival. By co-culturing leukemia cells with adipocytes, one of the main bone marrow (BM) cell components, we observed an enhanced FA release, suggesting a link between FA, microenvironmental BM cells, and the maintenance of leukemic stem cells (LSC). Methods K562 and KCL22 cell lines were subjected to Glc and/or Gln deprivation under hypoxic conditions (96 h at 0.1% O2). Metabolic profiling was conducted through the Seahorse XFe96 analyzer, and the contribution of L-Glutamine-13C5 to FA de novo synthesis was determined via GC/MS. Intracellular neutral LD were measured using BODIPY 493/503 in confocal microscopy and flow cytometry, with their presence and morphology further examined via transmission electron microscopy. BCR::ABL1 as well as several FA-related markers were evaluated via Western Blotting, whilst CD36 was determined through flow cytometry. LC2 assay was used for measuring leukemia stem cell potential by inhibiting FA uptake via the usage of the Sulfo-N-Succinimidyl Oleate, a CD36 inhibitor. qPCR was exploited to detect markers of FA secretion in CML-adipocytes co-culture together with Nile Red staining to assess free FA in the media. Conclusions These findings underscore the central role of FA in the regulation of the LSC compartment of CML, highlighting the importance of Gln in facilitating CML cell survival under restrictive metabolic conditions and preparing the cell population for expansion upon the release of these restrictions.
format Article
id doaj-art-a51f08a5b32b4b96971cdb1ff9ad8e3d
institution OA Journals
issn 1475-2867
language English
publishDate 2025-05-01
publisher BMC
record_format Article
series Cancer Cell International
spelling doaj-art-a51f08a5b32b4b96971cdb1ff9ad8e3d2025-08-20T02:25:12ZengBMCCancer Cell International1475-28672025-05-0125111610.1186/s12935-025-03805-yBCR::ABL1 expression in chronic myeloid leukemia cells in low oxygen is regulated by glutamine via CD36-mediated fatty acid uptakeCaterina Mancini0Giulio Menegazzi1Silvia Peppicelli2Giampaolo Versienti3Daniele Guasti4Giuseppe Pieraccini5Elisabetta Rovida6Matteo Lulli7Laura Papucci8Persio Dello Sbarba9Alessio Biagioni10Department of Experimental and Clinical Medicine, Università degli Studi di FirenzeDepartment of Experimental and Clinical Biomedical Sciences “Mario Serio”, Università degli Studi di FirenzeDepartment of Experimental and Clinical Biomedical Sciences “Mario Serio”, Università degli Studi di FirenzeDepartment of Experimental and Clinical Biomedical Sciences “Mario Serio”, Università degli Studi di FirenzeDepartment of Experimental and Clinical Medicine, Università degli Studi di FirenzeCISM Mass Spectrometry Center, Università degli Studi di FirenzeDepartment of Experimental and Clinical Biomedical Sciences “Mario Serio”, Università degli Studi di FirenzeDepartment of Experimental and Clinical Biomedical Sciences “Mario Serio”, Università degli Studi di FirenzeDepartment of Experimental and Clinical Biomedical Sciences “Mario Serio”, Università degli Studi di FirenzeDepartment of Experimental and Clinical Biomedical Sciences “Mario Serio”, Università degli Studi di FirenzeDepartment of Experimental and Clinical Biomedical Sciences “Mario Serio”, Università degli Studi di FirenzeAbstract Background Chronic myeloid leukemia (CML) is influenced by microenvironmental nutrients, glucose (Glc), and glutamine (Gln) which regulate cell proliferation, viability, and the expression of the driver oncoprotein (BCR::ABL1). Results Our study revealed that Glc, while partially supporting alone cell growth in normoxia, is essential in low oxygen conditions, whereas Gln is ineffective. Under low oxygen, Gln reduced oxidative respiratory activity while enhancing glycolysis. In these conditions, fatty acid (FA) metabolism becomes crucial, as evidenced by increased lipid droplets (LD) accumulation when Glc was absent. Gln, in particular, drives CD36-mediated FA uptake, suppressing the BCR::ABL1 oncoprotein and facilitating cell survival. By co-culturing leukemia cells with adipocytes, one of the main bone marrow (BM) cell components, we observed an enhanced FA release, suggesting a link between FA, microenvironmental BM cells, and the maintenance of leukemic stem cells (LSC). Methods K562 and KCL22 cell lines were subjected to Glc and/or Gln deprivation under hypoxic conditions (96 h at 0.1% O2). Metabolic profiling was conducted through the Seahorse XFe96 analyzer, and the contribution of L-Glutamine-13C5 to FA de novo synthesis was determined via GC/MS. Intracellular neutral LD were measured using BODIPY 493/503 in confocal microscopy and flow cytometry, with their presence and morphology further examined via transmission electron microscopy. BCR::ABL1 as well as several FA-related markers were evaluated via Western Blotting, whilst CD36 was determined through flow cytometry. LC2 assay was used for measuring leukemia stem cell potential by inhibiting FA uptake via the usage of the Sulfo-N-Succinimidyl Oleate, a CD36 inhibitor. qPCR was exploited to detect markers of FA secretion in CML-adipocytes co-culture together with Nile Red staining to assess free FA in the media. Conclusions These findings underscore the central role of FA in the regulation of the LSC compartment of CML, highlighting the importance of Gln in facilitating CML cell survival under restrictive metabolic conditions and preparing the cell population for expansion upon the release of these restrictions.https://doi.org/10.1186/s12935-025-03805-yChronic myeloid leukemiaHypoxiaBCR:ABL1Fatty acids
spellingShingle Caterina Mancini
Giulio Menegazzi
Silvia Peppicelli
Giampaolo Versienti
Daniele Guasti
Giuseppe Pieraccini
Elisabetta Rovida
Matteo Lulli
Laura Papucci
Persio Dello Sbarba
Alessio Biagioni
BCR::ABL1 expression in chronic myeloid leukemia cells in low oxygen is regulated by glutamine via CD36-mediated fatty acid uptake
Cancer Cell International
Chronic myeloid leukemia
Hypoxia
BCR:ABL1
Fatty acids
title BCR::ABL1 expression in chronic myeloid leukemia cells in low oxygen is regulated by glutamine via CD36-mediated fatty acid uptake
title_full BCR::ABL1 expression in chronic myeloid leukemia cells in low oxygen is regulated by glutamine via CD36-mediated fatty acid uptake
title_fullStr BCR::ABL1 expression in chronic myeloid leukemia cells in low oxygen is regulated by glutamine via CD36-mediated fatty acid uptake
title_full_unstemmed BCR::ABL1 expression in chronic myeloid leukemia cells in low oxygen is regulated by glutamine via CD36-mediated fatty acid uptake
title_short BCR::ABL1 expression in chronic myeloid leukemia cells in low oxygen is regulated by glutamine via CD36-mediated fatty acid uptake
title_sort bcr abl1 expression in chronic myeloid leukemia cells in low oxygen is regulated by glutamine via cd36 mediated fatty acid uptake
topic Chronic myeloid leukemia
Hypoxia
BCR:ABL1
Fatty acids
url https://doi.org/10.1186/s12935-025-03805-y
work_keys_str_mv AT caterinamancini bcrabl1expressioninchronicmyeloidleukemiacellsinlowoxygenisregulatedbyglutamineviacd36mediatedfattyaciduptake
AT giuliomenegazzi bcrabl1expressioninchronicmyeloidleukemiacellsinlowoxygenisregulatedbyglutamineviacd36mediatedfattyaciduptake
AT silviapeppicelli bcrabl1expressioninchronicmyeloidleukemiacellsinlowoxygenisregulatedbyglutamineviacd36mediatedfattyaciduptake
AT giampaoloversienti bcrabl1expressioninchronicmyeloidleukemiacellsinlowoxygenisregulatedbyglutamineviacd36mediatedfattyaciduptake
AT danieleguasti bcrabl1expressioninchronicmyeloidleukemiacellsinlowoxygenisregulatedbyglutamineviacd36mediatedfattyaciduptake
AT giuseppepieraccini bcrabl1expressioninchronicmyeloidleukemiacellsinlowoxygenisregulatedbyglutamineviacd36mediatedfattyaciduptake
AT elisabettarovida bcrabl1expressioninchronicmyeloidleukemiacellsinlowoxygenisregulatedbyglutamineviacd36mediatedfattyaciduptake
AT matteolulli bcrabl1expressioninchronicmyeloidleukemiacellsinlowoxygenisregulatedbyglutamineviacd36mediatedfattyaciduptake
AT laurapapucci bcrabl1expressioninchronicmyeloidleukemiacellsinlowoxygenisregulatedbyglutamineviacd36mediatedfattyaciduptake
AT persiodellosbarba bcrabl1expressioninchronicmyeloidleukemiacellsinlowoxygenisregulatedbyglutamineviacd36mediatedfattyaciduptake
AT alessiobiagioni bcrabl1expressioninchronicmyeloidleukemiacellsinlowoxygenisregulatedbyglutamineviacd36mediatedfattyaciduptake