[en] Oxazolidinones (linezolid and tedizolid) adverse reactions include thrombocytopenia, the mechanism of which is still largely unknown. In cultured cells, oxazolidinones impair mitochondrial protein synthesis and oxidative metabolism. As mitochondrial activity is essential for megakaryocyte differentiation and maturation into platelets, we examined whether oxazolidinones impair these processes ex vivo and alter, in parallel, the activity of mitochondrial cytochrome c-oxidase (CYTOX; enzyme partly encoded by the mitochondrial genome) and cell morphology. Human CD34+ cells were isolated, incubated with cytokines (up to 14 days) and clinically relevant oxazolidinone concentrations or in control conditions, and used for (i) clonogenic assays [counting of megakaryocyte (CFU-Mk), granulocyte-monocyte (CFU-GM), burst-forming unit-erythroid (BFU-E) colonies]; (ii) the measure of the expression of megakaryocyte surface antigens (CD34 to CD41 and CD42); (iii) counting of proplatelets; (iv) the measurement of CYTOX activity; and (v) cell morphology (optic and electron microscopy). Oxazolidinones caused a significant decrease in BFU-E but not CFU-Mk or CFU-GM colonies. Yet, the megakaryocytic lineage was markedly affected, with a decreased differentiation of CD34+ into CD41+/CD42+ cells, an abolition of proplatelet formation and striking decrease in the numbers of large polylobulated nucleus megakaryocytes, with a complete loss of intracellular demarcation membrane system, disappearance of mitochondria, and suppression of CYTOX activity. These alterations were more marked in cells incubated with tedizolid than linezolid. These data suggest that oxazolidinones may induce thrombocytopenia by impairing megakaryocytic differentiation through mitochondrial dysfunction. Pharmacological interventions to prevent this toxicity might therefore be difficult as mitochondrial toxicity is most probably inherently linked to their antibacterial activity.
Disciplines :
Hematology
Author, co-author :
Milosevic, Tamara V; Pharmacologie cellulaire et moléculaire, Louvain Drug Research Institute, Université catholique de Louvain (UCLouvain), Brussels, Belgium
VERTENOEIL, Gaëlle ✱; Centre Hospitalier Universitaire de Liège - CHU > > Service d'hématologie clinique ; Signal Transduction and Molecular Hematology Unit (SIGN), de Duve Institute, Université catholique de Louvain (UCLouvain), Brussels, Belgium ; Ludwig Institute for Cancer Research, Brussels, Belgium
Vainchenker, William; UMR 1170, Institut National de la Santé et de la Recherche Médicale, Université de Paris-Sud & Institut Gustave Roussy, Villejuif, France
Tulkens, Paul M ; Pharmacologie cellulaire et moléculaire, Louvain Drug Research Institute, Université catholique de Louvain (UCLouvain), Brussels, Belgium
Constantinescu, Stefan N; Signal Transduction and Molecular Hematology Unit (SIGN), de Duve Institute, Université catholique de Louvain (UCLouvain), Brussels, Belgium ; Ludwig Institute for Cancer Research, Brussels, Belgium ; WELBIO Department, WEL Research Institute, Wavre, Belgium ; Nuffield Department of Medicine, Ludwig Institute for Cancer Research, University of Oxford, Oxford, United Kingdom
Van Bambeke, Françoise ; Pharmacologie cellulaire et moléculaire, Louvain Drug Research Institute, Université catholique de Louvain (UCLouvain), Brussels, Belgium
✱ These authors have contributed equally to this work.
Language :
English
Title :
Oxazolidinone antibiotics impair ex vivo megakaryocyte differentiation from hematopoietic progenitor cells and their maturation into platelets.
LICR - Ludwig Institute for Cancer Research Stichting Tegen Kanker Salus Sanguinis Les Avions de Sébastien ASBL WRI. WELBIO - Wel Research Institute. Walloon Excellence in Life Sciences and Biotechnology Ligue Contre le Cancer INCa - Institut National du Cancer INSERM - Institut National de la Santé et de la Recherche Médicale
Funding text :
P.M.T. and F.V.B. have received speaker's and advisory boards' honoraria from Bayer and Merck (holders of marketing rights of tedizolid) and research grants from Trius Pharmaceuticals (now part of Merck) for preclinical studies of tedizolid. These companies were not involved in the design and performance of the studies presented here and did not take any part in their interpretation and/or decision to submit them to publication. The other authors have no conflict of interest to disclose in relation to the present work.We thank Prof. Carine Michiels (Universit\u00E9 de Namur, Namur, Belgium) for help in the preparation of samples for electron microscopy and Novartis Pharma AG, Basel, Switzerland, for providing us the sample of eltrombopag used in our studies. Virginie Mohymont and Katia Santos-Saial (Universit\u00E9 catholique de Louvain), and Lidvine Genet and Caroline De Bona (Universit\u00E9 de Namur) provided dedicated technical assistance. We thank Dr. Nicolas Dauguet for flow cytometry assistance. T.V.M. was supported by the Universit\u00E9 catholique de Louvain and G.V. by a PhD Mandat Aspirant of the FRS-FNRS Belgium. F.V.B. is Research Director of FRS-FNRS Belgium. This work was supported by the general budget awarded from various sources to F.V.B. Funding to S.N.C. is acknowledged from Ludwig Institute for Cancer Research, Fondation contre le cancer, Salus Sanguinis, and Fondation \u201CLes avions de S\u00E9bastien\u201D, projects Actions de recherche concert\u00E9e (ARC) 16/21\u2013073 and Wel Research Institute WelBio F 44/8/5 - MCF/UIG \u2013 10955. Funding of W.V. is acknowledged from Ligue Nationale Contre le Cancer (\u201CEquipe labellis\u00E9e 2016\u201D, H.R.), Institut National du Cancer (INCA-PLBIO-2015, I.P.), and Institut National de la Sant\u00E9 et de la Recherche M\u00E9dicale (Inserm). T.V.M. and G.V. designed and performed all experiments and analyzed the results. W.V. examined and interpreted the electron microscopy images presented here and made substantial contributions to the interpretation of all data. S.N.C. provided guidance throughout the study and made essential contributions in the interpretation of all data. P.M.T. and F.V.B. initiated the work, supervised all steps of the studies, and actively participated in the interpretation of all data. T.M., P.M.T., and F.V.B. wrote the paper. All authors commented, made corrections, and approved the submitted version. Funder Grant(s) Author(s) Ludwig Institute for Cancer Research Stefan N. Constantinescu Stichting Tegen Kanker (Fondation Contre le Cancer) Stefan N. Constantinescu Salus Sanguinis Stefan N. Constantinescu Les avions de S\u00E9bastien Stefan N. Constantinescu Actions de recherche Concert\u00E9e 16/21-073 Stefan N. Constantinescu Wel research Institute Welbio F 44/8/5 - MCF/UIG - 10955 Stefan N. Constantinescu Ligue Contre le Cancer (French League Against Cancer) William Vainchenker Institut National Du Cancer (INCa) INCA-PLBIO-2015 William Vainchenker Institut National de la Sant\u00E9 et de la Recherche M\u00E9dicale (Inserm) William VainchenkerT.V.M. was supported by the Universit\u00E9 catholique de Louvain and G.V. by a PhD Mandat Aspirant of the FRS-FNRS Belgium. F.V.B. is Research Director of FRS-FNRS Belgium. This work was supported by the general budget awarded from various sources to F.V.B. Funding to S.N.C. is acknowledged from Ludwig Institute for Cancer Research, Fondation contre le cancer, Salus Sanguinis, and Fondation \u201CLes avions de S\u00E9bastien\u201D, projects Actions de recherche concert\u00E9e (ARC) 16/21\u2013073 and Wel Research Institute WelBio F 44/8/5 - MCF/UIG \u2013 10955. Funding of W.V. is acknowledged from Ligue Nationale Contre le Cancer (\u201CEquipe labellis\u00E9e 2016\u201D, H.R.), Institut National du Cancer (INCA-PLBIO-2015, I.P.), and Institut National de la Sant\u00E9 et de la Recherche M\u00E9dicale (Inserm).
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