Abstract
Mitochondrial metabolism is a tightly regulated process that plays a central role throughout the lifespan of hematopoietic cells. Herein, we analyze the consequences of the mitochondrial oxidative phosphorylation (OXPHOS)/metabolism disorder associated with the cell-specific hematopoietic ablation of apoptosis-inducing factor (AIF). AIF-null (AIF−/Y) mice developed pancytopenia that was associated with hypocellular bone marrow (BM) and thymus atrophy. Although myeloid cells were relatively spared, the B-cell and erythroid lineages were altered with increased frequencies of precursor B cells, pro-erythroblasts I, and basophilic erythroblasts II. T-cell populations were dramatically reduced with a thymopoiesis blockade at a double negative (DN) immature state, with DN1 accumulation and delayed DN2/DN3 and DN3/DN4 transitions. In BM cells, the OXPHOS/metabolism dysfunction provoked by the loss of AIF was counterbalanced by the augmentation of the mitochondrial biogenesis and a shift towards anaerobic glycolysis. Nevertheless, in a caspase-independent process, the resulting excess of reactive oxygen species compromised the viability of the hematopoietic stem cells (HSC) and progenitors. This led to the progressive exhaustion of the HSC pool, a reduced capacity of the BM progenitors to differentiate into colonies in methylcellulose assays, and the absence of cell-autonomous HSC repopulating potential in vivo. In contrast to BM cells, AIF−/Y thymocytes compensated for the OXPHOS breakdown by enhancing fatty acid β-oxidation. By over-expressing CPT1, ACADL and PDK4, three key enzymes facilitating fatty acid β-oxidation (e.g., palmitic acid assimilation), the AIF−/Y thymocytes retrieved the ATP levels of the AIF+/Y cells. As a consequence, it was possible to significantly reestablish AIF−/Y thymopoiesis in vivo by feeding the animals with a high-fat diet complemented with an antioxidant. Overall, our data reveal that the mitochondrial signals regulated by AIF are critical to hematopoietic decision-making. Emerging as a link between mitochondrial metabolism and hematopoietic cell fate, AIF-mediated OXPHOS regulation represents a target for the development of new immunomodulatory therapeutics.
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Acknowledgements
We thank G Delespesse, M Rubio, K Waahara, and N Baba (CRCHUM, Montréal, Canada) for support and helpful tips and discussions, Sophie Ezine and Victoria Michaels (INEM UMR-S1151, CNRS UMR 8253 Université Paris Descartes-Site Broussais, Paris, France) for invaluable help in thymocyte labeling and flow cytometry analysis, the animal facility staff (CEF Cordeliers, Paris, France) for mice housing support, Christophe Klein for help in confocal image acquisition (CICC, Cordeliers, Paris, France), V Della Valle and D Roos-Weil (U1170 INSERM, Villejuif, France) for advice in transplantation and colony-forming assays, A Prola and C Lemaire (UMRS 1180, Châtenay-Malabry, France) for help in the set-up of Clark electrode oxygen consumption assays and for providing amytal, and M Segade (Houston, US) for proofreading. This work was supported by Fondation ARC (PJA20151203407), Ligue Contre le Cancer-Comité de Paris (RS15/75-63 and RS16/75-50), Association Laurette Fugain (ALF 15/09), Fondation de France-Comité Cancer, Fondation pour la Recherche Medicale, French National Research Agency (ANR-09-BLAN-0247 and ANR-12-EMMA-0045), and French National Cancer Institute (INCa-5839) (to SA Susin). LC received PhD fellowship support from ENS-Cachan, Société Française d’Hématologie, and Fondation ARC, as well as travel grants from Boehringer Ingelheim and Journal of Cell Science. AB received PhD fellowships from the French Research Ministry and Société Française d’Hématologie.
Author Contributions
LC and AB designed experiments, performed in vitro and in vivo studies, interpreted the data and helped to write the manuscript. M-NB-N, LS, LD, LV, and MB, performed in vitro and/or in vivo experiments. IN designed and performed the Seahorse metabolic assessment. CL performed histological analyses. KG and DC carried out electron microscopy. FG, performed metabolomic approach. VQV, MS, TM, and OAB provided reagents, analyzed data, and/or contributed to the experimental design and analysis. SAS supervised the study, designed experiments, interpreted the data, and wrote the manuscript.
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Lauriane Cabon and Audrey Bertaux share first authorship.
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Cabon, L., Bertaux, A., Brunelle-Navas, MN. et al. AIF loss deregulates hematopoiesis and reveals different adaptive metabolic responses in bone marrow cells and thymocytes. Cell Death Differ 25, 983–1001 (2018). https://doi.org/10.1038/s41418-017-0035-x
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DOI: https://doi.org/10.1038/s41418-017-0035-x
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