Abstract
B-cell development in the bone marrow comprises proliferative and resting phases in different niches. We asked whether B-cell metabolism relates to these changes. Compared to pro B and small pre B cells, large pre B cells revealed the highest glucose uptake and ROS but not mitochondrial mass, whereas small pre B cells exhibited the lowest mitochondrial membrane potential. Small pre B cells from Rag1−/−;33.C9 μ heavy chain knock-in mice revealed decreased glycolysis (ECAR) and mitochondrial spare capacity compared to pro B cells from Rag1−/− mice. We were interested in the step regulating this metabolic switch from pro to pre B cells and uncovered that Swiprosin-2/EFhd1, a Ca2+-binding protein of the inner mitochondrial membrane involved in Ca2+-induced mitoflashes, is expressed in pro B cells, but downregulated by surface pre B-cell receptor expression. Knockdown and knockout of EFhd1 in 38B9 pro B cells decreased the oxidative phosphorylation/glycolysis (OCR/ECAR) ratio by increasing glycolysis, glycolytic capacity and reserve. Prolonged expression of EFhd1 in EFhd1 transgenic mice beyond the pro B cell stage increased expression of the mitochondrial co-activator PGC-1α in primary pre B cells, but reduced mitochondrial ATP production at the pro to pre B cell transition in IL-7 cultures. Transgenic EFhd1 expression caused a B-cell intrinsic developmental disadvantage for pro and pre B cells. Hence, coordinated expression of EFhd1 in pro B cells and by the pre BCR regulates metabolic changes and pro/pre B-cell development.
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Acknowledgements
We thank Dr. Carlo Croce for the VHP-EμH vector used to generate the transgene cassette, Dr. Roberta Pelanda for pIVHL2neo, Dr. Irmgard Förster for Rag1−/− mice, Dr. Chris Paige for R5B cells and Dr. Jan Tuckermann for support. We also thank Drs. Mark Shlomcik and Florian Weisel for helpful comments during preparation of this manuscript. We thank Uwe Appelt and Markus Mroz for cell sorting and Stefan Hofmann as well as Domenica Saul for technical assistance. This work was supported by grants from the Deutsche Forschungsgemeinschaft (DFG) (TRR130, to DM, H-MJ, THW; SPP1468, to DM and AB, IRTG1660, to DM, H-MJ, THW).
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MS, SD and DM designed experiments, performed experiments, analyzed data and wrote the paper. DM designed experiments and wrote the paper. MB and THW designed and performed the experiments. KF, DR, SU and TS performed experiments. AB, WS and H-MJ provided important conceptual input and wrote the paper.
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Stein, M., Dütting, S., Mougiakakos, D. et al. A defined metabolic state in pre B cells governs B-cell development and is counterbalanced by Swiprosin-2/EFhd1. Cell Death Differ 24, 1239–1252 (2017). https://doi.org/10.1038/cdd.2017.52
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DOI: https://doi.org/10.1038/cdd.2017.52
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