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Sensitivity and Resistance to Therapy

NF-κB-dependent DNA damage-signaling differentially regulates DNA double-strand break repair mechanisms in immature and mature human hematopoietic cells

Leukemia volume 29, pages 1543–1554 (2015)Cite this article

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Abstract

Hematopoietic stem and progenitor cells (HSPC), that is, the cell population giving rise not only to all mature hematopoietic lineages but also the presumed target for leukemic transformation, can transmit (adverse) genetic events, such as are acquired from chemotherapy or ionizing radiation. Data on the repair of DNA double-strand-breaks (DSB) and its accuracy in HSPC are scarce, in part contradictory, and mostly obtained in murine models. We explored the activity, quality and molecular components of DSB repair in human HSPC as compared with mature peripheral blood lymphocytes (PBL). To consider chemotherapy/radiation-induced compensatory proliferation, we established cycling HSPC cultures. Comparison of pathway-specific repair activities using reporter systems revealed that HSPC were severely compromised in non-homologous end joining and homologous recombination but not microhomology-mediated end joining. We observed a more pronounced radiation-induced accumulation of nuclear 53BP1 in HSPC relative to PBL, despite evidence for comparable DSB formation from cytogenetic analysis and γH2AX signal quantification, supporting differential pathway usage. Functional screening excluded a major influence of phosphatidylinositol-3-OH-kinase (ATM/ATR/DNA-PK)- and p53-signaling as well as chromatin remodeling. We identified diminished NF-κB signaling as the molecular component underlying the observed differences between HSPC and PBL, limiting the expression of DSB repair genes and bearing the risk of an inaccurate repair.

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Acknowledgements

We are indebted to Jeremy M. Stark, Beckman Research Institute of the City of Hope, Duarte, CA, USA for providing plasmid EJ5SceGFP. We thank Marco Durante, Darmstadt, Germany, for his continuous support. This work was supported by the European/German Space Agency (ESA/DLR) and German Ministry of Economy (BMWi): A0-10-IBER-2 funding 50WB1225, the German Research Foundation (DFG, PA3 in the Research Training Group GRK1789 ‘Cellular and Molecular Mechanisms in aging’, CEMMA and Research Training Group GRK1657) as well as by a postdoc fellowship of the Alexander von Humboldt Foundation to DS. HB is a member of the LOEWE Cell and Gene Therapy Frankfurt faculty funded by Hessian Ministry of Higher Education, Research and the Arts ref.no.: III L 4-518/17.004 (2010).

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Author notes

  1. E Nasonova

    Present address: 6Present address: Joint Institute for Nuclear Research, 141980 Dubna, Russia.,

  2. D Kraft, M Rall and M Volcic: These authors contributed equally to this work.

  3. C Fournier and L Wiesmüller: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biophysics, GSI Helmholtz Center for Heavy Ion Research, Planckstrasse, Darmstadt, Germany

    D Kraft, E Metzler, A Groo, L Bauer, E Nasonova, G Taucher-Scholz & C Fournier

  2. Department of Obstetrics and Gynecology, Ulm University, Prittwitzstrasse, Ulm, Germany

    M Rall, M Volcic, A Stahl, D Salles & L Wiesmüller

  3. Department of Cellular Therapeutics/Cell Processing, DRK-Blutspendedienst Baden-Württemberg – Hessen, Sandhofstrasse, Frankfurt, Germany

    H Bönig

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  1. D Kraft
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Kraft, D., Rall, M., Volcic, M. et al. NF-κB-dependent DNA damage-signaling differentially regulates DNA double-strand break repair mechanisms in immature and mature human hematopoietic cells. Leukemia 29, 1543–1554 (2015). https://doi.org/10.1038/leu.2015.28

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