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Alloreactive microenvironment after human hematopoietic cell transplantation induces genomic alterations in epithelium through an ROS-mediated mechanism: in vivo and in vitro study and implications to secondary neoplasia

Leukemia volume 24, pages 536–543 (2010)Cite this article

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Abstract

We hypothesized that chronic tissue stress due to interaction of alloreactive donor cells with host epithelium after allogeneic hematopoietic cell transplantation (allo-HCT) may cause genomic alterations. We therefore analyzed 176 buccal samples obtained from 71 unselected allotransplanted patients for microsatellite instability (MSI). MSI was observed in 52% of allotransplanted patients but never in 31 healthy or autotransplanted controls. The patient age, the donor age, a female-to-male transplantation and a low number of CD34+ cells in the graft were significantly correlated with genomic instability. There was a trend for increasing risk of MSI for patients who experienced severe graft-vs-host disease. Secondary malignancy was diagnosed in five (14%) of the MSI+ and only in one (3%) MSI− patient. In an in vitro model of mutation analysis we found significant induction of frameshift mutations and DNA strand breaks in HaCaT keratinocytes co-cultured with mixed lymphocyte cultures (MLCs) but not after their exposure to interferon-γ, tumor necrosis factor-α, transforming growth factor-β (TGF-β), MLC supernatant, peripheral blood mononuclear cells (PBMCs) or phytohemagglutinin-stimulated PBMC. A reactive oxygen species-mediated mechanism is implicated. The in vivo and in vitro data of our study show that alloreactions after allo-HCT may induce genomic alterations in epithelium. Progress in understanding DNA damage and repair after allo-HCT can potentially provide molecular biomarkers and therapeutic targets.

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Acknowledgements

We acknowledge P Faber for excellent work in setting the in vitro system, C Zouvelou for help in p53 mutation analysis and A Klamargias for help in statistical analysis. We thank S Krüger (Freiburg), G Oikonomopoulou, N Zoudiari and D Kokkinou (Patras) for excellent technical assistance and to the transplantation teams at Freiburg and Patras University for the dedicated patient care. We also thank Prof R Mertelsmann (Freiburg) for continuous support. This work was supported by grants from the Landesstiftung Baden Württemberg and the IKY- DAAD program to AS and JF.

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Authors and Affiliations

  1. Hematology Division, Department of Internal Medicine, Patras University Medical School, Patras, Greece

    M Themeli, E Lagadinou, N Zoumbos & A Spyridonidis

  2. Department of Hematology/Oncology, Albert Ludwigs University of Freiburg, Freiburg, Germany

    L Petrikkos, M Waterhouse, H Bertz & J Finke

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  1. M Themeli
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Correspondence to A Spyridonidis.

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Themeli, M., Petrikkos, L., Waterhouse, M. et al. Alloreactive microenvironment after human hematopoietic cell transplantation induces genomic alterations in epithelium through an ROS-mediated mechanism: in vivo and in vitro study and implications to secondary neoplasia. Leukemia 24, 536–543 (2010). https://doi.org/10.1038/leu.2009.284

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