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Chronic Lymphocytic Leukemia

Loss-of-function mutations within the IL-2 inducible kinase ITK in patients with EBV-associated lymphoproliferative diseases

Leukemia volume 26, pages 963–971 (2012)Cite this article

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Abstract

The purpose of this study was the appraisal of the clinical and functional consequences of germline mutations within the gene for the IL-2 inducible T-cell kinase, ITK. Among patients with Epstein-Barr virus-driven lymphoproliferative disorders (EBV-LPD), negative for mutations in SH2D1A and XIAP (n=46), we identified two patients with R29H or D500T,F501L,M503X mutations, respectively. Human wild-type (wt) ITK, but none of the mutants, was able to rescue defective calcium flux in murine Itk−/− T cells. Pulse-chase experiments showed that ITK mutations lead to varying reductions of protein half-life from 25 to 69% as compared with wt ITK (107 min). The pleckstrin homology domain of wt ITK binds most prominently to phosphatidylinositol monophosphates (PI(3)P, PI(4)P, PI(5)P) and to lesser extend to its double or triple phosphorylated derivates (PIP2, PIP3), interactions which were dramatically reduced in the patient with the ITKR29H mutant. ITK mutations are distributed over the entire protein and include missense, nonsense and indel mutations, reminiscent of the situation in its sister kinase in B cells, Bruton's tyrosine kinase.

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Acknowledgements

We thank all the patients, their parents and their treating physicians for sharing the clinical data with us. The cell lines HEK 293 and HT-1080, as well as the basic bicistronic expression vectors (pMC) and the coding sequences for ECFP and EYFP were kindly provided by F Boege, Dusseldorf, Germany. We thank S Furlan, S Bellert, M Oellers and U Wiczorek in Dusseldorf, as well as I Mueller-Fleckenstein and M Schmidt in Erlangen, for excellent technical assistance. The ALPS samples were kindly provided by G Lahr. The HLH samples were kindly provided by A Meindl and U zur Stadt. The EBV LPD samples were kindly provided by: J Richter, A Gennery, V Hazar, D K Uygun, P Vorwerk, E Mejstrikova, D Schwabe, M Seidel, C Prada, T Bernig, H von Bernuth, R Schneppenheim, S Choo, M van der Burg, P S Palacin and H Ören. This work was generously supported by the Elterninitative ‘Kinderkrebsklinik e.V.’. SLR, RDv and MRA were supported by the grants of the research committee of the medical faculty of the Heinrich-Heine-University Dusseldorf and the E-Rare project NSEuroNet. AH was supported by the Deutsche Forschungsgemeinschaft (He 2526/7-2). AB was also supported by Grants from the BMBF and the German-Israeli-Foundation.

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Authors and Affiliations

  1. Department of Pediatric Oncology, Hematology and Clinical Immunology, Centre for Child and Adolescent Health,

    R M Linka, K Bienemann, Y Linka, F Krux, S Ginzel, M Gombert, H-J Laws & A Borkhardt

  2. Medical Faculty, Heinrich-Heine University, Dusseldorf, Germany

    R M Linka, K Bienemann, Y Linka, F Krux, S Ginzel, M Gombert, H-J Laws & A Borkhardt

  3. Institute of Biochemistry and Molecular Biology II, Medical Faculty, Heinrich Heine University, Dusseldorf, Germany

    S L Risse, R Dvorsky & M R Ahmadian

  4. Faculty of Biology, Albert-Ludwigs University of Freiburg and Max-Planck-Institute for Immunobiology and Epigenetics, Freiburg, Germany

    M Werner & H Jumaa

  5. INSERM Unité 768, Laboratoire du Développement Normal et Pathologique du Système Immunitaire, Hôpital Necker-Enfants Malades, Paris, France

    C Synaeve, A Fischer & S Latour

  6. Université René Descartes, Paris, France

    C Synaeve, A Fischer & S Latour

  7. Hôpital Necker-Enfants Malades, AP-HP, Paris, France

    C Synaeve, A Fischer & S Latour

  8. Biological and Medical Research Center (BMFZ), Heinrich-Heine University, Dusseldorf, Germany

    R Deenen & K Köhrer

  9. Institute of Virology, Heinrich-Heine University, Dusseldorf, Germany

    A Halenius

  10. Institute of Molecular and Clinical Immunology, Otto-von-Guericke University of Magdeburg, Magdeburg, Germany

    R Hartig & B Schraven

  11. Paediatric Research Centre, University Hospital of Tampere, Tampere, Finland

    M Helminen & K Vettenranta

  12. Pediatric Hematology-Oncology, Hadassah University Hospital, Jerusalem, Israel

    P Stepensky

  13. Institute of Virology, Friedrich-Alexander-University Erlangen-Nurnberg, Erlangen, Germany

    B Fleckenstein

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  1. R M Linka
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Correspondence to A Borkhardt.

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Linka, R., Risse, S., Bienemann, K. et al. Loss-of-function mutations within the IL-2 inducible kinase ITK in patients with EBV-associated lymphoproliferative diseases. Leukemia 26, 963–971 (2012). https://doi.org/10.1038/leu.2011.371

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