A dominant gain-of-function mutation in universal tyrosine kinase SRC causes thrombocytopenia, myelofibrosis, bleeding, and bone pathologies

E. Turro, D. Greene, A. Wijgaerts, C. Thys, C. Lentaigne, T.K. Bariana, S.K. Westbury, A.M. Kelly, D. Selleslag, J.C. Stephens, S. Papadia, I. Simeoni, C.J. Penkett, S. Ashford, A. Attwood, S. Austin, T. Bakchoul, P. Collins, S.V.V. Deevi, R. Favier & 23 others M. Kostadima, M.P. Lambert, M. Mathias, C.M. Millar, K. Peerlinck, D.J. Perry, S. Schulman, D. Whitehorn, C. Wittevrongel, M. De Maeyer, A. Rendon, K. Gomez, Wendy N. Erber, A.D. Mumford, P. Nurden, K. Stirrups, J.R. Bradley, F.L. Raymond, M.A. Laffan, C. Van Geet, S. Richardson, K. Freson, H. Ouwehand

    Research output: Contribution to journalArticle

    36 Citations (Scopus)

    Abstract

    © 2016 by the American Association for the Advancement of Science; all rights reserved. The Src family kinase (SFK)member SRC is amajor target in drug development because it is activated in many human cancers, yet deleterious SRC germline mutations have not been reported. We used genome sequencing and Human Phenotype Ontology patient coding to identify a gain-of-function mutation in SRC causing thrombocytopenia, myelofibrosis, bleeding, and bone pathologies in nine cases. Modeling of the E527K substitution predicts loss of SRC's self-inhibitory capacity, whichwe confirmedwith in vitro studies showing increased SRC kinase activity and enhanced Tyr419 phosphorylation in COS-7 cells overexpressing E527K SRC. The active form of SRC predominates in patients' platelets, resulting in enhanced overall tyrosine phosphorylation. Patientswith myelofibrosis have hypercellular bone marrow with trilineage dysplasia, and their stem cells grown in vitro form more myeloid and megakaryocyte (MK) colonies than control cells. These MKs generate platelets that are dysmorphic, low in number, highly variable in size, and have a paucity of a-granules. Overactive SRC in patient-derived MKs causes a reduction in proplatelet formation, which can be rescued by SRC kinase inhibition. Stem cells transduced with lentiviral E527K SRC formMKs with a similar defect and enhanced tyrosine phosphorylation levels. Patient-derived and E527K-transduced MKs show Y419 SRC- positive stained podosomes that induce altered actin organization. Expression of mutated src in zebrafish recapitulates patients' blood and bone phenotypes. Similar studies of platelets andMKs may reveal the mechanism underlying the severe bleeding frequently observed in cancer patients treated with next-generation SFK inhibitors.
    Original languageEnglish
    Article number328ra30
    Pages (from-to)1-15
    Number of pages15
    JournalScience Translational Medicine
    Volume8
    Issue number328
    DOIs
    Publication statusPublished - 2016

    Fingerprint

    Primary Myelofibrosis
    Thrombocytopenia
    Protein-Tyrosine Kinases
    Pathology
    Hemorrhage
    Bone and Bones
    Mutation
    Blood Platelets
    src-Family Kinases
    Phosphorylation
    Tyrosine
    Phosphotransferases
    Stem Cells
    Phenotype
    Megakaryocytes
    Germ-Line Mutation
    COS Cells
    Zebrafish
    Human Genome
    Actins

    Cite this

    Turro, E. ; Greene, D. ; Wijgaerts, A. ; Thys, C. ; Lentaigne, C. ; Bariana, T.K. ; Westbury, S.K. ; Kelly, A.M. ; Selleslag, D. ; Stephens, J.C. ; Papadia, S. ; Simeoni, I. ; Penkett, C.J. ; Ashford, S. ; Attwood, A. ; Austin, S. ; Bakchoul, T. ; Collins, P. ; Deevi, S.V.V. ; Favier, R. ; Kostadima, M. ; Lambert, M.P. ; Mathias, M. ; Millar, C.M. ; Peerlinck, K. ; Perry, D.J. ; Schulman, S. ; Whitehorn, D. ; Wittevrongel, C. ; De Maeyer, M. ; Rendon, A. ; Gomez, K. ; Erber, Wendy N. ; Mumford, A.D. ; Nurden, P. ; Stirrups, K. ; Bradley, J.R. ; Raymond, F.L. ; Laffan, M.A. ; Van Geet, C. ; Richardson, S. ; Freson, K. ; Ouwehand, H. / A dominant gain-of-function mutation in universal tyrosine kinase SRC causes thrombocytopenia, myelofibrosis, bleeding, and bone pathologies. In: Science Translational Medicine. 2016 ; Vol. 8, No. 328. pp. 1-15.
    @article{ebdaa267e8fd4f87966bcc8e9b2f6c2e,
    title = "A dominant gain-of-function mutation in universal tyrosine kinase SRC causes thrombocytopenia, myelofibrosis, bleeding, and bone pathologies",
    abstract = "{\circledC} 2016 by the American Association for the Advancement of Science; all rights reserved. The Src family kinase (SFK)member SRC is amajor target in drug development because it is activated in many human cancers, yet deleterious SRC germline mutations have not been reported. We used genome sequencing and Human Phenotype Ontology patient coding to identify a gain-of-function mutation in SRC causing thrombocytopenia, myelofibrosis, bleeding, and bone pathologies in nine cases. Modeling of the E527K substitution predicts loss of SRC's self-inhibitory capacity, whichwe confirmedwith in vitro studies showing increased SRC kinase activity and enhanced Tyr419 phosphorylation in COS-7 cells overexpressing E527K SRC. The active form of SRC predominates in patients' platelets, resulting in enhanced overall tyrosine phosphorylation. Patientswith myelofibrosis have hypercellular bone marrow with trilineage dysplasia, and their stem cells grown in vitro form more myeloid and megakaryocyte (MK) colonies than control cells. These MKs generate platelets that are dysmorphic, low in number, highly variable in size, and have a paucity of a-granules. Overactive SRC in patient-derived MKs causes a reduction in proplatelet formation, which can be rescued by SRC kinase inhibition. Stem cells transduced with lentiviral E527K SRC formMKs with a similar defect and enhanced tyrosine phosphorylation levels. Patient-derived and E527K-transduced MKs show Y419 SRC- positive stained podosomes that induce altered actin organization. Expression of mutated src in zebrafish recapitulates patients' blood and bone phenotypes. Similar studies of platelets andMKs may reveal the mechanism underlying the severe bleeding frequently observed in cancer patients treated with next-generation SFK inhibitors.",
    author = "E. Turro and D. Greene and A. Wijgaerts and C. Thys and C. Lentaigne and T.K. Bariana and S.K. Westbury and A.M. Kelly and D. Selleslag and J.C. Stephens and S. Papadia and I. Simeoni and C.J. Penkett and S. Ashford and A. Attwood and S. Austin and T. Bakchoul and P. Collins and S.V.V. Deevi and R. Favier and M. Kostadima and M.P. Lambert and M. Mathias and C.M. Millar and K. Peerlinck and D.J. Perry and S. Schulman and D. Whitehorn and C. Wittevrongel and {De Maeyer}, M. and A. Rendon and K. Gomez and Erber, {Wendy N.} and A.D. Mumford and P. Nurden and K. Stirrups and J.R. Bradley and F.L. Raymond and M.A. Laffan and {Van Geet}, C. and S. Richardson and K. Freson and H. Ouwehand",
    year = "2016",
    doi = "10.1126/scitranslmed.aad7666",
    language = "English",
    volume = "8",
    pages = "1--15",
    journal = "Science Translational Medicine",
    issn = "1946-6234",
    publisher = "AMER ASSOC ADVANCEMENT SCIENCE",
    number = "328",

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    Turro, E, Greene, D, Wijgaerts, A, Thys, C, Lentaigne, C, Bariana, TK, Westbury, SK, Kelly, AM, Selleslag, D, Stephens, JC, Papadia, S, Simeoni, I, Penkett, CJ, Ashford, S, Attwood, A, Austin, S, Bakchoul, T, Collins, P, Deevi, SVV, Favier, R, Kostadima, M, Lambert, MP, Mathias, M, Millar, CM, Peerlinck, K, Perry, DJ, Schulman, S, Whitehorn, D, Wittevrongel, C, De Maeyer, M, Rendon, A, Gomez, K, Erber, WN, Mumford, AD, Nurden, P, Stirrups, K, Bradley, JR, Raymond, FL, Laffan, MA, Van Geet, C, Richardson, S, Freson, K & Ouwehand, H 2016, 'A dominant gain-of-function mutation in universal tyrosine kinase SRC causes thrombocytopenia, myelofibrosis, bleeding, and bone pathologies' Science Translational Medicine, vol. 8, no. 328, 328ra30, pp. 1-15. https://doi.org/10.1126/scitranslmed.aad7666

    A dominant gain-of-function mutation in universal tyrosine kinase SRC causes thrombocytopenia, myelofibrosis, bleeding, and bone pathologies. / Turro, E.; Greene, D.; Wijgaerts, A.; Thys, C.; Lentaigne, C.; Bariana, T.K.; Westbury, S.K.; Kelly, A.M.; Selleslag, D.; Stephens, J.C.; Papadia, S.; Simeoni, I.; Penkett, C.J.; Ashford, S.; Attwood, A.; Austin, S.; Bakchoul, T.; Collins, P.; Deevi, S.V.V.; Favier, R.; Kostadima, M.; Lambert, M.P.; Mathias, M.; Millar, C.M.; Peerlinck, K.; Perry, D.J.; Schulman, S.; Whitehorn, D.; Wittevrongel, C.; De Maeyer, M.; Rendon, A.; Gomez, K.; Erber, Wendy N.; Mumford, A.D.; Nurden, P.; Stirrups, K.; Bradley, J.R.; Raymond, F.L.; Laffan, M.A.; Van Geet, C.; Richardson, S.; Freson, K.; Ouwehand, H.

    In: Science Translational Medicine, Vol. 8, No. 328, 328ra30, 2016, p. 1-15.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - A dominant gain-of-function mutation in universal tyrosine kinase SRC causes thrombocytopenia, myelofibrosis, bleeding, and bone pathologies

    AU - Turro, E.

    AU - Greene, D.

    AU - Wijgaerts, A.

    AU - Thys, C.

    AU - Lentaigne, C.

    AU - Bariana, T.K.

    AU - Westbury, S.K.

    AU - Kelly, A.M.

    AU - Selleslag, D.

    AU - Stephens, J.C.

    AU - Papadia, S.

    AU - Simeoni, I.

    AU - Penkett, C.J.

    AU - Ashford, S.

    AU - Attwood, A.

    AU - Austin, S.

    AU - Bakchoul, T.

    AU - Collins, P.

    AU - Deevi, S.V.V.

    AU - Favier, R.

    AU - Kostadima, M.

    AU - Lambert, M.P.

    AU - Mathias, M.

    AU - Millar, C.M.

    AU - Peerlinck, K.

    AU - Perry, D.J.

    AU - Schulman, S.

    AU - Whitehorn, D.

    AU - Wittevrongel, C.

    AU - De Maeyer, M.

    AU - Rendon, A.

    AU - Gomez, K.

    AU - Erber, Wendy N.

    AU - Mumford, A.D.

    AU - Nurden, P.

    AU - Stirrups, K.

    AU - Bradley, J.R.

    AU - Raymond, F.L.

    AU - Laffan, M.A.

    AU - Van Geet, C.

    AU - Richardson, S.

    AU - Freson, K.

    AU - Ouwehand, H.

    PY - 2016

    Y1 - 2016

    N2 - © 2016 by the American Association for the Advancement of Science; all rights reserved. The Src family kinase (SFK)member SRC is amajor target in drug development because it is activated in many human cancers, yet deleterious SRC germline mutations have not been reported. We used genome sequencing and Human Phenotype Ontology patient coding to identify a gain-of-function mutation in SRC causing thrombocytopenia, myelofibrosis, bleeding, and bone pathologies in nine cases. Modeling of the E527K substitution predicts loss of SRC's self-inhibitory capacity, whichwe confirmedwith in vitro studies showing increased SRC kinase activity and enhanced Tyr419 phosphorylation in COS-7 cells overexpressing E527K SRC. The active form of SRC predominates in patients' platelets, resulting in enhanced overall tyrosine phosphorylation. Patientswith myelofibrosis have hypercellular bone marrow with trilineage dysplasia, and their stem cells grown in vitro form more myeloid and megakaryocyte (MK) colonies than control cells. These MKs generate platelets that are dysmorphic, low in number, highly variable in size, and have a paucity of a-granules. Overactive SRC in patient-derived MKs causes a reduction in proplatelet formation, which can be rescued by SRC kinase inhibition. Stem cells transduced with lentiviral E527K SRC formMKs with a similar defect and enhanced tyrosine phosphorylation levels. Patient-derived and E527K-transduced MKs show Y419 SRC- positive stained podosomes that induce altered actin organization. Expression of mutated src in zebrafish recapitulates patients' blood and bone phenotypes. Similar studies of platelets andMKs may reveal the mechanism underlying the severe bleeding frequently observed in cancer patients treated with next-generation SFK inhibitors.

    AB - © 2016 by the American Association for the Advancement of Science; all rights reserved. The Src family kinase (SFK)member SRC is amajor target in drug development because it is activated in many human cancers, yet deleterious SRC germline mutations have not been reported. We used genome sequencing and Human Phenotype Ontology patient coding to identify a gain-of-function mutation in SRC causing thrombocytopenia, myelofibrosis, bleeding, and bone pathologies in nine cases. Modeling of the E527K substitution predicts loss of SRC's self-inhibitory capacity, whichwe confirmedwith in vitro studies showing increased SRC kinase activity and enhanced Tyr419 phosphorylation in COS-7 cells overexpressing E527K SRC. The active form of SRC predominates in patients' platelets, resulting in enhanced overall tyrosine phosphorylation. Patientswith myelofibrosis have hypercellular bone marrow with trilineage dysplasia, and their stem cells grown in vitro form more myeloid and megakaryocyte (MK) colonies than control cells. These MKs generate platelets that are dysmorphic, low in number, highly variable in size, and have a paucity of a-granules. Overactive SRC in patient-derived MKs causes a reduction in proplatelet formation, which can be rescued by SRC kinase inhibition. Stem cells transduced with lentiviral E527K SRC formMKs with a similar defect and enhanced tyrosine phosphorylation levels. Patient-derived and E527K-transduced MKs show Y419 SRC- positive stained podosomes that induce altered actin organization. Expression of mutated src in zebrafish recapitulates patients' blood and bone phenotypes. Similar studies of platelets andMKs may reveal the mechanism underlying the severe bleeding frequently observed in cancer patients treated with next-generation SFK inhibitors.

    U2 - 10.1126/scitranslmed.aad7666

    DO - 10.1126/scitranslmed.aad7666

    M3 - Article

    VL - 8

    SP - 1

    EP - 15

    JO - Science Translational Medicine

    JF - Science Translational Medicine

    SN - 1946-6234

    IS - 328

    M1 - 328ra30

    ER -