Whole-exome sequencing identifies novel MPL and JAK2 mutations in triple-negative myeloproliferative neoplasms

Jelena D. Milosevic Feenstra, Harini Nivarthi, Heinz Gisslinger, Emilie Leroy, Elisa Rumi, Ilyas Chachoua, Klaudia Bagienski, Blanka Kubesova, Daniela Pietra, Bettina Gisslinger, Chiara Milanesi, Roland J. Ager, Doris Chen, Tiina Berg, Martin Schalling, Michael Schuster, Christoph Bock, Stefan N. Constantinescu, Mario Cazzola, Robert Kralovics

Research output: Contribution to journalArticle

Abstract

Essential thrombocythemia (ET) and primary myelofibrosis (PMF) are chronic diseases characterized by clonal hematopoiesis and hyperproliferation of terminally differentiated myeloid cells. The disease is driven by somatic mutations in exon 9 of CALR or exon 10 of MPL or JAK2-V617F in >90% of the cases, whereas the remaining cases are termed "triple negative." We aimed to identify the disease-causing mutations in the triple-negative cases of ET and PMF by applying whole-exome sequencing (WES) on paired tumor and controlsamples from8 patients.Wefound evidence of clonal hematopoiesis in 5 of 8 studied cases based on clonality analysis and presence of somatic genetic aberrations. WES identified somatic mutations in 3 of 8 cases. We did not detect any novel recurrent somatic mutations. In 3 patientswith clonal hematopoiesis analyzed byWES, we identified a somatic MPL-S204P, a germline MPL-V285E mutation, and a germline JAK2-G571S variant. We performed Sanger sequencing of the entire coding region of MPL in 62, and of JAK2 in 49 additional triple-negative cases of ET or PMF. New somatic (T119I, S204F, E230G, Y591D) and 1germline(R321W)MPLmutationweredetected.All of theidentifiedMPLmutationswere gain-of-functionwhenanalyzedin functionalassays. JAK2 variants were identified in 5 of 57 triple-negative cases analyzed by WES and Sanger sequencing combined.We could demonstrate that JAK2-V625F andJAK2-F556V are gain-of-function mutations. Our results suggest that triple-negative casesofETandPMFdo not represent a homogenous disease entity. Cases with polyclonal hematopoiesis might represent hereditary disorders.

Original languageEnglish
Pages (from-to)325-332
Number of pages8
JournalBlood
Volume127
Issue number3
DOIs
Publication statusPublished - Jan 21 2016

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Exome
Hematopoiesis
Essential Thrombocythemia
Primary Myelofibrosis
Mutation
Exons
Neoplasms
Aberrations
Germ-Line Mutation
Tumors
Myeloid Cells
Chronic Disease

ASJC Scopus subject areas

  • Hematology
  • Biochemistry
  • Cell Biology
  • Immunology

Cite this

Milosevic Feenstra, J. D., Nivarthi, H., Gisslinger, H., Leroy, E., Rumi, E., Chachoua, I., ... Kralovics, R. (2016). Whole-exome sequencing identifies novel MPL and JAK2 mutations in triple-negative myeloproliferative neoplasms. Blood, 127(3), 325-332. https://doi.org/10.1182/blood-2015-07-661835

Whole-exome sequencing identifies novel MPL and JAK2 mutations in triple-negative myeloproliferative neoplasms. / Milosevic Feenstra, Jelena D.; Nivarthi, Harini; Gisslinger, Heinz; Leroy, Emilie; Rumi, Elisa; Chachoua, Ilyas; Bagienski, Klaudia; Kubesova, Blanka; Pietra, Daniela; Gisslinger, Bettina; Milanesi, Chiara; Ager, Roland J.; Chen, Doris; Berg, Tiina; Schalling, Martin; Schuster, Michael; Bock, Christoph; Constantinescu, Stefan N.; Cazzola, Mario; Kralovics, Robert.

In: Blood, Vol. 127, No. 3, 21.01.2016, p. 325-332.

Research output: Contribution to journalArticle

Milosevic Feenstra, JD, Nivarthi, H, Gisslinger, H, Leroy, E, Rumi, E, Chachoua, I, Bagienski, K, Kubesova, B, Pietra, D, Gisslinger, B, Milanesi, C, Ager, RJ, Chen, D, Berg, T, Schalling, M, Schuster, M, Bock, C, Constantinescu, SN, Cazzola, M & Kralovics, R 2016, 'Whole-exome sequencing identifies novel MPL and JAK2 mutations in triple-negative myeloproliferative neoplasms', Blood, vol. 127, no. 3, pp. 325-332. https://doi.org/10.1182/blood-2015-07-661835
Milosevic Feenstra, Jelena D. ; Nivarthi, Harini ; Gisslinger, Heinz ; Leroy, Emilie ; Rumi, Elisa ; Chachoua, Ilyas ; Bagienski, Klaudia ; Kubesova, Blanka ; Pietra, Daniela ; Gisslinger, Bettina ; Milanesi, Chiara ; Ager, Roland J. ; Chen, Doris ; Berg, Tiina ; Schalling, Martin ; Schuster, Michael ; Bock, Christoph ; Constantinescu, Stefan N. ; Cazzola, Mario ; Kralovics, Robert. / Whole-exome sequencing identifies novel MPL and JAK2 mutations in triple-negative myeloproliferative neoplasms. In: Blood. 2016 ; Vol. 127, No. 3. pp. 325-332.
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AU - Milosevic Feenstra, Jelena D.

AU - Nivarthi, Harini

AU - Gisslinger, Heinz

AU - Leroy, Emilie

AU - Rumi, Elisa

AU - Chachoua, Ilyas

AU - Bagienski, Klaudia

AU - Kubesova, Blanka

AU - Pietra, Daniela

AU - Gisslinger, Bettina

AU - Milanesi, Chiara

AU - Ager, Roland J.

AU - Chen, Doris

AU - Berg, Tiina

AU - Schalling, Martin

AU - Schuster, Michael

AU - Bock, Christoph

AU - Constantinescu, Stefan N.

AU - Cazzola, Mario

AU - Kralovics, Robert

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N2 - Essential thrombocythemia (ET) and primary myelofibrosis (PMF) are chronic diseases characterized by clonal hematopoiesis and hyperproliferation of terminally differentiated myeloid cells. The disease is driven by somatic mutations in exon 9 of CALR or exon 10 of MPL or JAK2-V617F in >90% of the cases, whereas the remaining cases are termed "triple negative." We aimed to identify the disease-causing mutations in the triple-negative cases of ET and PMF by applying whole-exome sequencing (WES) on paired tumor and controlsamples from8 patients.Wefound evidence of clonal hematopoiesis in 5 of 8 studied cases based on clonality analysis and presence of somatic genetic aberrations. WES identified somatic mutations in 3 of 8 cases. We did not detect any novel recurrent somatic mutations. In 3 patientswith clonal hematopoiesis analyzed byWES, we identified a somatic MPL-S204P, a germline MPL-V285E mutation, and a germline JAK2-G571S variant. We performed Sanger sequencing of the entire coding region of MPL in 62, and of JAK2 in 49 additional triple-negative cases of ET or PMF. New somatic (T119I, S204F, E230G, Y591D) and 1germline(R321W)MPLmutationweredetected.All of theidentifiedMPLmutationswere gain-of-functionwhenanalyzedin functionalassays. JAK2 variants were identified in 5 of 57 triple-negative cases analyzed by WES and Sanger sequencing combined.We could demonstrate that JAK2-V625F andJAK2-F556V are gain-of-function mutations. Our results suggest that triple-negative casesofETandPMFdo not represent a homogenous disease entity. Cases with polyclonal hematopoiesis might represent hereditary disorders.

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