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CHRONIC MYELOPROLIFERATIVE NEOPLASMS

Synergistic effect of concurrent high molecular risk mutations and lower JAK2 mutant variant allele frequencies on prognosis in patients with myelofibrosis—insights from a multicenter study

Leukemia volume 39, pages 144–154 (2025)Cite this article

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Abstract

In addition to high-molecular risk (HMR) mutations (ASXL1, EZH2, SRSF2, IDH, and U2AF1Q157), lower JAK2V617F variant allele frequencies (VAF) have been demonstrated to be associated with poor prognosis of myelofibrosis (MF) patients. Nevertheless, the relationship between JAK2V617F VAF and HMR mutations remains inconclusive. To address this, we analyzed the mutation status of 54 myeloid neoplasm-relevant genes using targeted next-generation sequencing in 124 MF patients. Three cohorts from multiple international centers were analyzed for external validation. Among JAK2-mutated patients, the presence of HMR mutations drove poor prognosis in patients with lower JAK2V617F VAF but not in those with higher JAK2V617F VAF. Survival analyses showed consistent results across validation cohorts. In multivariable analysis, concurrent HMR and a lower JAK2V617F VAF was identified as an independent adverse prognostic factor for survival, irrespective of age, MIPSS70, MIPSS70 + v2, and GIPSS risk groups. Mutation co-occurrence tests revealed no shared mutational pattern over different cohorts, excluding potential confounding effect from other concurrent mutations. Importantly, the integration of HMR/JAK2V617F VAF (≤50%) status significantly enhanced existing prognostic models, as evidenced by higher c-indexes and time-dependent ROC analyses. Single-cell studies with sequential follow-ups are warranted to decipher the clonal evolution of MF and how it relates to JAK2V617F VAF dynamics.

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Fig. 1: Survival outcomes of PMF patients in the discovery cohort.
Fig. 2: Survival outcomes of JAK2-mutated MF patients in validation cohorts with lower or higher JAK2V617F variant allele frequencies (VAF), based on the median value of JAK2V617F VAF in each cohort as the cutoff.
Fig. 3: Survival outcomes of JAK2-mutated MF patients pooled from the discovery and validation cohorts.
Fig. 4: Mutation interaction and co-occurrence analysis.
Fig. 5: Re-stratification of existing prognostic models to the updated models incorporating high molecular risk (HMR) mutation and JAK2V617F VAF status for 292 JAK2-mutated patients pooled from discovery and validation cohorts 1–3 and their survival outcomes.
Fig. 6: Time-dependent ROC curve analysis in pooled cohorts (discovery and validation cohorts 1–3).

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Requests for additional information not provided in the main text or Supplementary Material should be sent to the corresponding authors. The data are not publicly available due to privacy and ethical restrictions.

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Acknowledgements

We would like to acknowledge the service provided by the Department of Laboratory Medicine, Division of Hematology, Department of Internal Medicine, and Department of Pathology, National Taiwan University Hospital.

Funding

The study was partially supported by grants from the Ministry of Health and Welfare, Taiwan (MOHW 112-TDU-B-211-124001) and the Ministry of Science and Technology, Taiwan (MOST 107-2314-B-002-013). The University of Manchester’s Epigenetics of Haematopoiesis Laboratory is core funded by grants from The Oglesby Charitable Trust. DHW is also supported by a Cancer Research UK Advanced Clinician Scientist Fellowship (RCCASF-Nov22/100001) and the University of Manchester Sybil Mary Pilkington Leukaemia Research Fellowship.

Author information

Author notes

  1. These authors contributed equally: Yu-Hung Wang, Chao-Hung Wei.

  2. These authors jointly supervised this work: Jean-Jacques Kiladjian, Hwei-Fang Tien.

Authors and Affiliations

  1. Division of Hematology, Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan

    Yu-Hung Wang, Chien-Chin Lin, Chi-Yuan Yao, Hsin-An Hou, Wen-Chien Chou & Hwei-Fang Tien

  2. Epigenetics of Haematopoiesis Laboratory, Division of Cancer Sciences, The University of Manchester, Manchester, UK

    Yu-Hung Wang, Kristian Gurashi, Kiran Batta & Daniel H. Wiseman

  3. Graduate Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei, Taiwan

    Chao-Hung Wei

  4. Department of Internal Medicine, National Taiwan University Hospital Hsin-Chu Branch, Hsinchu, Taiwan

    Chao-Hung Wei

  5. Department of Laboratory Medicine, National Taiwan University Hospital, Taipei, Taiwan

    Chien-Chin Lin, Chi-Yuan Yao & Wen-Chien Chou

  6. Department of Translational Hematology and Oncology Research, Taussig Cancer Institute, Cleveland Clinic, Cleveland, OH, USA

    Carmelo Gurnari, Hussein Awada & Jaroslaw P. Maciejewski

  7. Department of Biomedicine and Prevention, University of Tor Vergata, Rome, Italy

    Carmelo Gurnari

  8. Centre d’Investigations Cliniques, Hospital Saint Louis and University Paris Cité, Paris, France

    Lina Benajiba, Rafael Daltro de Oliveira, Juliette Soret-Dulphy & Jean-Jacques Kiladjian

  9. Laboratoire de Biologie Cellulaire, Hospital Saint Louis and University Paris Cité, Paris, France

    Bruno Cassinat

  10. Hematology, Oncology and Transfusion Medicine Center, Vilnius University Hospital Santaros Klinikos, Vilnius, Lithuania

    Andrius Zucenka

  11. Hematology and Oncology Department, Faculty of Medicine, Vilnius University, Vilnius, Lithuania

    Andrius Zucenka

  12. Hematology Department, University Hospital of Santiago de Compostela, Santiago de Compostela, Spain

    Adrián Mosquera Orgueira & Manuel Mateo Pérez Encinas

  13. Group of Computational Hematology and Genomics, Instituto de Investigación Sanitaria de Santiago de Compostela (IDIS), Santiago de Compostela, Spain

    Adrián Mosquera Orgueira

  14. Department of Pathology, National Taiwan University Hospital, Taipei, Taiwan

    Chang-Tsu Yuan

  15. Division of Cellular Therapy, Department of Integrated Diagnostics and Therapeutics, National Taiwan University Hospital, Taipei, Taiwan

    Sze-Hwei Lee

  16. Department of Internal Medicine, Far-Eastern Memorial Hospital, New Taipei City, Taiwan

    Hwei-Fang Tien

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  1. Yu-Hung Wang
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  2. Chao-Hung Wei
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Contributions

YHW and CHW contribute equally to this study. YHW and CHW were responsible for data collection and management, statistical analysis and interpretation, literature research, and manuscript writing; CG, HA, LB, RDO, JSD, BC, AZ, AMO, CTY, SHL, CYY, KG, HAH, KB, WCC, MMPE, and JPM helped data collection, analysis, and interpretation; CCL, DHW, JJK, and HFT helped revise the manuscript and gave recommendation; and YHW, CCL, and HFT planned, designed, and coordinated the study.

Corresponding authors

Correspondence to Yu-Hung Wang or Hwei-Fang Tien.

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This retrospective study was approved by the National Taiwan University Hospital Research Ethics Committee (reference number #201709072RINC) and institutional review boards of each participating hospital, with informed consent obtained from all patients in accordance with the Declaration of Helsinki. All methods were performed in accordance with the relevant guidelines and regulations.

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Wang, YH., Wei, CH., Lin, CC. et al. Synergistic effect of concurrent high molecular risk mutations and lower JAK2 mutant variant allele frequencies on prognosis in patients with myelofibrosis—insights from a multicenter study. Leukemia 39, 144–154 (2025). https://doi.org/10.1038/s41375-024-02422-4

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