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Next-generation sequencing in the diagnosis of non-cirrhotic splanchnic vein thrombosis

  • Marta Magaz
    Affiliations
    Barcelona Hepatic Hemodynamic Laboratory, Liver Unit, Hospital Clinic, Institut de Investigacions Biomèdiques August Pi i Sunyer, University of Barcelona, Centro de Investigación Biomédica en Red Enfermedades Hepáticas y Digestivas, Health Care Provider of the European Reference Network on Rare Liver Disorders, Barcelona, Spain
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  • Alberto Alvarez-Larrán
    Affiliations
    Hematology Department, Hospital Clinic, Institut de Investigacions Biomèdiques August Pi i Sunyer, University of Barcelona, Barcelona, Spain
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  • Dolors Colomer
    Affiliations
    Hematopathology Section, Pathology Department, Hospital Clinic, Institut de Investigacions Biomèdiques August Pi i Sunyer, Centro de Investigación Biomédica en Red en Cancer, University of Barcelona, Barcelona, Spain
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  • Mónica López-Guerra
    Affiliations
    Hematopathology Section, Pathology Department, Hospital Clinic, Institut de Investigacions Biomèdiques August Pi i Sunyer, Centro de Investigación Biomédica en Red en Cancer, University of Barcelona, Barcelona, Spain
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  • M. Ángeles García-Criado
    Affiliations
    Abdominal Radiology Section, Radiology Department, Hospital Clinic, University of Barcelona, Barcelona, Spain
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  • Gabriel Mezzano
    Affiliations
    Barcelona Hepatic Hemodynamic Laboratory, Liver Unit, Hospital Clinic, Institut de Investigacions Biomèdiques August Pi i Sunyer, University of Barcelona, Centro de Investigación Biomédica en Red Enfermedades Hepáticas y Digestivas, Health Care Provider of the European Reference Network on Rare Liver Disorders, Barcelona, Spain
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  • Ernest Belmonte
    Affiliations
    Abdominal Radiology Section, Radiology Department, Hospital Clinic, University of Barcelona, Barcelona, Spain
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  • Pol Olivas
    Affiliations
    Barcelona Hepatic Hemodynamic Laboratory, Liver Unit, Hospital Clinic, Institut de Investigacions Biomèdiques August Pi i Sunyer, University of Barcelona, Centro de Investigación Biomédica en Red Enfermedades Hepáticas y Digestivas, Health Care Provider of the European Reference Network on Rare Liver Disorders, Barcelona, Spain
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  • Guillem Soy
    Affiliations
    Barcelona Hepatic Hemodynamic Laboratory, Liver Unit, Hospital Clinic, Institut de Investigacions Biomèdiques August Pi i Sunyer, University of Barcelona, Centro de Investigación Biomédica en Red Enfermedades Hepáticas y Digestivas, Health Care Provider of the European Reference Network on Rare Liver Disorders, Barcelona, Spain
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  • Francisco Cervantes
    Affiliations
    Hematology Department, Hospital Clinic, Institut de Investigacions Biomèdiques August Pi i Sunyer, University of Barcelona, Barcelona, Spain
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  • Anna Darnell
    Affiliations
    Abdominal Radiology Section, Radiology Department, Hospital Clinic, University of Barcelona, Barcelona, Spain
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  • José Ferrusquía-Acosta
    Affiliations
    Barcelona Hepatic Hemodynamic Laboratory, Liver Unit, Hospital Clinic, Institut de Investigacions Biomèdiques August Pi i Sunyer, University of Barcelona, Centro de Investigación Biomédica en Red Enfermedades Hepáticas y Digestivas, Health Care Provider of the European Reference Network on Rare Liver Disorders, Barcelona, Spain
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  • Anna Baiges
    Affiliations
    Barcelona Hepatic Hemodynamic Laboratory, Liver Unit, Hospital Clinic, Institut de Investigacions Biomèdiques August Pi i Sunyer, University of Barcelona, Centro de Investigación Biomédica en Red Enfermedades Hepáticas y Digestivas, Health Care Provider of the European Reference Network on Rare Liver Disorders, Barcelona, Spain
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  • Fanny Turon
    Affiliations
    Barcelona Hepatic Hemodynamic Laboratory, Liver Unit, Hospital Clinic, Institut de Investigacions Biomèdiques August Pi i Sunyer, University of Barcelona, Centro de Investigación Biomédica en Red Enfermedades Hepáticas y Digestivas, Health Care Provider of the European Reference Network on Rare Liver Disorders, Barcelona, Spain
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  • Virginia Hernández-Gea
    Affiliations
    Barcelona Hepatic Hemodynamic Laboratory, Liver Unit, Hospital Clinic, Institut de Investigacions Biomèdiques August Pi i Sunyer, University of Barcelona, Centro de Investigación Biomédica en Red Enfermedades Hepáticas y Digestivas, Health Care Provider of the European Reference Network on Rare Liver Disorders, Barcelona, Spain
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  • Juan Carlos García-Pagán
    Correspondence
    Corresponding author. Address: Barcelona Hepatic Hemodynamic Laboratory, Liver Unit, Hospital Clinic, Villarroel 170, Barcelona 08036, Spain. Tel.: +34-93-2275790.
    Affiliations
    Barcelona Hepatic Hemodynamic Laboratory, Liver Unit, Hospital Clinic, Institut de Investigacions Biomèdiques August Pi i Sunyer, University of Barcelona, Centro de Investigación Biomédica en Red Enfermedades Hepáticas y Digestivas, Health Care Provider of the European Reference Network on Rare Liver Disorders, Barcelona, Spain
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      Highlights

      • NGS allows the simultaneous evaluation of multiple genes associated with myeloproliferative neoplasms, even at low mutational levels.
      • NGS could identify JAK2-exon12 mutations not previously detected by the conventional techniques.
      • NGS also identified high-molecular-risk HMR variants associated with clonal haematopoiesis.
      • Patients with idiopathic/local factor NC-SVT harbouring HRM variants seem to be at a higher risk of recurrent splanchnic thrombosis.

      Background & Aims

      Myeloproliferative neoplasms (MPNs) are the most frequent cause of non-tumoural non-cirrhotic splanchnic vein thrombosis (NC-SVT). Diagnosis of MPN is based on blood cell count alterations, bone marrow histology, and detection of specific gene mutations. Next-generation sequencing (NGS) allows the simultaneous evaluation of multiple genes implicated in myeloid clonal pathology. The aim of this study was to evaluate the potential role of NGS in elucidating the aetiology of NC-SVT.

      Methods

      DNA samples from 80 patients (75 with idiopathic or exclusively local factor [Idiop/loc-NC-SVT] and 5 with MPN and NC-SVT [SVT-MPN] negative for Janus kinase 2 gene [JAK2] [V617F and exon 12], calreticulin gene [CALR], and thrombopoietin gene [MPL] mutations by classic techniques) were analysed by NGS. Mutations involved in myeloid disorders different from JAK2, CALR, and MPL genes were categorised as high-molecular-risk (HMR) variants or variants of unknown significance.

      Results

      In 2/5 triple-negative SVT-MPN cases (40%), a mutation in exon 12 of JAK2 was identified. JAK2-exon 12 mutation was also identified in 1/75 patients with Idiop/loc-NC-SVT. Moreover, 28/74 (37.8%) of the remaining Idiop/loc-NC-SVT had at least 1 HMR variant. Sixty-two patients with Idiop/loc-NC-SVT were not receiving long-term anticoagulation and 5 of them (8.1%) had recurrent NC-SVT. This cumulative incidence was significantly higher in patients with HMR variants than in those without.

      Conclusions

      NGS identified JAK2-exon12 mutations not previously detected by conventional techniques. In addition, NGS detected HMR variants in approximately one-third of patients with Idiop/loc-NC-SVT. These patients seem to have a higher risk of splanchnic rethrombosis. NGS might be a useful diagnostic tool in NC-SVT.

      Lay summary

      Next-generation sequencing (NGS) performs massive sequencing of DNA allowing the simultaneous evaluation of multiple genes even at very low mutational levels. Application of this technique in a cohort of patients with non-cirrhotic non-tumoral portal vein thrombosis (NC-SVT) and a negative study for thrombophilic disorders was able to identify patients with a mutation in exon 12 not previously detected by conventional techniques. Moreover, NGS detected High Molecular Risk (HMR)-variants (Mutations involved in myeloid disorders different from JAK2, CALR and MPL genes) in approximately one third of patients. These patients appear to be at increased risk of rethrombosis. All these findings supports NGS as a potential useful tool in the management of NC-SVT.

      Graphical abstract

      Keywords

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