Establishing optimal quantitative-polymerase chain reaction assays for routine diagnosis and tracking of minimal residual disease in JAK2-V617F-associated myeloproliferative neoplasms: a joint European LeukemiaNet/MPN&MPNr-EuroNet (COST action BM0902) study

There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

Abstract

Reliable detection of JAK2-V617F is critical for accurate diagnosis of myeloproliferative neoplasms (MPNs); in addition, sensitive mutation-specific assays can be applied to monitor disease response. However, there has been no consistent approach to JAK2-V617F detection, with assays varying markedly in performance, affecting clinical utility. Therefore, we established a network of 12 laboratories from seven countries to systematically evaluate nine different DNA-based quantitative PCR (qPCR) assays, including those in widespread clinical use. Seven quality control rounds involving over 21 500 qPCR reactions were undertaken using centrally distributed cell line dilutions and plasmid controls. The two best-performing assays were tested on normal blood samples ( n=100) to evaluate assay specificity, followed by analysis of serial samples from 28 patients transplanted for JAK2-V617F-positive disease. The most sensitive assay, which performed consistently across a range of qPCR platforms, predicted outcome following transplant, with the mutant allele detected a median of 22 weeks (range 6–85 weeks) before relapse. Four of seven patients achieved molecular remission following donor lymphocyte infusion, indicative of a graft vs MPN effect. This study has established a robust, reliable assay for sensitive JAK2-V617F detection, suitable for assessing response in clinical trials, predicting outcome and guiding management of patients undergoing allogeneic transplant.

Related collections

Most cited references 40

Record: found
Abstract: found
Article: not found

Safety and efficacy of INCB018424, a JAK1 and JAK2 inhibitor, in myelofibrosis.

Srdan Verstovsek, Hagop Kantarjian, Ruben Mesa … (2010)

Myelofibrosis is a Philadelphia chromosome–negative myeloproliferative neoplasm associated with cytopenias, splenomegaly, poor quality of life, and shortened survival. About half of patients with myelofibrosis carry a gain-of-function mutation in the Janus kinase 2 gene (JAK2 V617F) that contributes to the pathophysiology of the disease. INCB018424 is a potent and selective Janus kinase 1 (JAK1) and JAK2 inhibitor. We conducted a phase 1−2 trial of INCB018424 in patients with JAK2 V617F−positive or JAK2 V617F−negative primary myelofibrosis, post–essential thrombocythemia myelofibrosis, or post–polycythemia vera myelofibrosis. A total of 153 patients received INCB018424 for a median duration of more than 14.7 months. The initial dose-escalation phase established 25 mg twice daily or 100 mg once daily as maximum tolerated doses, on the basis of reversible thrombocytopenia. A dose-dependent suppression of phosphorylated signal transducer and activator of transcription 3 (STAT3), a marker of JAK signaling, was demonstrated in patients with wild-type JAK2 and in patients with the JAK2 V617F mutation. We studied additional doses and established that a 15-mg twice-daily starting dose, followed by individualized dose titration, was the most effective and safest dosing regimen. At this dose, 17 of 33 patients (52%) had a rapid objective response (≥50% reduction of splenomegaly) lasting for 12 months or more, and this therapy was associated with grade 3 or grade 4 adverse events (mainly myelosuppression) in less than 10% of patients. Patients with debilitating symptoms, including weight loss, fatigue, night sweats, and pruritus, had rapid improvement. Clinical benefits were associated with a marked diminution of levels of circulating inflammatory cytokines that are commonly elevated in myelofibrosis. INCB018424 was associated with marked and durable clinical benefits in patients with myelofibrosis for whom no approved therapies existed. (Funded by Incyte; ClinicalTrials.gov number, NCT00509899.)

0 comments Cited 322 times – based on 0 reviews      Review now

Bookmark

Record: found
Abstract: found
Article: not found

Safety and efficacy of TG101348, a selective JAK2 inhibitor, in myelofibrosis.

Moshe Talpaz, Andrés J. Cortés, A Pardanani … (2011)

Myelofibrosis is a myeloid malignancy associated with anemia, splenomegaly, and constitutional symptoms. Patients frequently harbor JAK-STAT activating mutations that are sensitive to TG101348, a selective small-molecule Janus kinase 2 (JAK2) inhibitor. In a multicenter phase I trial, oral TG101348 was administered once a day to patients with high- or intermediate-risk primary or post-polycythemia vera/essential thrombocythemia myelofibrosis. Fifty-nine patients were treated, including 28 in the dose-escalation phase. The maximum-tolerated dose was 680 mg/d, and dose-limiting toxicity was a reversible and asymptomatic increase in the serum amylase level. Forty-three patients (73%) continued treatment beyond six cycles; the median cumulative exposure to TG101348 was 380 days. Adverse events included nausea, vomiting, diarrhea, anemia, and thrombocytopenia; corresponding grades 3 to 4 incidence rates were 3%, 3%, 10%, 35%, and 24%. TG101348 treatment had modest effect on serum cytokine levels, but greater than half of the patients with early satiety, night sweats, fatigue, pruritus, and cough achieved rapid and durable improvement in these symptoms. By six and 12 cycles of treatment, 39% and 47% of patients, respectively, had achieved a spleen response per International Working Group criteria. The majority of patients with leukocytosis or thrombocytosis at baseline (n = 28 and n = 10, respectively) achieved normalization of blood counts after six (57% and 90%, respectively) and 12 (56% and 88%, respectively) cycles. A significant decrease in JAK2 V617F allele burden was observed at 6 months in mutation-positive patients (n = 51; P = .04), particularly in the subgroup with allele burden greater than 20% (n = 23; P < .01); the decrease was durable at 12 months. TG101348 is well tolerated and produces significant reduction in disease burden and durable clinical benefit in patients with myelofibrosis.

0 comments Cited 102 times – based on 0 reviews      Review now

Bookmark

Record: found
Abstract: found
Article: not found

A prospective study of 338 patients with polycythemia vera: the impact of JAK2 (V617F) allele burden and leukocytosis on fibrotic or leukemic disease transformation and vascular complications.

E Rumi, E Roncoroni, M Cazzola … (2010)

We studied the relationship between JAK2 (V617F) mutant allele burden and clinical phenotype, disease progression and survival in patients with polycythemia vera (PV). The percentage of granulocyte mutant alleles was evaluated using a quantitative real-time polymerase chain reaction-based allelic discrimination assay. Of the 338 patients enrolled in this prospective study, 320 (94.7%) carried the JAK2 (V617F) mutation. Direct relationships were found between mutant allele burden and hemoglobin concentration (P=0.001), white blood cell count (P=0.001), spleen size (P=0.001) and age-adjusted bone marrow cellularity (P=0.002), while an inverse relationship was found with platelet count (P 50% mutant alleles), while 10 patients developed acute myeloid leukemia (AML). The mutant allele burden was significantly related to the risk of developing myelofibrosis (P=0.029) and retained its significant effect also in multivariable analysis (P=0.03). By contrast, the risk of developing AML as well as that of thrombosis was not significantly related to mutant allele burden. Leukocytosis did not affect thrombosis, MF, leukemia or survival. In conclusion, a JAK2 (V617F) allele burden >50% represents a risk factor for progression to MF in PV.

0 comments Cited 97 times – based on 0 reviews      Review now

Bookmark

All references

Author and article information

Journal

Journal ID (nlm-ta): Leukemia

Journal ID (iso-abbrev): Leukemia

Title: Leukemia

Publisher: Nature Publishing Group

ISSN (Print): 0887-6924

ISSN (Electronic): 1476-5551

Publication date (Print): October 2013

Publication date (Electronic preprint): 17 July 2013

Publication date (Electronic): 13 August 2013

Volume: 27

Issue: 10

Pages: 2032-2039

Affiliations

[1 ]King's College London School of Medicine, Cancer Genetics Lab, Department of Medical and Molecular Genetics , London, UK

[2 ]Molecular Oncology Diagnostics Unit, GSTS Pathology , London, UK

[3 ]Department of Experimental and Clinical Medicine, University of Florence , Florence, Italy

[4 ]Laboratoire d'Hématologie, Centre Hospitalier Universitaire , Bordeaux, France

[5 ]Department of Hematology, University Hospital and University of Bern , Bern, Switzerland

[6 ]AP-HP, Unité de Biologie Cellulaire, Hôpital Saint-Louis , Paris, France

[7 ]Klinisk Biokemi, Vejle Sygehus , Vejle, Denmark

[8 ]Qiagen Marseille , Marseille, France

[9 ]Centre Hospitalier Universitaire, Université de Nantes, Inserm U892 , Nantes, France

[10 ]Department of Laboratory Medicine, Laboratory of Hematology, Radboud University Medical Centre , Nijmegen, The Netherlands

[11 ]Munich Leukemia Laboratory , Munich, Germany

[12 ]Molecular Malignancy Laboratory and Haemato-Oncology Diagnostic Service, Addenbrooke's Hospital , Cambridge, UK

[13 ]Department of Histopathology, Guys and St Thomas' NHS Foundation Trust , London, UK

[14 ]King's College Hospital, Department of Haematology , London, UK

[15 ]West Midlands Regional Genetics Laboratory , Birmingham, UK

[16 ]Wessex Regional Genetics Laboratory , Salisbury, UK

[17 ]Department of Haematology, Belfast City Hospital , Belfast, UK

[18 ]Department of Molecular Hematology, University Hospital Freiburg , Freiburg, Germany

[19 ]Department of Pathology, University of Utah , Salt Lake City, UT, USA

[20 ]ARUP Laboratories , Salt Lake City, UT, USA

[21 ]Queen's University Belfast , Belfast, UK

[22 ]Department of Haematology, Guy's and St Thomas' NHS Foundation Trust , London, UK

[23 ]MPD-Research Consortium, Hematology Division, University of Utah School of Medicine , Salt Lake City, UT, USA

[24 ]Inserm UMR-S940, Institut Universitaire d'Hématologie , Paris, France

[25 ]Clinical Investigation Center (CIC), Hôpital Saint-Louis and French Intergroup of Myeloproliferative Disorders (FIM) , Paris, France

[26 ]Ospedali Riuniti di Bergamo , Bergamo, Italy

Author notes

[* ]King's College London School of Medicine, Cancer Genetics Lab, Department of Medical and Molecular Genetics , 8th Floor, Tower Wing, Guy's Hospital, London SE1 9RT, UK. E-mail: david.grimwade@ 123456genetics.kcl.ac.uk

[27]

These authors contributed equally to this work.

Article

Publisher Item ID: leu2013219

DOI: 10.1038/leu.2013.219

PMC ID: 3806250

PubMed ID: 23860450

SO-VID: c7a6ca3e-e18a-429a-bf0f-03d0d7e2d607

License:

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/

History

Date received : 07 March 2013

Date revision received : 30 June 2013

Date accepted : 02 July 2013

Comments

Comment on this article

scite_

Cited by 48

See all cited by

Most referenced authors 987

See all reference authors

Establishing optimal quantitative-polymerase chain reaction assays for routine diagnosis and tracking of minimal residual disease in JAK2-V617F-associated myeloproliferative neoplasms: a joint European LeukemiaNet/MPN&MPNr-EuroNet (COST action BM0902) study

Read this article at

Abstract

Related collections

Karger: Oncology

Most cited references 40

Safety and efficacy of INCB018424, a JAK1 and JAK2 inhibitor, in myelofibrosis.

Safety and efficacy of TG101348, a selective JAK2 inhibitor, in myelofibrosis.

A prospective study of 338 patients with polycythemia vera: the impact of JAK2 (V617F) allele burden and leukocytosis on fibrotic or leukemic disease transformation and vascular complications.

Author and article information

Journal

Affiliations

Author notes

Article

History

Categories

Comments

Comment on this article

Similar content 329

Cited by 48

Most referenced authors 987