Ropeginterferon alpha-2b targets JAK2V617F-positive polycythemia vera cells in vitro and in vivo

Emmanuelle Verger, Juliette Soret-Dulphy, Nabih Maslah, Lydia Roy, Jerome Rey, Zineb Ghrieb, Robert Kralovics, Heinz Gisslinger, Barbara Grohmann-Izay, Christoph Klade, Christine Chomienne, Stéphane Giraudier, Bruno Cassinat, Jean-Jacques Kiladjian, Emmanuelle Verger, Juliette Soret-Dulphy, Nabih Maslah, Lydia Roy, Jerome Rey, Zineb Ghrieb, Robert Kralovics, Heinz Gisslinger, Barbara Grohmann-Izay, Christoph Klade, Christine Chomienne, Stéphane Giraudier, Bruno Cassinat, Jean-Jacques Kiladjian

Abstract

Polycythemia vera is characterized by the acquisition of the JAK2V617F mutation. Recommended treatments include hydroxyurea and interferon-alpha. Several groups have reported a reduction in the JAK2 mutant allele burden in interferon-treated patients, but significance of this observation is questioned. We characterized the activity of ropeginterferon alpha-2b, a novel form of interferon-alpha recently shown to be safe and efficacious in polycythemia vera. Ropeginterferon was able to inhibit the proliferation of the HEL, UKE-1, and UT-7 JAK2-mutant cell lines while sparing JAK2-wild-type UT-7 and normal CD34+ cells growth. In vitro treatment of erythroid progenitors derived from PV patients showed that ropeginterferon could considerably inhibit the growth of endogenous erythroid colonies, a hallmark of polycythemia vera. Finally, we could study in sequential samples the clonal architecture of erythroid progenitors derived from patients included in a randomized study comparing hydroxyurea to ropeginterferon. After 1 year of treatment with ropeginterferon, the ratio of JAK2-mutated to wild-type colonies grown from bone marrow progenitors was reduced by 64%, compared to 25% in patients receiving hydroxyurea. This study shows that ropeginterferon has a potent targeted activity against JAK2-mutant cells and is able to drastically reduce the proportion of malignant progenitors in patients treated with this drug.

Trial registration: ClinicalTrials.gov NCT01949805.

Conflict of interest statement

Conflict of interest

Dr Cassinat and Dr Kiladjian received institutional research grants from Novartis and AOP Orphan; Dr Kiladjian participated in advisory boards for Novartis and AOP Orphan.

Ethics approval and consent to participate

The study was approved by the local ethics committee (IRB0006477). All patients participating in the PROUD-PV clinical trial (NCT01949805) provided informed consent.

Figures

Fig. 1. Antiproliferative effect of Ropeg in…
Fig. 1. Antiproliferative effect of Ropeg in MPN-derived human cell lines.
A and B The JAK2V617F positive UKE1 and HEL cell lines. C and D The UT-7 cell line expressing a wild type or a mutant form of JAK2. Cells were treated with the indicated drugs and the living cells were counted every day. Results are expressed as the fold increase compared to day 0
Fig. 2. Targeted inhibition of JAK2 V617F…
Fig. 2. Targeted inhibition of JAK2V617F progenitors in vitro and in vivo
A Clonogenic assays on primary peripheral blood mononuclear cells from 4 PV patients. Median percentages and standard deviations of residual erythroid colonies in treated conditions compared to untreated are presented. B and C JAK2V617F allele burden evolution in patients included in the PROUD-PV trial in France. B median of the %JAK2V617F in HU (N = 5) or Ropeg (N = 3) treatment arms. C Comparison of the % reduction of JAK2V617F allele burden in patients after 1 year of treatment in each arm. Median and standard deviations in both groups of treatment are represented. (*p < 0.02; unpaired t-test). D and E Clonogenic assays performed on bone marrow samples from patients included in the PROUD-PV trial in France. Percentages of JAK2V617F positive erythroid colonies before and after 1 year of treatment in patients treated with Ropeg or HU. UPN of each individual patient is given

References

    1. James C, et al. A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera. Nature. 2005;434:1144–1148. doi: 10.1038/nature03546.
    1. Vainchenker W, Kralovics R. Genetic basis and molecular pathophysiology of classical myeloproliferative neoplasms. Blood. 2017;129:667–679. doi: 10.1182/blood-2016-10-695940.
    1. Tefferi A. Myeloproliferative neoplasms: a decade of discoveries and treatment advances. Am. J. Hematol. 2016;91:50–58. doi: 10.1002/ajh.24221.
    1. Verger E, et al. Clinical and molecular response to interferon alpha therapy in essential thrombocythemia patients with CALR mutations. Blood. 2015;126:2585–91. doi: 10.1182/blood-2015-07-659060.
    1. Vannucchi AM, et al. Philadelphia chromosome-negative chronic myeloproliferative neoplasms: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up†. Ann. Oncol. 2015;26(suppl 5):v85–v99. doi: 10.1093/annonc/mdv203.
    1. Barbui T, et al. Philadelphia chromosome-negative classical myeloproliferative neoplasms: revised management recommendations from European Leukemia Net. Leukemia. 2018;32:1057–1069. doi: 10.1038/s41375-018-0077-1.
    1. Tefferi A, Vannucchi AM, Barbui T. Polycythemia vera treatment algorithm 2018. Blood Cancer J. 2018;8:3. doi: 10.1038/s41408-017-0042-7.
    1. Silver RT. Recombinant interferon-alpha for treatment of polycythaemia vera. Lancet. 1988;2:403. doi: 10.1016/S0140-6736(88)92881-4.
    1. Kiladjian JJ, Giraudier S, Cassinat B. Interferon-alpha for the therapy of myeloproliferative neoplasms: targeting the malignant clone. Leukemia. 2016;30:776–781. doi: 10.1038/leu.2015.326.
    1. Kiladjian JJ. Long-term treatment with interferon alfa for myeloproliferative neoplasms. Lancet Haematol. 2017;4:e150–e151. doi: 10.1016/S2352-3026(17)30039-X.
    1. Kiladjian JJ, et al. Pegylated interferon-alfa-2a induces complete hematologic and molecular responses with low toxicity in polycythemia vera. Blood. 2008;112:3065–3072. doi: 10.1182/blood-2008-03-143537.
    1. Quintás-Cardama A, et al. Pegylated interferon alfa-2a yields high rates of hematologic and molecular response in patients with advanced essential thrombocythemia and polycythemia vera. J. Clin. Oncol. 2009;27:5418–5424. doi: 10.1200/JCO.2009.23.6075.
    1. Hasan S, et al. Use of the 46/1 haplotype to model JAK2(V617F) clonal architecture in PV patients: clonal evolution and impact of IFNα treatment. Leukemia. 2014;28:460–463. doi: 10.1038/leu.2013.303.
    1. Mullally A, et al. Depletion of Jak2V617F myeloproliferative neoplasm-propagating stem cells by interferon-α in a murine model of polycythemia vera. Blood. 2013;121:3692–3702. doi: 10.1182/blood-2012-05-432989.
    1. Hasan S, et al. JAK2V617F expression in mice amplifies early hematopoietic cells and gives them a competitive advantage that is hampered by IFNα. Blood. 2013;122:1464–1477. doi: 10.1182/blood-2013-04-498956.
    1. Masarova L, et al. Pegylated interferon alfa-2a in patients with essential thrombocythaemia or polycythaemia vera: a post-hoc, median 83 month follow-up of an open-label, phase 2 trial. Lancet Haematol. 2017;4:e165–e175. doi: 10.1016/S2352-3026(17)30030-3.
    1. Them NC, et al. Molecular responses and chromosomal aberrations in patients with polycythemia vera treated with peg-proline-interferon alpha-2b. Am. J. Hematol. 2015;90:288–294. doi: 10.1002/ajh.23928.
    1. Gisslinger H, et al. Ropeginterferon alfa-2b, a novel IFNα-2b, induces high response rates with low toxicity in patients with polycythemia vera. Blood. 2015;126:1762–9. doi: 10.1182/blood-2015-04-637280.
    1. Mahon FX. Hematol. Am. Soc. Hematol. Educ. Program. 2017. Treatment-free remission in CML: who, how, and why? pp. 102–109.

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