Page 91 - 2019_05-HaematologicaMondo-web

Myeloproliferative Neoplasms
Ruxolitinib in combination with prednisone and nilotinib exhibit synergistic effects in human cells lines and primary cells from myeloproliferative neoplasms
Alicia Arenas Cortés,1 Rosa Ayala Diaz,1 Pilar Hernández-Campo,2
Julián Gorrochategui,2 Daniel Primo,2 Alicia Robles,2 María Luz Morales,1,3 Joan Ballesteros,2 Inmaculada Rapado,1 Miguel Gallardo,*3
María Linares*1,3,4 and Joaquín Martínez-López*1,3,4
ABSTRACT
Ruxolitinib is the front-line non-palliative treatment for myelofibro- sis (MF). However, a significant number of patients lose or present suboptimal response, are resistant or have unacceptable toxicity. In an attempt to improve response and avoid the adverse effects of this drug, we evaluated the combination of 17 drugs with ruxolitinib in ex vivo models of peripheral blood mononuclear cells from MF patients and cell lines. We found that the combination ruxolitinib and nilotinib had a synergistic effect against MF cells (ΔEC50 nilotinib, -21.6%). Moreover, the addition of prednisone to combined ruxolitinib/nilotinib improved the synergistic effect in all MF samples studied. We evaluated the molecular mechanisms of combined ruxolitinib/nilotinib/pred- nisone and observed inhibition of JAK/STAT (STAT5, 69.2+11.8% inhi- bition) and MAPK (ERK, 29.4+4.5% inhibition) signaling pathways. Furthermore, we found that the triple therapy combination inhibited collagen protein and COL1A1 gene expression in human bone marrow mesenchymal cells. Taken together, we provide evidence that combined ruxolitinib/nilotinib/prednisone is a potential therapy for MF, possibly through the anti-fibrotic effect of nilotinib, the immunomodulatory effect of ruxolitinib and prednisone, and the anti-proliferative effect of ruxolitinib. This combination will be further investigated in a phase Ib/II clinical trial in MF.
Introduction
Myelofibrosis (MF) is a Philadelphia chromosome-negative chronic myeloprolif- erative neoplasm (MPN) clinically characterized by stem cell-derived clonal myelo- proliferation and a reactive cytokine-driven increase in bone marrow (BM) fibrosis.1,2 Patients with MF have a poor prognosis and a median survival of 5.8 years.1
Dysregulation of JAK/STAT signaling is the main cause of MPN and, accordingly, inhibitors of the JAK/STAT signal transduction pathway are currently the best clin- ical approach to treat this disease. Discovered in 2005,3-7 a mutation in the JAK2 gene resulting in a substitution of valine for phenylalanine (V617F) was found in approximately 90% of patients with polycythemia vera (PV), and in 50-60% of patients with essential thrombocythemia (ET) and primary myelofibrosis (PMF).8 In addition, mutations in the MPL gene, which encodes the thrombopoietin recep- tor, were found in approximately 1% of patients with MPN,9 and 12% of patients with MPN (35-50% of MF) have mutations in calreticulin (CALR).10-12 Interestingly, the mutated forms of CALR acquire the ability to activate the thrombopoietin
Ferrata Storti Foundation
*MG, ML and JM-L contributed equally to this work
1Hematology Service, Hospital Universitario 12 de Octubre; 2Vivia Biotech, Tres Cantos; 3H12O-CNIO Haematological Malignancies Clinical Research Unit, Centro Nacional de Investigaciones Oncológicas (CNIO) and 4Universidad Complutense de Madrid, Madrid, Spain.
Haematologica 2019 Volume 104(5):937-946
Correspondence:
MIGUEL GALLARDO miguelgallardodelgado@gmail.com
Received: July 10, 2018.
Accepted: December 10, 2018. Pre-published: December 13, 2018.
doi:10.3324/haematol.2018.201038
Check the online version for the most updated information on this article, online supplements, and information on authorship & disclosures: www.haematologica.org/content/104/5/937
©2019 Ferrata Storti Foundation
Material published in Haematologica is covered by copyright. All rights are reserved to the Ferrata Storti Foundation. Use of published material is allowed under the following terms and conditions: https://creativecommons.org/licenses/by-nc/4.0/legalcode. Copies of published material are allowed for personal or inter- nal use. Sharing published material for non-commercial pur- poses is subject to the following conditions: https://creativecommons.org/licenses/by-nc/4.0/legalcode, sect. 3. Reproducing and sharing published material for com- mercial purposes is not allowed without permission in writing from the publisher.
haematologica | 2019; 104(5)
937
ARTICLE