title: Inhibition of related JAK/STAT pathways with molecular targeted drugs shows strong synergy with ruxolitinib in chronic myeloproliferative neoplasm.
abstract: This study aimed to assess the antitumour effects, molecular mechanisms of action, and potential synergy of ruxolitinib with sorafenib, KNK437, dasatinib, and perifosine, in Philadelphia-negative chronic myeloproliferative neoplasms (MPN). Cytotoxic and cytostatic effects of the different compounds were determined in the JAK2 V617F-positive cell lines, HEL and Ba/F3 (JAK2V617F EPOR) , and in primary mononuclear and bone marrow CD34-positive cells from 19 MPN patients. Ruxolitinib [50% inhibitory concentration (IC50 )(PV)  = 15 nmol/l], as well as sorafenib (IC50 PV=8μmol/l), KNK437 (IC50 PV=100μmol/l ), and perifosine (IC50 PV=15μmol/l ), were able to inhibit proliferation in cell line models and in primary cells from MPN patients. Dasatinib, KNK437, and sorafenib showed a strong synergistic effect in combination with ruxolitinib [combination index (CI)(PV)  < 0·3]. Western blot confirmed that ruxolitinib blocked ERK, and consequently STAT5 activation, sorafenib inhibited ERK, P38 and STAT5, dasatinib blocked SRC and STAT5, and KNK437 decreased the stability of the JAK2 protein, reducing its expression. Inhibiting JAK2-related proliferative pathways has the potential to inhibit cell proliferation in MPNs. Furthermore, the combination of ruxolitinib with inhibitors that target these pathways has a strong synergistic effect, which may be due to decreased activation of the common effector, STAT5.
authors.affiliation: Haematology Service, Hospital Universitario 12 de Octubre, Madrid, Spain.
authors.forename: Santiago
authors.initials: S
authors.lastname: Barrio
authors.affiliation: Haematology Service, Hospital Universitario 12 de Octubre, Madrid, Spain.|Department of Leukemia, MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: Miguel
authors.initials: M
authors.lastname: Gallardo
authors.affiliation: Haematology Service, Hospital Universitario 12 de Octubre, Madrid, Spain.
authors.forename: Alicia
authors.initials: A
authors.lastname: Arenas
authors.affiliation: Haematology Service, Hospital Universitario 12 de Octubre, Madrid, Spain.
authors.forename: Rosa
authors.initials: R
authors.lastname: Ayala
authors.affiliation: Haematology Service, Hospital Universitario 12 de Octubre, Madrid, Spain.
authors.forename: Inmaculada
authors.initials: I
authors.lastname: Rapado
authors.affiliation: Haematology Service, Hospital Universitario 12 de Octubre, Madrid, Spain.
authors.forename: Daniel
authors.initials: D
authors.lastname: Rueda
authors.affiliation: Haematology Service, Hospital Universitario 12 de Octubre, Madrid, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Jiménez-Ubieto
authors.affiliation: Haematology Service, Hospital Universitario 12 de Octubre, Madrid, Spain.|Haematology Service, Hospital Virgen de la Salud, Toledo, Spain.
authors.forename: Enriqueta
authors.initials: E
authors.lastname: Albizua
authors.affiliation: Hospital Principe de Asturias, Alcala de Henares, Madrid, Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Burgaleta
authors.affiliation: Haematology Service, Hospital Universitario 12 de Octubre, Madrid, Spain.
authors.forename: Florinda
authors.initials: F
authors.lastname: Gilsanz
authors.affiliation: Haematology Service, Hospital Universitario 12 de Octubre, Madrid, Spain.
authors.forename: Joaquin
authors.initials: J
authors.lastname: Martinez-Lopez
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001559:Benzhydryl Compounds / Q000494:pharmacology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002908:Chronic Disease; D000069439:Dasatinib; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004357:Drug Synergism; D005260:Female; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics / Q000378:metabolism; D053612:Janus Kinases / Q000037:antagonists & inhibitors*; D008297:Male; D008875:Middle Aged; D009196:Myeloproliferative Disorders / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology; D009536:Niacinamide / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000494:pharmacology; D009570:Nitriles; D010671:Phenylurea Compounds / Q000008:administration & dosage / Q000494:pharmacology; D010767:Phosphorylcholine / Q000031:analogs & derivatives / Q000494:pharmacology; D011087:Polycythemia Vera / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011720:Pyrazoles / Q000008:administration & dosage / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology; D011760:Pyrrolidinones / Q000494:pharmacology; D050799:STAT5 Transcription Factor / Q000037:antagonists & inhibitors / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D000077157:Sorafenib; D013844:Thiazoles / Q000494:pharmacology; D013920:Thrombocythemia, Essential / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology; D014407:Tumor Cells, Cultured / Q000187:drug effects
chemical_list: D001559:Benzhydryl Compounds; C410027:KNK 437; D009570:Nitriles; D010671:Phenylurea Compounds; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D011743:Pyrimidines; D011760:Pyrrolidinones; D050799:STAT5 Transcription Factor; D013844:Thiazoles; D010767:Phosphorylcholine; D009536:Niacinamide; C105905:perifosine; C540383:ruxolitinib; D000077157:Sorafenib; C507924:JAK2 protein, human; D053614:Janus Kinase 2; D053612:Janus Kinases; D000069439:Dasatinib
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