JAK the Raf Kinase Inhibitor Sorafenib Inhibits
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The Raf Kinase Inhibitor Sorafenib Inhibits JAK−STAT Signal Transduction in Human Immune Cells This information is current as Sara E. Martin del Campo, Kala M. Levine, Bethany L. of October 2, 2021. Mundy-Bosse, Valerie P. Grignol, Ene T. Fairchild, Amanda R. Campbell, Prashant Trikha, Thomas A. Mace, Bonnie K. Paul, Alena Cristina Jaime-Ramirez, Joseph Markowitz, Sri Vidya Kondadasula, Kristan D. Guenterberg, Susan McClory, Volodymyr I. Karpa, Xueliang Pan, Thomas E. Olencki, J. Paul Monk, Amir Mortazavi, Susheela Downloaded from Tridandapani, Gregory B. Lesinski, John C. Byrd, Michael A. Caligiuri, Manisha H. Shah and William E. Carson III J Immunol published online 3 August 2015 http://www.jimmunol.org/content/early/2015/08/01/jimmun ol.1400084 http://www.jimmunol.org/ Supplementary http://www.jimmunol.org/content/suppl/2015/08/01/jimmunol.140008 Material 4.DCSupplemental Why The JI? Submit online. by guest on October 2, 2021 • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2015 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published August 3, 2015, doi:10.4049/jimmunol.1400084 The Journal of Immunology The Raf Kinase Inhibitor Sorafenib Inhibits JAK–STAT Signal Transduction in Human Immune Cells Sara E. Martin del Campo,* Kala M. Levine,† Bethany L. Mundy-Bosse,† Valerie P. Grignol,*,† Ene T. Fairchild,‡ Amanda R. Campbell,x Prashant Trikha,† Thomas A. Mace,† Bonnie K. Paul,{ Alena Cristina Jaime-Ramirez,‖ Joseph Markowitz,† Sri Vidya Kondadasula,# Kristan D. Guenterberg,** Susan McClory,†† Volodymyr I. Karpa,‡‡ Xueliang Pan,xx Thomas E. Olencki,{{ J. Paul Monk,{{ Amir Mortazavi,{{ Susheela Tridandapani,†,‖‖ Gregory B. Lesinski,†,{{ John C. Byrd,†,## Michael A. Caligiuri,†,##,*** Manisha H. Shah,{{ and William E. Carson, III*,†,*** Sorafenib is an oral multikinase inhibitor that was originally developed as a Raf kinase inhibitor. We hypothesized that sorafenib Downloaded from would also have inhibitory effects on cytokine signaling pathways in immune cells. PBMCs from normal donors were treated with varying concentrations of sorafenib and stimulated with IFN-a or IL-2. Phosphorylation of STAT1 and STAT5 was measured by flow cytometry and confirmed by immunoblot analysis. Changes in IFN-a– and IL-2–stimulated gene expression were measured by quantitative PCR, and changes in cytokine production were evaluated by ELISA. Cryopreserved PBMCs were obtained from cancer patients before and after receiving 400 mg sorafenib twice daily. Patient PBMCs were thawed, stimulated with IL-2 or IFN-a, and evaluated for phosphorylation of STAT1 and STAT5. Pretreatment of PBMCs with 10 mM sorafenib decreased STAT1 http://www.jimmunol.org/ and STAT5 phosphorylation after treatment with IFN-a or IL-2. This inhibitory effect was observed in PBMCs from healthy donors over a range of concentrations of sorafenib (5–20 mM), IL-2 (2–24 nM), and IFN-a (101–106 U/ml). This effect was observed in immune cell subsets, including T cells, B cells, NK cells, regulatory T cells, and myeloid-derived suppressor cells. Pretreatment with sorafenib also inhibited PBMC expression of IFN-a– and IL-2–regulated genes and inhibited NK cell produc- tion of IFN-g, RANTES, MIP1-a, and MIG in response to IFN-a stimulation. PBMCs from patients receiving sorafenib therapy showed decreased responsiveness to IL-2 and IFN-a treatment. Sorafenib is a Raf kinase inhibitor that could have off-target effects on cytokine-induced signal transduction in immune effector cells. The Journal of Immunology, 2015, 195: 000–000. by guest on October 2, 2021 orafenib (BAY43-9006, Nexavar) is an oral multikinase tumor-induced angiogenesis (3). Activating mutations in B-RAF inhibitor that was originally developed as a Raf kinase in- and genetic rearrangements in RET are critical to the development S hibitor (1). It was shown to have inhibitory effects on the of thyroid cancers, and phase II trials have shown that sorafenib wild-type B-Raf and oncogenic b-raf V600E serine/threonine leads to stabilization of disease in this setting (4–6). Sorafenib kinases, proangiogenic receptor tyrosine kinases such as vascular leads to reduced tumor cell growth and vascularization in renal endothelial growth factor receptor-1, -2, and -3, platelet-derived cell carcinoma (7) and induces apoptosis in imatinib mesylate– growth factor receptor-b, and fibroblast growth factor receptor-1. resistant human leukemia cells by inhibiting STAT5 in myeloid Sorafenib also has inhibitory effects on other receptor tyrosine cell leukemia-1 (8). In human glioblastoma, medulloblastoma, and kinases such as c-Kit, FLT-3, p38, and RET (1, 2). These molecular neuroblastoma, sorafenib contributes to growth arrest and apo- targets are implicated in the etiology of several human cancers (1). ptosis by inhibition of STAT signaling (9–11). In hepatocellular carcinoma, sorafenib has been shown to inhibit Sorafenib is the only Food and Drug Administration–approved the RAF/MEK/ERK pathway, inducing apoptosis and reducing medication for use in advanced hepatocellular carcinoma (HCC). *Department of Surgery, The Ohio State University, Columbus, OH 43210; This work was supported by National Institutes of Health Grants P01 CA095426 †Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210; (to M.A.C.), P30 CA16058 (to M.A.C.), T32 CA090223 (to W.E.C.), T32 ‡Department of General Pediatrics, Nationwide Children’s Hospital, Columbus, OH CA009338 (to M.A.C.), T32 GM068412 (to A.C.J.-R. and B.L.M.-B.), K24 CA093670 43205; xMedical Scientist Training Program and Biomedical Sciences Graduate Program, (to W.E.C.), K22 CA134551 (to G.B.L.), and by a Valvano Foundation for Cancer The Ohio State University, Columbus, OH 43210; {Cardiovascular Medicine, The Ohio Research award (to G.B.L.). State University, Columbus, OH 43210; ‖Department of Neurological Surgery, The Ohio Address correspondence and reprint requests to Dr. William E. Carson, III, Depart- State University, Arthur G. James Cancer Hospital and Richard J. Solove Research Insti- ment of Surgery, The Ohio State University, N924 Doan Hall, 410 West 10th Avenue, tute, Columbus, OH 43210; #Department of Oncology, Karmanos Cancer Institute, Detroit, Columbus, OH 43210. E-mail address: [email protected] MI 48201; **Kitsap General Surgery, Silverdale, WA 98383; ††Department of Internal Medicine, Barnes-Jewish Hospital, St. Louis, MO 63110; ‡‡Department of Psychiatry, The online version of this article contains supplemental material. University of Hawaii, Honolulu, HI 96813; xxDepartment of Biomedical Informatics, {{ Abbreviations used in this article: CIS, cytokine-inducible SH2 domain–con- The Ohio State University, Columbus, OH 43210; Medical Oncology, The Ohio State taining protein; F , specific fluorescence; HCC, hepatocellular carcinoma; University, Arthur G. James Cancer Hospital and Richard J. Solove Research Institute, sp ‖‖ IFIT2, IFN-induced protein with tetratricopeptide repeats 2; IFNAR, IFN-a Columbus, OH 43210; Department of Pulmonary, Allergy, Critical Care and Sleep, The receptor; MDSC, myeloid-derived suppressor cell; OAS1, 29,59-oligoadenylate Ohio State University, Columbus, OH 43210; ##Department of Internal Medicine, The synthetase 1; RCC, renal cell carcinoma; SOCS1, suppressor of cytokine sig- Ohio State University, Columbus, OH 43210; and ***Department of Molecular Virology, naling 1. Immunology and Medical Genetics, The Ohio State University, Arthur G. James Cancer Hospital and Richard J. Solove Research Institute, Columbus, OH 43210 Copyright Ó 2015 by The American Association of Immunologists, Inc. 0022-1767/15/$25.00 Received for publication January 31, 2014. Accepted for publication July 7, 2015. www.jimmunol.org/cgi/doi/10.4049/jimmunol.1400084 2 SORAFENIB INHIBITS CYTOKINE SIGNALING IN IMMUNE CELLS The phase III Sorafenib Hepatocellular Carcinoma Assessment Cell lines Randomized Protocol trial demonstrated that sorafenib signifi- The lymphoma cell lines 697 and Ramos were obtained from Dr. John Byrd cantly prolonged time to progression and overall survival in patients (The Ohio State University). The SK-RC-45 human renal cell carcinoma with unresectable HCC (12, 13). Sorafenib has also been approved cell lines were obtained from Dr. Charles Tannenbaum (Cleveland Clinic 2 + by the Food and Drug Administration for use in renal cell carci- and Foundation, Cleveland, OH). NK-92 is a CD3 CD56 NK cell leu- noma (RCC) based on its ability to increase progression-free sur- kemia cell line (15) provided by Dr. Hans G. Klingermann (Vancouver, BC, Canada). The Caki human renal cell carcinoma, A375 and Hs294T vival in patients with metastatic disease. The activity of sorafenib in human melanoma cell lines, and K562 human chronic myelogenous leu- this setting is attributed to its ability to abrogate the effects of se- kemia cell line were