The contribution of tumor and host tissue factor expression to oncogene-driven gliomagenesis Nathalie Magnus a , Brian Meehan a , Delphine Garnier a , Maryam Hashemi a , Laura Montermini a , Tae Hoon Lee a , Chloe Milsom b , Rafal Pawlinski c , John Ohlfest d , G. Mark Anderson e , Nigel Mackman c , Janusz Rak a,⇑ a Montreal Children’s Hospital, RI MUHC, McGill University, Montreal, Quebec, Canada b Sunnybrook Research Institute, Toronto, Ontario, Canada c McAllister Heart Institute, Department of Medicine, University of North Carolina at Chapel Hill, USA d Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA e Biologics Research, Janssen Research & Development, Radnor, PA, USA article info Article history: Received 6 October 2014 Available online 23 October 2014 Keywords: Cancer Tissue factor Oncogene Brain tumor EGFR RAS abstract Glioblastoma multiforme (GBM) is an aggressive form of glial brain tumors, associated with angiogenesis, thrombosis, and upregulation of tissue factor (TF), the key cellular trigger of coagulation and signaling. Since TF is upregulated by oncogenic mutations occurring in different subsets of human brain tumors we investigated whether TF contributes to tumourigenesis driven by oncogenic activation of EGFR (EGFR- vIII) and RAS pathways in the brain. Here we show that TF expression correlates with poor prognosis in glioma, but not in GBM. In situ, the TF protein expression is heterogeneously expressed in adult and pedi- atric gliomas. GBM cells harboring EGFRvIII (U373vIII) grow aggressively as xenografts in SCID mice and their progression is delayed by administration of monoclonal antibodies blocking coagulant (CNTO 859) and signaling (10H10) effects of TF in vivo. Mice in which TF gene is disrupted in the neuroectodermal lineage exhibit delayed progression of spontaneous brain tumors driven by oncogenic N-ras and SV40 large T antigen (SV40LT) expressed under the control of sleeping beauty transposase. Reduced host TF lev- els in low-TF/SCID hypomorphic mice mitigated growth of glioma subcutaneously but not in the brain. Thus, we suggest that tumor-associated TF may serve as therapeutic target in the context of oncogene- driven disease progression in a subset of glioma. Ó 2014 Elsevier Inc. All rights reserved. 1. Introduction Astrocytic malignancies (gliomas) comprise a diverse cluster of primary brain tumors, of which glioblastoma multiforme (GBM) is especially aggressive [1]. GBM is also amongst the most vascular and procoagulant human malignancies [1–3] where microvascular vaso-occlusive thrombi occur regularly within tumor masses, and patients are at high risk of peripheral venous thromboembolism (VTE) [2,4]. These exclusively intracranial and lethal tumors have recently been subdivided into at least four molecular forms, including: proneural, neural, classical and mesenchymal GBM. Each of these subtypes is associated with a unique mutational and gene expression signatures, which are indicative of divergent pathogenetic mechanisms [5,6]. This diversity also includes dis- tinctive profiles of genes related to coagulation and fibrinolytic systems (coagulome), many of which are expressed by cancer cells ectopically (e.g., FVII) [7]. In this context, the key receptor trigger- ing the coagulation cascade, tissue factor (TF) is especially highly expressed in the classical subtype of GBM, which is also character- ized by the upregulation of the oncogenic epidermal growth factor receptor (EGFR) and the expression of its transforming mutant (EGFRvIII), along with activation of the RAS signaling pathway [7,8]. This is consistent with findings suggesting that the EGFR/ RAS pathways, regulate TF expression in cancer cells, including in GBM [9–12]. http://dx.doi.org/10.1016/j.bbrc.2014.10.041 0006-291X/Ó 2014 Elsevier Inc. All rights reserved. Abbreviations: EGFR, epidermal growth factor receptor; EGFRvIII, EGFR variant III; GBM, glioblastoma multiforme; PAI-1, plasminogen activator inhibitor 1; PAR (1–4), protease activated receptor (1–4); RAS, rat sarcoma oncogene; SCID, severe combined immunodeficiency (in mice); TF, tissue factor; VTE, venous thromboembolism. ⇑ Corresponding author at: Montreal Children’s Hospital Research Institute, McGill University, 4060 St. Catherine West, Montreal, QC H3Z 2Z3, Canada. Fax: +1 (514) 412 4331. E-mail address: janusz.rak@mcgill.ca (J. Rak). Biochemical and Biophysical Research Communications 454 (2014) 262–268 Contents lists available at ScienceDirect Biochemical and Biophysical Research Communications journal homepage: www.elsevier.com/locate/ybbrc