Expression of angiopoietin-like 4 fibrinogen-like domain (cANGPTL4) increases risk of brain metastases in women with breast cancer

// Tu Dao 1 , 2 , 3 , 9 , * , Guillaume Gapihan 1 , * , Christophe Leboeuf 1 , Diaddin Hamdan 1 , Jean-Paul Feugeas 4 , 10 , Hanene Boudabous 5 , Laurent Zelek 5 , 6 , Catherine Miquel 1 , 7 , Thuan Tran 2 , 3 , Catherine Monnot 8 , Stéphane Germain 8 , Anne Janin 1 , 7 , # and Guilhem Bousquet 1 , 5 , 6 , # 1 Université Paris Diderot, Inserm, UMR_S942, Paris, France 2 Medical Oncology Department, National Cancer Hospital, Ha Noi, Vietnam 3 Ha Noi Medical University, Oncology Department, Ha Noi, Vietnam 4 INSERM, U722-Paris, Paris, France 5 Oncology Department, Hôpital Avicenne, APHP, Bobigny, France 6 Université Paris 13, Villetaneuse, Paris, France 7 Pathology Department, Hôpital St Louis, APHP, Paris, France 8 Center for Interdisciplinary Research in Biology (CIRB), College de France, CNRS, INSERM, PSL Research University, Paris, France 9 Cancer Research and Clinical Trial Center, National Cancer Hospital, Ha Noi, Vietnam 10 Université de Franche Comté, Besançon, France * These authors contributed equally to this work # These authors are co-senior authors Correspondence to: Guilhem Bousquet, email: guilhem.bousquet@aphp.fr Anne Janin, email: anne.janin1165@gmail.com Keywords: cANGPTL4; brain metastases; breast cancer; blood-brain barrier; vascular permeability Received: November 27, 2019     Accepted: March 19, 2020     Published: May 05, 2020 ABSTRACT Background: Brain metastases challenge daily clinical practice, and the mechanisms by which cancer cells cross the blood-brain barrier remain largely undeciphered. Angiopoietin-like 4 (ANGPTL4) proteolytic fragments have controversial biological effects on endothelium permeability. Here, we studied the link between ANGPTL4 and the risk of brain metastasis in cancer patients. Materials and Methods: From June 2015 to June 2016, serum samples from 113 cancer patients were prospectively collected, and ANGPTL4 concentrations were assessed. Using a murine model of brain metastases, we investigated the roles of nANGPTL4 and cANGPTL4, the two cleaved fragments of ANGPTL4, in the occurrence of brain metastases. Results: An ANGPTL4 serum concentration over 0.1 ng/mL was associated with decreased overall-survival. Multivariate analyses found that only breast cancer brain metastases were significantly associated with elevated ANGPTL4 serum concentrations. 4T1 murine breast cancer cells were transfected with either nANGPTL4- or cANGPTL4 -encoding cDNAs. Compared to mice injected with wild-type 4T1 cells, mice injected with nANGPTL4 cells had shorter median survival ( p < 0.05), while mice injected with cANGPTL4 had longer survival ( p < 0.01). On tissue sections, compared to wild-type mice, mice injected with nANGPTL4 cells had significantly larger surface areas of lung metastases ( p < 0.01), and mice injected with cANGPTL4 had significantly larger surface areas of brain metastases ( p < 0.01). Conclusions: In this study, we showed that a higher expression of Angiopoietin-like 4 Fibrinogen-Like Domain (cANGPTL4) was associated with an increased risk of brain metastases in women with breast cancer.