ET-13 CONTROL OF ACTIVATED MICROGLIA THROUGH P2X4 RECEPTOR IN RADIATION BRAIN NECROSIS

Abstract INTRODUCTION Brain radiation necrosis (RN) is severe adverse event after radiation therapy for brain tumor patients, especially in case of re-irradiation. Although corticosteroids or vitamin E, etc. are clinically used for RN, the effect is limited and underlying mechanism is to be cleared. Therefore, we established RN mouse model with irradiating right hemisphere of mouse brain using proton beam at dose of 60 Gy [Kondo et al., 2015]. In this study, we investigated change of phospholipids and lipid mediators after irradiation using this RN model in correlation with microglia activation. METHODS After irradiation, change of phospholipids and lipid mediators in mouse brain was investigated using imaging mass spectrometry and LC-MS. Immunohistochemistry on microglia and P2X4 receptor, a receptor for lysophosphatidylcholine (LPC) was performed. RESULTS In imaging mass spectrometry, 1 and 4 months after irradiation, phosphatidylcholine (PC): (16:0/20:4), (18:0/20:4) decreased in irradiated area compared non-irradiated area. On the other hand, LPC: (16:0) increased in irradiated area compared to non-irradiated area after 1 month and 4 months irradiation. PC (16:0/20:4) is a precursor of LPC (16:0) and arachidonic acid (20:4). By LC-MS, LPC was twice higher in irradiated area compared to non-irradiated, 6 months after irradiation. Microglia was highly activated in irradiated area compared to non-irradiated from 3 months after irradiation to 8 months and strongly co-expressed P2X4 receptor was confirmed in irradiated area after 6 months. Preliminary P2X4 receptor agonist administration test prolonged the RN to 12 months after irradiation. CONCLUSION In RN, LPC may continuously activated microglia through P2X4 receptor and cause chronic inflammation after irradiation. P2X4 agonist administration test including action resolution and immunohistochemistry is ongoing.