P10.17.B Androgen receptor signalling has independent prognostic value and is associated with resistance to temozolomide and tumour growth in glioblastoma

Abstract

Abstract Background The spectrum of sex differences in glioblastoma (GBM) is a rising topic. GBM is more frequent (sex ratio M:F=1.6) and seem to have worse prognosis in males (M) compared to females. Androgen receptor (AR) is expressed in GBM but its role in GBM biology and clinics is not yet fully understood. Material and Methods We have selected from our tumour tissue bank: (i) GBM samples with clinical, pathological and molecular annotations and (ii) GBM-patient derived cell lines -GBM-PDCL- with transcriptomic profiling and available tissue from parental tumours.AR expression was investigated using multiplex IHC Opal stained-FFPE sections. Sensitivity of PDCL to hormonotherapy (dihydrotestosterone or enzalutamide EZT) alone or combined with temozolomide (TMZ) was tested using WST1 viability assay. Response to drugs were correlated to AR expression (high: log2 [FPKM] >2 or low: log2 [FPKM] <2). PDCLs dihydrotestosterone secretion was examined by ELISA in both supernatants and cell lysates. Results In our cohort of 1188 IDH muted GBM patients, the median follow-up was 27.7 (25.8-31.3). The sex ratio M:F was 1.68 and the median age at diagnosis was 57 years [18-69]. Compared to males, females have better prognosis with a median overall survival of 26.1 months vs 20.8 months (p<0.0001). In multivariate analysis, male gender was an independent prognostic factor with a HR of 1.43 CI95%(1.21-1.69) (p<0.0001) adjusted on age, MGMT status and Karnofsky indice . Although interaction between age and sex was not significant (p = 0.854), HR of gender varied through age category 1.53 CI95%( 1.01-2.33 ), 1.51 CI95%( 0.888-2.55 ) and 1.38 CI95%( 0.857-2.21) for males <45 years, between 46-55 years and > 55 years respectively. In GBM tumours, AR staining was positive in 5- 30% tumour cells. After 72h exposure to 10nM dihydrotestosterone, ARhigh GBM-PDCLs exhibit AR nuclear translocation and c-myc expression upregulation, suggesting functional activation of the receptor. AR pharmacological blockade increased temozolomide sensitivity in ARhigh GBM-PDCLs regardless of the patient’s sex. Moreover, ARhigh PDCLs secreted DHT, suggesting autocrine and paracrine AR stimulation in tumour microenvironment. Conclusion Our study highlights the independent prognostic significance of sex in GBM patients, where males exhibit a worse prognosis. In GBM-PDCL, we show that androgen receptor is (i) functional in vitro and (ii) associated with resistance to temozolomide. Further studies are needed to investigate the role of AR in tumour tissue microenvironment.