Greg Brooke Molecular Oncology

Prostate Cancer - Leading Cause of Cancer- related death in men - Growth dependent on androgens - Therapy targets androgen action - Therapy failure is a significant clinical problem Breast Cancer - Leading Cause of Cancer- related death in women - Growth often dependent upon oestrogens - Therapy targets oestrogen action - Therapy failure is a significant clinical problem

Prostate Cancer growth is dependent upon the Androgen Receptor Pathway The Androgen Receptor Pathway ARE HSP complex Corepressors DHTAntiandrogen The AR Pathway Androgen Blockade Coactivators GROWTH Potential Biomarkers/Therapeuti c Targets Characterising proteins that regulate AR action - Inhibition of critical interactions Mechanisms of therapy resistance

FUS is an Important Regulator of Prostate Cancer Growth EtOH MIB Time (hrs) FUS  -actin FUS  -actin blot Time (days) Relative Growth * *** LNCaP FUS - DOX + EtOH + DOX +EtOH - DOX + MIB + DOX + MIB Time (days) Tumour size Increasing FUS Decreasing FUS Increasing FUS IN VIVO Brooke et al Cancer Research. 71;

ER ERE E2Antiestrogen AR ER Corepressors The ERα Pathway Hormone therapy Coactivators GROWTH The Androgen Receptor Pathway Can Drive Therapy Resistance in Breast Cancer GROWTH

MCF7 TAMR LTED E2 (nM) MIB (nM) *** ** *** ** Androgens stimulate the growth of endocrine resistant breast cancer Inhibition of AR may be a valid method to inhibit endocrine resistance

Confocal imaging and FRET (with Phillippe Laissue) RNA-seq and ChIP-seq Characterisation of AR/ER α crosstalk

AF-1 LBD AF-1 AR 1-54 Repressor Motif ARE AF-1 LBD Coactivators Corepressors Engineered Repressors are potent inhibitors of Androgen Receptor Activity Brooke et al. Submitted [AR 1-54 ] ng [MAD 7-35 AR 1-54 ] ng Isolated domain Fused domains Repression domain / interaction motif: Mock REP2-IM ** Inhibition of AR activity % activity Repressor Inhibition of cell growth

Techniques and Collaborations In cell and in vivo models (Honorary Contract with Imperial) – Transcription assays, 2-hybrid assays, confocal, ChIP, growth assays, FACS, DOX-inducible cell lines, motility assays…… Collaborations – Common interests in Breast and Prostate Cancer e.g. FUS and CTCF/BORIS (Elena) – Peptide inhibitors of AR (Jody) – Fluorescent Microscopy – AR/ER in BCa (Phillipe) – Proteomics (Metodi) Future Collaborations? – Protein aggregations FUS (Amyotrophic Lateral Sclerosis) Androgen Receptor (Kennedy’s Disease) – Metabolomics