TY - JOUR
T1 - Androgen receptor-modulatory microRNAs provide insight into therapy resistance and therapeutic targets in advanced prostate cancer
AU - Fletcher, Claire E.
AU - Sulpice, Eric
AU - Combe, Stephanie
AU - Shibakawa, Akifumi
AU - Leach, Damien A.
AU - Hamilton, Mark P.
AU - Chrysostomou, Stelios L.
AU - Sharp, Adam
AU - Welti, Jon
AU - Yuan, Wei
AU - Dart, Dafydd A.
AU - Knight, Eleanor
AU - Ning, Jian
AU - Francis, Jeffrey C.
AU - Kounatidou, Evangelia E.
AU - Gaughan, Luke
AU - Swain, Amanda
AU - Lupold, Shawn E.
AU - de Bono, Johann S.
AU - McGuire, Sean E.
AU - Gidrol, Xavier
AU - Bevan, Charlotte L.
N1 - Funding Information:
Acknowledgements We are grateful for the assistance of Laurent Guyon and Kristina Skare with statistical analysis of screening data. This work was supported by Prostate Cancer UK and the Movember Foundation through the London Movember Centre of Excellence (CEO13_2-002). We also gratefully acknowledge funding from Movember, Prostate Cancer UK and also the Prostate Cancer Foundation, and infrastructure support from the Cancer Research UK Imperial Centre, the Imperial Experimental Cancer Medicine Centre and the National Institute for Health Research Imperial Biomedical Research Centre.
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/7/11
Y1 - 2019/7/11
N2 - Androgen receptor (AR) signalling is a key prostate cancer (PC) driver, even in advanced ‘castrate-resistant’ disease (CRPC). To systematically identify microRNAs (miRs) modulating AR activity in lethal disease, hormone-responsive and -resistant PC cells expressing a luciferase-based AR reporter were transfected with a miR inhibitor library; 78 inhibitors significantly altered AR activity. Upon validation, miR-346, miR-361-3p and miR-197 inhibitors markedly reduced AR transcriptional activity, mRNA and protein levels, increased apoptosis, reduced proliferation, repressed EMT, and inhibited PC migration and invasion, demonstrating additive effects with AR inhibition. Corresponding miRs increased AR activity through a novel and anti-dogmatic mechanism of direct association with AR 6.9 kb 3′UTR and transcript stabilisation. In addition, miR-346 and miR-361-3p modulation altered levels of constitutively active AR variants, and inhibited variant-driven PC cell proliferation, so may contribute to persistent AR signalling in CRPC in the absence of circulating androgens. Pathway analysis of AGO-PAR-CLIP-identified miR targets revealed roles in DNA replication and repair, cell cycle, signal transduction and immune function. Silencing these targets, including tumour suppressors ARHGDIA and TAGLN2, phenocopied miR effects, demonstrating physiological relevance. MiR-346 additionally upregulated the oncogene, YWHAZ, which correlated with grade, biochemical relapse and metastasis in patients. These AR-modulatory miRs and targets correlated with AR activity in patient biopsies, and were elevated in response to long-term enzalutamide treatment of patient-derived CRPC xenografts. In summary, we identified miRs that modulate AR activity in PC and CRPC, via novel mechanisms, and may represent novel PC therapeutic targets.
AB - Androgen receptor (AR) signalling is a key prostate cancer (PC) driver, even in advanced ‘castrate-resistant’ disease (CRPC). To systematically identify microRNAs (miRs) modulating AR activity in lethal disease, hormone-responsive and -resistant PC cells expressing a luciferase-based AR reporter were transfected with a miR inhibitor library; 78 inhibitors significantly altered AR activity. Upon validation, miR-346, miR-361-3p and miR-197 inhibitors markedly reduced AR transcriptional activity, mRNA and protein levels, increased apoptosis, reduced proliferation, repressed EMT, and inhibited PC migration and invasion, demonstrating additive effects with AR inhibition. Corresponding miRs increased AR activity through a novel and anti-dogmatic mechanism of direct association with AR 6.9 kb 3′UTR and transcript stabilisation. In addition, miR-346 and miR-361-3p modulation altered levels of constitutively active AR variants, and inhibited variant-driven PC cell proliferation, so may contribute to persistent AR signalling in CRPC in the absence of circulating androgens. Pathway analysis of AGO-PAR-CLIP-identified miR targets revealed roles in DNA replication and repair, cell cycle, signal transduction and immune function. Silencing these targets, including tumour suppressors ARHGDIA and TAGLN2, phenocopied miR effects, demonstrating physiological relevance. MiR-346 additionally upregulated the oncogene, YWHAZ, which correlated with grade, biochemical relapse and metastasis in patients. These AR-modulatory miRs and targets correlated with AR activity in patient biopsies, and were elevated in response to long-term enzalutamide treatment of patient-derived CRPC xenografts. In summary, we identified miRs that modulate AR activity in PC and CRPC, via novel mechanisms, and may represent novel PC therapeutic targets.
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U2 - 10.1038/s41388-019-0823-5
DO - 10.1038/s41388-019-0823-5
M3 - Article
C2 - 31043708
AN - SCOPUS:85065237759
SN - 0950-9232
VL - 38
SP - 5700
EP - 5724
JO - Oncogene
JF - Oncogene
IS - 28
ER -