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Amplification of androgen receptor ( AR ) is a common genomic event in metastatic castration-resistant prostate cancer (mCRPC). To evaluate the prognostic value of the amplifications of specific loci in the AR gene in cell-free DNA, we developed a multiplex digital PCR (dPCR) assay that targeted AR enhancer ( AR -En), AR exon 1 ( AR -E1), AR exon 8 ( AR -E8) and OPHN1 (downstream of AR ). We selected three relatively stable genes, C2orf16 , FAM111B , and GRIA3, as reference controls for copy number normalization. One hundred and eight mCRPC patients were recruited to test the association of specific AR loci amplification with clinical outcome. Using a normalized ratio ≥ 1.92 as cutoff, amplification of AR -En, AR- E1, AR -E8 and OPHN1 was observed in 28, 25, 24 and 19 of 108 mCRPC patients, respectively. Among the 41 patients with AR region amplification, 9 (21.9%) showed amplification at all four selected regions and 15 (36.6%) showed amplification at AR -En, AR-E1, and AR- E8. Six (14.6%) patients showed independent AR -En amplification, while the remaining 3 (7.3%) demonstrated AR -E8 amplification only. Kaplan-Meier analysis showed overall survival's association with the amplification of AR -En ( p = 0.02, HR = 1.68 (1.07-2.65)), AR -E8 ( p = 0.02, HR = 1.78 (1.08-2.92)) and AR -En-E8 (the combination of AR -En and AR -E8 ( p = 0.009, HR = 1.77 (1.15-2.73)). Multivariate models that included AR -En-E8 amplification and clinical factors significantly improved prognostic performance ( p = 0.0001). With further validation, the multiplex dPCR assay may assist in prognostication of mCRPC patients.

cancers

Multiplex Digital PCR to Detect Amplifications of Specific Androgen Receptor Loci in Cell-Free DNA for Prognosis of Metastatic Castration-Resistant Prostate Cancer