High throughput multiplex image analyses for androgen receptor function

Abstract

Evidence suggest that a subgroup of patients affected by either prostate cancer or androgen insensitivity syndrome harbor mutations within the androgen receptor that may contribute to the disease phenotype. To characterize the effects of these AR mutations, we have developed a high content screening assay able to determine AR transcriptional activity, cellular distribution, and cellular patterning simultaneously at the single cell level. We demonstrate that two mutations (F764L, R840C) isolated from AIS patients retain the ability to achieve similar levels of transcriptional activity, nuclear translocation, and nuclear hyperspeckling as wild type receptor, but require significantly higher levels of agonist. Differences in responses seen between the different compounds tested also suggest that the assay could be amendable to agonist screening for personalized patient drug selection.