Bioluminescence Microscopy as a Method to Measure Single Cell Androgen Receptor Activity Heterogeneous Responses to Antiandrogens.

Pallavi Jain,Frédéric Pouliot,Lily Wu,Yves Fradet,Lauriane Velot,Bertrand Neveu

doi:10.1038/srep33968

Pallavi Jain, Frédéric Pouliot + Show 4 more

Open Access

https://doi.org/10.1038/srep33968

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Abstract

Cancer cell heterogeneity is well-documented. Therefore, techniques to monitor single cell heterogeneous responses to treatment are needed. We developed a highly translational and quantitative bioluminescence microscopy method to measure single cell androgen receptor (AR) activity modulation by antiandrogens from fluid biopsies. We showed that this assay can detect heterogeneous cellular response to drug treatment and that the sum of single cell AR activity can mirror the response in the whole cell population. This method may thus be used to monitor heterogeneous dynamic treatment responses in cancer cells.

Highlights

In addition to tumor heterogeneity, another barrier to predict drug response is the number of possible resistance mechanisms used by cancer cells to escape anti-cancer-drug inhibitory effects[6]
With the goal of imaging primary prostate cancer (PCa) single cell response to antiandrogens, we first had to develop conditions for an appropriate imaging system driven by a promoter containing the androgen response elements sequence (ARE), which could be delivered into PCa cells
We determined the optimal conditions for viral transduction and the amount of infectious viral particles required to enable the detection of more than 90% of the cells, as the ultimate goal was to enable the detection of primary PCa cells in fluid biopsies (Fig. 1b and Supplementary Fig. 2)

Summary

Introduction

In addition to tumor heterogeneity, another barrier to predict drug response is the number of possible resistance mechanisms used by cancer cells to escape anti-cancer-drug inhibitory effects[6]. As an alternative to culture, we hypothesized that dynamic monitoring of a drug target modulation upon drug exposure in single cells could predict cell responsiveness and better differentiate resistant cells to drugs within a single output[10,11]. Bioluminescence microscopy is a novel technique that uses the ability of reporter enzymes, named luciferases, to emit light with high energy after substrate addition Because this enzymatic reaction needs ATP and substrate, only live cells expressing the reporter gene will produce light. Our overall findings showed that a single cell bioluminescence microscopy could be performed to assess drug sensitivity with high accuracy, opening the door to the development of dynamic drug response assays in live circulating tumor cells from patients

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