Quantitative image based analysis of endocrine disruptor effects on mitochondria morphology‐function in prostate cancer cells.

National audiencePersistent organic pollutants (POPs) are environmental contaminants that interfere with normal hormonal homeostasis and act as endocrine disrupting compounds (EDC). These molecules can mimic hormone effects on metabolism. The links between metabolism and cancer are now well established. Metabolism generates reactive oxygen species (ROS), which contribute to mutations and induces oncogenic transformation. In turn, cancer cells display high metabolic flexibility allowing them to grow in various cellular environments and favoring their proliferative and invasive capacities. Mitochondria are key players in this complex interplay since they produce ROS, generate energy and participate in nucleotide synthesis and in glutamine metabolism of cancer cells. Regarding the importance of hormones on prostate cancer risk and outcomes, we are developing a multiple parameters in vitro assay conducted in a high-throughput screening format relevant for prostate cancer metabolism and aggressiveness. This screening method includes a microscopy based analysis of mitochondria structure and functions as well as flow cytometry analysis. We analyzed the effects of five EDCs (Aldrin, BDE28, TCDD, PCB153, PFOA) identified in the plasma of patients on two prostate cancer cell lines, 22RV1 (androgen-responsive) and DU145 (androgen-unresponsive). Each compound was tested in a dose dependent manner to determine its effects on ROS production, mitochondrial membrane potential, mitochondrial biogenesis and mitophagy. In addition, we performed an image based computational analysis of the mitochondrial network morphology and dynamics. This strategy allows us to extract some quantitative parameters on the mitochondrial network as fragmentation index, compactness, average volume, etc. When combined, morphological and functional parameters allow us to discriminate subtle perturbations of the mitochondrial structure-function induced by EDCs in prostate cancer cells. We are confident that this multiparameter analysis strategy could represent a new perspective in identification and characterization of EDCs based on their effects on cell metabolism (phenoscore) in order to estimate their potential risk on human health.This work is supported by ITMO Cancer