The project will address the role of mitochondrial function and metabolism and its impact on cancer resistance. It is clear that these organelles are vital in the process of carcinogenesis, but is becoming evident that they could also participate in the resistance to treatment.
The main aim of this project is to delineate the role of metabolic alterations coupled with mitochondria and their participation in the development or maintenance of cancer resistance. The role of experimental targets, which affect mitochondrial function and metabolism, will be tested by gain of function (overexpression) and loss of function (knockout, CRISPR/Cas system) studies. Genetically modified cell lines will be tested by live cell imaging using Etaluma720, enabling simultateous determination of viable and dead cells in real time. We will then assess the impact of such modifications on resistance of these cell lines to cell death induction. We also plan to investigate the role of post-translational regulatory sites of the prospective targets governing the mitochondrial metabolism, this will be assesed by site-directed mutagenesis, that would either result in loss of function or lead to a constitutively active or inactive mutants.
A possible specific objective derived from the main aim is the study of lipids in cancer resistance, with main focus on cardiolipin and its metabolism. Cardiolipin is a specific mitochondrial lipid that enables proper assembly of respiratory supercomplexes, thus leading to efficient functioning of the OXPHOS inside mitochondria. Recently, it has been proposed that the mitochondrial functions is not only regulated by the quantity of this lipid, but also its quality and the composition of its acyl chains and oxidation status, which greatly affects its biological role. However, the participation of cardiolipin in the context of cancer resistance has not not been studied in detail.
The experimental model that will primarily be explored is a model of tamoxifen resistant breast cancer cells, due to its high clinical relevance and our previous characterization of its particular metabolic phenotype.