Discerning MCT1s role in chromatin remodelling and metabolomics changes in cancer progression

Monocarboxylate transporter 1 (MCT1) is a transmembrane protein involved in cell metabolism and mediates transport of monocarboxylates (such as pyruvate {a metabolic substrate} and lactate {a metabolic by product}) in and out of the cell. Increased plasma membrane (PM) expression of MCT1 has been reported in several cancer types including endometrial cancer (EC) and is associated with reduced patient survival. Although nucleus is not a usual location for MCTs (based on current knowledge on their function), we and others reported nuclear MCT1 (nMCT1) expression in endometrial [2] and soft sarcoma [3] biopsies. Both studies showed that patients with nMCT1 survive longer than the patients without it. Further, a number of studies showed that metabolic proteins found in the nucleus can regulate transcription and chromatin remodelling by acting as co-factors or by providing the substrates (such as lactate, acetyl-coA) for these processes (reviewed in [4, 5]). Interestingly, we also found that when in nucleus, MCT1 interacts with other proteins that modify chromatin remodelling and regulate gene expression. Therefore, understanding nMCT1's novel role in nucleus is vital to identify and exploit novel approaches in cancer treatment to enhance life-expectancy amongst patients. This project aims to identify nMCT1s role in relation to potential chromatin remodelling and/or metabolomic changes involved in cancer progression and patient survival. For this purpose, we will first combine CRISPR-Cas9 technology with molecular cloning to engineer cancer cell lines (Ishikawa and HEC1A) with MCT1 protein targeted specifically to the nucleus (nMCT1) or plasma membrane (PM MCT1). We will then subject these cell lines to a number of in vitro methods (e.g. immunofluorescence, western blotting, electron microscopy) to characterise subcellular localisation and expression levels in engineered cell lines. Metabolic profile of the engineered cell lines with nMCT1 wand PM MCT1 will be performed using mass spectroscopy and data analysis will be performed using R package. Changes in chromatin remodelling will be assessed using ATAC-sequencing. Results of metabolomics and chromatin remodelling will be compared and correlated to evaluate correlation between MCT1 subcellular localisation, changes in metabolism and chromatin structure by using R or any other relevant bioinformatics tools. By the end of this interdisciplinary project, the selected student will be equipped in sought after wet lab and bioinformatics skills for post-doctoral positions.