The role of AKR1C1 in metabolic liver disease and hepatocellular cancer
Ismael da Conceicao, University of Oxford
The incidence of non-alcoholic fatty liver disease (NAFLD) and hepatocellular cancer (HCC) continue to increase, yet the clinical tools to aid diagnosis and treatment remain limited.
Our preliminary data, in which we have examined steroid hormone metabolites in clinical samples from more than 250 patients with NAFLD, have demonstrated that the products of a reaction mediated by the enzyme AKR1C1 (20a-hydroxylated steroid metabolites) are highly predictive of the stage of NAFLD and offers the potential as both a biomarker of disease as well as a therapeutic target. Further, AKR1C1 is upregulated in HCC compared to healthy livers; datamining studies have shown higher AKR1C1 expression in HCC population, in addition, increased AKR1C1 expression is associated with lower rates of survival.
We aim to uncover metabolic, inflammatory and proliferative pathways affected by AKR1C1 in hepatoma cell lines which is so far entirely unexplored. Due to the COVID-19 pandemic leading to lab closure and repurposing and relocation of our qPCR machine for COVID-19 testing by the NHS my research has been severely impacted. Therefore, I request money for RNA sequencing of my hepatoma AKR1C1 manipulation models. This data will be used for specific target validation by qPCR and western blotting.
According to my plan, I prepared and sent both AKR1C1 siRNA and CRISPR/Cas9 transfected HepG2 with validated knock down for RNA sequencing. After sequencing I performed the computational analysis of differential expression with featureCount and DESeq2. The results were incredibly helpful as they showed very few differentially expressed genes in liver cell lines which is in contrast to cell lines from other tissues. This helped me quickly focus on explorative functional experiments looking at proliferation, chemoresistance and motility based on the few dysregulated gene we observed (e.g., CDK2NA, ESAM, L1CAM). These further underlined that AKR1C1 seems to have no effect on signaling pathways in hepatoma cell lines and in its absence is most likely compensated for by other enzymes in the very active metabolic milieu of the liver. It is likely that AKR1C1 is correlating with HCC and might serve as a biomarker but has no regulatory function in progression or development and therefore does not provide any benefit as a therapeutic target.
Grant awarded: £4,795.07