Investigating the effect of KEAP1 knockout and lactate treatment on histone lysine lactylation in KRAS-driven lung adenocarcinoma
Typ
Examensarbete för masterexamen
Master's Thesis
Master's Thesis
Program
Biotechnology (MPBIO), MSc
Publicerad
2024
Författare
Karlsson, Elin
Modellbyggare
Tidskriftstitel
ISSN
Volymtitel
Utgivare
Sammanfattning
Lung cancer is a major global health concern, responsible for the highest number of deaths among all cancer types. Whereof, the subtype lung adenocarcinoma is the most common form, accounting for 40% of all lung cancer cases. Loss-of-function mutations in the gene KEAP1, found in 20% of lung adenocarcinoma patients, are associated with treatment resistance and lower overall survival, highlighting the importance of investigating this mutation in research. Additionally, KEAP1 loss of function are linked to metabolic reprogramming, a known hallmark of cancer. The Warburg effect, a metabolic reprogramming in cancer cells characterized by high rates of aerobic glycolysis , leads to elevated lactate concentrations in cancerous tissue compared to healthy tissue. Lactate, earlier considered to be a waste product, is now recognized for its role as a signaling molecule and its function in coordinating metabolism. Recent studies has identified histone lysine lactylation, a post-translational modification induced by lactate, which affects gene expression and chromatin structure.
The aim of this thesis project was to investigate the effect of KEAP1 knockout and lactate treatment on histone lysine lactylation levels in lung adenocarcinoma. This study utilized mouse lung adenocarcinoma cell lines derived from genetically engineered mouse models with an activating mutation in the oncogene Kras. Using the protein detection method Western blot, the results demonstrated that histone lysine lactylation levels on H3 and H4 were lower in Keap1 knockout cells compared to Keap1 wild-type cells in one of the cell lines, while no definitive conclusion could be drawn for the other cell line. Inhibition of the KEAP1 protein without affecting the Keap1 gene directly, showed the same results. Additionally, lactate treatment induced histone lysine lactylation levels on H3 and H4 in Keap1 wild-type cells, but no increase was observed in Keap1 knockout cells. By measuring intracellular lactate concentration it was observed that the lactate treatment induced intracellular lactate levels for both Keap1 wild-type cells and Keap1 knockout cells. Indicating that there is no direct correlation between the intracellular lactate levels and histone lysine lactylation in cells with a Keap1 knockout.
Beskrivning
Ämne/nyckelord
lung cancer , lung adenocarcinoma , Kras , Keap1 , lactate , histone lysine lactylation.