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Rešetanje ligandov aminotransferaze razvejanih aminokislin kot zaviralcev proliferacije glioblastomskih matičnih celic = Screening of branched-chain amino acid transferase ligands as proliferation inhibitors of glioblastoma stem cells : magistrski študijski program Laboratorijska biomedicina
Kocijančič, Anja, farmacevtka
In recent decades, the number of cases and deaths caused by genetic diseases such as cancer has increased. Although conventional therapy works for many, there are people who have a very aggressive ...type of cancer or relapse after a few years - for these people, novel treatment options are the only way to treat them. Glioblastoma (GBM) is one of the cancers that, although it mainly affects older people, can also affect young people, and has a very poor prognosis, with a median 5-year survival of only 5-10%. Classical therapy has proven ineffective in many cases and a possible explanation for this lies in the cells responsible for the tumor and the progression of the disease. These cells are called cancer stem cells and should be the main target in the treatment of glioblastoma. Therapies can target many different processes and properties of the cell, and one that is commonly researched is metabolism. The reason for this is that metabolism connects everything in the cell together and can adapt to the environment so that the cell can survive. Cancer cells have shown these adaptations in many cancers and in many different processes, including branched-chain amino acid metabolism. Branched-chain amino acids are essential amino acids obtained through diet intake and protein breakdown in humans. GBM and other cancer cells rely heavily on these amino acids, so targeting this metabolic pathway may yield promising results. This study as part of Master’s thesis has aimed to test twelve ligands targeting branched-chain amino acid transferases, BCAT1 and BCAT2, as potential inhibitors of GBM, using glioblastoma patient-derived neural stem cells and other non-glioblastoma cell lines - HeLa, RPE1, HCT116, and fibroblasts - as control cell lines. BCATs are step-one enzymes in the metabolism of branched-chain amino acids. The main part of the project was the evaluation to identify the most promising of these ligands as potential inhibitors of GBM. The evaluation identified two ligands as the promising candidates- ligand 4 and ligand 8 showed favorable results out of all twelve ligands tested. It was performed using the PrestoBlue cell viability assay and approximate IC50 values were determined to compare the efficacy of the ligands on the cells. For the two ligands that showed promising results, clonogenic survival assay was performed, and the cell viability assay was repeated on an extended set of cell lines. Further experiments confirmed that ligand 4 and ligand 8 could potentially be successful in inhibiting GBMs, but since both also affected control cell lines, the optimal concentration range should be determined for their potential use in specific inhibition of GBMs. Future experiments are required to gain further insights into how exactly these ligands act on the cell and whether they inhibit the enzyme of interest. To clarify this, additional in-depth studies using an extended range of methods, including RNA sequencing, will be useful.
Type of material - master's thesis
; adult, serious
Publication and manufacture - Ljubljana : [A. Kocijančič], 2023
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