Cancer metabolism and its role in the sensitivity to ASNase of leukemic cells to L-asparaginase

• Thesis title in Czech: Nádorový metabolismus a jeho role v citlivosti leukemických buněk k L-asparagináze
• Thesis title in English: Cancer metabolism and its role in the sensitivity to ASNase of leukemic cells to L-asparaginase
• English key words: Asparagine, PI3K/Akt pathway, cellular metabolism, childhood acute lymphoblastic leukemia, glutamine, l-asparaginase, mesenchymal stromal cells, metabolic profile, mTOR signaling pathway.
• Academic year of topic announcement: 2016/2017
• Thesis type: dissertation
• Thesis language: angličtina
• Department: Department of Paediatric Haematology and Oncology (13-462)
• Supervisor: RNDr. Júlia Starková, Ph.D.
• Author: hidden - assigned and confirmed by the Study Dept.
• Date of registration: 28.10.2016
• Date of assignment: 28.10.2016
• Confirmed by Study dept. on: 28.10.2016
• Date and time of defence: 12.09.2023 13:00
• Date of electronic submission: 22.06.2023
• Date of submission of printed version: 23.06.2023
• Date of proceeded defence: 12.09.2023
• Opponents: doc. MUDr. Vít Procházka, Ph.D.

Preliminary scope of work in English

Introduction
Acute lymphoblastic Leukemica (ALL) is the most common type of cancer in children. This disease treatment is one of best achieves in cancer field. It produces a remission of 98% of ALL patients. Unfortunately, relapses occur in approximately 15 - 20% cases. L-asparaginase (ASNase), the principal component of therapy, manages to deplete glutamine and asparagine from extracellular environment. This depletion is disastrous to leukemic cells while it has almost no effect in healthy cells. The underlying mechanism of sensitivity to ASNase is not completely understood; one of the assumptions is the capability of leukemic cells to adapt to nutrition stress conditions. Cancer cells use as a substrate glucose as well as glutamine and fatty acids. These cells have higher demand for energy and biomass/building blocks to keep constant growth and proliferation. Recent data show that cancer cells preferentially choose glycolysis for its carbon pool used further in biosynthetic processes rather than for ATP production. Moreover, glycolysis is favoured dependently on tumor type and nutrient availability.

Project objectives and aims
The main target of our project is to understand the action mechanism of ASNase. This knowledge could then be applied in the new treatment strategy for ALL patients non-responsive to the current therapy. Optimization and employment of new technologies will allow their broader use in other projects as is the study of resistance to ASNase or cause of unfavourable side effects associated with ASNase administration.

Methodology
1. Characterization of metabolic pathways affected by ASNase in leukemic cells
Use Liquid chromatography - mass spectrometry and High Performance Liquid Chromatography, Sea horse analyser to check bioenergetic pathways. Leukemia cell culture as our research model.

2. Impact of the microenvironment
Mesenchymal stem cells culture, leukemia cell culture, multi-cell culture. Flow cytometry to analyse the cytotoxic effect and PCR+sequencing. Sea horse analyser to bioenergetic pathways research.