B121
The effects of L-glutamine and L-asparagine deprivation on paediatric sarcomas
Jodie Allsup1, Ian Podmore1, Edward Estlin2, Alan McGown1, Nicholas Coe1
1The University of Salford, Manchester, UK, 2Royal Manchester Children's Hospital, UK
Background
L-glutamine (gln) is vital to many cellular biosynthetic pathways including nucleotide and protein biosynthesis. Gln is involved in cellular energy generation and is a major respiratory fuel in many different cell lines, playing a large role in the Warburg effect in cancer cells. Previous unpublished research in this group has been performed on the effects of Asparaginase in paediatric cancers. Asparaginase has slight activity as a Glutaminase which may play a crucial role in its anti-cancer effects.
Method
Effects of gln and asparagine (asn) deprivation in paediatric sarcomas has been investigated. A panel of cell lines (Ewings sarcoma and osteosarcoma) has been screened for sensitivity to deprivation. Apoptosis and cell cycle analyses have been performed in sensitive cells using flow cytometry and intracellular gln and asn quantification has been performed using GC-MS. Different depths and durations of gln and asn deprivation in combination with anti-cancer drugs such as Temozolomide and Irinotecan have been investigated using cytotoxicity assays in an attempt to chemosensitise these cells.
Results
Initial screening experiments have shown TC32 Ewings sarcoma cells to be particularly sensitive. Simultaneous gln and asn deprivation resulted in a ten-fold decrease in cell growth in these cells. Preliminary flow cytometric analyses show apoptosis to be induced in these cells. Cytotoxicity has also been observed in HOS and 791T Osteosarcoma cells. A673 Ewings sarcoma cells have shown no response. Further flow cytometric analyses will be performed on these cell lines. Cytotoxicity assays performed on sensitive cell lines using Temozolomide and Irinotecan are yielding promising results.
Conclusion
Gln and asn deprivation is clinically relevant over a short period of time. Synergistic relationships found between gln and asn deprivation and selected anti-cancer agents in these sarcomas show potential for a new chemotherapeutic regime.
