Quantification of Ceramides & Sphingosine 1-phosphate Levels In vitro in U87MG cells after Administration of ASAH1 Inhibitors Using High-Performance Liquid Chromatography Mass Spectrometry
Mentor 1
Shama Mirza
Start Date
10-5-2022 10:00 AM
Description
Glioblastoma is the most common and an aggressive form of cancer that occurs in the nervous system and either forms de novo or from nerve support cells called astrocytes (astrocytoma). Treatment includes but is not limited to surgical removal of the tumor, radiation therapy, and chemotherapy. Furthermore, targeted drug therapy is directed towards abnormalities that allow cancer cells to grow and proliferate such as inhibitors of enzymes in metabolic pathways. Proteomic analysis of over 600 glioblastoma-specific proteins revealed that expression of acid ceramidase (ASAH1) is associated with poor glioblastoma survival (Mirza, S.P. et al). ASAH1 converts ceramide into sphingososine-1-phospate (Sph-1P), and its inhibition increases cellular ceramide levels which promotes senescence and apoptosis. In addition to this, preliminary data in our lab indicates that carmofur increases ceramide levels in U87MG glioblastoma cells and mouse tissues. Mouse tissue samples were homogenized, ceramides were extracted, and quantification was done using High-Performance Liquid Chromatography Mass Spectrometry (HPLC-MS) revealing that approximately 30% of the cells underwent apoptosis. Overall, glioblastoma patients have a life expectancy of only 15 months with interventions and less than 25% of patients survive longer, therefore it is beneficial to develop targeted drug therapies that improve patient outcome.
Quantification of Ceramides & Sphingosine 1-phosphate Levels In vitro in U87MG cells after Administration of ASAH1 Inhibitors Using High-Performance Liquid Chromatography Mass Spectrometry
Glioblastoma is the most common and an aggressive form of cancer that occurs in the nervous system and either forms de novo or from nerve support cells called astrocytes (astrocytoma). Treatment includes but is not limited to surgical removal of the tumor, radiation therapy, and chemotherapy. Furthermore, targeted drug therapy is directed towards abnormalities that allow cancer cells to grow and proliferate such as inhibitors of enzymes in metabolic pathways. Proteomic analysis of over 600 glioblastoma-specific proteins revealed that expression of acid ceramidase (ASAH1) is associated with poor glioblastoma survival (Mirza, S.P. et al). ASAH1 converts ceramide into sphingososine-1-phospate (Sph-1P), and its inhibition increases cellular ceramide levels which promotes senescence and apoptosis. In addition to this, preliminary data in our lab indicates that carmofur increases ceramide levels in U87MG glioblastoma cells and mouse tissues. Mouse tissue samples were homogenized, ceramides were extracted, and quantification was done using High-Performance Liquid Chromatography Mass Spectrometry (HPLC-MS) revealing that approximately 30% of the cells underwent apoptosis. Overall, glioblastoma patients have a life expectancy of only 15 months with interventions and less than 25% of patients survive longer, therefore it is beneficial to develop targeted drug therapies that improve patient outcome.
