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Presented by: Allison Anguren and Priya Vasudeva

Presented by: Allison Anguren and Priya Vasudeva. Introduction. Metabolomics the quantitative measurement of the dynamic metabolics response of living systems to pathophysiological stimuli, environmental modulation, or genetic modifications

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Presented by: Allison Anguren and Priya Vasudeva

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  1. Presented by: Allison Anguren and Priya Vasudeva

  2. Introduction • Metabolomics • the quantitative measurement of the dynamic metabolics response of living systems to pathophysiological stimuli, environmental modulation, or genetic modifications • Monitoring of small molecule concentrations can provide information about disease progression or treatment response • Cancer cells have a number of specific metabolic features • Ex) Warburg effect

  3. Introduction • Agents that promote apoptosis are powerful tools for cancer therapeutics • Detection and monitoring of early apoptosis biomarkers are urgently needed in clinical trials • Monitoring apoptosis is challenging

  4. Aim of the study: • To design fast, robust, reliable, and affordable method of measuring metabolites in cells • To find early biomarkers of apoptosis in cancer cell lines

  5. Methods • Human cancer cell lines • HEK 293, HepG2, PC3, MCF7 • MTT Assay • Caspase 3/7 assay

  6. Methods • NBS Assay • Newborn Screening assay detect levels of 42 amino acids • Tandem Mass Spectrometry • Quantified the amount of 42 amino acids and acylcarnitines

  7. Results-MTT Assay • Tested the viability of cell lines to treatments of staurosporine and heating. • Decreased viability after 24hrs of staurosporine treatment.

  8. Results-Apoptosis Validation • Caspase 3 and 7 activity were analyzed to assess induced apoptosis. • Treatment with 4um of staurosporine had greater effect.

  9. Results-Metabolic Signatures • Used NBS to detect metabolites. • Analyzed 42 amino acids by and acylcarnitines by MS/MS • Differences in the concentrations of metabolites in various cell lines.

  10. Results-Table 1

  11. Results-Table 2 • In HEK 293, HepG2, and PC3, Asp, Glu, Met, Gly, Ala, C3-carnitine, and C3DC-carnitine up regulated in apoptotic vs necrotic cells • MCF7 cells display very different metabolic signature

  12. Conclusions • Used metabolics as markers of genomic changes due to apoptotic processes • Discriminated between necrosis and apoptosis by metabolics signatures • NBS and MS/MS can be used for high-throughput quantification of metabolites. • Distinct changes in Asp, Glu, Met, Ala, Gly, C3-carnitine, and C3DC-carnitine.

  13. Big Picture • Able to use targeted metabolics to assay a known group of molecules • Use metabolic signatures to test the efficiency of agents causing apoptosis • Can monitor apoptosis metabolic biomarkers in cancer therapeutics and anticancer therapies. • Cons: Can only detect known metabolites

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