1 / 15

Metabolic flux analysis of mantle cell lymphoma cells upon Bruton tyrosine kinase inhibition

Metabolic flux analysis of mantle cell lymphoma cells upon Bruton tyrosine kinase inhibition. Seung-Cheol Lee, PhD University of Pennsylvania School of Medicine Department of Radiology. 13C Metabolic Flux Analysis (MFA). Chance (1984) – heart, NMR Malloy (1988) – heart, NMR

ileclair
Download Presentation

Metabolic flux analysis of mantle cell lymphoma cells upon Bruton tyrosine kinase inhibition

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Metabolic flux analysis of mantle cell lymphoma cells upon Bruton tyrosine kinase inhibition Seung-CheolLee, PhD University of Pennsylvania School of Medicine Department of Radiology

  2. 13C Metabolic Flux Analysis (MFA) • Chance (1984) – heart, NMR • Malloy (1988) – heart, NMR • Mason, Gruetter, Rothman (1995) – brain, NMR • Wiechert (1997) – microorganism, NMR, MS • Stephanopoulos (2009) – cancer cells, MS • Shestov (2016) – cancer cells, NMR

  3. Melanoma cells, Dynamic MFA Shestov, Mancuso and Lee et al., Journal of Biological Chemistry 2016

  4. Application of MFA to lymphoma cells and BTK inhibitor • Bruton tyrosine kinase (BTK) inhibitor • Mantle cell lymphoma (MCL) • Cytostatic drug • Find which pathways are affected by the inhibitor and determine metabolic pathways to be further targeted

  5. Differential response of MCL cells to BTK inhibitor Jeko RL

  6. 1H NMR p=0.003 p=0.006 p=0.0003 p=0.01 p=0.0006 48 hr treatment with 500 nMibrutnib

  7. Metabolite 13C enrichment determination(Proton Observe Carbon Edited NMR) 12C Lac p=0.01 p=0.02 RL cells 12C Glu 48 hr treatment + incubation with [1,6]glucose for 8 hr 12C Ala 13C Lac 2.4 2.2 2.0 1.8 1.6 1.4 1.2 PPM 13C Ala 13C Glu Jeko cells 2.6 2.4 2.2 2.0 1.8 1.6 1.4 1.2 PPM

  8. Glutamate mulitiplet ratio determination 13C NMR p=0.03 RL cells 4s Glu4 Glu3 3s 3d43 4d34 4d34 3d43 Jeko cells 34.8 34.6 34.4 34.2 34.0 33.8 PPM 28.2 28.0 27.8 27.6 PPM

  9. Mass balance equations • Chemical mass balance • 13C isotope balance

  10. Metabolic network

  11. Determined metabolic fluxes

  12. Correlation with RNA-seq analysis * data not shown due to very low gene expression of this LDH isoform in JeKo-1 cells

  13. Treatment of MCL cells with a glutaminase inhibitor Cell growth (MTT assay)

  14. Summary • We have determined various metabolic fluxes of MCL cells (RL and Jeko) upon perturbation with BTK inhibition by using 13C NMR metabolic flux analysis. • Glutaminolytic pathway was found to be unaffected by BTK inhibition in Jeko cells . • Treatment of Jeko cells with a glutaminase inhibitor resulted in profound growth inhibition.

  15. Acknowledgement • NIH R01-CA172820 R01-CA129544

More Related