RT Journal Article SR Electronic T1 Systems level profiling of arginine starvation reveals MYC and ERK adaptive metabolic reprogramming JF bioRxiv FD Cold Spring Harbor Laboratory SP 2020.01.13.904011 DO 10.1101/2020.01.13.904011 A1 Caitlyn B. Brashears A1 Richa Rathore A1 Matthew Schultze A1 William R. Ehrhardt A1 Shin-Chen Tzeng A1 Brian A. Van Tine A1 Jason M. Held YR 2020 UL http://biorxiv.org/content/early/2020/01/13/2020.01.13.904011.abstract AB Arginine auxotrophy due to the silencing of argininosuccinate synthetase 1 (ASS1) occurs in many cancers, especially sarcomas. Arginine deiminase (ADI-PEG20) therapy exploits this metabolic vulnerability by depleting extracellular arginine, causing arginine starvation. ASS1-negative cells develop resistance to ADI-PEG20 through a metabolic adaptation that includes re-expressing ASS1. As arginine-based multiagent therapies are being developed, further characterization of the changes induced by arginine starvation is needed. In order to develop a systems-level understanding of these changes, activity-based proteomic profiling (ABPP) and phosphoproteomic profiling were performed before and after ADI-PEG20 treatment in ADI-PEG20-sensitive and resistant sarcoma cells. When integrated with previous metabolomic profiling (Kremer et al, 2017a), this multi-omic analysis reveals that cellular response to arginine starvation is mediated by adaptive ERK signaling, driving a Myc-Max transcriptional network. Concomitantly, these data elucidate proteomic changes that facilitate oxaloacetate production by enhancing glutamine and pyruvate anaplerosis, and altering lipid metabolism to recycle citrate for oxidative glutaminolysis. Based on the complexity of metabolic and cellular signaling interactions, these multi-omic approaches could provide valuable tools for evaluating response to metabolically targeted therapies.αKGalphaketoglutarateABPPActivity-based proteomic profilingAcCoAAcetyl-CoAACNacetonitrileADI-PEG20Arginine diminasePEG-2000 AspaspartateATPadenosine triphosphateBMEB-mercaptoethanolCitcitrateCScitrate synthaseDDAData-dependent acquisition mass spectrometryDIA-MSData-independent acquisition mass spectrometry, aka SWATHDTTdithiothreitolFASNfatty acid synthaseFDRFalse discovery rateGBMglioblastoma multiformeIsoCitisocitrateIDHisocitrate dehydrogenaseMalmalateMAPKmitogen-activated protein kinaseMDHmalate dehydrogenaseMitomitochondriaMycMyc-COAAoxaloacetatePCpyruvate carboxylasePDHApyruvate dehydrogenase, Alpha-1PMSFphenylmethylsulfonyl fluorideTFAtrifluoroacetic acidVehvehicle