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Precision Nutrition/Nutrient-Gene Interactions (Poster Session)

(P21-014-25) Effects of Intermittent Fasting on Global Metabolomics and Metabolic Pathways Using High-Resolution Mass Spectrometry and Predictive Models in Muscle Cell Lines (C2C12)

Sunday, June 1, 2025
11:45 AM - 12:45 PM ET
Location: Poster Board A-92

    Presenting Author(s)

  • Ghada A. Soliman, MBBCh, PhD, RDN, CDN

    Professor of Nutrition and Doctoral Program Director
    City University of New York, Graduate School of Public Health and Health Policy
    NEW YORK, New York, United States

    • Co-Author(s)

  • RA

    Rinat Abzalimov, PhD

    CUNY Advanced Science Research Center (ASRC-GC)

  • LX

    Lele Xu, MS

    CUNY-ASRC
    New York, New York, United States

  • YH

    Ye He, PhD

    Director, Live Imaging and Bioenergetics Facilities, Research Assistant Professor | Neuroscience Ini
    CUNY GC Advanced Science Research Center
    New York, New York, United States

Disclosure(s):
Ghada A. Soliman, MBBCh, PhD, RDN, CDN: No relevant financial relationship(s) with ineligible companies to disclose.


Objectives: Determine the effects of intermittent fasting (IF) for 6 or 18 hrs on global metabolomics and amino acid levels and apply predictive models in C2C12 muscle cell lines to explain the altered metabolic pathways.

Methods: C2C12 muscle cell lines were used to determine the metabolic changes during IF. To mimic IF, we cultured the C2C12 cell lines under 6 different conditions: (1) Starving serum-free media for 6 hours; (2) Control-complete media for 6 hours; (3) Starving serum-free media for 18 hours; (4) Control complete media for 18 hours; (5) Rapamycin (mTORC1 inhibitor) for 18 hrs; (6) Rapalink (mTORC1/mTORC2) for 18 hours. C2C12 muscle cells were harvested and extracted. The high-resolution untargeted metabolomics LC-MS/MS analysis was performed using Bruker's maXis-II ESI-Q-q-TOF. We utilized the MetaboAnalyst 6 and the XCMS plus platforms to determine the impact of fasting at 6 hrs or 18 hrs on amino acids and untargeted metabolites to predict biological pathways using the Mummichog python algorithm. Statistical significance (p < 0.001) was determined by the Kruskal-Wallis test, followed by the Welch t-test.

Results: C2C12 cell lines incubated with starving media for 6 hrs showed decreased levels of L-serine, L-proline, L-alanine, and L-glutamine compared to the Control. Similarly, cells incubated with the starving media for 18 hrs displayed decreased levels of L-glutamate, L-proline, L-alanine and L-arginine compared to Control. Incubation with RAPA-18hrs and RL-18hrs, which inhibit mTOR complexes, showed a different inhibition pattern compared to starving at 6 and 18hrs, suggesting that there is an input from other metabolic pathways. Mapping the metabolite changes to predictive pathways and activity networks revealed that the top pathways affected by starve-6hrs included gluconeogenesis, glycolysis, sucrose degradation, serine, glutamine, and aspartate pathways compared to the control group. Starve-18-hrs affected heme degradation, purine and pyrimidine, and arginine biosynthesis pathways compared to the Control.

Conclusions: Intermittent fasting (6 and 18 hours) has a differential metabolic effect on metabolic pathways and amino acid metabolism. Future studies will examine the effects of intermittent fasting on bioenergetic dynamics and cellular metabolism.

Funding Sources: PSC-CUNY Grant 54-101 and CUNY-ASRC Seed Grant Award # 95649-00