Hypermetabolic state is associated with circadian rhythm disruption in mouse and human cancer cells

• Type of Material: Article
• Authors: Daniel Maxim Iascone, University of Pennsylvania; Xue Zhang, University of Pennsylvania; Patricia Bafford, University of Pennsylvania; Clementina Mesaros, University of Pennsylvania; Yogev Sela, University of Pennsylvania; Samuel Hofbauer, University of Pennsylvania; Shirley Zhang, Emory University; Kieona Cook, University of Pennsylvania; Pavel Pivarshev, University of Pennsylvania; Ben Z. Stanger, University of Pennsylvania; Stewart Anderson, University of Pennsylvania; Chi V. Dang, University of Pennsylvania; Amita Sehgal, University of Pennsylvania
• Language: English
• Date: 2023-11-13
• Publisher: NIH
• Publication Version: Preprint (Prior to Peer Review)
• Copyright Statement: The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.
• License: Creative Commons Attribution-NonCommercial 4.0 International
• Final Published Version (URL): https://doi.org/10.1101/2023.11.08.566310
• Title of Journal or Parent Work: bioRxiv
• Start Page: 566310
• Grant/Funding Information: We thank the Translational Biomarker Core at the University of Pennsylvania for their support with LC-HRMS experiments. This work was supported by grants from the NIH: F32MH125600 (to DMI), P30ES013508 (to CM), K99HL147212 (to SLZ), R01MH066912 (to SA), R01CA051497 (to CVD), R37NS048471 (to AS), and from the Howard Hughes Medical Institute (to AS).
• Keywords: circadian rhythms; degenerative disease; ATP levels; cellular metabolism; multicellular life
• Research Categories: Health Sciences, Oncology; Biology, Cell; Biology, Molecular

Abstract

Crosstalk between cellular metabolism and circadian rhythms is a fundamental building block of multicellular life, and disruption of this reciprocal communication could be relevant to degenerative disease, including cancer. Here, we investigated whether maintenance of circadian rhythms depends upon specific metabolic pathways, particularly in the context of cancer. We found that in adult mouse fibroblasts, ATP levels were a major contributor to overall levels of a clock gene luciferase reporter, although not necessarily to the strength of circadian cycling. In contrast, we identified significant metabolic control of circadian function in an in vitro mouse model of pancreatic adenocarcinoma. Metabolic profiling of a library of congenic tumor cell clones revealed significant differences in levels of lactate, pyruvate, ATP, and other crucial metabolites that we used to identify candidate clones with which to generate circadian reporter lines. Despite the shared genetic background of the clones, we observed diverse circadian profiles among these lines that varied with their metabolic phenotype: the most hypometabolic line had the strongest circadian rhythms while the most hypermetabolic line had the weakest rhythms. Treatment of these tumor cell lines with bezafibrate, a peroxisome proliferator-activated receptor (PPAR) agonist shown to increase OxPhos, decreased the amplitude of circadian oscillation in a subset of tumor cell lines. Strikingly, treatment with the Complex I antagonist rotenone enhanced circadian rhythms only in the tumor cell line in which glycolysis was also low, thereby establishing a hypometabolic state. We further analyzed metabolic and circadian phenotypes across a panel of human patient-derived melanoma cell lines and observed a significant negative association between metabolic activity and circadian cycling strength. Together, these findings suggest that metabolic heterogeneity in cancer directly contributes to circadian function, and that high levels of glycolysis or OxPhos independently disrupt circadian rhythms in these cells.