Urinary signatures are associated with calorie restriction-mediated weight loss in obese Diversity Outbred mice

Authors: PAULES, EVAN - University Of North Carolina; TRUJILLO-GONZALEZ, ISIS - University Of North Carolina; VERHAGUE, MELISSA - University Of North Carolina; ALBRIGHT, JODY - University Of North Carolina; STEWART, DELISHA - Howard University; SUMNER, SUSAN - University Of North Carolina; MCRITCHIE, SUSAN - University Of North Carolina; KIRCHNER, DAVID - University Of North Carolina; COLEMAN, MICHAEL - University Of North Carolina; Bennett, Brian; HOWARD, ANNIE - University Of North Carolina; GORDON-LARSEN, PENNY - University Of North Carolina; FRENCH, JOHN - University Of North Carolina; HURSTING, STEPHEN - University Of North Carolina

Submitted to: PLOS ONE
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/21/2025
Publication Date: 12/9/2025
Citation: Paules, E.M., Trujillo-Gonzalez, I., VerHague, M., Albright, J., Stewart, D., Sumner, S.J., McRitchie, S.L., Kirchner, D., Coleman, M.F., Bennett, B.J., Howard, A.G., Gordon-Larsen, P., French, J.E., Hursting, S.D. 2025. Urinary signatures are associated with calorie restriction-mediated weight loss in obese Diversity Outbred mice. PLOS ONE. 20(12). Article e0329422. https://doi.org/10.1371/journal.pone.0329422.
DOI: https://doi.org/10.1371/journal.pone.0329422

Interpretive Summary: This study explored whether urinary metabolite profiles could predict weight loss response to calorie restriction (CR) in genetically diverse, obese Diversity Outbred (DO) mice. After inducing obesity with a high-fat diet, researchers implemented an 8-week CR regimen and categorized mice into responders or nonresponders based on weight loss. Metabolomic analysis of urine revealed distinct metabolite signatures—such as glutamic acid, dopamine, and hydroxyproline—that differed between groups, with additional sex-specific markers. Predictive models incorporating these metabolites showed moderate accuracy for males and high accuracy overall. Pathway analysis indicated that metabolic reprogramming may underlie improved insulin sensitivity and energy metabolism in responders.

Technical Abstract: Predictive analytics encompassing metabolomic profiles are increasingly being used to forecast responders to dietary interventions. Advances using this approach are particularly needed to personalize and enhance the effectiveness of dietary weight loss interventions. Using obese Diversity Outbred (DO) mice that model genetic and phenotypic heterogeneity of human populations, we aimed to identify urinary metabolite signatures predictive of responsiveness to calorie restriction (CR)-mediated weight loss. DO mice (150 males, 150 females) were fed a high-fat diet for 12 weeks to induce obesity, then urine was collected and an 8-week CR regimen (30% decrease in energy intake) initiated. At study completion, mice were rank-ordered according to their percent body weight change, with mice in the extreme quartiles deemed CR responders versus nonresponders. Targeted semi-quantitative metabolomics identified elevated glutamic acid and hydroxyproline as key urinary metabolites that distinguish CR responders from nonresponders, independent of sex. Other distinguishing urinary metabolites included putrescine in males, and dopamine, histamine, lysine, and spermine in females. Multivariate receiver operating characteristic analyses integrated these metabolites into a robust prediction model of CR-mediated weight loss. Further, pathway analysis identified several metabolic pathways, including arginine and proline metabolism, and alanine, aspartate, and glutamate biosynthesis, that distinguished CR responders from nonresponders and were indicative of metabolic reprogramming to enhance insulin sensitivity and energy metabolism.