A prometabolite strategy inhibits cardiometabolic disease in an ApoE–/– murine model of atherosclerosis
Taryn N. Beckman, … , Eugene B. Chang, Jeffrey A. Hubbell
Taryn N. Beckman, … , Eugene B. Chang, Jeffrey A. Hubbell
Published August 8, 2025
Citation Information: JCI Insight. 2025;10(15):e191090. https://doi.org/10.1172/jci.insight.191090.
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Research Article Inflammation Therapeutics

A prometabolite strategy inhibits cardiometabolic disease in an ApoE–/– murine model of atherosclerosis

Abstract

Butyrate, a microbiome-derived short-chain fatty acid with pleiotropic effects on inflammation and metabolism, has been shown to significantly reduce atherosclerotic lesions, rectify routine metabolic parameters such as low-density lipoprotein cholesterol (LDL-C), and reduce systemic inflammation in murine models of atherosclerosis. However, its foul odor, rapid metabolism in the gut and thus low systemic bioavailability limit its therapeutic effectiveness. Our laboratory has engineered an ester-linked L-serine conjugate to butyrate (SerBut) to mask its taste and odor and to coopt amino acid transporters in the gut to increase its systemic bioavailability, as determined by tissue measurements of free butyrate, produced by hydrolysis of SerBut. In an apolipoprotein E–knockout (ApoE)–/– mouse model of atherosclerosis, SerBut reduced systemic LDL-C, proinflammatory cytokines, and circulating neutrophils. SerBut enhanced inhibition of plaque progression and reduced monocyte accumulation in the aorta compared with sodium butyrate. SerBut suppressed liver injury biomarkers alanine transaminase and aspartate aminotransferase and suppressed steatosis in the liver. SerBut overcomes several barriers to the translation of butyrate and shows superior promise in slowing atherosclerosis and liver injury compared with equidosed sodium butyrate.

Authors

Taryn N. Beckman, Lisa R. Volpatti, Salvador Norton de Matos, Anna J. Slezak, Joseph W. Reda, Ada Weinstock, Leah Ziolkowski, Alex Turk, Erica Budina, Shijie Cao, Gustavo Borjas, Jung Woo Kwon, Orlando deLeon, Kirsten C. Refvik, Abigail L. Lauterbach, Suzana Gomes, Eugene B. Chang, Jeffrey A. Hubbell

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Figure 1

SerBut reduces cytotoxicity as a carrier of inflammatory NF-κB pathway–suppressing butyrate in vitro.

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SerBut reduces cytotoxicity as a carrier of inflammatory NF-κB pathway–s...
(A) Viability of RAW 264.7 cells treated with 0.15, 0.44, 1.33, 4, and 12 mM SerBut or NaBut for 24 hours. (B) LC-MS/MS analysis of free butyrate in solution of aged SerBut (incubated at 37°C in FBS-supplemented media for 1 month) and freshly dissolved SerBut and NaBut. (C) NF-κB activity of RAW Blue reporter cells upon treatment with the indicated concentrations of SerBut or NaBut for 24 hours followed by stimulation with 200 ng/mL LPS for 6 hours. (D–J) RAW 264.7 cells treated for 24 hours with the indicated concentrations of SerBut or NaBut followed by stimulation with 200 ng/mL LPS for 6 hours. (D–E) Mean fluorescence intensity (MFI) of iNOS+ or CD80+ RAW 264.7 cells analyzed by flow cytometry. (F–J) Cytokine concentrations in the cell culture supernatant of RAW 264.7 cells. “x” indicates untreated group. In A, n = 3 per group representing technical replicates. In B, n = 3 freshly prepared samples per 2 distinct experiments pooled for aged SerBut and NaBut groups, while fresh the SerBut group represents a single experiment. One sample in the aged SerBut group and 2 samples in the NaBut group were lost due to filtering malfunctions or LC-MS/MS autosampling malfunctions. In C, n = 6 technical replicates. Data represent mean ± SEM. Experiments were performed 2 or more times with similar results. In A, 2-way ANOVA was performed. Statistical analyses were performed using a 1-way ANOVA with Tukey’s, Welch’s (if standard deviations were significantly different by Bartlett and Brown-Forsyth tests), or Kruskal-Wallis (if data were not normally distributed determined by Shapiro-Wilk test) post hoc test in C–J. P values less than 0.10 are shown.