Danielle Goodspeed

Essential nutrient supplementation prevents heritable metabolic disease in multigenerational intrauterine growth-restricted rats

Intrauterine growth restriction (IUGR) confers heritable alterations in DNA methylation, rendering risk of adult metabolic syndrome (MetS). Because CpG methylation is coupled to intake of essential nutrients along the one‐carbon pathway, we reasoned that essential nutrient supplementation (ENS) may abrogate IUGR‐conferred multigenerational MetS. Pregnant Sprague‐Dawley rats underwent bilateral uterine artery ligation causing IUGR in F1. Among the F2 generation, IUGR lineage rats were underweight at birth (6.7 vs. 8.0 g, P < 0.0001) and obese by adulthood (p160: 613 vs. 510 g; P < 0.0001). Dual energy X‐ray absorptiometry studies revealed increased central fat mass (Δ+40 g), accompanied by dyslipidemic (>30% elevated, P < 0.05) serum triglycerides (139 mg/dl), very‐LDLs (27.8 mg/dl), and fatty acids (632 μM). Hyperglycemic‐euglycemic clamp studies and glucose tolerance testing revealed insulin resistance. Conversely, IUGR lineage ENS‐fed rats did not manifest MetS, with significantly lower body weight (p160: 410 g), >5‐fold less central fat mass, normal hepatic glucose efflux, and >70% reduced circulating triglycerides and very‐LDLs compared with IUGR control‐fed F2 offspring (P < 0.01). Moreover, increased methylation of the IGF‐1 P2 transcriptional start site among IUGR lineage F2 offspring was reversed in ENS (P < 0.04). This is an initial demonstration that supplementation along the one‐carbon pathway abrogates adult morbidity and associated epigenomic modifications of IGF‐1 in a rodent model of multigenerational MetS.—Goodspeed, D., Seferovic, M. D., Holland, W., Mcknight, R. A., Summers, S. A., Branch, D. W., Lane, R. H., Aagaard, K. M., Essential nutrient supplementation prevents heritable metabolic disease in multigenerational intrauterine growth‐restricted rats. FASEB J. 29, 807–819 (2015). www.fasebj.org

Heritable IUGR and adult metabolic syndrome are reversible and associated with alterations in the metabolome following dietary supplementation of 1-carbon intermediates

Metabolic syndrome (MetS), following intrauterine growth restriction (IUGR), is epigenetically heritable. Recently, we abrogated the F2 adult phenotype with essential nutrient supplementation (ENS) of intermediates along the 1‐carbon pathway. With the use of the same grandparental uterine artery ligation model, we profiled the F2 serum metabolome at weaning [postnatal day (d)21; n = 76] and adulthood (d160; n = 12) to test if MetS is preceded by alterations in the metabolome. Indicative of developmentally programmed MetS, adult F2, formerly IUGR rats, were obese (621 vs. 461 g; P< 0.0001), dyslipidemic (133 vs. 67 mg/dl; P< 0.001), and glucose intolerant (26 vs. 15 mg/kg/min; P< 0.01). Unbiased gas chromatography‐mass spectrometry (GC‐MS) profiling revealed 34 peaks corresponding to 12 nonredundant metabolites and 9 unknowns to be changing at weaning [false discovery rate (FDR) < 0.05]. Markers of later‐in‐life MetS included citric acid, glucosamine, myoinositol, and proline (P < 0.03). Hierarchical clustering revealed grouping by IUGR lineage and supplementation at d21 and d160. Weanlings grouped distinctly for ENS and IUGR by partial least‐squares discriminate analysis (PLS‐DA; P< 0.01), whereas paternal andmaternal IUGR (IUGRpat/IUGRmat, respectively) control‐fed rats, destined for MetS, had a distinct metabolome at weaning (randomForest analysis; class error < 0.1) and adulthood (PLS‐DA; P< 0.05). In sum, we have found that alterations in the metabolome accompany heritable IUGR, precede adult‐onset MetS, and are partially amenable to dietary intervention.— Seferovic, M. D., Goodspeed, D. M., Chu, D. M., Krannich, L. A., Gonzalez‐Rodriguez, P. J., Cox, J. E., Aagaard, K. M. Heritable IUGR and adult metabolic syndrome are reversible and associated with alterations in the metabolome following dietary supplementation of one‐carbon intermediates. FASEB J. 29, 2640‐2652 (2015). www.fasebj.org

Conditional postnatal deletion of the neonatal murine hepatic circadian gene, Npas2, alters the gut microbiome following restricted feeding

BACKGROUND: We have recently shown in both non‐human primates and in rodents that fetal and neonatal hepatic expression of the circadian transcription factor, Npas2, is modulated by a high fat maternal diet and plays a critical role in establishing life‐long metabolic homeostasis. Similarly, we and others have also established the importance of the maternal and early postnatal diet on establishment of the early gut microbiome. OBJECTIVE: We hypothesized that altered circadian gene expression solely in the neonatal liver would result in gut microbiome dysbiosis, especially with diet‐induced metabolic stress (ie, restricted feeding). Using a murine model in which we conditionally knock out Npas2 in the neonatal liver, we aimed to determine the role of the circadian machinery in gut dysbiosis with restricted feeding. STUDY DESIGN: We collected fecal samples from liver Npas2 conditional knockout (n = 11) and wild‐type (n = 13) reproductive‐aged mice before (study day 0) and after the restricted feeding study (study day 17). Extracted DNA was sequenced using the MiSeq Illumina platform using primers specific for the V4 region of the 16S ribosomal DNA gene. The resulting sequences were quality filtered, aligned, and assigned taxonomy. Principal coordinate analysis was performed on unweighted and weighted UniFrac distances between samples with a permutation analysis of variance to assess clustering significance between groups. Microbial taxa that significantly differ between groups of interest was determined using linear discriminate analysis effect size and randomForrest. RESULTS: Principal coordinate analysis performed on weighted UniFrac distances between male conditional knockout and wild‐type cohorts revealed that the gut microbiome of the mice did not differ by genotype at the start of the restricted feeding study but did differ by virtue of genotype at the end of the study (P = .001). Moreover, these differences could be at least partially attributed to restricted feeding–associated alterations in relative abundance of the Bacteroides genus, which has been implicated as crucial to establishing a healthy gut microbiome early in development. CONCLUSION: Here we have provided an initial key insight into the interplay between neonatal establishment of the peripheral circadian clock in the liver and the ability of the gut microbiome to respond to dietary and metabolic stress. Because Npas2 expression in the liver is a target of maternal high‐fat diet–induced metabolic perturbations during fetal development, we speculate that these findings have potential implications in the long‐term metabolic health of their offspring.