Early-life ketone body signaling promotes beige fat biogenesis through changes in histone acetylome and ß-hydroxybutyrylome [ChIP-seq]

Infants experience distinct ketogenesis during the preweaning period, but its physiological significance remains unclear. Here, we show that preweaning ketosis enhances beige fat biogenesis and improves metabolic health in adulthood. To elucidate the underlying mechanisms, we performed multi-omics analyses of inguinal white adipose tissue (iWAT) and its stromal vascular fraction (SVF). Bulk RNA-seq of iWAT was conducted in 8-week-old Hmgcs2 knockout (KO) and control mice housed at room temperature (23 °C) or exposed to cold (4 °C). To further examine the effects of elevated ketosis, we administered oral 1,3-butanediol (1,3BD) or water to pups from postnatal day 2 to 21, and analyzed the iWAT SVF using bulk RNA-seq, single-cell RNA-seq, and ChIP-seq targeting H3K9ac, H3K14ac, and H3K9bhb. These integrative analyses reveal how early-life ketosis programs adipose progenitors to promote beige adipocyte development. Overall design: To investigate how both loss and enhancement of ketogenesis influence histone modifications in adipose progenitors, SVF was isolated from iWAT depots of P21 control, Hmgcs2 KO, and 1,3BD-supplemented (1,3BD) male mice and cultured overnight. Sonicated chromatin was immunoprecipitated with antibodies against H3K9ac, H3K14ac, and H3K9bhb and subjected to ChIP-seq.