Histone Acetylation Landscape Changes During Adipocyte Differentiation
DOI:
https://doi.org/10.5281/zenodo.22760019Keywords:
adipogenesis, histone acetylation, H3K27ac, CEBPA, PPARgamma, super-enhancer, ChIP-seqAbstract
Terminal differentiation of preadipocytes into mature, lipid-storing adipocytes requires coordinated activation of a
small set of master transcription factors acting on a chromatin landscape that is remodeled in successive waves, yet
the genome-wide dynamics of histone acetylation across this process, and their relationship to super-enhancer
formation at adipogenic master regulator loci, remain incompletely mapped at high temporal resolution. Here we
profiled H3K27ac and H3K9ac by chromatin immunoprecipitation sequencing (ChIP-seq), chromatin accessibility by
ATAC-seq, and gene expression by RNA-sequencing across six time points spanning confluent growth arrest,
mitotic clonal expansion and terminal differentiation of 3T3-L1 preadipocytes induced with a standard hormonal
cocktail. H3K27ac signal at a defined set of 1,842 genomic regions increased more than four-fold during the first 48
hours of differentiation, preceding peak expression of the early response factors CEBPB and CEBPD, and a
substantially larger, largely non-overlapping set of regions gained H3K27ac only after 72 hours, coincident with
induction of the terminal adipogenic regulators PPARgamma and CEBPA. Clustering of dynamic H3K27ac regions
by their acetylation trajectory identified a coherent late-gained cluster strongly enriched for super-enhancers
overlapping PPARgamma and CEBPA target genes central to lipid storage and adipokine secretion, and CRISPR
interference-mediated silencing of three constituent enhancers within the Fabp4 super-enhancer individually
reduced Fabp4 induction by 35-58% without eliminating it, consistent with additive rather than fully redundant
enhancer contributions. Pharmacological inhibition of the acetyltransferases p300 and CBP during the mitotic clonal
expansion window, but not after its completion, blunted lipid accumulation and terminal marker gene induction,
indicating a temporally restricted requirement for acetyltransferase activity during early adipogenic commitment.
These results define a two-wave histone acetylation program during adipocyte differentiation and identify a
temporally restricted, p300/CBP-dependent window during which the terminal adipogenic super-enhancer
landscape is established.
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