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TLE3 CUT&Tag-Seq to characterize TLE3-binding sites in colonic macrophages

ABSTRACT: We found that TLE3 in colonic macrophages mainly occupied distal intergenic regions (48%) and gene body regions (47%) with limited occupancy at the gene proximal regions (5%).

ORGANISM(S): Mus musculus

PROVIDER: GSE200645 | GEO | 2023/04/07

REPOSITORIES: GEO

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TLE3 CUT&Tag-Seq to characterize TLE3-binding sites in colonic macrophages

Project description:TLE3 CUT&Tag-Seq to characterize TLE3-binding sites in colonic macrophages

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Adipose subtype-selective recruitment of TLE3 in thermogenic gene programs

Project description:Transcriptional effectors of white adipocyte-selective gene expression have not been described. TLE3 is a white-selective cofactor that acts reciprocally with the brown-selective cofactor Prdm16 to specify lipid storage and thermogenic gene programs. When expressed at elevated levels in brown fat, TLE3 counters Prdm16, suppressing brown-selective genes and inducing white-selective genes, resulting in impaired fatty acid oxidation and thermogenesis. To further test the functional consequences of the changes in BAT phenotype in aP2-TLE3 Tg mice, we challenged them with cold exposure at 4C. aP2-TLE3 Tg mice had an impaired ability to respond to cold exposure compared to littermate control mice, as evidenced by a marked drop in core body temperature. Transgenic mice expressing TLE3 from the aP2 enhancer and wild type mice were housed individually without food and bedding immediately before the start of experiments. Mice were allowed free access to water and placed at 4C for 5 hours while core body temperature was monitored every hour using a rectal probe. For microarray experiments of BAT, RNA was extracted and pooled from 4 mice in each group.

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We performed ChIP-seq for TLE3 in MDA-MB-453 cells with and without FOXA1 transient silencing (siFOXA1 vs siNS) to determine the requirement of FOXA1 for TLE3 binding.

Project description:The identification of mechanisms controlling breast tumor progression from less aggressive to highly metastatic disease should provide novel therapeutic targets for patients. Here we report the transcriptional corepressor, TLE3, as a critical regulator of cellular plasticity in breast cancer. TLE3 facilitates repression of genes associated with the highly aggressive basal-like breast cancer (BLBC) subtype within luminal cells. Additionally, maintenance of the luminal lineage is dependent on appropriate localization of TLE3 to its transcriptional targets which is mediated through interaction with FOXA1. Furthermore, the ability of TLE3 to repress BLBC transcription results in reduced metastatic capacity and aggressive cellular behaviors. Together our findings establish TLE3 as a critical transcriptional mediator of breast cancer aggressiveness, with TLE3 sustaining the more differentiated and less metastatic nature of luminal tumors through suppression of gene expression programs associated with BLBC.

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Adipose subtype-selective recruitment of TLE3 in thermogenic gene programs

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Determination of the genome wide binding pattern of TLE3 in luminal breast cancer

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TLE3 drives luminal identity of breast cancer through transcriptional repression of FOXA1-target genes

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H3K27Ac CUT&Tag-Seq to characterize the epigenetic profiles in colonic macrophages from Tle4KO (Tle4fl/fl, Lyz2-Cre) and WT (Lyz2-Cre) mice

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Loss of TLE3 promotes the mitochondrial program in beige adipocytes and improves glucose metabolism

Project description:Prolonged cold exposure stimulates the recruitment of beige adipocytes within white adipose tissue. Beige adipocytes depend on mitochondrial oxidative phosphorylation to drive thermogenesis. The transcriptional mechanisms that promote remodeling in adipose tissue during the cold are not well understood. Here we demonstrate that the transcriptional coregulator transducin-like enhancer of split 3 (TLE3) inhibits mitochondrial gene expression in beige adipocytes. Conditional deletion of TLE3 in adipocytes promotes mitochondrial oxidative metabolism and increases energy expenditure, thereby improving glucose control. Using chromatin immunoprecipitation and deep sequencing, we found that TLE3 occupies distal enhancers in proximity to nuclear-encoded mitochondrial genes and that many of these binding sites are also enriched for early B-cell factor (EBF) transcription factors. TLE3 interacts with EBF2 and blocks its ability to promote the thermogenic transcriptional program. Collectively, these studies demonstrate that TLE3 regulates thermogenic gene expression in beige adipocytes through inhibition of EBF2 transcriptional activity. Inhibition of TLE3 may provide a novel therapeutic approach for obesity and diabetes.

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