Transcriptional determinants of lipid mobilization in human adipocytes

Defects in adipocyte lipolysis drive multiple aspects of cardiometabolic disease but the transcriptional framework controlling this process has not been established. To address this, we performed a targeted perturbation screen in primary human adipocytes. Our analyses identified 37 transcriptional regulators of lipid mobilization, which we classified as: i) transcription factors, ii) histone chaperones, and iii) mRNA processing proteins. Based on its strong relationship with multiple readouts of lipolysis in patient samples, we performed mechanistic studies on one hit, ZNF189, which encodes the Zinc Finger Protein 189. Using mass-spectrometry and chromatin profiling techniques, we show that ZNF189 interacts with the tripartite motif family member TRIM28 and represses the transcription of an adipocyte-specific isoform of Phosphodiesterase 1B (PDE1B2). The regulation of lipid mobilization by ZNF189 requires PDE1B2 and overexpression of PDE1B2 is sufficient to attenuate hormone-stimulated ...,

RNAi Screen - Glycerol Release: RNA interference screening experiment, in adipocytes differentiated from CD55+/DPP4+ progenitor cells, was performed with the siGENOME® SMARTpool® siRNA Human Transcription Factor Library (Dharmacon, Lafayette, CO, USA). Lipolysis was assessed by fluorometric detection of glycerol in phenol-red-free media (ThermoFisher Scientific, 21041-033) collected from cells after 72 hours after siRNA transfection. Cytotoxicity was assessed by measuring LDH activity using the Cytotoxicity Detection KitPLUS 548 (Roche, 4744926001) according to the manufacturer’s instructions. Glycerol release and LDH data were analyzed using R v4.2.2. Outliers were identified using the IQR method, additionally triplicates with CV > 20% were excluded from further analysis. The reproducibility of the glycerol release assays was assessed by calculation of the Pearson correlation coefficient between the experimental replicates. Data were normalized and scaled using the robust z-score ..., , # Transcriptional determinants of lipid mobilization in human adipocytes

This data set contains the following:

1. **RNAi Screen - Glycerol Release:** RNA interference screening experiment, in adipocytes differentiated from CD55+/DPP4+ progenitor cells, was performed with the siGENOME® SMARTpool® siRNA Human Transcription Factor Library (Dharmacon, Lafayette, CO, USA). Lipolysis was assessed by fluorometric detection of glycerol in phenol-red-free media (ThermoFisher Scientific, 21041-033) collected from cells after 72 hours after siRNA transfection. Cytotoxicity was assessed by measuring LDH activity using the Cytotoxicity Detection KitPLUS 548 (Roche, 4744926001) according to the manufacturer’s instructions. Glycerol release and LDH data were analyzed using R v4.2.2. Outliers were identified using the IQR method, additionally triplicates with CV > 20% were excluded from further analysis. The reproducibility of the glycerol release assays was assessed by calculation of the Pears...