Dataset Information

Endogenous aryl hydrocarbon receptor ligands-dysregulated phosphoproteomes and proteomes in human fetal endothelial cells

ABSTRACT: Preeclampsia (PE) is a leading cause of maternal and fetal morbidity and mortality and is characterized by a wide spectrum of impaired maternal and fetal vascular function. Aryl hydrocarbon receptor (AhR, a ligand-activated transcription factor) plays a critical role in regulating vascular development and function. Endogenous AhR ligands can induce endothelial dysfunction. However, the underlying protein phospho-signaling mechanisms remain unknown. To determine if endogenous AhR ligands dysregulate the phosphoproteomes and proteomes in endothelial cells, primary human umbilical vein endothelial cells (HUVECs) (n = 4; 2 cell preparations/cell sex) were cultured in endothelial cell media (ECM). After 16 hr serum starvation, subconfluent cells were treated with 1 µM 2-(1’H-indole-3’-carbonyl)-thiazole-4-carboxylic acid methyl ester (ITE, an endogenous AhR ligand) or DMSO (vehicle) for 4 and 24 hr. The cell proteins were subjected to a bottom-up phosphoproteomic analysis to determine acute and prolonged effects of ITE on protein phosphorylation.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Endothelial Cell Of Umbilical Vein, Cell Culture

DISEASE(S): Disease Free

SUBMITTER: Si-yan Zhang

LAB HEAD: Jing Zheng

PROVIDER: PXD047834 | Pride | 2025-02-10

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	221219_ITE_TMT_TiO2-ZrIMAC_Zheng.mzML	Mzml
	221219_ITE_TMT_TiO2-ZrIMAC_Zheng.raw	Raw
	221219_ITE_TMT_Total_Zheng.mzML	Mzml
	221219_ITE_TMT_Total_Zheng.raw	Raw
	Lumos_Phospho_ITE.mzid.gz	Mzid

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Publications

An endogenous aryl hydrocarbon receptor ligand dysregulates endothelial functions, transcriptome, and phosphoproteome.

Zhao Ying-Jie YJ Zhang Si-Yan SY Wei Ying-Ying YY Li Hui-Hui HH Lei Wei W Wang Kai K Kumar Sathish S Zhou Chi C Zheng Jing J

American journal of physiology. Cell physiology 20250205 3

We have reported that an endogenous aryl hydrocarbon receptor (AhR) ligand, 2-(1'H-indole-3'-carbonyl)-thiazole-4-carboxylic acid methyl ester (ITE), inhibits functions of human umbilical vein endothelial cells (HUVECs) and induces preeclampsia (PE)-like symptoms in rats. Herein, we tested the hypothesis that ITE impairs endothelial functions via disturbing transcriptome and phosphoproteome in HUVECs. We measured AhR activity in human maternal and umbilical vein sera from PE and normotensive (NT ...[more]

PMID: 39907700

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Project description:Background: The Aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor that is activated by xenobiotic chemicals that function as AHR ligands. The response to xenobiotic AHR ligands is toxicity and the induction of drug metabolizing enzymes. The impact of AHR knockdown on gene expression and pathways in human breast cancer cells in absence of xenobiotic AHR ligands has not been investigated on a genome-wide scale. Methods: MCF-7 cells were used as a model of human breast cancer. The AHR was silenced with short interfering RNA against AHR (siAHR). RNA-sequencing coupled with Ingenuity pathway analysis (IPA) was used to determine the impact of AHR knockdown on gene expression and pathways in the absence of xenobiotic AHR ligands. Western blot analysis and recombinant tumor necrosis factor (TNF) was used to investigate the impact of AHR knockdown on TNF induction of MNSOD expression. Results: We found that the AHR is transcriptionally active in MCF-7 breast cancer cells in the absence of xenobiotic AHR ligands. In total, the expression of 634 genes was significantly changed in AHR knockdown cells compared to controls at a false discovery rate of < 10%. The analysis confirmed that drug metabolizing enzymes were AHR targets; however, we found that AHR also promoted the expression of genes that were not directly related to the metabolism of xenobiotics, such as those involved in lipid and eicosanoid synthesis. Gene pathway analysis of the AHR regulated gene dataset predicted TNF activity to be reduced in AHR knockdown cells. Our finding that AHR knockdown inhibited TNF-stimulated increases in MNSOD expression confirmed this IPA prediction. Conclusions: This is the first gene expression profiling study of AHR knockdown breast cancer cells. Several known and novel AHR targets were identified. The results suggest that endogenous AHR regulation impacts eicosanoid synthesis by regulating gene expression. The IPA prediction of reduced TNF activity in AHR knockdown cells was confirmed by showing that TNF-induced increases in MNSOD expression was inhibited in AHR knockdown cells. The requirement of AHR for MNSOD activation is novel and provides a new link by which AHR may impact reactive oxidative species (ROS) signaling. Expression profiles by mRNA sequencing were generated for human MCF-7 cells transfected with cRNAi (6 replicates) or AHR-siRNA (5 replicates) for 36hr. Sequencing was performed on an Illumina HiSeq 1000 using a 2x100 base paired-end strategy.