Integrated Transcriptome and Metabolome Analyses Uncover Cholesterol-Responsive Gene Networks

Area of Science:

• Cellular Biology
• Metabolomics
• Systems Biology

Background:

• Cholesterol stress significantly affects cellular functions, but the precise mechanisms are not fully understood.
• Investigating cellular responses to cholesterol is crucial for understanding metabolic disorders.

Purpose of the Study:

• To elucidate the molecular mechanisms underlying cellular responses to cholesterol stress.
• To identify key genes and pathways involved in cholesterol metabolism and cellular adaptation.

Main Methods:

• Integrated transcriptome (RNA-seq) and metabolome (LC-MS) analyses were performed on HeLa cells treated with cholesterol.
• Co-expression network analysis was used to identify functional modules.
• RT-qPCR, Western blot, and analysis of clinical datasets were employed for validation.

Main Results:

• Cholesterol stress induced stage-specific transcriptomic changes, including early stress responses and late immune-metabolic coordination.
• Metabolomic analysis revealed extensive lipid remodeling and identified GFPT1-driven UDP-GlcNAc accumulation and increased taurine.
• GFPT1 upregulation and ANGPTL4 downregulation were confirmed, correlating with cholesterol levels in NASH liver cancer patients.

Conclusions:

• This study establishes a chronological model of cholesterol sensing.
• GFPT1 and ANGPTL4 are identified as critical regulators connecting glycosylation and lipid metabolism.
• These findings provide mechanistic insights into cholesterol-associated metabolic diseases like NASH.