NAT10-mediated N4-acetylcytidine modification in KLF9 mRNA promotes adipogenesis

Area of Science:

• Molecular Biology
• Cell Biology
• Metabolic Diseases

Background:

• Dysfunctional adipogenesis is a key driver of obesity.
• N-acetyltransferase 10 (NAT10) regulates RNA modifications (ac4C), impacting mRNA stability and translation.
• The role of NAT10 in adipogenesis and obesity remains largely unexplored.

Purpose of the Study:

• To investigate the role of NAT10 in adipogenesis and obesity.
• To identify NAT10's downstream targets and molecular mechanisms in adipose tissue.
• To evaluate NAT10 as a potential therapeutic target for obesity.

Main Methods:

• Analysis of NAT10 expression in obese individuals and mice.
• Overexpression and silencing studies in human adipose tissue-derived stem cells (hADSCs) and 3T3-L1 cells.
• Combined acRIP-sequencing (acRIP-seq) and RNA-sequencing (RNA-seq) to identify NAT10 targets.
• acRIP-PCR, dual-luciferase reporter assays, and in vivo studies using AAV-NAT10 and a NAT10 inhibitor (Remodelin).

Main Results:

• NAT10 is significantly upregulated in adipose tissue of obese subjects and mice.
• NAT10 overexpression promotes adipogenesis, while silencing inhibits it.
• NAT10 enhances KLF9 mRNA stability and activates the CEBPA/B-PPARG pathway.
• NAT10 directly binds to KLF9 mRNA motifs.
• In vivo inhibition of NAT10 (AAV-shRNA-mNAT10 or Remodelin) reduced adipose tissue expansion and body weight in diet-induced obese mice.

Conclusions:

• NAT10 plays a critical role in promoting adipogenesis and obesity.
• NAT10 regulates adipogenesis through KLF9 mRNA stabilization and subsequent pathway activation.
• Targeting NAT10 and its downstream effectors presents a promising therapeutic strategy for obesity prevention and treatment.