NAT10-mediated ac⁴C modification of TNFRSF1A promotes acute kidney injury by activating NF-κB pathway

Acute kidney injury (AKI) is closely linked to inflammatory responses driven by the NF-κB pathway, yet the epigenetic mechanisms underlying this association remain poorly understood. This study investigates the role of N-acetyltransferase 10 (NAT10)-mediated RNA N⁴-acetylcytidine (ac⁴C) modification in regulating NF-κB signaling during AKI.

Clinical AKI specimens and experimental models (in vivo and in vitro) were analyzed to assess NAT10 expression in renal tubular epithelial cells (RTECs). Genetic knockdown and overexpression of NAT10 (including the catalytically inactive NAT10-G641E mutant) were performed in vitro to evaluate NF-κB pathway activity and hypoxia/reoxygenation (H/R)-induced inflammatory responses. In murine ischemia/reperfusion (I/R)-induced AKI models, genetic ablation of NAT10 in RTECs and pharmacological inhibition using Remodelin were employed to study renal inflammation and injury. Mechanistic insights were derived from integrated ac⁴C acetylated RNA immunoprecipitation sequencing (acRIP-seq) and RNA sequencing (RNA-seq) analyses to identify NAT10-mediated ac⁴C modification targets and their effects on RNA stability, translation efficiency, and signaling pathways.

NAT10 was significantly upregulated in RTECs of clinical AKI specimens and experimental models. In vitro, NAT10 knockdown suppressed NF-κB activation, mitigating H/R-induced inflammation and cellular damage, while NAT10 overexpression (but not the G641E mutant) activated NF-κB, exacerbating RTECs injury. In murine I/R-AKI models, genetic or pharmacological inhibition of NAT10 attenuated NF-κB-mediated renal inflammation and tubular damage. Mechanistically, NAT10-mediated ac⁴C modification enhanced RNA stability and translation efficiency of tumor necrosis factor receptor superfamily member 1A (TNFRSF1A), leading to aberrant NF-κB activation and subsequent inflammatory cascades.

This study identifies the NAT10-ac⁴C-TNFRSF1A axis as a novel epigenetic regulator of NF-κB-driven renal inflammation in AKI. NAT10 promotes inflammatory signaling and tubular damage by enhancing TNFRSF1A mRNA stability and translation efficiency, highlighting its potential as a therapeutic target. These findings provide a theoretical foundation for developing AKI treatments targeting NAT10-mediated ac⁴C modification to mitigate NF-κB-dependent pathology.