Identification of key mitochondria-related genes in necrotizing enterocolitis using single-cell hdWGCNA and...

Necrotizing enterocolitis (NEC) is a severe gastrointestinal disorder in preterm infants. The interplay between mitochondrial metabolism and immune inflammation in its development is not fully understood.

Single-cell data were analyzed using dimensionality reduction, clustering, and the high-dimensional weighted gene co-expression network analysis (hdWGCNA) algorithm to identify key gene modules in monocytes. GSE46619 was integrated with the MitoCarta3.0 to identify mitochondria-associated differentially expressed genes (MitoDEGs). Acyl-CoA synthetase long-chain family member 1 (ACSL1) was selected as a candidate. Immune infiltration was evaluated via the CIBERSORT algorithm, and a competing endogenous RNA (ceRNA) regulatory network was constructed using Cytoscape. The expression and function of ACSL1 were validated both in vivo and in vitro, using immunohistochemistry (IHC), qRT-PCR, western blot, and siRNA knockdown.

A key monocyte subset was identified in NEC. Integrated analysis revealed three MitoDEGs (ACSL1, SOD2, SLC25A37) were linked to NEC, with ACSL1 showing the most significant upregulation. ACSL1 expression correlated strongly with immune cell infiltration and was confirmed to be elevated in vivo and in vitro models. Knocking down ACSL1 suppressed lipopolysaccharide (LPS)-induced inflammatory factor expression and ROS production.

ACSL1 plays a critical role in the pathogenesis of NEC, suggesting its potential as a novel biomarker.

Our study reveals massive monocyte infiltration and identifies the mitochondria-related gene ACSL1 as highly expressed and functionally significant in NEC. This is the first integrated analysis (single-cell, hdWGCNA, MitoCarta3.0) pinpointing ACSL1 as a novel immunometabolic hub specific to NEC pathophysiology. ACSL1 provides a crucial mechanistic link between mitochondrial function and NEC development. It significantly correlates with key immune cells (neutrophils/mast cells/macrophages/T cells), highlighting its role in NEC immune dysregulation. These findings reveal a critical role of ACSL1 in NEC pathogenesis, highlighting its potential as a novel biomarker.