Alterations of telomere-associated genes and underlying mechanisms in hypoxic-ischemic encephalopathy

Hypoxic-ischemic encephalopathy (HIE) is a severe neurological disorder with complex pathogenesis. The role of telomere-associated genes in HIE remains unclear. This study aims to investigate their alterations and mechanisms to provide new therapeutic insights.

Telomere-associated differentially expressed genes (Telomere-DEGs) were identified from neonatal mouse HIE transcriptome datasets (GSE23317, GSE144456) in GEO. GO and KEGG enrichment analyses were performed. A protein-protein interaction network was constructed using STRING, and the best Telomere-DEGs (BTDGs) were identified via CytoHubba, followed by validation in GSE144456. Immune infiltration was assessed with CIBERSORT. Potential drugs targeting BTDGs were predicted using DGIdb. HT22 cell OGD/R and neonatal mouse HIE models were established. qRT-PCR, Western blot, immunofluorescence, flow cytometry, ROS detection, and CCK-8 assays were used to validate BTDG expression and explore the regulatory role of the SOCS3/NF-κB axis in telomere dysfunction and oxidative stress.

Eighteen HIE-related Telomere-DEGs were identified, with ten defined as BTDGs. Enrichment analyses revealed involvement in glial cell regulation, inflammation, and immune cell migration. Immune infiltration analysis showed significantly increased infiltration of Th1 and mature dendritic cells in the HIE group. BTDG expression was positively associated with neutrophil infiltration, negatively correlated with M1 macrophage infiltration, and positively correlated with M2 macrophage infiltration. Drug prediction identified 19 FDA-approved drugs targeting BTDGs, including Phenobarbital, Baclofen, and Bromocriptine. In vivo, SOCS3 and Ccl4 mRNA and protein were significantly upregulated in the HIE mouse hippocampus. In vitro, OGD/R upregulated SOCS3 in HT22 cells, accompanied by decreased TRF2, increased γ-H2AX and ROS, and reduced cell viability. SOCS3 knockdown inhibited NF-κB activation, upregulated TRF2, reduced γ-H2AX and ROS, and decreased apoptosis, which were reversed by the NF-κB activator PMA.

The study systematically identified Telomere-DEGs in neonatal HIE, characterized the disease's inflammatory immune microenvironment, and predicted potential targeted therapeutic agents. SOCS3 upregulation in HIE was confirmed both in vivo and in vitro. In vitro mechanistic studies suggest that SOCS3 aggravates telomere damage and oxidative stress through NF-κB activation, leading to neuronal injury, thus providing experimental evidence for HIE mechanism research and targeted therapy.