Title:Core Neuroinflammatory Pathways Contributing to Delayed
Encephalopathy After Acute Carbon Monoxide Poisoning Revealed
by Multi-omics and Single Nucleus RNA-Seq
Volume: 33
Issue: 1
Author(s): Jinlan Li, Jun Li, Junming Fu, Chuying Huang, Wenqi Lv, Chunlong Zhang, Tianjie Tian, Qunhui Liu, Shijun Yang, Yong Tan, Guogen Sun, Ying Xiang, Guoquan Huang*Ning Wang*
Affiliation:
- Hubei Selenium and Human Health Institute, The Central Hospital of Enshi Tujia and Miao Autonomous Prefecture, Enshi, China
- Present Address: Department of Colorectal
and Anal Surgery, Central Hospital of Enshi Tujia and Miao Autonomous Prefecture, Enshi, 445000, China
- Department of Neurology, The First Affiliated Hospital of Fujian Medical University, Fuzhou, China
Keywords:
DEACMP, carbon monoxide poisoning, microglia, neuroinflammation, multi-omics, snRNA-seq.
Abstract:
Introduction: The pathogenesis of Delayed Encephalopathy After Acute Carbon Monoxide
Poisoning (DEACMP) remains mysterious, and specific predictive markers are lacking. This study
aimed to elucidate the molecular underpinnings and identify predictive biomarkers of DEACMP
through multi-omics and single-nucleusRNA sequencing (snRNA-seq).
Methods: Clinical data and blood samples were collected from 105 participants. Untargeted
metabolomics sequencing was employed to profile serum metabolites across these participants. Additionally,
individuals from the Healthy Controls (HCs), Acute Carbon Monoxide Poisoning patients (ACOP),
Non-Delayed Encephalopathy After ACOP (DEACMP-N), and DEACMP groups (n=3 each)
were randomly selected for transcriptome sequencing to identify potential predictive targets and pivotal
signaling pathways associated with DEACMP. Furthermore, Severe DEACMP and Control rat models
were established. Three rats from the Control, DEACMP, and DEACMP + Dexamethasone + Selenomethionine
groups were selected for snRNA-seq. Immunofluorescence multiplexing and qRT-PCR (quantitative
Reverse Transcription Polymerase Chain Reaction) were then performed to validate the identified
predictive targets.
Results: Analysis of clinical data from 105 participants highlights the pivotal role of inflammation in influencing
the prognosis of carbon monoxide poisoning. Metabolomics analysis identified 19 metabolites
that significantly differed between the DEACMP-N and DEACMP groups. Transcriptomics analysis of
12 participants indicated that DEACMP is primarily associated with six signaling pathways, including
lysosome and tuberculosis. Considering that microglia are central nervous system immune effectors, the
snRNA-seq analysis revealed altered gene expression and signaling pathways in microglia during
DEACMP, with KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis highlighting neutrophil
extracellular trap formation, lysosome, and tuberculosis as the predominant pathways. Differential gene
analysis from transcriptome and snRNA-seq identified 28 genes differentially expressed in DEACMP.
The STRING (Search Tool for the Retrieval of Interacting Genes/Proteins) database, immune multiplexing,
and qRT-PCR confirmed the pivotal role of the Ifngr1/Stat1/Ctss axis in DEACMP.
Discussion: This research identifies the Ifngr1/Stat1/Ctss axis as a key inflammatory mechanism in the
pathogenesis of DEACMP, thereby clarifying previous uncertainties regarding the sequelae of carbon
monoxide poisoning. The intersection of lysosomal and tuberculosis pathways, as revealed through
metabolomic, transcriptomic, and single-nucleus RNA sequencing analyses—especially within microglia—
offers novel mechanistic insights that could inform therapeutic interventions. While the integration
of multiple omics methodologies enhances the robustness of these findings, their biological relevance to
the pathogenesis of DEACMP requires rigorous validation through independent cohort verification approaches.
Conclusion: This study provides a comprehensive overview of serum metabolite expression, differential
gene expression, and signaling pathways in DEACMP, offering a theoretical foundation for understanding
the pathogenesis of DEACMP.
