ORIGINAL RESEARCH article
Front. Immunol.
Sec. Molecular Innate Immunity
Volume 16 - 2025 | doi: 10.3389/fimmu.2025.1587225
This article is part of the Research TopicInnovative Insights into Pattern Recognition and Signaling in Innate ImmunityView all 9 articles
Exploring the Biological Functions and Immune Regulatory Roles of IRAK3, TNFRSF1A, CX3CR1, and JUNB in T2DM Combined with MAFLD: Integrated Bioinformatics and Single-cell Analysis
Provisionally accepted
- 1Second Affiliated Hospital, Xinjiang Medical University, Urumqi, China
- 2Xinjiang Medical University, Ürümqi, Xinjiang Uyghur Region, China
- 3Fourth Affiliated Hospital of Xinjiang Medical University, Urumqi, China
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Objective: T2DM combined with MAFLD is a prevalent and complex metabolic disorder with significant global health implications. This study aims to identify and validate autophagy-related biomarkers for T2DM-associated MAFLD, explore regulatory mechanisms in disease progression, and investigate cellular diversity within the same tissue using single-cell sequencing data.Methods: This study utilized four datasets retrieved from the Gene Expression Omnibus (GEO) database: GSE15653, GSE89632, GSE24807and GSE23343. The analysis involved variance analysis, WGCNA analysis, PPI network construction, machine learning, and ROC analysis to identify autophagy-related biomarkers in T2DM combined with MAFLD within an independent external dataset. Enrichment analysis and immune infiltration analysis were conducted to understand the biological functions and immunomodulatory roles. Pathological changes of the disease were observed through animal experiments, while simultaneously validating key genes. Cellular diversity within liver tissues was characterized at the single-cell level, exploring interrelationships, differentiation, and developmental trajectories among cell populations through cellular communication and pseudo-temporal analyses.The study identified four key biomarkers (IRAK3, TNFRSF1A, CX3CR1, JUNB). RT-PCR analysis in animal experiments demonstrated significantly higher mRNA expression levels of IRAK3, TNFRSF1A, CX3CR1, and JUNB in T2DM and MAFLD rat liver tissues compared to the control group. Quantitative immunohistochemical analysis revealed notably elevated protein expression levels of IRAK3, TNFRSF1A, CX3CR1, and JUNB in liver tissues of rats with T2DM and MAFLD when contrasted with the control group (P < 0.05). Enrichment analysis indicated associations of T2DM combined with MAFLD pathogenesis with pathways such as the NFkappa B signaling pathway, MAPK signaling pathway and Insulin resistance. Correlative analysis uncovered connections between immune infiltration and the identified genes. Single-cell transcriptomic analysis highlighted the differentiation of CX3CR1, JUNB, and TFRC in various single-cell-annotated populations. The pseudo-temporal analysis of epithelial cells identified enriched genes at crucial nodes related to "Lipid and atherosclerosis", and "Type II diabetes mellitus" signaling pathways. Additionally, four cellular communication signaling pathways (TNF, CXCL, VEGF, and MIF) potentially significant in T2DM combined with MAFLD progression were identified through cell communication analysis.This study unveiled potential associations and key biomarkers concerning T2DM combined with MAFLD and relevant pathways, offering novel insights for the investigation of these two conditions.
Keywords: T2DM, MAFLD, single-cell RNA sequencing, biomarkers, Immune signaling pathway
Received: 04 Mar 2025; Accepted: 21 Jul 2025.
Copyright: © 2025 Wang, Li, Wushoulaji, Wang, Wan, Yang and Gong. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Ye Yang, Second Affiliated Hospital, Xinjiang Medical University, Urumqi, China
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