Medical Journal of the Islamic Republic Of Iran

en Peripheral IL-6/IL-17/NF-κB1 and IL-10 Signaling in Children with Autism Spectrum Disorder: Integrative Transcriptomic Analysis and qRT-PCR Validation Medical Genetics Medical Genetics Original Research Original Research     Background: Autism spectrum disorder (ASD) is associated with immune and inflammatory dysregulation. However, the molecular networks linking peripheral immune signatures to neuroinflammatory processes remain poorly understood. This study aimed to explore inflammation-related molecular pathways in ASD through integrated transcriptomic network analysis and to validate key cytokine genes (IL6, IL10, IL17, NF-κB1) using quantitative real-time polymerase chain reaction (qRT-PCR).    Methods: This was an integrative computational-experimental study. We analyzed 4 gene expression Omnibus (GEO) transcriptomic datasets (GSE18123, GSE111176, GSE87847, GSE6575), constructed protein-protein interaction (PPI) networks, and identified inflammation-related modules. Selected inflammatory genes (IL6, IL10, IL17, NF-ΚB1) were validated by qRT-PCR in peripheral blood samples from ASD (n = 15) and healthy controls (n = 5). Statistical analyses were conducted in R. Data normality was assessed using the Shapiro-Wilk test, and normally distributed variables were compared using t-tests.    Results: Integration of datasets revealed core differentially expressed genes (DEGs) and a connected PPI network (26 nodes, 88 edges), with hub genes such as PUM1, TRRAP, ILF3, INO80, and PTBP1. Functional enrichment indicated cytokine-mediated signaling, leukocyte activation, and neuroinflammation processes. Network analysis highlighted central regulators linking chromatin remodeling, ribonucleic acid (RNA) processing, and immune signaling. qRT-PCR confirmed dysregulation of IL6 (fold change ≈ 12.8, P = 0.049), IL17 (≈ 21.3, P = 0.048), NF-ΚB1 (≈ 42.4, P = 0.039), and IL10 (≈ 0.101, P = 0.038).    Conclusion: The findings suggest an IL-6/IL-17/NF-κB1–centric proinflammatory axis and reduced IL-10–mediated regulation in ASD, implicating peripheral immune activation and transcriptional regulators in neuroinflammatory processes. The identified hub genes and pathways may serve as biomarkers and therapeutic targets for an inflammation-associated ASD subtype. Limitations include small qRT-PCR sample size and lack of protein-level validation; future studies should explore longitudinal and multiomics approaches.   Autism Spectrum Disorder, Neuroinflammation, Inflammatory Factors, Protein–Protein Interaction, Transcriptomics, Computational Biology 1486 1498 http://mjiri.iums.ac.ir/browse.php?a_code=A-10-9678-1&slc_lang=en&sid=1 Yoosra Najim Abed Al-Saadi najimyusra0@gmail.com 200319475328460095746 200319475328460095746 No Ibn Sina University of Medical and Pharmaceutical Sciences, Baghdad, Iraq Zainab Basim Mohammed Dr.zainab.basim@ibnsina.edu.iq 200319475328460095747 200319475328460095747 No Ibn Sina University of Medical and Pharmaceutical Sciences, Baghdad, Iraq Mithal Abdulkareem Abdoun Mithalaon@gmail.com 200319475328460095748 200319475328460095748 No Ibn Sina University of Medical and Pharmaceutical Sciences, Baghdad, Iraq Ali Mahmoudi alimahmoudi68@yahoo.com 200319475328460095749 200319475328460095749 Yes Department of Basic Medical Sciences, Faculty of Medicine, Abadan University of Medical Sciences, Abadan, Iran