Osama Aya, Anwar Ali Mostafa, Ezzeldin Shahd, Ahmed Eman Ali, Mahgoub Sebaey, Ibrahim Omneya, Ibrahim Sherif Abdelaziz, Abdelhamid Ismail Abdelshafy, Bakry Usama, Diab Aya A, A Sayed Ahmed, Magdeldin Sameh
Proteomics and Metabolomics Unit, Basic Research Department, Children's Cancer Hospital, 57357 Cairo, (CCHE-57357), Egypt.
Proteomics and Metabolomics Unit, Basic Research Department, Children's Cancer Hospital, 57357 Cairo, (CCHE-57357), Egypt; Department of Pharmacology, Faculty of Veterinary Medicine, Suez Canal University, 41522 Ismailia, Egypt.
J Adv Res. 2025 Jan 25. doi: 10.1016/j.jare.2025.01.036.
Gut microbiota alterations have been implicated in Autism Spectrum Disorder (ASD), yet the mechanisms linking these changes to ASD pathophysiology remain unclear.
This study utilized a multi-omics approach to uncover mechanisms linking gut microbiota to ASD by examining microbial diversity, bacterial metaproteins, associated metabolic pathways and host proteome.
The gut microbiota of 30 children with severe ASD and 30 healthy controls was analyzed. Microbial diversity was assessed using 16S rRNA V3 and V4 sequencing. A novel metaproteomics pipeline identified bacterial proteins, while untargeted metabolomics explored altered metabolic pathways. Finally, multi-omics integration was employed to connect macromolecular changes to neurodevelopmental deficits.
Children with ASD exhibited significant alterations in gut microbiota, including lower diversity and richness compared to controls. Tyzzerella was uniquely associated with the ASD group. Microbial network analysis revealed rewiring and reduced stability in ASD. Major metaproteins identified were produced by Bifidobacterium and Klebsiella (e.g., xylose isomerase and NADH peroxidase). Metabolomics profiling identified neurotransmitters (e.g., glutamate, DOPAC), lipids, and amino acids capable of crossing the blood-brain barrier, potentially contributing to neurodevelopmental and immune dysregulation. Host proteome analysis revealed altered proteins, including kallikrein (KLK1) and transthyretin (TTR), involved in neuroinflammation and immune regulation. Finally, multi-omics integration supported single-omics findings and reinforced the hypothesis that gut microbiota and their macromolecular products may contribute to ASD-associated symptoms.
The integration of multi-omics data provided critical evidence that alteration in gut microbiota and associated macromolecule production may play a role in ASD-related symptoms and co-morbidities. Key bacterial metaproteins and metabolites were identified as potential contributors to neurological and immune dysregulation in ASD, underscoring possible novel targets for therapeutic intervention.
肠道微生物群的改变与自闭症谱系障碍（ASD）有关，然而，将这些变化与ASD病理生理学联系起来的机制仍不清楚。
本研究采用多组学方法，通过检查微生物多样性、细菌元蛋白质、相关代谢途径和宿主蛋白质组，揭示肠道微生物群与ASD之间的联系机制。
分析了30名重度ASD儿童和30名健康对照儿童的肠道微生物群。使用16S rRNA V3和V4测序评估微生物多样性。一种新的元蛋白质组学流程鉴定细菌蛋白质，而非靶向代谢组学探索改变的代谢途径。最后，采用多组学整合将大分子变化与神经发育缺陷联系起来。
ASD儿童的肠道微生物群表现出显著改变，包括与对照组相比多样性和丰富度较低。泰泽氏菌与ASD组独特相关。微生物网络分析显示ASD中存在重新布线和稳定性降低。鉴定出的主要元蛋白质由双歧杆菌和克雷伯氏菌产生（如木糖异构酶和NADH过氧化物酶）。代谢组学分析确定了能够穿过血脑屏障的神经递质（如谷氨酸、3,4-二羟基苯乙酸）、脂质和氨基酸，可能导致神经发育和免疫失调。宿主蛋白质组分析揭示了改变的蛋白质，包括参与神经炎症和免疫调节的激肽释放酶（KLK1）和转甲状腺素蛋白（TTR）。最后，多组学整合支持单组学研究结果，并强化了肠道微生物群及其大分子产物可能导致ASD相关症状的假设。
多组学数据的整合提供了关键证据，表明肠道微生物群的改变和相关大分子产物可能在ASD相关症状和共病中起作用。关键细菌元蛋白质和代谢产物被确定为ASD神经和免疫失调的潜在促成因素，强调了可能的新型治疗干预靶点。
