Department of Gastroenterology, Guangdong Provincial Key Laboratory of Gastroenterology, State Key Laboratory of Organ Failure Research, Nanfang Hospital, Southern Medical University, Guangzhou, China.
The First School of Clinical Medicine, Southern Medical University, Guangzhou, China.
J Transl Med. 2023 Jul 24;21(1):497. doi: 10.1186/s12967-023-04359-1.
Celiac disease (CeD) is a primary malabsorption syndrome with no specific therapy, which greatly affects the quality of life. Since the pathogenesis of CeD remains riddled, based on multiple transcriptome profiles, this study aimed to establish an immune interaction network and elucidated new mechanisms involved in the pathogenesis of CeD, providing potentially new evidence for the diagnosis and treatment of CeD.
Three microarray and three RNA sequencing datasets of human duodenal tissue with or without CeD were included in Gene Expression Omnibus and respectively merged into derivation and validation cohorts. Differential expression gene and functional enrichment analysis were developed, then pyroptosis enrichment score (PES) model was established to quantify pyroptosis levels. Immune infiltration and co-expression network were constructed based on Xcell database. Protein-protein interaction and weighted gene co-expression network analysis were determined to identify pyroptosis relative hub genes, whose predictive efficiency were tested using a least absolute shrinkage and selection operator (LASSO) regression model. CeD animal and in vitro cell line models were established to verify the occurrence of pyroptosis and molecules expression employing immunofluorescence, western blotting, cell counting kit-8 assay and enzyme-linked immunosorbent assay. Analysis of single-cell RNAseq (scRNAseq) was performed using "Seurat" R package.
Differentially expressed genes (DEGs) (137) were identified in derivation cohort whose function was mainly enriched in interferon response and suppression of metabolism. Since an enrichment of pyroptosis pathway in CeD was unexpectedly discovered, a PES model with high efficiency was constructed and verified with two external databases, which confirmed that pyroptosis was significantly upregulated in CeD epithelia. γδT cells exhibited high expression of IFN-γ were the most relevant cells associated with pyroptosis and occupied a greater weight in the LASSO predictive model of CeD. An accumulation of GSDMD expressed in epithelia was identified using scRNAseq, while animal model and in vitro experiments confirmed that epithelium cells were induced to become "pre-pyroptotic" status via IFN-γ/IRF1/GSDMD axis. Furthermore, gluten intake triggered pyroptosis via caspase-1/GSDMD/IL-1β pathway.
Our study demonstrated that pyroptosis was involved in the pathogenesis of CeD, and elucidated the novel role of γδT cells in mediating epithelial cell pyroptosis.
乳糜泻（CeD）是一种无特定疗法的原发性吸收不良综合征，极大地影响了生活质量。由于 CeD 的发病机制仍然存在许多未解之谜，本研究基于多个转录组谱，旨在建立一个免疫相互作用网络，并阐明 CeD 发病机制中涉及的新机制，为 CeD 的诊断和治疗提供潜在的新证据。
纳入基因表达综合数据库中包含乳糜泻患者和非乳糜泻患者人十二指肠组织的三个微阵列和三个 RNA 测序数据集，并分别合并为推导和验证队列。进行差异表达基因和功能富集分析，然后建立焦亡富集评分（PES）模型以量化焦亡水平。基于 Xcell 数据库构建免疫浸润和共表达网络。确定蛋白质-蛋白质相互作用和加权基因共表达网络分析，以识别焦亡相关的枢纽基因，使用最小绝对收缩和选择算子（LASSO）回归模型测试其预测效率。建立乳糜泻动物和体外细胞系模型，通过免疫荧光、western blot、细胞计数试剂盒-8 检测和酶联免疫吸附试验验证焦亡和分子表达的发生。使用“Seurat”R 包对单细胞 RNAseq（scRNAseq）进行分析。
在推导队列中鉴定出差异表达基因（DEGs）（137 个），其功能主要富集在干扰素反应和代谢抑制中。由于出乎意料地发现 CeD 中焦亡途径的富集，构建了一个高效的 PES 模型，并通过两个外部数据库进行验证，证实 CeD 上皮中焦亡明显上调。γδT 细胞表现出高表达的 IFN-γ，是与焦亡最相关的细胞，在 CeD 的 LASSO 预测模型中占有更大的权重。使用 scRNAseq 鉴定出上皮细胞中 GSDMD 的积累，而动物模型和体外实验证实，上皮细胞通过 IFN-γ/IRF1/GSDMD 轴被诱导至“前焦亡”状态。此外，谷朊摄入通过 caspase-1/GSDMD/IL-1β 途径触发焦亡。
本研究表明焦亡参与了 CeD 的发病机制，并阐明了 γδT 细胞在介导上皮细胞焦亡中的新作用。
