Yao Lu, Liu Zuqiang, Chen Weifeng, Xu Jiaqi, Xu Xiaoyue, Xu Jiaxin, Ma Liyun, Li Xiaoqing, Li Quanlin, Zhou Pinghong
Endoscopy Center and Endoscopy Research Institute, Zhongshan Hospital, Fudan University, Shanghai, China.
J Neurogastroenterol Motil. 2023 Oct 30;29(4):486-500. doi: 10.5056/jnm21246. Epub 2023 Aug 16.
BACKGROUND/AIMS: Previous studies reveal that immune-mediated neuroinflammation plays a key role in the etiology of esophageal achalasia. However, the understanding of leucocyte phenotype and proportion is limited. This study aim to evaluate the phenotypes of leukocytes and peripheral blood mononuclear cells transcriptomes in esophageal achalasia.
We performed high-dimensional flow cytometry to identified subsets of peripheral leukocytes, and further validated in lower esophageal sphincter histologically. RNA sequencing was applied to investigate the transcriptional changes in peripheral blood mononuclear cells of patients with achalasia. Cell-type Identification by Estimating Relative Subsets of RNA Transcripts (CIBERSORT) was used for estimating the immune cell types. A differential gene expression analysis was performed and the differential expressed genes were subjected to gene ontology, Kyoto Encyclopedia of Genes and Genomes network, protein-protein interaction network construction.
An imbalance between innate and adaptive immune cells occurred in achalasia. Specifically, neutrophils and CD8+ T cells increased both in peripheral blood and lower esophageal sphincter in achalasia. Eosinophils decreased in peripheral blood but massively infiltrated in lower esophageal sphincter. CIBERSORT analysis of peripheral blood mononuclear cells RNA sequencing displayed an increased prevalence of CD8+ T cells. 170 dysregulated genes were identified in achalasia, which were enriched in immune cells migration, immune response, etc. Proton pump inhibitor analysis revealed the intersections and gained 7 hub genes in achalasia, which were IL-6, Toll-like receptor 2, IL-1β, tumor necrosis factor, complement C3, and complement C1q A chain.
Patients with achalasia exhibited an imbalance of systematic innate and adaptive immunity, which may play an important role in the development of achalasia.
背景/目的：既往研究表明，免疫介导的神经炎症在贲门失弛缓症的病因中起关键作用。然而，对白细胞表型和比例的了解有限。本研究旨在评估贲门失弛缓症患者白细胞的表型及外周血单核细胞转录组。
我们进行了高维流式细胞术以鉴定外周白细胞亚群，并在食管下括约肌进行组织学进一步验证。应用RNA测序研究贲门失弛缓症患者外周血单核细胞的转录变化。通过估计RNA转录本相对亚群进行细胞类型鉴定（CIBERSORT）来估计免疫细胞类型。进行差异基因表达分析，并对差异表达基因进行基因本体论、京都基因与基因组百科全书网络、蛋白质-蛋白质相互作用网络构建。
贲门失弛缓症患者先天性和适应性免疫细胞之间出现失衡。具体而言，贲门失弛缓症患者外周血和食管下括约肌中的中性粒细胞和CD8 + T细胞均增加。外周血中嗜酸性粒细胞减少，但在食管下括约肌中大量浸润。外周血单核细胞RNA测序的CIBERSORT分析显示CD8 + T细胞的患病率增加。在贲门失弛缓症中鉴定出170个失调基因，这些基因在免疫细胞迁移、免疫反应等方面富集。质子泵抑制剂分析揭示了交集并在贲门失弛缓症中获得了7个枢纽基因，即IL-6、Toll样受体2、IL-1β、肿瘤坏死因子、补体C3和补体C1q A链。
贲门失弛缓症患者表现出系统性先天性和适应性免疫失衡，这可能在贲门失弛缓症的发展中起重要作用。
