Ye Rongchen, Ma Sige, Chen Yan, Shan Jiarou, Tan Ledong, Su Liang, Tong Yanlu, Zhao Ziyang, Chen Hongjiao, Fu Ming, Guo Zhipeng, Zuo Xiaoyu, Yu Jiakang, Zhong Wei, Zeng Jixiao, Liu Fei, Chai Chenwei, Guan Xisi, Wang Zhe, Liu Tao, Liang Jiankun, Zhang Yan, Shi Hongguang, Wen Zhe, Xia Huimin, Zhang Ruizhong
Guangdong Provincial Key Laboratory of Research in Structure Birth Defect Disease and Department of Pediatric Surgery, Guangzhou Women and Children's Medical Center, Guangzhou Medical University, Guangdong Provincial Clinical Research Center for Child Health, Guangzhou, 510623, China.
Faculty of Medicine, Macau University of Science and Technology, Macau, 999078, China.
JHEP Rep. 2023 Sep 12;5(11):100908. doi: 10.1016/j.jhepr.2023.100908. eCollection 2023 Nov.
BACKGROUND & AIMS: Our previous study indicated that CD177 neutrophil activation has a vital role in the pathogenesis of biliary atresia (BA), which is partially ameliorated by -acetylcysteine (NAC) treatment. Here, we evaluated the clinical efficacy of NAC treatment and profiled liver-resident immune cells via single cell RNA-sequencing (scRNA-seq) analysis to provide a comprehensive immune landscape of NAC-derived immune regulation.
A pilot clinical study was conducted to evaluate the potential effects of intravenous NAC treatment on infants with BA, and a 3-month follow-up was carried out to assess treatment efficacy. scRNA-seq analysis of liver CD45 immune cells in the control (n = 4), BA (n = 6), and BA + NAC (n = 6) groups was performed and the effects on innate cells, including neutrophil and monocyte-macrophage subsets, and lymphoid cells were evaluated.
Intravenous NAC treatment demonstrated beneficial efficacy for infants with BA by improving bilirubin metabolism and bile acid flow. Two hepatic neutrophil subsets of innate cells were identified by scRNA-seq analysis. NAC treatment suppressed oxidative phosphorylation and reactive oxygen species production in immature neutrophils, which were transcriptionally and functionally similar to CD177 neutrophils. We also observed the suppression of hepatic monocyte-mediated inflammation, decreased levels of oxidative phosphorylation, and M1 polarisation in Kupffer-like macrophages by NAC. In lymphoid cells, enhancement of humoral immune responses and attenuation of cellular immune responses were observed after NAC treatment. Moreover, cell-cell interaction analysis showed that innate/adaptive proinflammatory responses were downregulated by NAC.
Our clinical and scRNA-seq data demonstrated that intravenous NAC treatment partially reversed liver immune dysfunction, alleviated the proinflammatory responses in BA by targeting innate cells, and exhibited beneficial clinical efficacy.
BA is a serious liver disease that affects newborns and has no effective drug treatment. In this study, scRNA-seq showed that NAC treatment can partially reverse the immune dysfunction of neutrophil extracellular trap-releasing CD177+ neutrophils and Kupffer cells, and lower the inflammatory responses of other innate immune cells in BA. In consequence, intravenous NAC treatment improved the clinical outcomes of patients with BA in term of bilirubin metabolism.
我们之前的研究表明，CD177中性粒细胞活化在胆道闭锁（BA）的发病机制中起关键作用，而N - 乙酰半胱氨酸（NAC）治疗可部分改善这一情况。在此，我们评估了NAC治疗的临床疗效，并通过单细胞RNA测序（scRNA - seq）分析对肝脏驻留免疫细胞进行了剖析，以全面了解NAC介导的免疫调节的免疫格局。
开展了一项初步临床研究，以评估静脉注射NAC治疗对BA婴儿的潜在影响，并进行了为期3个月的随访以评估治疗效果。对对照组（n = 4）、BA组（n = 6）和BA + NAC组（n = 6）的肝脏CD45免疫细胞进行scRNA - seq分析，并评估其对包括中性粒细胞和单核细胞 - 巨噬细胞亚群在内的固有细胞以及淋巴细胞的影响。
静脉注射NAC治疗通过改善胆红素代谢和胆汁酸流动，对BA婴儿显示出有益的疗效。通过scRNA - seq分析鉴定出了固有细胞的两种肝脏中性粒细胞亚群。NAC治疗抑制了未成熟中性粒细胞中的氧化磷酸化和活性氧生成，这些未成熟中性粒细胞在转录和功能上与CD177中性粒细胞相似。我们还观察到NAC抑制了肝脏单核细胞介导的炎症，降低了氧化磷酸化水平，并使库普弗样巨噬细胞中的M1极化减弱。在淋巴细胞中，NAC治疗后观察到体液免疫反应增强和细胞免疫反应减弱。此外，细胞 - 细胞相互作用分析表明，NAC下调了固有/适应性促炎反应。
我们的临床和scRNA - seq数据表明，静脉注射NAC治疗可部分逆转肝脏免疫功能障碍，通过靶向固有细胞减轻BA中的促炎反应，并显示出有益的临床疗效。
BA是一种影响新生儿的严重肝脏疾病，目前尚无有效的药物治疗方法。在本研究中，scRNA - seq表明NAC治疗可部分逆转释放中性粒细胞胞外陷阱的CD177 + 中性粒细胞和库普弗细胞的免疫功能障碍，并降低BA中其他固有免疫细胞的炎症反应。因此，静脉注射NAC治疗在胆红素代谢方面改善了BA患者的临床结局。
