Department of Biochemistry, All India Institute of Medical Sciences, Lab 3002, 3rd floor Teaching Block, New Delhi, 110029, India.
Disease Biology Lab, Department of Biosciences, Sri Sathya Sai Institute of Higher Learning, Puttaparthi, Andhra Pradesh, India.
Sci Rep. 2021 Sep 22;11(1):18849. doi: 10.1038/s41598-021-98086-6.
Acute-on-Chronic Liver Failure (ACLF) is associated with innate immune dysfunction and high short-term mortality. Neutrophils have been identified to influence prognosis in ACLF. Neutrophil biology is under-evaluated in ACLF. Therefore, we investigated neutrophil-specific genes and their association with ACLF outcomes. This is an observational study. Enriched granulocytes, containing neutrophils, isolated from study participants in three groups- ACLF(n = 10), chronic liver disease (CLD, n = 4) and healthy controls (HC, n = 4), were analysed by microarray. Differentially expressed genes were identified and validated by qRT-PCR in an independent cohort of ACLF, CLD and HC (n = 30, 15 and 15 respectively). The association of confirmed overexpressed genes with ACLF 28-day non-survivors was investigated. The protein expression of selected neutrophil genes was confirmed using flow cytometry and IHC. Differential gene expression analysis showed 1140 downregulated and 928 upregulated genes for ACLF versus CLD and 2086 downregulated and 1091 upregulated genes for ACLF versus HC. Significant upregulation of neutrophilic inflammatory signatures were found in ACLF compared to CLD and HC. Neutrophil enriched genes ELANE, MPO and CD177 were highly upregulated in ACLF and their expression was higher in ACLF 28-day non-survivors. Elevated expression of CD177 protein on neutrophil surface in ACLF was confirmed by flow cytometry. IHC analysis in archival post mortem liver biopsies showed the presence of CD177 neutrophils in the liver tissue of ACLF patients. Granulocyte genes ELANE, MPO and CD177 are highly overexpressed in ACLF neutrophils as compared to CLD or HC. Further, this three-gene signature is highly overexpressed in ACLF 28-day non-survivors.
急性肝衰竭（ACLF）与固有免疫功能障碍和高短期死亡率相关。中性粒细胞已被确定为影响 ACLF 预后的因素。ACLF 中对中性粒细胞生物学的评估不足。因此，我们研究了中性粒细胞特异性基因及其与 ACLF 结局的关系。这是一项观察性研究。从 ACLF（n=10）、慢性肝病（CLD，n=4）和健康对照（HC，n=4）三组研究参与者中分离出富含中性粒细胞的富集粒细胞，并通过微阵列进行分析。在 ACLF、CLD 和 HC 的独立队列中通过 qRT-PCR 鉴定和验证差异表达基因（n=30、15 和 15）。研究了证实过表达基因与 ACLF 28 天非幸存者的关系。使用流式细胞术和免疫组化证实了选定中性粒细胞基因的蛋白表达。差异基因表达分析显示，与 CLD 相比，ACLF 中有 1140 个下调基因和 928 个上调基因，与 HC 相比，ACLF 中有 2086 个下调基因和 1091 个上调基因。与 CLD 和 HC 相比，ACLF 中发现中性粒细胞炎症特征明显上调。ELANE、MPO 和 CD177 等中性粒细胞富集基因在 ACLF 中高度上调，并且在 ACLF 28 天非幸存者中表达更高。通过流式细胞术证实了 ACLF 中性粒细胞表面 CD177 蛋白的升高表达。在 ACLF 患者的肝组织中通过免疫组化分析证实了 CD177 中性粒细胞的存在。与 CLD 或 HC 相比，ACLF 中性粒细胞中粒细胞基因 ELANE、MPO 和 CD177 的表达高度上调。此外，该三基因特征在 ACLF 28 天非幸存者中高度上调。
