N-myc and STAT interactor correlates with severity and prognosis in acute-on-chronic liver failure of hepatitis B virus

Abstract

Background and aim: Hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) is characterized by acute deterioration of chronic liver disease with excessive inflammation. N-myc and STAT interactor (NMI), an inflammation-mediated protein, involves in various inflammatory-related diseases, but the role of NMI in development and prognosis in HBV-ACLF remains to be elucidated.

Methods: Serum NMI from healthy controls (HCs, n = 20), chronic hepatitis B (CHB, n = 50) patients, and HBV-ACLF patients (n = 50) was determined using ELISA. NMI from peripheral blood mononuclear cells and liver was confirmed using quantitative real-time polymerase chain reaction, Western blot, and immunofluorescence.

Results: Serum NMI was increased 1.9-fold or 2.2-fold from HBV-ACLF patients compared with that from HCs (P < 0.01) or CHB patients (P < 0.01). Consistently, NMI from peripheral blood mononuclear cells was upregulated significantly from HBV-ACLF patients compared with that from HCs and CHB patients at mRNA and protein levels. Hepatic NMI from HBV-ACLF patients was 2.8-fold higher than that from HCs. Serum NMI was correlated with Model for End-stage Liver Disease, Chronic Liver Failure Consortium ACLF score, and ACLF grades. In contrast, serum NMI was significantly decreased in HBV-ACLF ameliorated patients during follow-up, whereas serum NMI was sustained at high levels in non-ameliorated patients. Elevated serum NMI (≥ 198.5 pg/mL) was correlated with poor survival rate of HBV-ACLF patients. Using receiver operating characteristics curves, it was suggested that serum NMI was a potential biomarker in predicting 3-month mortality of HBV-ACLF patients.

Conclusions: Our study highlights the potential role of NMI in assessing the development and prognosis of HBV-ACLF.

Keywords: N-myc and STAT interactor; acute-on-chronic liver failure; chronic hepatitis B; prognosis.

Figure 1

Serum and hepatic N‐myc and STAT interactor (NMI) increased in hepatitis B virus‐related acute‐on‐chronic liver failure (HBV‐ACLF) patients. (a) Comparison of serum NMI from healthy controls (HCs, n = 20), chronic hepatitis B (CHB) patients (n = 50), and HBV‐ACLF patients (n = 50) at baseline. (b) Hepatic NMI protein from HCs (n = 3) and HBV‐ACLF patients (n = 6). The bars represented bands density ratios of NMI/glyceraldehyde phosphate dehydrogenase (GAPDH). (c) The representative images of immunofluorescence analysis showed co‐staining of NMI (red) and CD68 (green) in the liver from HCs (n = 5), CHB patients (n = 5), and HBV‐ACLF patients (n = 6). Percentage of NMI⁺CD68⁺cells/CD68⁺cells was presented. Immunofluorescence double staining revealed that NMI was expressed by Kupffer cells, which were increased in HBV‐ACLF patients. Arrows indicate positive cells. The median with interquartile ranges is shown. ^** P < 0.01; ^*** P < 0.001. [Color figure can be viewed at http://wileyonlinelibrary.com]

Figure 2

N‐myc and STAT interactor (NMI) in peripheral blood mononuclear cells (PBMCs) from hepatitis B virus‐related acute‐on‐chronic liver failure (HBV‐ACLF) patients was upregulated. (a) NMI protein in PBMCs from healthy controls (HCs, n = 10), chronic hepatitis B (CHB) patients (n = 10), and HBV‐ACLF patients (n = 10). The bars represented bands density ratios of NMI/glyceraldehyde phosphate dehydrogenase (GAPDH). (b) NMI mRNA in PBMCs from HCs (n = 10), CHB patients (n = 10), and HBV‐ACLF patients (n = 10). The median with interquartile ranges is shown. ^* P < 0.05; ^** P < 0.01; ^*** P < 0.001.

Figure 3

Serum N‐myc and STAT interactor (NMI) was correlated with prognostic scores. The correlation between serum NMI and Chronic Liver Failure Consortium Acute‐on‐Chronic Liver Failure score (CLIF‐C ACLFs) (a), Model for End‐stage Liver Disease (MELD) (b), total bilirubin (TBil) (c), or prothrombin time (PT) (d). (e) The comparison of serum NMI at baseline between the hepatitis B virus‐related acute‐on‐chronic liver failure (HBV‐ACLF) patients grade < 2 and grade ≥ 2. The median with interquartile ranges is shown. ^** P < 0.01.

Figure 4

Serum N‐myc and STAT interactor (NMI) was inversely correlated with prognosis in hepatitis B virus‐related acute‐on‐chronic liver failure (HBV‐ACLF) patients. (a) The comparison of serum NMI between HBV‐ACLF ameliorated patients (n = 26) and HBV‐ACLF non‐ameliorated patients (n = 24) at baseline. Horizontal bars indicate the median values and interquartile ranges in each group. (b and c) The kinetic changes of serum NMI in HBV‐ACLF ameliorated and non‐ameliorated patients during follow‐up. (b) ▪▪▪, Ameliorated. (c) ▪▪▪, Non‐ameliorated. (d) Kaplan–Meier graphs showing the survival probability of two groups of HBV‐ACLF patients stratified by serum NMI of 198.5 pg/mL. (e) Area under the receiver operating characteristic curve (AUROC) of serum NMI, Chronic Liver Failure Consortium ACLF score (CLIF‐C ACLFs), Model for End‐stage Liver Disease (MELD), combination of serum NMI and CLIF‐C ACLFs (NMI–CLIF‐C ACLFs), or combination of serum NMI and MELD (NMI–MELD) in predicting 3‐month mortality of HBV‐ACLF patients. , NMI‐CLIF‐C ACLFs; , NMI‐MELD; , CLIF‐C ACLFs; , MELD; , NMI; , reference line. ^** P < 0.01; ^*** P < 0.001. [Color figure can be viewed at http://wileyonlinelibrary.com]