Acetaminophen-induced reduction of NIMA-related kinase 7 expression exacerbates acute liver injury

Zhenzhen Sun; Qian Wang; Le Sun; Mengying Wu; Shuzhen Li; Hu Hua; Ying Sun; Tong Ni; Chunlei Zhou; Songming Huang; Aihua Zhang; Yue Zhang; Zhanjun Jia

doi:10.1016/j.jhepr.2022.100545

Acetaminophen-induced reduction of NIMA-related kinase 7 expression exacerbates acute liver injury

JHEP Rep. 2022 Jul 20;4(10):100545. doi: 10.1016/j.jhepr.2022.100545. eCollection 2022 Oct.

Authors

Zhenzhen Sun^{1

2

3}, Qian Wang^{1

2

3}, Le Sun^{1

2

3}, Mengying Wu^{1

2

3}, Shuzhen Li^{1

2

3}, Hu Hua^{1

2

3}, Ying Sun⁴, Tong Ni^{1

2

3}, Chunlei Zhou⁵, Songming Huang^{1

2

3}, Aihua Zhang^{1

2

3}, Yue Zhang^{1

2

3}, Zhanjun Jia^{1

2

3}

Affiliations

¹ Nanjing Key Laboratory of Pediatrics, Children's Hospital of Nanjing Medical University, Nanjing, China.
² Department of Nephrology, Children's Hospital of Nanjing Medical University, Nanjing, China.
³ Jiangsu Key Laboratory of Pediatrics, Nanjing Medical University, Nanjing, China.
⁴ Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, Jiangsu, China.
⁵ Department of Pathology, Children's Hospital of Nanjing Medical University, Nanjing, China.

Abstract

Background & aims: Acetaminophen (APAP)-induced acute liver injury (ALI) is a global health issue characterised by an incomplete understanding of its pathogenesis and unsatisfactory therapies. NEK7 plays critical roles in both cell cycle regulation and inflammation. In the present study, we investigated the role and mechanism of NEK7 in APAP-induced ALI.

Methods: In mice with NEK7 overexpression (hydrodynamic tail vein injection of NEK7 plasmids), hepatocyte-specific NEK7 knockout (cKO), and inducible NEK7 knockout (iKO), an overdose of APAP was administered to induce ALI. Liver injury was determined by an analysis of serum liver enzymes, pathological changes, inflammatory cytokines, and metabonomic profiles. In vitro, hepatocyte damage was evaluated by an analysis of cell viability, the reactive oxygen species levels, and mitochondrial function in different cell lines. Hepatocyte proliferation and the cell cycle status were determined by Ki-67 staining, EdU staining, and the cyclin levels.

Results: NEK7 was markedly downregulated in APAP-induced injured liver and damaged hepatocytes. NEK7 overexpression in the liver significantly alleviated APAP-induced liver injury, as shown by the restored liver function, reduced pathological injury, and decreased inflammation and oxidative stress, which was confirmed in a hepatocyte cell line. Moreover, both NEK7 cKO and iKO mice exhibited exacerbation of APAP-induced ALI. Finally, we determined that cyclin B1-mediated cell cycle progression could mediate the protective effect of NEK7 against APAP-induced ALI.

Conclusions: Reduced NEK7 contributes to APAP-induced ALI, possibly by dysregulating cyclins and disturbing cell cycle progression.

Lay summary: Acetaminophen-induced acute liver injury is one of the major global health issues, owing to its high incidence, potential severity, and limited therapeutic options. Our current understanding of its pathogenesis is incomplete. Herein, we have shown that reduced NEK7 (a protein with a key role in the cell cycle) exacerbates acetaminophen-induced acute liver injury. Hence, NEK7 could be a possible therapeutic target for the prevention or treatment of this condition.

Keywords: ALF, acute liver failure; ALI, acute liver injury; ALT, alanine aminotransferase; APAP, acetaminophen; AST, aspartate aminotransferase; Apoptosis; CCK-8, cell counting kit 8; CYP2E1; Ctrl, control; GSH, glutathione; Het, heterozygous; LDH, lactate dehydrogenase; MCP-1, monocyte chemoattractant protein-1; MDA, malondialdehyde; NAPQI, N-acetyl-p-benzoquinone imine; ND1, NADH dehydrogenase 1; NEK7, NIMA-related kinase 7; NLRP3; PI, propidium iodide; ROS, reactive oxygen species; TNF-α, tumour necrosis factor-alpha; TPM, transcripts per kilobase of exon model per million mapped reads; WT, wild-type; cKO mice, conditional knockout mice; iKO mice, inducible knockout mice; si, small interfering.