Genetic and Functional Evaluation of the Role of FOXO1 in Antituberculosis Drug-Induced Hepatotoxicity.

Jingwei Zhang,Tao Wu,Jiajia Song,Hao Bai,Tangyuheng Liu,Binwu Ying,Lin Jiao,Xuejiao Hu,Zhenzhen Zhao,Teh Lay Kek

doi:10.1155/2021/3185874

Abstract

Background The accumulation of the hepatotoxic substance protoporphyrin IX (PPIX) induced by aminolevulinate synthase 1 (ALAS1) activation is one of the important mechanisms of antituberculosis drug-induced hepatotoxicity (ATDH). Forkhead box protein O1 (FOXO1) may activate ALAS1 transcription. However, little is known about their roles in ATDH; we performed a study to determine the association between polymorphisms in the two genes and ATDH susceptibility. Then, we verified this possible association by cellular functional experiments. Materials and Methods Tag single-nucleotide polymorphisms (TagSNPs) in the two genes were genotyped in 746 tuberculosis patients. The frequencies of the alleles, genotypes, genetic models, and haplotype distribution of the variants were compared between the case and control groups. L-02 cells and HepG2 cells were incubated with the indicated concentration of isoniazid (INH) and rifampicin (RIF) for the desired times, and then the expression levels of ALAS1 and FOXO1 mRNAs and proteins were detected. HepG2 cells were transiently transfected with FOXO1 siRNA to observe the effect of changes in the FOXO1 expression on the cell survival rate and ALAS1 expression. Results The C allele at rs2755237 and the T allele at rs4435111 in the FOXO1 gene were associated with a decreased risk of ATDH. The expression of ALAS1 in both L-02 cells and HepG2 cells was increased by the coadministration of INH/RIF (600/200 μM) for 24 h. Although FOXO1 expression was reduced slightly by the same treatment, its content in the nucleus was significantly increased. However, the cell survival rate and ALAS1 expression level were not significantly altered by the downregulation of FOXO1 in HepG2 cells. Conclusions Variants of the rs4435111 and rs2755237 loci in the FOXO1 gene were associated with susceptibility to ATDH. Coadministration of INH/RIF promoted the transfer of FOXO1 from the cytoplasm to the nucleus, but the functional significance of its nuclear translocation requires further verification.

Highlights

Jingwei Zhang,1 Lin Jiao,2 Jiajia Song,2 Tao Wu,2 Hao Bai,2 Tangyuheng Liu,2 Zhenzhen Zhao,2 Xuejiao Hu,2 and Binwu Ying 2
Results. e C allele at rs2755237 and the T allele at rs4435111 in the Forkhead box protein O1 (FOXO1) gene were associated with a decreased risk of antituberculosis drug-induced hepatotoxicity (ATDH). e expression of aminolevulinate synthase 1 (ALAS1) in both L-02 cells and HepG2 cells was increased by the coadministration of INH/RIF (600/200 μM) for 24 h
Ethical approval for this study was obtained from the Institutional Review Board of the West China Hospital of Sichuan University. e definition of drug-induced hepatotoxicity used in this study was based on the National Institutes of Health and Common Toxicity Criteria for Adverse Events v5.0 (CTCAE v5.0) [24, 25, 27]. e definition of the severity of hepatotoxicity was based on the WHO Adverse Drug Reaction Terminology: mild liver injury, ALT level

Summary

Introduction

Jingwei Zhang ,1 Lin Jiao ,2 Jiajia Song ,2 Tao Wu ,2 Hao Bai ,2 Tangyuheng Liu ,2 Zhenzhen Zhao, Xuejiao Hu ,2 and Binwu Ying 2. E efficiency of this protocol is high as 85%, but the adverse reactions caused by coadministration of INH and RIF cannot be ignored Among these adverse reactions, antituberculosis drug-induced hepatotoxicity (ATDH) is the most common and serious, with an incidence of 5.0–28.0% and mortality of 0.042–0.07‰ [2,3,4]. E PHARMGKB database recommends important NAT2 genetic variants as clinical markers for predicting the risk of ATDH caused by INH in patients with tuberculosis [8]. PPIX accumulation caused by PXR/ALAS1 axis activation induced by the coadministration of INH/RIF was proposed to be one of the important mechanisms of ATDH. It has milestone significance in the research history of the ATDH mechanism [9, 10]. The relative lack of haem caused by the increased demand for cytochrome enzyme synthesis will activate ALAS transcription; excessive haem directly reduces ALAS1 mRNA stability and inhibits ALAS1 mRNA transcription

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