Integration of transcriptomics, proteomics and metabolomics identifies biomarkers for pulmonary injury by polyhexamethylene guanidine phosphate (PHMG-p), a humidifier disinfectant, in rats.
Jung Dae LeeHyang Yeon KimKeunsoo KangHye Gwang JeongMi-Kyung SongIn Hwan TaeSu Hyun LeeHae Ri KimKyuhong LeeSehyun ChaeDaehee HwangSuhkmann KimHyung Sik KimKyu-Bong KimByung-Mu LeePublished in: Archives of toxicology (2020)
Polyhexamethylene guanidine phosphate (PHMG-p) was used as a humidifier disinfectant in Korea. PHMG induced severe pulmonary fibrosis in Koreans. The objective of this study was to elucidate mechanism of pulmonary toxicity caused by PHMG-p in rats using multi-omics analysis. Wistar rats were intratracheally instilled with PHMG-p by single (1.5 mg/kg) administration or 4-week (0.1 mg/kg, 2 times/week) repeated administration. Histopathologic examination was performed with hematoxylin and eosin staining. Alveolar macrophage aggregation and granulomatous inflammation were observed in rats treated with single dose of PHMG-p. Pulmonary fibrosis, chronic inflammation, bronchiol-alveolar fibrosis, and metaplasia of squamous cell were observed in repeated dose group. Next generation sequencing (NGS) was performed for transcriptome profiling after mRNA isolation from bronchiol-alveoli. Bronchiol-alveoli proteomic profiling was performed using an Orbitrap Q-exactive mass spectrometer. Serum and urinary metabolites were determined using 1H-NMR. Among 418 differentially expressed genes (DEGs) and 67 differentially expressed proteins (DEPs), changes of 16 mRNA levels were significantly correlated with changes of their protein levels in both single and repeated dose groups. Remarkable biological processes represented by both DEGs and DEPs were defense response, inflammatory response, response to stress, and immune response. Arginase 1 (Arg1) and lipocalin 2 (Lcn2) were identified to be major regulators for PHMG-p-induced pulmonary toxicity based on merged analysis using DEGs and DEPs. In metabolomics study, 52 metabolites (VIP > 0.5) were determined in serum and urine of single and repeated-dose groups. Glutamate and choline were selected as major metabolites. They were found to be major factors affecting inflammatory response in association with DEGs and DEPs. Arg1 and Lcn2 were suggested to be major gene and protein related to pulmonary damage by PHMG-p while serum or urinary glutamate and choline were endogenous metabolites related to pulmonary damage by PHMG-p.
Keyphrases
- oxidative stress
- inflammatory response
- pulmonary hypertension
- pulmonary fibrosis
- single cell
- mass spectrometry
- ms ms
- genome wide
- diabetic rats
- immune response
- high resolution
- drug induced
- magnetic resonance
- high glucose
- gene expression
- binding protein
- adipose tissue
- lps induced
- dna methylation
- randomized controlled trial
- copy number
- early onset
- dendritic cells
- nitric oxide
- clinical trial
- genome wide identification
- liver fibrosis
- idiopathic pulmonary fibrosis
- heat stress
- cell free