Regulation of transforming growth factor-beta1 (TGF-β1)-induced pro-fibrotic activities by circadian clock gene BMAL1.

Chunmin Dong,Cecilia G Sanchez,Fayong Luo,Rafael Gongora,Meredith L Sosulski

doi:10.1186/s12931-016-0320-0

Chunmin Dong, Cecilia G Sanchez + Show 3 more

Open Access

https://doi.org/10.1186/s12931-016-0320-0

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Journal: Respiratory research	Publication Date: Jan 12, 2016
Citations: 133	License type: cc-by

Affiliation: Tulane University

Abstract

BackgroundBMAL1 is a transcriptional activator of the molecular clock feedback network. Besides its role in generating circadian rhythms, it has also been shown to be involved in the modulation of cell proliferation, autophagy and cancer cell invasion. However, the role of BMAL1 in pulmonary fibrogenesis is still largely unknown. In this study, we investigated the crosstalk between BMAL1 and the signaling transduction and cellular activities of TGF-β1, a key player in lung fibrogenesis.MethodsLungs from wild type and TGF-β1-adenovirus-infected mice were harvested and homogenized for isolation of RNA and protein. RT-PCR and Western Blotting were employed to measure the expression level of clock genes and TGF-β1-induced downstream target genes. siRNA against human BMAL1 gene was transfected by using lipofectamine RNAiMAX to knockdown the endogenous BMAL1 in both lung epithelial cells and fibroblasts.ResultsOur results showed that TGF-β1 is able to up-regulate BMAL1 expression in both lung epithelial cells and normal lung fibroblasts. In animal models of pulmonary fibrosis, BMAL1 expression was also significantly higher in adenovirus-TGF-β1-infected mice than in the control group. Interestingly, BMAL1 was mostly found in a deacetylated form in the presence of TGF-β1. Importantly, siRNA–mediated knockdown of BMAL1 significantly attenuated the canonical TGF-β1 signaling pathway and altered TGF-β1-induced epithelial-mesenchymal transition and MMP9 production in lung epithelial cells. In addition, BMAL1 knockdown inhibited the fibroblast to myofibroblast differentiation of normal human lung fibroblasts.ConclusionsOur results indicate that activation of TGF-β1 promotes the transcriptional induction of BMAL1. Furthermore, BMAL1 is required for the TGF-β1-induced signaling transduction and pro-fibrotic activities in the lung.

Highlights

Brain and muscle Arnt-like protein 1 (BMAL1) is a transcriptional activator of the molecular clock feedback network
Our results indicate that activation of TGF-β1 promotes the transcriptional induction of BMAL1
BMAL1 is required for the TGF-β1-induced signaling transduction and pro-fibrotic activities in the lung

Summary

Introduction

BMAL1 is a transcriptional activator of the molecular clock feedback network. Besides its role in generating circadian rhythms, it has been shown to be involved in the modulation of cell proliferation, autophagy and cancer cell invasion. Idiopathic pulmonary fibrosis is a chronic, progressive, irreversible, and age-related lung disease that has no known cause and few treatment options. This disease was once thought to be a chronic inflammatory process, but current evidence indicates that the fibrotic response is driven by abnormally activated alveolar epithelial cells [1,2,3]. It stimulates the fibroblast to myofibroblast differentiation (FMD), the epithelial to mesenchymal transition (EMT) and the production of matrix metalloproteinases (MMPs) to promote the formation of the fibroblast and myofibroblast foci These foci produce excessive amounts of extracellular matrix, like collagen, resulting in scarring and destruction of the lung architecture [4, 5].

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