RNA sequencing of LX-2 cells treated with TGF-β1 identifies genes associated with hepatic stellate cell activation

Abstract

Background Hepatic stellate cells (HSCs) are liver-resident myofibroblast precursors responsible for the production of collagen and maintenance of the hepatic extracellular matrix (ECM). As such, they are generally associated with fibrotic liver diseases. HSCs become “activated” in response to tissue damage or pathogen invasion, a process most commonly driven by transforming growth factor-β1 (TGF-β1). Despite this, the full extent of TGF-β1 signalling in these cells is poorly understood. Clarifying the range and diversity of this signalling will further improve our understanding of the process of HSC activation. Methods and results RNA sequencing was used to quantitate the transcriptomic changes induced in LX-2 cells, an activated human HSC line, following TGF-b1 treatment. In total, 5,258 genes were found to be significantly differentially expressed with a false discovery rate cut-off of < 0.1. The topmost deregulated of these genes included those with no currently characterised role in either HSC activation or fibrotic processes, including CIITA and SERPINB2. In silico analysis revealed the prominent signalling pathways downstream of TGF-β1 in LX-2 cells. Conclusions In this study, we describe the genes and signalling pathways significantly deregulated in LX-2 cells following TGF-β1 treatment. We identified several highly deregulated genes with no currently characterised role in HSC activation, which may represent novel mediators of fibrotic responses in HSCs or the liver macroenvironment. This work may be of use in the identification of new markers of liver fibrosis and could provide insight into prospective genes or pathways that might be targeted for the amelioration of fibrotic liver disease in the future.

FOXS1 is increased in liver fibrosis and regulates TGFβ responsiveness and proliferation pathways in human hepatic stellate cells

“…Carson et al. showed that TGFβ treatment in human LX2 HSCs increased several genes as they measured by RNA-seq, and one of their top-induced genes was FOXS1 (56.49-fold) with the TGFβ treatments ( 25 ). Our study is the first to determine the role of FOXS1 in the TGFβ activation of HSCs, its signaling mechanisms, and the kinase pathways it controls.…”

“…However, we did not observe that FOXS1 is increased in patients with HCC. Others have shown that TGFβ induces FOXS1 expression ( 11 , 25 ). Carson et al.…”

Formation and Investigation of Cell‐Derived Nanovesicles as Potential Therapeutics against Chronic Liver Disease

“…[42] To generate perpetuated, highly fibrogenic LX-2 cells, TGF-𝛽 1 was employed, as previously described. [35,43] Afterward, cdNVs were produced by subjecting differently pretreated LX-2 cells (antifibrotic/fibrotic) to serial extrusion. cdNVs were then purified using a combination of density-gradient ultracentrifugation steps, followed by standard ultracentrifugation.…”