Abstract
The insulin receptor (INSR) has been shown to be hyperactive in hepatic stellate cells (HSCs) in humans and rodents with liver fibrosis. To explore HSC cellular mechanisms that INSR regulates during pro-fibrotic stimulation, we used CRISPR-Cas9 technology. We knocked out a portion of the INSR gene in human LX2 HSC cells (INSRe5- 8 KO) that regulates insulin responsiveness but not the insulin-like growth factor (IGF) or transforming growth factor-β (TGFβ) signaling. The INSRe5- 8 KO HSCs had significantly higher cell growth, BrdU incorporation, and lower TP53 expression that suppresses growth, and they also exhibited increased migration compared to the Scramble control. We treated the scramble control and INSRe5- 8 KO HSCs with insulin or TGFβ and profiled hundreds of kinase activities using the PamGene PamStation kinome technology. Our analysis showed that serine/threonine kinase (STK) activities were reduced, and most of the protein-tyrosine kinase (PTK) activities were increased in the INSRe5- 8 KO compared to the Scramble control HSCs. To study gene transcripts altered in activated Scramble control and INSRe5- 8 KO HSCs, we treated them with TGFβ for 24 h. We isolated RNA for sequencing and found that the INSRe5- 8 KO cells, compared to control HSCs, had altered transcriptional responsiveness to TGFβ stimulation, collagen-activated signaling, smooth muscle cell differentiation pathways, SMAD protein signaling, collagen metabolic process, integrin-mediated cell adhesion, and notch signaling. This study demonstrates that reduced INSR responsiveness enhances HSC growth and selectively mediates TGFβ- induced HSC activation. These findings provide new insights into the development of more effective treatments for liver fibrosis.
Document Type
Article
Publication Date
3-2025
Digital Object Identifier (DOI)
https://doi.org/10.1096/fj.202402169R
Funding Information
HHS | NIH | NIDDK | Division of Diabetes, Endocrinology, and Metabolic Diseases (DEM), Grant/Award Number: R01DK121797; HHS | NIH | National Institute on Drug Abuse (NIDA), Grant/Award Number: R01DA058933; HHS | NIH | National Heart, Lung, and Blood Institute (NHLBI), Grant/ Award Number: F31HL170972 and F31HL175979; American Heart Association (AHA), Grant/Award Number: 25PRE1374495
Repository Citation
Lee, Wang-Hsin; Bates, Evelyn A.; Kipp, Zachary A.; Pauss, Sally; Martinez, Genesee J.; Blair, Cheavar A.; and Hinds, Terry D. Jr., "Insulin receptor responsiveness governs TGFβ-induced hepatic stellate cell activation: Insulin resistance instigates liver fibrosis" (2025). Markey Cancer Center Faculty Publications. 300.
https://uknowledge.uky.edu/markey_facpub/300
Included in
Biochemistry Commons, Biotechnology Commons, Genetics Commons, Molecular Biology Commons, Oncology Commons
Notes/Citation Information
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. © 2025 The Author(s). The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.