Understanding the mechanisms by which reactive oxygen species modulate both the expression and the alternative splicing of transcription factors such as KLF6, a tumor suppressor gene mutated in several carcinomas and likely involved in the development of alcoholic liver disease, is of great relevance for potential therapeutic intervention, and this constitutes the major goal of this application. We hypothesize that reactive oxygen species in general and reactive oxygen species derived from cytochrome P450 2E1 in particular may play a critical role in regulating KLF6 splicing and its biological actions. We propose: 1) To explore the significance of CYP2E1-derived reactive oxygen species modulation of KLF6 expression and alternative splicing in CYP2E1-expressing cells and in primary hepatocytes isolated from chronic alcohol fed mice. We plan to analyze the effects of prooxidants and of glutathione depletion and to analyze the contribution of each KLF6 splice isoform to the effects mediated by the prooxidants. 2) To establish the relevance in vivo of the spliced isoforms KLF6_V1 and KLF6_V2 in alcoholic liver disease using the following models: a) wild-type mice infected with lentivirus overexpressing either the full-length KLF6 or any of the spliced variants or the corresponding siRNAs before and after inducing liver injury by ethanol feeding;b) Sod1-/- mice whose compromised antioxidant defense is hoped to increase the responsiveness to reactive oxygen species in term of modulating KLF6 splicing and activity;and c) Ktf6+/- in which the potential heterozygosity may modify the responsiveness to the alcohol effects. 3) To evaluate whether stress-activated kinases modulate the up-regulation and the alternative splicing of KLF6 under reactive oxygen species production in CYP2E1-expresing cells and in hepatocytes from chronic ethanol-fed mice. Mechanistic studies will follow to understand the impact of oxidant stress on KLF6 expression, alternative splicing, and downstream targets such as TNFalpha, TGFbeta, iNOS, catalase, and NFkB. We hope that these models and the approach taken will help us to define how reactive oxygen species can modulate splicing and particularly the splicing of KLF6 and the actions of each isoform to dissect potential therapeutic targets for preventing liver disease.

National Institute of Health (NIH)
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Exploratory Grants (P20)
Project #
Application #
Study Section
Special Emphasis Panel (ZAA1)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Icahn School of Medicine at Mount Sinai
New York
United States
Zip Code
Ge, Xiaodong; Arriazu, Elena; Magdaleno, Fernando et al. (2018) High Mobility Group Box-1 Drives Fibrosis Progression Signaling via the Receptor for Advanced Glycation End Products in Mice. Hepatology 68:2380-2404
Magdaleno, Fernando; Blajszczak, Chuck C; Nieto, Natalia (2017) Key Events Participating in the Pathogenesis of  Alcoholic Liver Disease. Biomolecules 7:
Arriazu, Elena; Ge, Xiaodong; Leung, Tung-Ming et al. (2017) Signalling via the osteopontin and high mobility group box-1 axis drives the fibrogenic response to liver injury. Gut 66:1123-1137
Laitman, Benjamin M; Asp, Linnéa; Mariani, John N et al. (2016) The Transcriptional Activator Krüppel-like Factor-6 Is Required for CNS Myelination. PLoS Biol 14:e1002467
Renault, Thibaud T; Luna-Vargas, Mark P A; Chipuk, Jerry E (2016) Mouse Liver Mitochondria Isolation, Size Fractionation, and Real-time MOMP Measurement. Bio Protoc 6:
Kocabayoglu, Peri; Zhang, David Y; Kojima, Kensuke et al. (2016) Induction and contribution of beta platelet-derived growth factor signalling by hepatic stellate cells to liver regeneration after partial hepatectomy in mice. Liver Int 36:874-82
Magdaleno, Fernando; Arriazu, Elena; Ruiz de Galarreta, Marina et al. (2016) Cartilage oligomeric matrix protein participates in the pathogenesis of liver fibrosis. J Hepatol 65:963-971
Kocabayoglu, Peri; Lade, Abigale; Lee, Youngmin A et al. (2015) ?-PDGF receptor expressed by hepatic stellate cells regulates fibrosis in murine liver injury, but not carcinogenesis. J Hepatol 63:141-7
Renault, Thibaud T; Floros, Konstantinos V; Elkholi, Rana et al. (2015) Mitochondrial shape governs BAX-induced membrane permeabilization and apoptosis. Mol Cell 57:69-82
Hasegawa, Daisuke; Calvo, Veronica; Avivar-Valderas, Alvaro et al. (2015) Epithelial Xbp1 is required for cellular proliferation and differentiation during mammary gland development. Mol Cell Biol 35:1543-56

Showing the most recent 10 out of 90 publications