Acetaminophen (APAP) toxicity is the leading cause of acute liver failure. The mouse model of APAP hepatotoxicity is highly reproducible and widely utilized to elucidate the mechanisms of liver injury, due to drugs and toxins. Recently, a major role for signal transduction pathways in modulating toxicity downstream of APAP metabolism has been identified using this model. The activation of c-jun-N-terminal kinase (JNK) and its effects on mitochondria have been found to be critical in the signaling pathway leading to APAP-induced necrosis. The goal of this proposal is to elucidate the mechanism of APAP-induced activation of MAPK kinases and the significance, targets, and consequences of the targeting of activated JNK to mitochondria in carrying out the lethal effects of APAP in hepatocytes.
The aims are as follows: (1) Determine the role and significance of binding of P-JNK to specific mitochondrial target protein which has been identified in the pathogenesis of APAP hepatotoxicity: Binding partners will be identified by immunoprecipitation and proteomic approaches and the importance of the mitochondrial JNK target will be determined by adenovirus-shRNA knockdown;the effects of pure activated JNK on mitochondrial function and integrity will be assessed: (2) Determine the signal transduction pathways that lead to both early and sustained activation of JNK in APAP toxicity: the interplay of various upstream pathways such as GSK32, Ras/Raf-1, MLK-3, MEKK-1, PKC1, Akt 1+2, will be examined using antisense oligonucleotide silencing and/or knockout mice to elucidate the initial ASK-1 independent activation of JNK required for subsequent ASK-1 dependent sustained JNK activation;(3) Determine the role of ER stress in APAP toxicity: the plausible contribution of ER stress to the signal transduction pathways leading to mitochondrial collapse will be assessed by exploiting knockout or over expression of the key ER stress sensor, GRP78;(4) Determine the role of RIP kinases in APAP-induced necrosis and the relationship to the GSK/JNK/Sab pathway: RIP may play a key role in signal transduction pathways leading to APAP necrosis which will be assessed using specific inhibitors and antisense silencing of RIP-1. The elucidation of these four aims will lead to an improved understanding of the role and interplay of signal transduction pathways and mitochondria in hepatotoxicity and which will identify new therapeutic targets.

Public Health Relevance

Adverse effects of drugs such as acetaminophen on the liver, which is the leading cause of acute liver failure in the United States, are a major stumbling block in drug development. This proposal is aimed at developing a detailed understanding of the molecular pathways in the liver that lead to injury (killing of liver cells) and identifying new therapeutic targets to prevent injury form acetaminophen or other causes.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK067215-07
Application #
8238296
Study Section
Hepatobiliary Pathophysiology Study Section (HBPP)
Program Officer
Serrano, Jose
Project Start
2004-04-01
Project End
2015-03-31
Budget Start
2012-04-01
Budget End
2013-03-31
Support Year
7
Fiscal Year
2012
Total Cost
$356,700
Indirect Cost
$139,200
Name
University of Southern California
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
072933393
City
Los Angeles
State
CA
Country
United States
Zip Code
90089
Win, Sanda; Than, Tin Aung; Min, Robert Win Maw et al. (2016) c-Jun N-terminal kinase mediates mouse liver injury through a novel Sab (SH3BP5)-dependent pathway leading to inactivation of intramitochondrial Src. Hepatology 63:1987-2003
Suda, Jo; Dara, Lily; Yang, Luoluo et al. (2016) Knockdown of RIPK1 Markedly Exacerbates Murine Immune-Mediated Liver Injury through Massive Apoptosis of Hepatocytes, Independent of Necroptosis and Inhibition of NF-κB. J Immunol 197:3120-3129
Dara, Lily; Liu, Zhang-Xu; Kaplowitz, Neil (2016) Mechanisms of adaptation and progression in idiosyncratic drug induced liver injury, clinical implications. Liver Int 36:158-65
Dara, Lily; Liu, Zhang-Xu; Kaplowitz, Neil (2016) A murder mystery in the liver: who done it and how? J Clin Invest 126:4068-4071
Dara, Lily; Kaplowitz, Neil (2016) The many faces of RIPK3: What about NASH? Hepatology 64:1411-1413
Dara, Lily; Liu, Zhang-Xu; Kaplowitz, Neil (2016) Questions and controversies: the role of necroptosis in liver disease. Cell Death Discov 2:16089
Dara, Lily; Johnson, Heather; Suda, Jo et al. (2015) Receptor interacting protein kinase 1 mediates murine acetaminophen toxicity independent of the necrosome and not through necroptosis. Hepatology 62:1847-57
Baulies, Anna; Ribas, Vicent; Núñez, Susana et al. (2015) Lysosomal Cholesterol Accumulation Sensitizes To Acetaminophen Hepatotoxicity by Impairing Mitophagy. Sci Rep 5:18017
Kaplowitz, Neil; Win, Sanda; Than, Tin Aung et al. (2015) Targeting signal transduction pathways which regulate necrosis in acetaminophen hepatotoxicity. J Hepatol 63:5-7
Win, Sanda; Than, Tin Aung; Le, Bao Han Allison et al. (2015) Sab (Sh3bp5) dependence of JNK mediated inhibition of mitochondrial respiration in palmitic acid induced hepatocyte lipotoxicity. J Hepatol 62:1367-74

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