Protein aggregation occurs in association with a variety of liver diseases and is typically cytoplasmic. A common form of protein aggregates are Mallory-Denk bodies (MDBs) that are found in association with steatohepatitis, viral hepatitis and other forms of liver disease. Recent findings indicate that the presence of MDBs provides an independent poor clinical and histological outcome in patients with hepatitis C. The propensity to form MDBs is linked to the genetic background as supported by higher predisposition to their formation in specific mouse (C57BL >C3H strain) and human (Hispanic >White) genotypes. Among the molecular alterations that positively associate with experimental MDB formation are hypersumoylation and hypoacetylation of keratins (the major protein constituents of MDBs). Also, the overall liver sumoylation is impaired in the C57BL MDB-susceptible strain. Another potential genetic modifier we identified is the ecto-5'- nucleotidase, CD73, whereby a novel and hitherto unknown spliced variant is modulated differently in the MDB formation-prone versus formation-resistant strains. Aside from the cytoplasmic MDBs, a novel type of nuclear protein aggregation that we recently observed in cell culture, in mouse liver injury models undergoing oxidative stress, and in some human chronic liver disease explants involves the nuclear lamins. Formation of lamin- containing aggregates is rapid and is noted within days of exposure to the insult;and is observed prior to MDB formation, which requires weeks. Lamin aggregation could interfere with one or more nuclear functions, and its rapid onset may serve as an early marker of specific types of oxidative injury. Our hypothesis is that genetic variations unmask differences in hepatocyte responses to oxidative injury which manifest as nuclear and cytoplasmic protein aggregation. This hypothesis will be tested using 3 specific aims: (i) Characterize the role of CD73 in strain-selective MDB formation and liver injury;(ii) Characterize keratin acetylation and its relationship to keratin sumoylation and aggregation during oxidative stress;and (iii) Define the biochemical and functional alterations that associate with nuclear lamin disorganization in response to liver injury. Completion of our aims should provide fundamental knowledge regarding genetic modifiers and posttranslational regulation involved in MDB formation. In addition, the exciting finding of a novel CD73 isoform and its genetic background-related differences in response to liver injury may unfold a new paradigm for the role of CD73 in liver and other cell responses to injury. Furthermore, the newly appreciated aggregation of nuclear lamins in response to liver injury provides a potential novel mechanism for direct nuclear involvement that may protect from or promote liver injury, with consequent possible therapeutic, prognostic and diagnostic targets and approaches.

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The overarching goal of this proposal is to identify genetic influences on the predisposition to protein aggregation in response to oxidative liver injury. Understanding the role of genetic factors is highly beneficial and relevant since it provides potential diagnostic, prognostic and therapeutic targets in human liver disease, in addition to providing possible molecular explanations to the observed differences in susceptibility to a range of liver diseases in humans.

National Institute of Health (NIH)
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
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Hepatobiliary Pathophysiology Study Section (HBPP)
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Serrano, Jose
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University of Michigan Ann Arbor
Schools of Medicine
Ann Arbor
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Snider, Natasha T; Altshuler, Peter J; Omary, M Bishr (2015) Modulation of cytoskeletal dynamics by mammalian nucleoside diphosphate kinase (NDPK) proteins. Naunyn Schmiedebergs Arch Pharmacol 388:189-97
Snider, Natasha T; Park, Haewon; Omary, M Bishr (2013) A conserved rod domain phosphotyrosine that is targeted by the phosphatase PTP1B promotes keratin 8 protein insolubility and filament organization. J Biol Chem 288:31329-37
Snider, Natasha T; Griggs, Nicholas W; Singla, Amika et al. (2013) CD73 (ecto-5'-nucleotidase) hepatocyte levels differ across mouse strains and contribute to mallory-denk body formation. Hepatology 58:1790-800
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Snider, Natasha T; Leonard, Jessica M; Kwan, Raymond et al. (2013) Glucose and SIRT2 reciprocally mediate the regulation of keratin 8 by lysine acetylation. J Cell Biol 200:241-7
Kwan, Raymond; Hanada, Shinichiro; Harada, Masaru et al. (2012) Keratin 8 phosphorylation regulates its transamidation and hepatocyte Mallory-Denk body formation. FASEB J 26:2318-26
Singla, Amika; Moons, David S; Snider, Natasha T et al. (2012) Oxidative stress, Nrf2 and keratin up-regulation associate with Mallory-Denk body formation in mouse erythropoietic protoporphyria. Hepatology 56:322-31
Snider, Natasha T; Weerasinghe, Sujith V W; Singla, Amika et al. (2011) Energy determinants GAPDH and NDPK act as genetic modifiers for hepatocyte inclusion formation. J Cell Biol 195:217-29
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