Numerous animal and human studies have demonstrated an important hepatoprotective function of keratin intermediate filament (IF) proteins in several acute and chronic liver diseases. The hepatocyte IF cytoskeleton, which consists of keratin 8 and 18 (K8/K18) heterodimers, undergoes extensive physiological and disease- related reorganization that is mediated by post-translational modifications. Keratin-rich hepatocyte Mallory- Denk bodies (MDBs), and other histological alterations affecting the cytoskeleton, such as hepatocyte ballooning, are prominent features of alcoholic liver disease (ALD), nonalcoholic steatohepatitis (NASH), and other diseases characterized by oxidative injury and metabolic abnormalities. The objective of this application is to understand the regulation, functional significance, and liver disease relevance of keratin sumoylation. Sumoylation is a novel post-translational modification by Small Ubiquitin-like Modifier (SUMO) proteins with important implications to human disease pathogenesis. The central hypothesis of this proposal is that stress- induced keratin sumoylation participates in cross-talk with other post-translational modifications to regulate keratin properties and function during liver injury.
Three specific aims will be pursued: (i) Characterize the regulatory mechanisms behind stress-induced K8/K18 sumoylation;(ii) Determine the functional significance of K8/K18 sumoylation;and (iii) Examine the regulation and significance of K8/K18 sumoylation in metabolic liver disease. The goals that the candidate will achieve through the proposed research and training plan include: learning novel techniques to study protein function and regulation;becoming adept at analysis of human and animal pathology;gaining expertise in cellular metabolic signaling and animal models of metabolic liver disease;and becoming grounded with the skills needed to become a successful independent investigator. The candidate's long-term career goals are to become an expert on intermediate filament proteins and their roles in human diseases, obtain a tenure-track faculty position and become a successful extramurally-funded independent investigator leading a strong research program in digestive disease related research. The research will be carried out at a premier institution (University of Michigan) and will involve primary mentorship from a leader in digestive disease related research;co-mentorship by two experts of metabolic signaling, diabetes, and insulin resistance;and collaborations with three experts of liver pathology, proteomics and post- translational protein regulation. The proposed research will provide mechanistic understanding of keratin regulation during metabolic and oxidative liver injury and may serve as the basis for novel approaches to treat common liver diseases, like ALD and NASH, where pharmacologic interventions are critically needed. Importantly, these studies will create new opportunities for investigation that the candidate will pursue during her independent research career stage.
There are over 80 human diseases that are caused or worsened by mutations in genes that encode various members of the intermediate filament family of proteins. This proposal specifically addresses the regulation and significance of keratin intermediate filaments in metabolically-induced liver disease. The results will provide insights into potential novel treatments for common liver diseases, such as those caused by obesity or alcohol consumption.
|Snider, Natasha T; Portney, Daniel A; Willcockson, Helen H et al. (2016) Ethanol and Acetaminophen Synergistically Induce Hepatic Aggregation and TCH346-Insensitive Nuclear Translocation of GAPDH. PLoS One 11:e0160982|
|Snider, Natasha T; Omary, M Bishr (2016) Assays for Posttranslational Modifications of Intermediate Filament Proteins. Methods Enzymol 568:113-38|
|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|
|Kwan, Raymond; Chen, Lu; Looi, Koksun et al. (2015) PKC412 normalizes mutation-related keratin filament disruption and hepatic injury in mice by promoting keratin-myosin binding. Hepatology 62:1858-69|
|Snider, Natasha T; Altshuler, Peter J; Wan, Shanshan et al. (2014) Alternative splicing of human NT5E in cirrhosis and hepatocellular carcinoma produces a negative regulator of ecto-5'-nucleotidase (CD73). Mol Biol Cell 25:4024-33|
|Snider, Natasha T; Omary, M Bishr (2014) Post-translational modifications of intermediate filament proteins: mechanisms and functions. Nat Rev Mol Cell Biol 15:163-77|
|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|
|Singla, Amika; Griggs, Nicholas W; Kwan, Raymond et al. (2013) Lamin aggregation is an early sensor of porphyria-induced liver injury. J Cell Sci 126:3105-12|
|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|