Cytosolic accumulation of triglyceride (TG)-rich lipid droplets (LDs) is a hallmark of nonalcoholic fatty liver disease (NAFLD). Many LD-associated proteins are implicated in the pathogenesis of human and rodent NAFLD, including CGI-58 (Comparative Gene Identification-58). Mutations in human CGI-58 gene cause ichthyosis (thickened dry scaly skin) and TG-rich LD accumulation in most cell types. Patients display NAFLD ranging from simple steatosis to nonalcoholic steatohepatitis (NASH), fibrosis and cirrhosis. CGI-58 was shown to promote TG hydrolysis in vitro by activating Adipose Triglyceride Lipase (ATGL), but ATGL mutations in humans cause no ichthyosis. While liver-specific CGI-58 knockout (LivKO) mice develop severe hepatic steatosis, NASH and fibrosis even on a regular chow diet, liver-specific ATGL KO mice display only mild hepatic steatosis without NASH and fibrosis. These phenotypic differences between ATGL and CGI-58 mutations provide us a unique opportunity to exploit molecular mechanisms of NAFLD progression. Two major pathways are implicated in intracellular LD breakdown: 1) Cytosolic/Neutral Lipolysis mainly mediated by ATGL, and 2) Lysosomal/Acidic Lipolysis mediated by a lipid-specific macroautophagy (lipophagy). Lipophagy brings cytosolic LDs to lysosomes for degradation by acidic lipases, but its molecular details are unknown. Our preliminary data suggest that hepatic autophagy-related proteins are reduced, the major upstream autophagy inhibitory signaling pathway (mTORC1) is activated, and there is a defect in association between LDs and acidic organelles in CGI-58-deficient liver and/or hepatocytes. It was recently shown that perilipin 2, a major LD coat protein known to interact with CGI-58, also binds the heat shock cognate protein of 70 kDa (hsc70) for degradation via chaperone-mediated autophagy (CMA), and inhibition of CMA reduces both Neutral and Acidic Lipolysis, leading to severe hepatic steatosis and liver damage. Based on these observations and our preliminary data, we hypothesize that CGI-58 may coordinate with hsc70 to stimulate LD uncoating by promoting perilipin 2 degradation via CMA, thereby activating both ATGL lipolysis and lipophagy. Lipophagy failure may in turn inhibit autophagy by altering cellular energy balance. This hypothesis may explain why ATGL and CGI-58 mutations cause overlapping yet distinct phenotypes. We will test this central hypothesis by comparing liver-specific CGI-58 and ATGL KO mice to determine whether liver CGI-58 and ATGL differentially, regulates lipophagy and autophagy. We will then perform detailed protein-protein interaction studies to examine whether CGI-58 and hsc70 coordinate CMA-mediated LD uncoating (degradation of perilipin 2). Finally we will determine if mTORC1 inhibition and autophagy induction protect against NAFLD progression in CGI-58 LivKO mice. Given that CGI-58 resides at the critical crossroad of cellular fat breakdown, this project holds promise of revealing general mechanisms for progression of NAFLD, a major public health problem.

Public Health Relevance

Nonalcoholic fatty liver disease (NAFLD) is a major human disease with limited treatment options. Mutations of CGI-58, a gene critically implicated in cellular fat breakdown, cause advanced stages of NAFLD in humans and animals, but the underlying mechanisms remain unknown. We propose to molecularly define these mechanisms, and thus our work is directly relevant to human health.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
7R01DK111052-02
Application #
9335630
Study Section
Hepatobiliary Pathophysiology Study Section (HBPP)
Program Officer
Doo, Edward
Project Start
2016-07-18
Project End
2020-06-30
Budget Start
2017-01-01
Budget End
2017-06-30
Support Year
2
Fiscal Year
2016
Total Cost
Indirect Cost
Name
Georgia State University
Department
Miscellaneous
Type
Organized Research Units
DUNS #
837322494
City
Atlanta
State
GA
Country
United States
Zip Code
30302
Shin, Hyunsu; Ma, Yinyan; Chanturiya, Tatyana et al. (2017) Lipolysis in Brown Adipocytes Is Not Essential for Cold-Induced Thermogenesis in Mice. Cell Metab 26:764-777.e5