A Cholecystokinin Receptor Antagonist Halts Nonalcoholic Steatohepatitis and Prevents Hepatocellular Carcinoma
Background and Aims
Nonalcoholic steatohepatitis (NASH) is a common inflammatory liver condition that may lead to cirrhosis and hepatocellular carcinoma (HCC). Risk factors for NASH include a saturated fat diet, altered lipid metabolism, and genetic and epigenetic factors, including microRNAs. Serum levels of cholecystokinin (CCK) are elevated in mice and humans that consume a high-saturated fat diet. CCK receptors (CCK-Rs) have been reported on fibroblasts which when activated can induce fibrosis; however, their role in hepatic fibrosis remains unknown. We hypothesized that elevated levels of CCK acting on the CCK-Rs play a role in the development of NASH and in NASH-associated HCC.
We performed a NASH Prevention study and Reversal study in mice fed a saturated fat 75% choline-deficient–ethionine-supplemented (CDE) diet for 12 or 18 weeks. In each study, half of the mice received untreated drinking water, while the other half received water supplemented with the CCK-R antagonist proglumide. CCK-R expression was evaluated in mouse liver and murine HCC cells.
CCK receptor antagonist treatment not only prevented NASH but also reversed hepatic inflammation, fibrosis, and steatosis and normalized hepatic transaminases after NASH was established. Thirty-five percent of the mice on the CDE diet developed HCC compared with none in the proglumide-treated group. We found that CCK-BR expression was markedly upregulated in mouse CDE liver and HCC cells compared with normal hepatic parenchymal cells, and this expression was epigenetically regulated by microRNA-148a.
These results support the novel role of CCK receptors in the pathogenesis of NASH and HCC.
KeywordsHepatocellular carcinoma Nonalcoholic steatohepatitis Cholecystokinin Fibrosis Receptors
Serum aspartate aminotransferase
Choline-deficient–ethionine-supplemented diet and regular untreated water
Choline-deficient–ethionine-supplemented diet and proglumide-treated water
Control diet and regular untreated water
Control diet and proglumide-treated water
Drug-induced liver injury
Nonalcoholic fatty liver disease
Polymerase chain reaction
Pioglitazone, vitamin E, or placebo for nonalcoholic steatohepatitis
Quantitative real-time polymerase chain reaction
We appreciate the staff in the Lombardi Georgetown Histology Core facility and technical assistance of the Department of Comparative Medicine.
JPS conceived and designed the study. RT, VC, SN, MG, HC, MH, AS, NS, BK, and JPS carried out acquisition of data. RT, JPS, HC, AHKK, MG, and AJK analyzed and interpreted the data. JPS and RT drafted the manuscript. All authors critically revised and approved the manuscript. HC, MG, MH, NS, and JPS carried out statistical analysis. JPS obtained funding.
The authors were supported by the AGA Elsevier Pilot Research Award and also supported by the Ruesch Center for the Cure of Gastrointestinal Cancers, Georgetown University Medical Center. Postdoctoral support was provided by a NIH training grant to Martha Gay TL1TR001431. These studies were conducted in part at the Lombardi Comprehensive Cancer Center Histopathology & Tissue Shared resource and in the Preclinical Imaging Research Laboratory which is supported in part by NIH/NCI Grant P30-CA051008.
Compliance with Ethical Standards
Conflict of interest
Jill P. Smith has intellectual property on the use of CCK receptor antagonists in NASH. Sandeep Nadella and Jill P. Smith are co-inventors on a pending patent for proglumide and fibrosis. None of the other authors have conflicts to report.
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