A Critical Evaluation of Liver Pathology in Humans with Danon Disease and Experimental Correlates in a Rat Model of LAMP-2 Deficiency
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Danon disease is a genetic deficiency in lysosome-associated membrane protein 2 (LAMP-2), a highly glycosylated constituent of the lysosomal membrane and characterized by a cardiomyopathy, skeletal muscle myopathy, and cognitive impairment. Patients, however, often manifest hepatic abnormalities, but liver function has not been well evaluated and the syndrome is relatively uncommon. Hence, we have taken advantage of a rat that has been deleted of LAMP-2 to study the relative role of LAMP-2 on liver function. Interestingly, rats deficient in LAMP-2 develop a striking increase in serum alkaline phosphatase (ALP) and a decrease in bile flow compared with wild-type littermates. Importantly and by ultrastructural analysis, deficient rats manifest dilated canaliculi that lack microvilli with evidence of bile-containing bodies. Moreover, following bile duct ligation, LAMP-2-deficient rats develop rapid and severe evidence of advanced cholestasis, with an increase in serum bilirubin, as early as 6 h later. In wild-type control rats, multidrug resistance-associated protein 2 (Mrp2) normally concentrates at the bile canalicular membranes to secrete conjugated bilirubin into bile. However, in LAMP-2y/− rats, Mrp2 was detected in hepatocytes compared with other canalicular proteins including P-glycoproteins, dipeptidyl peptidase IV (CD26), and aminopeptidase (CD13). Our data further suggest that LAMP-2 interacts with the membrane cytoskeletal proteins radixin and F-actin in determining the localization of integral membrane proteins.
KeywordsLysosome-associated membrane protein 2 Intrahepatic cholestasis Multidrug resistance-associated protein 2
Bile duct ligation
Bile salt export pump
Glyceraldehyde 3-phosphate dehydrogenase
Lysosome-associated membrane protein 2
Multidrug resistance-associated protein 2
Real-time polymerase chain reaction
Transcription activator-like effector nuclease
We thank Wanjun Chen (Mucosal Immunology Section, National Institutes of Health) for the technical guidance during the experiment and reviewing the manuscript. We thank Professor Stieger (Department of Clinical Pharmacology and Toxicology, University Hospital, Zürich, Switzerland) for the gift of anti-BSEP antibodies. The study was funded by the National Natural Science Foundation of China (Nos. 81600451, 81470835, and 81370519), National Science and Technology Major Projects for New Drugs Innovation and Development (Nos. 2014ZX09508002-001 and 2014ZX09104002-003), and Natural Science Foundation of Shaanxi Province (Nos. 2013HM-01 and 2011KTCL03-09).
YH and EG conceived and designed the experiments and wrote the manuscript; LW performed the experiments and wrote the manuscript; JBW and WLC assisted in some experiments and revised the manuscript; YQS, XMZ, GYG, and CCG assisted in some experiments; LW, XFH, and ZYH collected samples and interpreted the data; and SZ, SYM, XZ, and DMF collected samples.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
This article contains studies with animals. All animal experimental protocols were approved by our local animal care and use committee according to criteria outlined in Guide for the Care and Use of Laboratory Animals from the National Academy of Sciences (National Institutes of Health publication 86-23, 1985 revision).
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