<i>In vivo</i>antisense oligonucleotide reduction of NPC1 expression as a novel mouse model for Niemann Pick type C- associated liver disease

Rimkunas, Victoria; Graham, Mark; Crooke, Rosanne M.; Liscum, Laura

doi:10.1002/hep.22327

Cited by 33 publications

(56 citation statements)

References 36 publications

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“…To determine how recovery from NPC1 knockdown affected liver tissue histology, H/E-stained tissue sections were examined. Mice lacking NPC1 had accumulation of lipid-laden macrophages in the liver, as shown previously (9)(10)(11)23 ) ( Fig. 4A , green arrows), accompanied by infi ltration of liver parenchyma by immune cells ( Fig.…”

Section: Lipid-laden Cell Infi Ltration Slowly Resolves Post-treatmentsupporting

confidence: 83%

“…Membranes were rinsed four times for 15 min each time in TBS/T, and then probed with a 1:1,000 dilu- kidneys. Liver knockdown of NPC1 caused mice to have hallmarks of NPC1 liver disease, including hepatomegaly, cholesterol storage, accumulation of lipid-laden cells, celldeath, infl ammation, and fi brosis ( 10 ). To determine whether it is possible to recover from NPC1 liver disease, two treatment groups of mice were evaluated.…”

Section: Western Blotting Of Npc1mentioning

confidence: 99%

“…To determine whether it is possible to recover from NPC1 liver disease, two treatment groups of mice were evaluated. One group was subjected to NPC1 knockdown for 9 weeks, representing a time point similar to one at which signifi cant liver disease occurs in the NPC nih mouse model ( 9,10,21 ). The second group of mice was subjected to NPC1 knockdown for 15 weeks to determine the ability of the liver to recover from prolonged NPC1 liver disease.…”

Section: Western Blotting Of Npc1mentioning

confidence: 99%

“…NPC1 liver disease results in hepatocyte apoptosis and an infl ammatory response ( 9,10 ). The trigger for hepatocyte apoptosis in NPC disease is unknown; however, higher levels of liver apoptosis are correlated with increased lysosomal free cholesterol ( 23 ).…”

Section: Cell Death Resolves Along the Same Time Line As Cholesterol mentioning

confidence: 99%

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Recovery from liver disease in a Niemann-Pick type C mouse model

Sayre

Rimkunas

Graham

et al. 2010

Journal of Lipid Research

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This article is available online at http://www.jlr.org clinical cases and NPC2 in 5% of clinical cases. NPC is characterized by storage of free cholesterol and glycosphingolipids in late endosomes and lysosomes. The clinical phenotype that arises, neurodegeneration, hepatosplenomegaly, ataxia, seizures, and dystonia, is similar regardless of whether NPC1 or NPC2 is the cause ( 1 ).Although most NPC patients die due to complications of their neurodegenerative disease, some patients also develop liver disease. Aside from hepatosplenomegaly, NPC patients often suffer from prolonged neonatal jaundice and ascites ( 2 ), as well as liver failure ( 3-7 ). NPC is the second most common cause of neonatal cholestasis ( 7 ), and 10% of these patients die because of liver failure ( 8 ). On the tissue level, livers from NPC-diseased mice exhibit increased hepatocyte apoptosis, infi ltration of foamy macrophages, infl ammation, proliferation of hepatic stellate cells, and fi brosis ( 9-11 ). To better understand liver disease in NPC, our laboratory has developed a mouse model using 2'-O -methoxyethyl modifi ed antisense oligonucleotides (ASO) to block expression of NPC1 specifi cally in the liver ( 10 ).In the present study, we aimed to determine whether recovery is possible in mice with NPC-associated liver disease by using ASOs to knock down hepatic NPC1 expression. After halting treatment for different lengths of time, liver disease was assessed. We hypothesized that extensive liver recovery was possible in NPC1 ASO-treated mice, because the liver has a remarkable capacity for regeneration ( 12 ). Here, we show that substantial reversal of the NPC disease phenotype occurred, including alleviation of hepatomegaly, loss of lipid-laden macrophage accumulations, and decreasing liver cell apoptosis. Much pathology Abstract Loss of function of Niemann-Pick C1 (NPC1) leads to lysosomal free cholesterol storage, resulting in the neurodegenerative disease Niemann-Pick disease type C (NPC). Signifi cant numbers of patients with NPC also suffer from liver disease. Currently, no treatments exist that alter patient outcome, and it is unknown if recovery from tissue damage can occur even if a treatment were found. Our laboratory developed a strategy to test whether mice can recover from NPC liver disease. We used antisense oligonucleotides to knock down hepatic expression of NPC1 in BALB/C mice for either 9 or 15 weeks. This recapitulated liver disease with hepatomegaly, cell death, and fi brosis. Then, antisense oligonucleotide treatment was halted for an additional 4, 9, or 15 weeks. We report that significant liver recovery occurred even when NPC1 protein expression only partially returned to normal. Several pathological phenotypes were alleviated, including hepatomegaly, cholesterol storage, and liver cell death. Histological examination revealed that foamy cell accumulation was relieved; however, liver fi brosis increased. Additionally, resolution of cholesterol storage and liver cell death took longer in mice with long-term knockd...

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Section: Lipid-laden Cell Infi Ltration Slowly Resolves Post-treatmentsupporting

confidence: 83%

Section: Western Blotting Of Npc1mentioning

confidence: 99%

Section: Western Blotting Of Npc1mentioning

confidence: 99%

Section: Cell Death Resolves Along the Same Time Line As Cholesterol mentioning

confidence: 99%

See 2 more Smart Citations

Recovery from liver disease in a Niemann-Pick type C mouse model

Sayre

Rimkunas

Graham

et al. 2010

Journal of Lipid Research

Self Cite

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“…NPC2 is a lysosomal glycoprotein involved in regulating intracellular cholesterol homeostasis through direct binding with free cholesterol (17,18). Besides, GNMT deficiency in mice results in phenotypes that resemble NPC disease, including hepatic free cholesterol accumulation, hepatomegaly, higher serum alanine aminotransferase and hypercholesterolemia (20,33,34). In this study, we also observed that NPC1 was decreased in Gnmt -/-mice.…”

Section: Discussionsupporting

confidence: 68%

Glycine N-Methyltransferase Deficiency Affects Niemann-Pick Type C2 Protein Stability and Regulates Hepatic Cholesterol Homeostasis

Liao¹,

Chen²,

Lee³

et al. 2011

Mol Med

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Nonalcoholic fatty liver disease (NAFLD) is associated with the development of metabolic syndromes and hepatocellular carcinoma (HCC). Cholesterol accumulation is related to NAFLD, whereas its detailed mechanism is not fully understood. Previously, we reported that glycine N-methyltransferase (GNMT) knockout (Gnmt ings and coimmunoprecipitation assays elucidated that the C conserved region (81-105 amino acids) of NPC2 interacts with the carboxyl-terminal fragment (171-295 amino acids) of GNMT. Confocal microscopy demonstrated that when cells were treated with low-density lipoprotein, NPC2 was released from lysosomes and interacts with GNMT in the cytosol. Overexpression of GNMT doubled the half-lives of both NPC2 isoforms and reduced cholesterol accumulation in cells. Furthermore, GNMT was downregulated in the liver tissues from patients suffering with NAFLD as well as from mice fed a high-fat diet, high-cholesterol diet or methionine/choline-deficient diet. In conclusion, our study demonstrated that GNMT regulates the homeostasis of cholesterol metabolism, and hepatic cholesterol accumulation may result from downregulation of GNMT and instability of its interactive protein NPC2. Novel therapeutics for steatohepatitis and HCC may be developed by using this concept.

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Niemann-Pick type C2 protein regulates liver cancer progressionviamodulating ERK1/2 pathway: Clinicopathological correlations and therapeutical implications

et al. 2015

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Primary hepatocellular carcinoma (HCC) is the fifth most common malignancy worldwide and the third leading cause of cancer-related death. It is important to identify new targets for early diagnosis and treatment of HCC. Niemann-Pick type C2 (NPC2) plays an important role in the regulation of intracellular cholesterol homeostasis via direct binding with free cholesterol. However, little is known about the significance of NPC2 in HCC tumorigenesis. In this study, we showed that NPC2 is abundantly expressed in normal liver, but is downregulated in human HCC tissues. The patients with NPC2 downregulation expressed much higher a-fetoprotein, multiple tumor type, vascular invasion, later pathological stage and shorter survival rate. Knockdown NPC2 in liver cancer cell lines promote cell proliferation, migration and xenograft tumorigenesis. In contrast, NPC2 overexpression inhibits HuH7 promoted tumor growth. Furthermore, administration of hepatotropic adeno-associated virus 8 (AAV8) delivered NPC2 decreased the inflammatory infiltration, the expression of two early HCC markers-glypican 3 and survivin and suppressed the spontaneous HCC development in mice. To identify the NPC2-dependent mechanism, we emphasized on the status of MAPK/ERK signaling. MEK1/2 inhibitor treatment demonstrated that the expression of NPC2 affected the activation of ERK1/2 but not MEK1/2. In addition, cholesterol trafficking inhibitor treatment did not alter the cell proliferation and the activation of MEK/ERK. In conclusion, our study demonstrates that NPC2 may play an important role in negatively regulate cell proliferation and ERK1/2 activation that were independent of cholesterol accumulation. AAV-NPC2 may thus represent a new treatment strategy for liver cancer.

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In vivoantisense oligonucleotide reduction of NPC1 expression as a novel mouse model for Niemann Pick type C- associated liver disease

Cited by 33 publications

References 36 publications

Recovery from liver disease in a Niemann-Pick type C mouse model

Recovery from liver disease in a Niemann-Pick type C mouse model

Glycine N-Methyltransferase Deficiency Affects Niemann-Pick Type C2 Protein Stability and Regulates Hepatic Cholesterol Homeostasis

Niemann-Pick type C2 protein regulates liver cancer progressionviamodulating ERK1/2 pathway: Clinicopathological correlations and therapeutical implications

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