Role of microRNA-122 in hepatocarcinogenesis using conditional knock out mice

Summary

Principal Investigator: Kalpana Ghoshal
Abstract: DESCRIPTION (provided by applicant): The objective of this new R01 application is to advance our understanding of the role of miR-122 in hepatocarcinogenesis. HCC is the fifth most common cancer and the third common cause of cancer related death. The incidence of HCC is on the rise in USA, with little hope for cure or treatment through chemotherapy, radiation or other traditional cancer treatments. Its major risk factors are infection with hepatitis B and C viruses, and exposure to toxic chemicals, including alcohol, all of which cause chronic liver injury and inflammation. Using an animal model for hepatocarcinogenesis we were the first to demonstrate down regulation of miR-122, the most abundant liver-specific microRNA (~70% of the total miRNA) in the liver, during the initiation and progression of HCC and also in human primary HCCs. Suppression of miR-122 is a signature of HCCs with poor prognosis and metastasis. Studies with HCC cells in culture have shown that miR-122 functions as a tumor suppressor in vitro and in nude mice. To understand the biological functions of miR-122, especially in hepatocarcinogenesis, we have generated conditional knockout mice (supported by an R21 grant to the PI). These mice express 100 fold less miR-122 when crossed to AlbCre mice and spontaneously develop hepatitis in the liver with age, which is facilitated after feeding choline-deficient diet that promotes hepatocarcinogenesis. More importantly, miR-122 deleted (KO) mice are more susceptible to HCCs when exposed to diethylnitrosamine, a potent liver carcinogen. Based on these observations we hypothesize that miR-122 plays a critical role in maintaining liver function, and loss of miR-122 predisposes to liver disease including cancer. The specific aims of the proposal are: Aim 1. Investigate the role of miR-122 in a mouse model of nonalcoholic fatty liver disease (NAFLD) related HCC induced by feeding choline-deficient diet. 1a) The susceptibility of miR-122 / (KO) and miR-122fl/fl (control) mice to CDAA diet will be examined by comparing liver damage (apoptosis, steatosis or fatty liver, inflammation, fibrosis) and liver tumors (formation of adenomas and carcinomas) between these mice, and 1b) the involvement of miR-122 targets will be assessed. Aim 2. Investigate the role of miR-122 in diethylnitrosamine (DEN)-induced hepatocarcinogenesis. Pathological/molecular changes of mice injected with DEN will be monitored as described in Aim 1. Aim 3. Examine the therapeutic potential of miR-122 alone or in combination with chemotherapeutic agents to inhibit tumor growth in vivo in the DEN model. Mice will be injected weekly for 4 weeks with miR-122 mimetics loaded in galactosylated nanoparticles (to specifically target it to HCC cells) at early stages of tumor development (visualized by MRI) and the regression in the tumor growth will be compared to those in mice treated with the scrambled RNA nanoparticles. This study will elucidate the function of the most abundant liver-specific microRNA in maintaining normal liver physiology and also its therapeutic efficacy against hepatocellular carcinomas in an animal model. PUBLIC HEALTH RELEVANCE: Layman's Abstract The urgent need for new treatments for hepatocellular cancer (HCC) is warranted because incidence of HCC is increasing considerably in the United States, their dismal prognosis and the poor response of this cancer to treatment regimen currently available. This project focuses on the role of the loss of function of miR-122 in the development and progression of hepatocellular carcinoma (HCC), one of the most common human malignancies. Our previous studies have shown downregulation of miR-122 in HCCs of both human and rodent origin. Now we intend to show that the loss of function of this liver specific miR plays a causal role in the initiation and progression of HCC. If this is the case, these studies will open the way to the development of novel miR-122 therapy for this incurable disease. Thus, establishing role of miR-122 mimetic in an animal model (in preclinical trial) would be a major milestone in the treatment of this deadly disease in the near future.
Funding Period: 2010-05-17 - 2015-03-31
more information: NIH RePORT

Top Publications

  1. pmc Stat3-mediated activation of microRNA-23a suppresses gluconeogenesis in hepatocellular carcinoma by down-regulating glucose-6-phosphatase and peroxisome proliferator-activated receptor gamma, coactivator 1 alpha
    Bo Wang
    Department of Molecular and Cellular Biochemistry, College of Medicine, Ohio State University, Columbus, OH 43210, USA
    Hepatology 56:186-97. 2012
  2. pmc Hepatic loss of miR-122 predisposes mice to hepatobiliary cyst and hepatocellular carcinoma upon diethylnitrosamine exposure
    Shu Hao Hsu
    Department of Molecular and Cellular Biochemistry, The Ohio State University, Columbus, Ohio Department of Molecular, Cellular and Developmental Biology Program, The Ohio State University, Columbus, Ohio
    Am J Pathol 183:1719-30. 2013
  3. pmc Reciprocal regulation of microRNA-122 and c-Myc in hepatocellular cancer: role of E2F1 and transcription factor dimerization partner 2
    Bo Wang
    Department of Molecular and Cellular Biochemistry, Ohio State University, Columbus, OH Molecular, Cellular and Developmental Biology Program, Ohio State University, Columbus, OH
    Hepatology 59:555-66. 2014
  4. pmc Cationic lipid nanoparticles for therapeutic delivery of siRNA and miRNA to murine liver tumor
    Shu Hao Hsu
    Molecular, Cellular, and Developmental Biology Program, The Ohio State University, Columbus, OH 43210, USA Department of Molecular and Cellular Biochemistry, The Ohio State University, Columbus, OH 43210, USA
    Nanomedicine 9:1169-80. 2013
  5. pmc Lactosylated gramicidin-based lipid nanoparticles (Lac-GLN) for targeted delivery of anti-miR-155 to hepatocellular carcinoma
    Mengzi Zhang
    Molecular, Cellular and Developmental Biology Program, Ohio State University, Columbus, OH 43210, USA
    J Control Release 168:251-61. 2013
  6. pmc Methylation of the PTPRO gene in human hepatocellular carcinoma and identification of VCP as its substrate
    Shu Hao Hsu
    Department of Molecular and Cellular Biochemistry, The Ohio State University, Columbus, Ohio 43210, USA
    J Cell Biochem 114:1810-8. 2013
  7. pmc Reduced susceptibility of DNA methyltransferase 1 hypomorphic (Dnmt1N/+) mice to hepatic steatosis upon feeding liquid alcohol diet
    Huban Kutay
    Department of Molecular and Cellular Biochemistry, College of Medicine, The Ohio State University, Columbus, Ohio, United States of America
    PLoS ONE 7:e41949. 2012
  8. pmc Essential metabolic, anti-inflammatory, and anti-tumorigenic functions of miR-122 in liver
    Shu Hao Hsu
    Molecular, Cellular, and Developmental Biology Program, The Ohio State University, Columbus, OH, USA
    J Clin Invest 122:2871-83. 2012
  9. pmc Lipid nanoparticles for hepatic delivery of small interfering RNA
    Bo Yu
    Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH, USA
    Biomaterials 33:5924-34. 2012
  10. ncbi Enhanced hepatic delivery of siRNA and microRNA using oleic acid based lipid nanoparticle formulations
    Xinmei Wang
    NSF Nanoscale Science and Engineering Center NSEC, The Ohio State University, Columbus, USA
    J Control Release 172:690-8. 2013

Detail Information

Publications11

  1. pmc Stat3-mediated activation of microRNA-23a suppresses gluconeogenesis in hepatocellular carcinoma by down-regulating glucose-6-phosphatase and peroxisome proliferator-activated receptor gamma, coactivator 1 alpha
    Bo Wang
    Department of Molecular and Cellular Biochemistry, College of Medicine, Ohio State University, Columbus, OH 43210, USA
    Hepatology 56:186-97. 2012
    ..G6PC expression also correlated with tumor grade in human primary HCCs. Finally, this study showed that the activation of interleukin (IL)-6-Stat3 signaling caused the up-regulation of miR-23a expression in HCC...
  2. pmc Hepatic loss of miR-122 predisposes mice to hepatobiliary cyst and hepatocellular carcinoma upon diethylnitrosamine exposure
    Shu Hao Hsu
    Department of Molecular and Cellular Biochemistry, The Ohio State University, Columbus, Ohio Department of Molecular, Cellular and Developmental Biology Program, The Ohio State University, Columbus, Ohio
    Am J Pathol 183:1719-30. 2013
    ..Collectively, miR-122 depletion facilitates cystogenesis and hepatocarcinogenesis in mice on DEN challenge by up-regulating several genes involved in proliferation, growth factor signaling, neovascularization, and metastasis...
  3. pmc Reciprocal regulation of microRNA-122 and c-Myc in hepatocellular cancer: role of E2F1 and transcription factor dimerization partner 2
    Bo Wang
    Department of Molecular and Cellular Biochemistry, Ohio State University, Columbus, OH Molecular, Cellular and Developmental Biology Program, Ohio State University, Columbus, OH
    Hepatology 59:555-66. 2014
    ....
  4. pmc Cationic lipid nanoparticles for therapeutic delivery of siRNA and miRNA to murine liver tumor
    Shu Hao Hsu
    Molecular, Cellular, and Developmental Biology Program, The Ohio State University, Columbus, OH 43210, USA Department of Molecular and Cellular Biochemistry, The Ohio State University, Columbus, OH 43210, USA
    Nanomedicine 9:1169-80. 2013
    ..These data demonstrate the potential of LNP-DP1-mediated microRNA delivery as a novel strategy for HCC therapy...
  5. pmc Lactosylated gramicidin-based lipid nanoparticles (Lac-GLN) for targeted delivery of anti-miR-155 to hepatocellular carcinoma
    Mengzi Zhang
    Molecular, Cellular and Developmental Biology Program, Ohio State University, Columbus, OH 43210, USA
    J Control Release 168:251-61. 2013
    ..5 mg/kg anti-miR-155 loaded Lac-GLN resulted in up-regulation of C/EBPβ and FOXP3 by 6.9- and 2.2-fold, respectively. These results suggest potential application of Lac-GLN as a liver-specific delivery vehicle for anti-miR therapy...
  6. pmc Methylation of the PTPRO gene in human hepatocellular carcinoma and identification of VCP as its substrate
    Shu Hao Hsu
    Department of Molecular and Cellular Biochemistry, The Ohio State University, Columbus, Ohio 43210, USA
    J Cell Biochem 114:1810-8. 2013
    ....
  7. pmc Reduced susceptibility of DNA methyltransferase 1 hypomorphic (Dnmt1N/+) mice to hepatic steatosis upon feeding liquid alcohol diet
    Huban Kutay
    Department of Molecular and Cellular Biochemistry, College of Medicine, The Ohio State University, Columbus, Ohio, United States of America
    PLoS ONE 7:e41949. 2012
    ..Aberrations in DNA methylation and Dnmts are linked to different diseases including cancer. However, their role in alcoholic liver disease (ALD) has not been elucidated...
  8. pmc Essential metabolic, anti-inflammatory, and anti-tumorigenic functions of miR-122 in liver
    Shu Hao Hsu
    Molecular, Cellular, and Developmental Biology Program, The Ohio State University, Columbus, OH, USA
    J Clin Invest 122:2871-83. 2012
    ....
  9. pmc Lipid nanoparticles for hepatic delivery of small interfering RNA
    Bo Yu
    Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH, USA
    Biomaterials 33:5924-34. 2012
    ..These data suggest potential therapeutic applications of TRENL3 mediated delivery of siRNA for liver diseases...
  10. ncbi Enhanced hepatic delivery of siRNA and microRNA using oleic acid based lipid nanoparticle formulations
    Xinmei Wang
    NSF Nanoscale Science and Engineering Center NSEC, The Ohio State University, Columbus, USA
    J Control Release 172:690-8. 2013
    ..LNPs containing OA is a promising nanocarrier system for the delivery of RNA-based therapeutics in liver diseases. ..