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Molecular Medicine

, Volume 18, Issue 3, pp 412–422 | Cite as

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

  • Yi-Jen Liao
  • Tzu-Lang Chen
  • Tzong-Shyuan Lee
  • Hsiang-An Wang
  • Chung-Kwe Wang
  • Li-Ying Liao
  • Ren-Shyan Liu
  • Shiu-Feng Huang
  • Yi-Ming Arthur Chen
Research Article

Abstract

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-/-) mice develop chronic hepatitis and HCC. In this study, we showed that Gnmt-/- mice had hyperlipidemia and steatohepatitis. Single photon emission computed tomography images of mice injected with 131I-labeled 6β-iodocholesterol demonstrated that Gnmt-/- mice had slower hepatic cholesterol uptake and excretion rates than wild-type mice. In addition, genes related to cholesterol uptake (scavenger receptor class B type 1 [SR-B1] and ATP-binding cassette A1 (ABCA1)), intracellular trafficking (Niemann-Pick type C1 protein [NPC1] and Niemann-Pick type C2 protein [NPC2]) and excretion (ATP-binding cassette G1 [ABCG1]) were downregulated in Gnmt-/- mice. Yeast two-hybrid screenings 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

Notes

Acknowledgments

This study was supported by a grant from the National Science Council of the Republic of China (NSC99-2628-B-010-010-MY3 and NSC100-2325-B-010-008) and a grant from the Genomic Research Center of the National Yang-Ming University given by the Ministry of Education of the Republic of China (Top University and Center Grant). The authors thank Fu-Hui Wang (Molecular and Genetic Imaging Core), Chi-Hung Lin (National Research Program for Genomic Medicine) and Tung-Wei Chen and Chia-Yen Chen (Institute of Microbiology and Immunology, School of Life Science), all at National Yang-Ming University, for technical support.

Supplementary material

10020_2012_1803412_MOESM1_ESM.pdf (2.2 mb)
Glycine N-Methyltransferase Deficiency Affects Niemann-Pick Type C2 Protein Stability and Regulates Hepatic Cholesterol Homeostasis

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Authors and Affiliations

  • Yi-Jen Liao
    • 1
  • Tzu-Lang Chen
    • 1
  • Tzong-Shyuan Lee
    • 2
  • Hsiang-An Wang
    • 1
  • Chung-Kwe Wang
    • 3
  • Li-Ying Liao
    • 4
  • Ren-Shyan Liu
    • 5
  • Shiu-Feng Huang
    • 6
  • Yi-Ming Arthur Chen
    • 1
    • 7
    • 8
  1. 1.AIDS Prevention and Research CenterNational Yang-Ming UniversityTaipeiTaiwan
  2. 2.Department and Institute of PhysiologyNational Yang-Ming UniversityTaipeiTaiwan
  3. 3.Department of International MedicineTaipei City Hospital Renai BranchTaipeiTaiwan
  4. 4.Department of Gastroenterology, Liver CenterTaipei City Hospital Renai BranchTaipeiTaiwan
  5. 5.Molecular and Genetic Imaging CoreNational Research Program for Genomic Medicine (NRPGM)TaipeiTaiwan
  6. 6.Division of Molecular and Genomic MedicineNational Health Research InstituteMiaoliTaiwan
  7. 7.Department of Microbiology, School of Medicine and Institute of Microbiology and Immunology, School of Life ScienceNational Yang-Ming UniversityTaipeiTaiwan
  8. 8.VYM Genome Research CenterNational Yang-Ming UniversityTaipeiTaiwan

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