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Open Access 01-09-2021 | Article

Non-alcoholic fatty liver disease in mice with hepatocyte-specific deletion of mitochondrial fission factor

Authors: Yukina Takeichi, Takashi Miyazawa, Shohei Sakamoto, Yuki Hanada, Lixiang Wang, Kazuhito Gotoh, Keiichiro Uchida, Shunsuke Katsuhara, Ryuichi Sakamoto, Takaya Ishihara, Keiji Masuda, Naotada Ishihara, Masatoshi Nomura, Yoshihiro Ogawa

Published in: Diabetologia | Issue 9/2021

Abstract

Aims/hypothesis

Mitochondria are highly dynamic organelles continuously undergoing fission and fusion, referred to as mitochondrial dynamics, to adapt to nutritional demands. Evidence suggests that impaired mitochondrial dynamics leads to metabolic abnormalities such as non-alcoholic steatohepatitis (NASH) phenotypes. However, how mitochondrial dynamics are involved in the development of NASH is poorly understood. This study aimed to elucidate the role of mitochondrial fission factor (MFF) in the development of NASH.

Methods

We created mice with hepatocyte-specific deletion of MFF (MffLiKO). MffLiKO mice fed normal chow diet (NCD) or high-fat diet (HFD) were evaluated for metabolic variables and their livers were examined by histological analysis. To elucidate the mechanism of development of NASH, we examined the expression of genes related to endoplasmic reticulum (ER) stress and lipid metabolism, and the secretion of triacylglycerol (TG) using the liver and primary hepatocytes isolated from MffLiKO and control mice.

Results

MffLiKO mice showed aberrant mitochondrial morphologies with no obvious NASH phenotypes during NCD, while they developed full-blown NASH phenotypes in response to HFD. Expression of genes related to ER stress was markedly upregulated in the liver from MffLiKO mice. In addition, expression of genes related to hepatic TG secretion was downregulated, with reduced hepatic TG secretion in MffLiKO mice in vivo and in primary cultures of MFF-deficient hepatocytes in vitro. Furthermore, thapsigargin-induced ER stress suppressed TG secretion in primary hepatocytes isolated from control mice.

Conclusions/interpretation

We demonstrated that ablation of MFF in liver provoked ER stress and reduced hepatic TG secretion in vivo and in vitro. Moreover, MffLiKO mice were more susceptible to HFD-induced NASH phenotype than control mice, partly because of ER stress-induced apoptosis of hepatocytes and suppression of TG secretion from hepatocytes. This study provides evidence for the role of mitochondrial fission in the development of NASH.

Graphical abstract

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Simoes ICM, Fontes A, Pinton P, Zischka H, Wieckowski MR (2018) Mitochondria in non-alcoholic fatty liver disease. Int J Biochem Cell Biol 95:93–99. https://doi.org/10.1016/j.biocel.2017.12.019CrossRefPubMed

Chalasani N, Younossi Z, Lavine JE et al (2012) The diagnosis and management of non-alcoholic fatty liver disease: practice Guideline by the American Association for the Study of Liver Diseases, American College of Gastroenterology, and the American Gastroenterological Association. Hepatology 55(6):2005–2023. https://doi.org/10.1002/hep.25762CrossRefPubMed

Rosso N, Chavez-Tapia NC, Tiribelli C, Bellentani S (2014) Translational approaches: from fatty liver to non-alcoholic steatohepatitis. World J Gastroenterol 20(27):9038–9049. https://doi.org/10.3748/wjg.v20.i27.9038CrossRefPubMedPubMedCentral

Archer SL (2013) Mitochondrial Dynamics — Mitochondrial Fission and Fusion in Human Diseases. N Engl J Med 369(23):2236–2251. https://doi.org/10.1056/NEJMra1215233CrossRefPubMed

Zemirli N, Morel E, Molino D (2018) Mitochondrial Dynamics in Basal and Stressful Conditions. Int J Mol Sci 19(2):564. https://doi.org/10.3390/ijms19020564CrossRefPubMedCentral

van der Bliek AM, Sedensky MM, Morgan PG (2017) Cell Biology of the Mitochondrion. Genetics 207(3):843–871. https://doi.org/10.1534/genetics.117.300262CrossRefPubMedPubMedCentral

Karbowski M, Youle RJ (2003) Dynamics of mitochondrial morphology in healthy cells and during apoptosis. Cell Death Differ 10(8):870–880. https://doi.org/10.1038/sj.cdd.4401260CrossRefPubMed

Chan DC (2012) Fusion and fission: interlinked processes critical for mitochondrial health. Annu Rev Genet 46:265–287. https://doi.org/10.1146/annurev-genet-110410-132529CrossRefPubMed

Liesa M, Shirihai OS (2013) Mitochondrial dynamics in the regulation of nutrient utilization and energy expenditure. Cell Metab 17(4):491–506. https://doi.org/10.1016/j.cmet.2013.03.002CrossRefPubMedPubMedCentral

10.

Otera H, Mihara K (2011) Molecular mechanisms and physiologic functions of mitochondrial dynamics. J Biochem 149(3):241–251. https://doi.org/10.1093/jb/mvr002CrossRefPubMed

11.

Ishihara N, Otera H, Oka T, Mihara K (2013) Regulation and physiologic functions of GTPases in mitochondrial fusion and fission in mammals. Antioxid Redox Signal 19(4):389–399. https://doi.org/10.1089/ars.2012.4830CrossRefPubMed

12.

Smirnova E, Griparic L, Shurland DL, van der Bliek AM (2001) Dynamin-related protein Drp1 is required for mitochondrial division in mammalian cells. Mol Biol Cell 12(8):2245–2256. https://doi.org/10.1091/mbc.12.8.2245CrossRefPubMedPubMedCentral

13.

Gandre-Babbe S, van der Bliek AM (2008) The novel tail-anchored membrane protein Mff controls mitochondrial and peroxisomal fission in mammalian cells. Mol Biol Cell 19(6):2402–2412. https://doi.org/10.1091/mbc.E07-12-1287CrossRefPubMedPubMedCentral

14.

Otera H, Wang C, Cleland MM et al (2010) Mff is an essential factor for mitochondrial recruitment of Drp1 during mitochondrial fission in mammalian cells. J Cell Biol 191(6):1141–1158. https://doi.org/10.1083/jcb.201007152CrossRefPubMedPubMedCentral

15.

Santel A, Fuller MT (2001) Control of mitochondrial morphology by a human mitofusin. J Cell Sci 114(Pt 5):867–874CrossRef

16.

Alexander C, Votruba M, Pesch UE et al (2000) OPA1, encoding a dynamin-related GTPase, is mutated in autosomal dominant optic atrophy linked to chromosome 3q28. Nat Genet 26(2):211–215. https://doi.org/10.1038/79944CrossRefPubMed

17.

Delettre C, Lenaers G, Griffoin JM et al (2000) Nuclear gene OPA1, encoding a mitochondrial dynamin-related protein, is mutated in dominant optic atrophy. Nat Genet 26(2):207–210. https://doi.org/10.1038/79936CrossRefPubMed

18.

Marchi S, Patergnani S, Pinton P (2014) The endoplasmic reticulum-mitochondria connection: one touch, multiple functions. Biochim Biophys Acta 1837(4):461–469. https://doi.org/10.1016/j.bbabio.2013.10.015CrossRefPubMed

19.

Phillips MJ, Voeltz GK (2016) Structure and function of ER membrane contact sites with other organelles. Nat Rev Mol Cell Biol 17(2):69–82. https://doi.org/10.1038/nrm.2015.8CrossRefPubMed

20.

Galloway CA, Lee H, Brookes PS, Yoon Y (2014) Decreasing mitochondrial fission alleviates hepatic steatosis in a murine model of nonalcoholic fatty liver disease. Am J Physiol Gastrointest Liver Physiol 307(6):G632–G641. https://doi.org/10.1152/ajpgi.00182.2014CrossRefPubMedPubMedCentral

21.

Du J, Zhang X, Han J et al (2017) Pro-Inflammatory CXCR3 Impairs Mitochondrial Function in Experimental Non-Alcoholic Steatohepatitis. Theranostics 7(17):4192–4203. https://doi.org/10.7150/thno.21400CrossRefPubMedPubMedCentral

22.

Caldwell SH, Swerdlow RH, Khan EM et al (1999) Mitochondrial abnormalities in non-alcoholic steatohepatitis. J Hepatol 31(3):430–434CrossRef

23.

Lotowska JM, Sobaniec-Lotowska ME, Bockowska SB, Lebensztejn DM (2014) Pediatric non-alcoholic steatohepatitis: the first report on the ultrastructure of hepatocyte mitochondria. World J Gastroenterol 20(15):4335–4340. https://doi.org/10.3748/wjg.v20.i15.4335CrossRefPubMedPubMedCentral

24.

Sanyal AJ, Campbell-Sargent C, Mirshahi F et al (2001) Nonalcoholic steatohepatitis: association of insulin resistance and mitochondrial abnormalities. Gastroenterology 120(5):1183–1192. https://doi.org/10.1053/gast.2001.23256CrossRefPubMed

25.

Kirsch R, Clarkson V, Shephard EG et al (2003) Rodent nutritional model of non-alcoholic steatohepatitis: species, strain and sex difference studies. J Gastroenterol Hepatol 18(11):1272–1282. https://doi.org/10.1046/j.1440-1746.2003.03198.xCrossRefPubMed

26.

Kathirvel E, Morgan K, French SW, Morgan TR (2013) Acetyl-L-carnitine and lipoic acid improve mitochondrial abnormalities and serum levels of liver enzymes in a mouse model of nonalcoholic fatty liver disease. Nutr Res 33(11):932–941. https://doi.org/10.1016/j.nutres.2013.08.001CrossRefPubMed

27.

Yamada T, Murata D, Adachi Y et al (2018) Mitochondrial Stasis Reveals p62-Mediated Ubiquitination in Parkin-Independent Mitophagy and Mitigates Nonalcoholic Fatty Liver Disease. Cell Metab 28(4):588–604 e585. https://doi.org/10.1016/j.cmet.2018.06.014CrossRefPubMedPubMedCentral

28.

Wang L, Ishihara T, Ibayashi Y et al (2015) Disruption of mitochondrial fission in the liver protects mice from diet-induced obesity and metabolic deterioration. Diabetologia 58(10):2371–2380. https://doi.org/10.1007/s00125-015-3704-7CrossRefPubMed

29.

Zhang Z, Li B, Meng X et al (2016) Berberine prevents progression from hepatic steatosis to steatohepatitis and fibrosis by reducing endoplasmic reticulum stress. Sci Rep 6:20848. https://doi.org/10.1038/srep20848CrossRefPubMedPubMedCentral

30.

Kleiner DE, Brunt EM, Van Natta M et al (2005) Design and validation of a histological scoring system for nonalcoholic fatty liver disease. Hepatology 41(6):1313–1321. https://doi.org/10.1002/hep.20701CrossRefPubMed

31.

Brunt EM, Janney CG, Di Bisceglie AM, Neuschwander-Tetri BA, Bacon BR (1999) Nonalcoholic steatohepatitis: a proposal for grading and staging the histological lesions. Am J Gastroenterol 94(9):2467–2474. https://doi.org/10.1111/j.1572-0241.1999.01377.xCrossRefPubMed

32.

Giacomello M, Pellegrini L (2016) The coming of age of the mitochondria-ER contact: a matter of thickness. Cell Death Differ 23(9):1417–1427. https://doi.org/10.1038/cdd.2016.52CrossRefPubMedPubMedCentral

33.

Severgnini M, Sherman J, Sehgal A et al (2012) A rapid two-step method for isolation of functional primary mouse hepatocytes: cell characterization and asialoglycoprotein receptor based assay development. Cytotechnology 64(2):187–195. https://doi.org/10.1007/s10616-011-9407-0CrossRefPubMed

34.

Wieckowski MR, Giorgi C, Lebiedzinska M, Duszynski J, Pinton P (2009) Isolation of mitochondria-associated membranes and mitochondria from animal tissues and cells. Nat Protoc 4(11):1582–1590. https://doi.org/10.1038/nprot.2009.151CrossRefPubMed

35.

Ishihara T, Ban-Ishihara R, Maeda M et al (2015) Dynamics of mitochondrial DNA nucleoids regulated by mitochondrial fission is essential for maintenance of homogeneously active mitochondria during neonatal heart development. Mol Cell Biol 35(1):211–223. https://doi.org/10.1128/MCB.01054-14CrossRefPubMed

36.

Itoh M, Kato H, Suganami T et al (2013) Hepatic crown-like structure: a unique histological feature in non-alcoholic steatohepatitis in mice and humans. PLoS One 8(12):e82163. https://doi.org/10.1371/journal.pone.0082163CrossRefPubMedPubMedCentral

37.

Itoh M, Suganami T, Kato H et al (2017) CD11c+ resident macrophages drive hepatocyte death-triggered liver fibrosis in a murine model of nonalcoholic steatohepatitis. JCI Insight 2(22):e92902. https://doi.org/10.1172/jci.insight.92902CrossRefPubMedCentral

38.

Jamil H, Dickson JK Jr, Chu CH et al (1995) Microsomal triglyceride transfer protein. Specificity of lipid binding and transport. J Biol Chem 270(12):6549–6554. https://doi.org/10.1074/jbc.270.12.6549CrossRefPubMed

39.

Alves-Bezerra M, Cohen DE (2017) Triglyceride Metabolism in the Liver. Compr Physiol 8(1):1–8. https://doi.org/10.1002/cphy.c170012CrossRefPubMedPubMedCentral

40.

Cianflone K, Dahan S, Monge JC, Sniderman AD (1992) Pathogenesis of Carbohydrate-Induced Hypertriglyceridemia Using HepG2 Cells as a Model System. Arterioscler Thromb 12(3):271–277. https://doi.org/10.1161/01.ATV.12.3.271CrossRefPubMed

41.

Nasca A, Nardecchia F, Commone A et al (2018) Clinical and Biochemical Features in a Patient With Mitochondrial Fission Factor Gene Alteration. Front Genet 9:625. https://doi.org/10.3389/fgene.2018.00625CrossRefPubMedPubMedCentral

42.

Li L, Martin-Levilain J, Jiménez-Sánchez C et al (2019) In vivo stabilization of OPA1 in hepatocytes potentiates mitochondrial respiration and gluconeogenesis in a prohibitin-dependent way. J Biol Chem 294(34):12581–12598. https://doi.org/10.1074/jbc.RA119.007601CrossRefPubMedPubMedCentral

43.

Tilg H, Moschen AR (2010) Evolution of inflammation in nonalcoholic fatty liver disease: the multiple parallel hits hypothesis. Hepatology 52(5):1836–1846. https://doi.org/10.1002/hep.24001CrossRefPubMed

44.

Hernandez-Alvarez MI, Sebastian D, Vives S et al (2019) Deficient Endoplasmic Reticulum-Mitochondrial Phosphatidylserine Transfer Causes Liver Disease. Cell 177(4):881–895 e817. https://doi.org/10.1016/j.cell.2019.04.010CrossRefPubMed

45.

Hammerschmidt P, Ostkotte D, Nolte H et al (2019) CerS6-Derived Sphingolipids Interact with Mff and Promote Mitochondrial Fragmentation in Obesity. Cell 177(6):1536–1552 e1523. https://doi.org/10.1016/j.cell.2019.05.008CrossRefPubMed

46.

Lee J, Homma T, Fujii J (2017) Mice in the early stage of liver steatosis caused by a high fat diet are resistant to thioacetamide-induced hepatotoxicity and oxidative stress. Toxicol Lett 277:92–103. https://doi.org/10.1016/j.toxlet.2017.06.005CrossRefPubMed

47.

Willy JA, Young SK, Stevens JL, Masuoka HC, Wek RC (2015) CHOP links endoplasmic reticulum stress to NF-kappaB activation in the pathogenesis of nonalcoholic steatohepatitis. Mol Biol Cell 26(12):2190–2204. https://doi.org/10.1091/mbc.E15-01-0036CrossRefPubMedPubMedCentral

48.

Han J, Kaufman RJ (2016) The role of ER stress in lipid metabolism and lipotoxicity. J Lipid Res 57(8):1329–1338. https://doi.org/10.1194/jlr.R067595CrossRefPubMedPubMedCentral

49.

Jo H, Choe SS, Shin KC et al (2013) Endoplasmic reticulum stress induces hepatic steatosis via increased expression of the hepatic very low-density lipoprotein receptor. Hepatology 57(4):1366–1377. https://doi.org/10.1002/hep.26126CrossRefPubMed

Title: Non-alcoholic fatty liver disease in mice with hepatocyte-specific deletion of mitochondrial fission factor
Authors: Yukina Takeichi
Takashi Miyazawa
Shohei Sakamoto
Yuki Hanada
Lixiang Wang
Kazuhito Gotoh
Keiichiro Uchida
Shunsuke Katsuhara
Ryuichi Sakamoto
Takaya Ishihara
Keiji Masuda
Naotada Ishihara
Masatoshi Nomura
Yoshihiro Ogawa
Publication date: 01-09-2021
Publisher: Springer Berlin Heidelberg
Published in: Diabetologia / Issue 9/2021
Print ISSN: 0012-186X
Electronic ISSN: 1432-0428
DOI: https://doi.org/10.1007/s00125-021-05488-2

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