JNK-mediated disruption of bile acid homeostasis promotes intrahepatic cholangiocarcinoma
Contributed by Roger J. Davis, May 14, 2020 (sent for review February 12, 2020; reviewed by Anton M. Bennett and J. Silvio Gutkind)
Significance
Obesity is associated with hepatic steatosis and activation of the cJun NH2-terminal kinase (JNK) stress-signaling pathway. Studies in mice demonstrate that JNK deficiency in the liver prevents the development of hepatic steatosis. This observation suggests that inhibition of JNK signaling may represent a possible treatment for hepatic steatosis. However, the long-term consequences of JNK inhibition are poorly understood. Here we demonstrate that loss of JNK causes changes in cholesterol and bile acid metabolism that promote cholestasis, bile duct proliferation, and intrahepatic cholangiocarcinoma. We identify PPARα activation as the molecular mechanism that accounts for this phenotype. Our analysis has important implications for the long-term use of JNK inhibitors for the treatment of obesity.
Abstract
Metabolic stress causes activation of the cJun NH2-terminal kinase (JNK) signal transduction pathway. It is established that one consequence of JNK activation is the development of insulin resistance and hepatic steatosis through inhibition of the transcription factor PPARα. Indeed, JNK1/2 deficiency in hepatocytes protects against the development of steatosis, suggesting that JNK inhibition represents a possible treatment for this disease. However, the long-term consequences of JNK inhibition have not been evaluated. Here we demonstrate that hepatic JNK controls bile acid production. We found that hepatic JNK deficiency alters cholesterol metabolism and bile acid synthesis, conjugation, and transport, resulting in cholestasis, increased cholangiocyte proliferation, and intrahepatic cholangiocarcinoma. Gene ablation studies confirmed that PPARα mediated these effects of JNK in hepatocytes. This analysis highlights potential consequences of long-term use of JNK inhibitors for the treatment of metabolic syndrome.
Data Availability
Acknowledgments
We thank S. Bartlett for English editing and David Garlick (University of Massachusetts Medical School) for pathological examination of tissue sections. We are grateful to the CNIC Advanced Imaging and Animal facility for technical support. G.S. (RYC-2009-04972) and F.J.C. (RYC-2014-15242) are investigators of the Ramón y Cajal Program. E.M. was awarded a La Caixa fellowship. C.F. was awarded a Sara Borrell contract (CD19/00078). This work was funded by grants supported in part by funds from the European Regional Development Fund: EU’s Seventh Framework Programme (FP7/2007-2013) ERC 260464, EFSD/Lilly European Diabetes Research Programme Dr. Sabio, 2017 Leonardo Grant for Researchers and Cultural Creators, BBVA Foundation (Investigadores-BBVA-2017) IN[17]_BBM_BAS_0066, MINECO-FEDER SAF2016-79126-R, and Comunidad de Madrid IMMUNOTHERCAN-CM S2010/BMD-2326 and B2017/BMD-3733 and La Asociación Española contra el Cáncer (to G.S.); EXOHEP-CM S2017/BMD-3727 and the European Cooperation in Science & Technology (COST) Action CA17112 (to F.J.C.); MINECO Retos SAF2016-78711, the AMMF Cholangiocarcinoma Charity 2018/117, NanoLiver-CM Y2018/NMT-4949, UCM-25-2019, ERAB EA/18-14 (to F.J.C.). F.J.C. is a Gilead Liver Research Scholar. Grant DK R01 DK107220 from the NIH (to R.J.D.); and PI16/00598 from Carlos III Institute of Health, Spain (to J.J.G.M.). The CNIC is supported by the Ministerio de Ciencia, Innovación y Universidades, and the Pro-CNIC Foundation and is a Severo Ochoa Center of Excellence (SEV-2015-0505).
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References
1
D. M. Parkin, F. Bray, J. Ferlay, P. Pisani, Global cancer statistics, 2002. CA Cancer J. Clin. 55, 74–108 (2005).
2
H. B. El-Serag, J. A. Davila, N. J. Petersen, K. A. McGlynn, The continuing increase in the incidence of hepatocellular carcinoma in the United States: An update. Ann. Intern. Med. 139, 817–823 (2003).
3
S. A. Khan, S. Tavolari, G. Brandi, Cholangiocarcinoma: Epidemiology and risk factors. Liver Int. 39 (suppl. 1), 19–31 (2019).
4
S. A. Khan et al.; British Society of Gastroenterology, Guidelines for the diagnosis and treatment of cholangiocarcinoma: An update. Gut 61, 1657–1669 (2012).
5
E. Manieri, G. Sabio, Stress kinases in the modulation of metabolism and energy balance. J. Mol. Endocrinol. 55, R11–R22 (2015).
6
G. Sabio, R. J. Davis, cJun NH2-terminal kinase 1 (JNK1): Roles in metabolic regulation of insulin resistance. Trends Biochem. Sci. 35, 490–496 (2010).
7
S. Vernia et al., The PPARα-FGF21 hormone axis contributes to metabolic regulation by the hepatic JNK signaling pathway. Cell Metab. 20, 512–525 (2014).
8
S. Vernia, J. Cavanagh-Kyros, T. Barrett, C. Tournier, R. J. Davis, Fibroblast growth factor 21 mediates glycemic regulation by hepatic JNK. Cell Rep. 14, 2273–2280 (2016).
9
T. Gulick, S. Cresci, T. Caira, D. D. Moore, D. P. Kelly, The peroxisome proliferator-activated receptor regulates mitochondrial fatty acid oxidative enzyme gene expression. Proc. Natl. Acad. Sci. U.S.A. 91, 11012–11016 (1994).
10
R. H. Unger, Y. T. Zhou, Lipotoxicity of beta-cells in obesity and in other causes of fatty acid spillover. Diabetes 50 (suppl. 1), S118–S121 (2001).
11
R. M. Evans, G. D. Barish, Y. X. Wang, PPARs and the complex journey to obesity. Nat. Med. 10, 355–361 (2004).
12
R. A. Memon et al., Up-regulation of peroxisome proliferator-activated receptors (PPAR-alpha) and PPAR-gamma messenger ribonucleic acid expression in the liver in murine obesity: Troglitazone induces expression of PPAR-gamma-responsive adipose tissue-specific genes in the liver of obese diabetic mice. Endocrinology 141, 4021–4031 (2000).
13
E. Ip et al., Central role of PPARalpha-dependent hepatic lipid turnover in dietary steatohepatitis in mice. Hepatology 38, 123–132 (2003).
14
N. C. Teoh et al., Short-term therapy with peroxisome proliferation-activator receptor-alpha agonist Wy-14,643 protects murine fatty liver against ischemia-reperfusion injury. Hepatology 51, 996–1006 (2010).
15
P. R. Holden, J. D. Tugwood, Peroxisome proliferator-activated receptor alpha: Role in rodent liver cancer and species differences. J. Mol. Endocrinol. 22, 1–8 (1999).
16
D. Panigrahy et al., PPARalpha agonist fenofibrate suppresses tumor growth through direct and indirect angiogenesis inhibition. Proc. Natl. Acad. Sci. U.S.A. 105, 985–990 (2008).
17
M. Maggiora, M. Oraldi, G. Muzio, R. A. Canuto, Involvement of PPARα and PPARγ in apoptosis and proliferation of human hepatocarcinoma HepG2 cells. Cell Biochem. Funct. 28, 571–577 (2010).
18
D. Yamasaki et al., Fenofibrate suppresses growth of the human hepatocellular carcinoma cell via PPARα-independent mechanisms. Eur. J. Cell Biol. 90, 657–664 (2011).
19
T. Li, J. Y. Chiang, Regulation of bile acid and cholesterol metabolism by PPARs. PPAR Res. 2009, 501739 (2009).
20
C. Yoo et al., Multiplexed gene expression profiling identifies the FGFR4 pathway as a novel biomarker in intrahepatic cholangiocarcinoma. Oncotarget 8, 38592–38601 (2017).
21
S. Miura et al., Fibroblast growth factor 19 expression correlates with tumor progression and poorer prognosis of hepatocellular carcinoma. BMC Cancer 12, 56 (2012).
22
H. Shapiro, A. A. Kolodziejczyk, D. Halstuch, E. Elinav, Bile acids in glucose metabolism in health and disease. J. Exp. Med. 215, 383–396 (2018).
23
P. C. de Groen, G. J. Gores, N. F. LaRusso, L. L. Gunderson, D. M. Nagorney, Biliary tract cancers. N. Engl. J. Med. 341, 1368–1378 (1999).
24
V. Keitel et al., The G-protein coupled bile salt receptor TGR5 is expressed in liver sinusoidal endothelial cells. Hepatology 45, 695–704 (2007).
25
P. A. Dawson, T. Lan, A. Rao, Bile acid transporters. J. Lipid Res. 50, 2340–2357 (2009).
26
D. A. Piccoli, N. B. Spinner, Alagille syndrome and the Jagged1 gene. Semin. Liver Dis. 21, 525–534 (2001).
27
J. Fan et al., Bone morphogenetic protein 4 mediates bile duct ligation induced liver fibrosis through activation of Smad1 and ERK1/2 in rat hepatic stellate cells. J. Cell. Physiol. 207, 499–505 (2006).
28
M. Yanai et al., FGF signaling segregates biliary cell-lineage from chick hepatoblasts cooperatively with BMP4 and ECM components in vitro. Dev. Dyn. 237, 1268–1283 (2008).
29
L. Yang et al., A single-cell transcriptomic analysis reveals precise pathways and regulatory mechanisms underlying hepatoblast differentiation. Hepatology 66, 1387–1401 (2017).
30
T. Li, J. Y. Chiang, Bile acids as metabolic regulators. Curr. Opin. Gastroenterol. 31, 159–165 (2015).
31
P. L. Jansen et al., The ascending pathophysiology of cholestatic liver disease. Hepatology 65, 722–738 (2017).
32
K. Allen, H. Jaeschke, B. L. Copple, Bile acids induce inflammatory genes in hepatocytes: A novel mechanism of inflammation during obstructive cholestasis. Am. J. Pathol. 178, 175–186 (2011).
33
W. Zhang et al., A weighted relative difference accumulation algorithm for dynamic metabolomics data: Long-term elevated bile acids are risk factors for hepatocellular carcinoma. Sci. Rep. 5, 8984 (2015).
34
S. Sombattheera et al., Total serum bile acid as a potential marker for the diagnosis of cholangiocarcinoma without jaundice. Asian Pac. J. Cancer Prev. 16, 1367–1370 (2015).
35
D. Yuan et al., Kupffer cell-derived Tnf triggers cholangiocellular tumorigenesis through JNK due to chronic mitochondrial dysfunction and ROS. Cancer Cell 31, 771–789.e6 (2017).
36
F. J. Cubero et al., Loss of c-Jun N-terminal kinase 1 and 2 function in liver epithelial cells triggers biliary hyperproliferation resembling cholangiocarcinoma. Hepatol. Commun 4, 834–851 (2020).
37
M. A. Abdelmegeed et al., PPARalpha expression protects male mice from high fat-induced nonalcoholic fatty liver. J. Nutr. 141, 603–610 (2011).
38
J. E. Yeon et al., Reduced expression of peroxisome proliferator-activated receptor-alpha may have an important role in the development of non-alcoholic fatty liver disease. J. Gastroenterol. Hepatol. 19, 799–804 (2004).
39
T. Inagaki et al., Fibroblast growth factor 15 functions as an enterohepatic signal to regulate bile acid homeostasis. Cell Metab. 2, 217–225 (2005).
40
V. J. Nies et al., Fibroblast growth factor signaling in metabolic regulation. Front. Endocrinol. (Lausanne) 6, 193 (2016).
41
M. Zhou et al., Mouse species-specific control of hepatocarcinogenesis and metabolism by FGF19/FGF15. J. Hepatol. 66, 1182–1192 (2017).
42
G. Cui et al., Up-regulation of FGF15/19 signaling promotes hepatocellular carcinoma in the background of fatty liver. J. Exp. Clin. Cancer Res. 37, 136 (2018).
43
J. M. Peters, R. C. Cattley, F. J. Gonzalez, Role of PPAR alpha in the mechanism of action of the nongenotoxic carcinogen and peroxisome proliferator Wy-14,643. Carcinogenesis 18, 2029–2033 (1997).
44
T. Hays et al., Role of peroxisome proliferator-activated receptor-alpha (PPARalpha) in bezafibrate-induced hepatocarcinogenesis and cholestasis. Carcinogenesis 26, 219–227 (2005).
45
J. Nishimura et al., Effect of fenofibrate on oxidative DNA damage and on gene expression related to cell proliferation and apoptosis in rats. Toxicol. Sci. 97, 44–54 (2007).
46
K. Takashima, Y. Ito, F. J. Gonzalez, T. Nakajima, Different mechanisms of DEHP-induced hepatocellular adenoma tumorigenesis in wild-type and Ppar alpha-null mice. J. Occup. Health 50, 169–180 (2008).
47
F. Heindryckx, I. Colle, H. Van Vlierberghe, Experimental mouse models for hepatocellular carcinoma research. Int. J. Exp. Pathol. 90, 367–386 (2009).
48
C. Cheung et al., Diminished hepatocellular proliferation in mice humanized for the nuclear receptor peroxisome proliferator-activated receptor alpha. Cancer Res. 64, 3849–3854 (2004).
49
K. Morimura, C. Cheung, J. M. Ward, J. K. Reddy, F. J. Gonzalez, Differential susceptibility of mice humanized for peroxisome proliferator-activated receptor alpha to Wy-14,643-induced liver tumorigenesis. Carcinogenesis 27, 1074–1080 (2006).
50
M. Das, D. S. Garlick, D. L. Greiner, R. J. Davis, The role of JNK in the development of hepatocellular carcinoma. Genes Dev. 25, 634–645 (2011).
51
M. Das et al., Induction of hepatitis by JNK-mediated expression of TNF-alpha. Cell 136, 249–260 (2009).
52
L. Ye, S. Liu, M. Wang, Y. Shao, M. Ding, High-performance liquid chromatography-tandem mass spectrometry for the analysis of bile acid profiles in serum of women with intrahepatic cholestasis of pregnancy. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 860, 10–17 (2007).
53
L. Barbier-Torres et al., Histone deacetylase 4 promotes cholestatic liver injury in the absence of prohibitin-1. Hepatology 62, 1237–1248 (2015).
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Copyright © 2020 the Author(s). Published by PNAS. This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND).
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Published online: June 29, 2020
Published in issue: July 14, 2020
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Acknowledgments
We thank S. Bartlett for English editing and David Garlick (University of Massachusetts Medical School) for pathological examination of tissue sections. We are grateful to the CNIC Advanced Imaging and Animal facility for technical support. G.S. (RYC-2009-04972) and F.J.C. (RYC-2014-15242) are investigators of the Ramón y Cajal Program. E.M. was awarded a La Caixa fellowship. C.F. was awarded a Sara Borrell contract (CD19/00078). This work was funded by grants supported in part by funds from the European Regional Development Fund: EU’s Seventh Framework Programme (FP7/2007-2013) ERC 260464, EFSD/Lilly European Diabetes Research Programme Dr. Sabio, 2017 Leonardo Grant for Researchers and Cultural Creators, BBVA Foundation (Investigadores-BBVA-2017) IN[17]_BBM_BAS_0066, MINECO-FEDER SAF2016-79126-R, and Comunidad de Madrid IMMUNOTHERCAN-CM S2010/BMD-2326 and B2017/BMD-3733 and La Asociación Española contra el Cáncer (to G.S.); EXOHEP-CM S2017/BMD-3727 and the European Cooperation in Science & Technology (COST) Action CA17112 (to F.J.C.); MINECO Retos SAF2016-78711, the AMMF Cholangiocarcinoma Charity 2018/117, NanoLiver-CM Y2018/NMT-4949, UCM-25-2019, ERAB EA/18-14 (to F.J.C.). F.J.C. is a Gilead Liver Research Scholar. Grant DK R01 DK107220 from the NIH (to R.J.D.); and PI16/00598 from Carlos III Institute of Health, Spain (to J.J.G.M.). The CNIC is supported by the Ministerio de Ciencia, Innovación y Universidades, and the Pro-CNIC Foundation and is a Severo Ochoa Center of Excellence (SEV-2015-0505).
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The authors declare no competing interest.
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JNK-mediated disruption of bile acid homeostasis promotes intrahepatic cholangiocarcinoma, Proc. Natl. Acad. Sci. U.S.A.
117 (28) 16492-16499,
https://doi.org/10.1073/pnas.2002672117
(2020).
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