Yes-associated protein is crucial for constitutive androstane receptor-driven hepatocyte proliferation but not for induction of drug metabolism genes in mice
Open accessAbstract
Background and Aims: Constitutive androstane receptor (CAR) agonists, such as 1,4‐bis [2‐(3,5‐dichloropyridyloxy)] benzene (TCPOBOP), are known to cause robust hepatocyte proliferation and hepatomegaly in mice along with induction of drug metabolism genes without any associated liver injury. Yes‐associated protein (Yap) is a key transcription regulator that tightly controls organ size, including that of liver. Our and other previous studies suggested increased nuclear localization and activation of Yap after TCPOBOP treatment in mice and the potential role of Yap in CAR‐driven proliferative response. Here, we investigated a direct role of Yap in CAR‐driven hepatomegaly and hepatocyte proliferation using hepatocyte‐specific Yap‐knockout (KO) mice. Approach and Results: Adeno‐associated virus 8‐thyroxine binding globulin promoter‐Cre recombinase vector was injected to Yap‐floxed mice for achieving hepatocyte‐specific Yap deletion followed by TCPOBOP treatment. Yap deletion did not decrease protein expression of CAR or CAR‐driven induction of drug metabolism genes (including cytochrome P450 [Cyp] 2b10, Cyp2c55, and UDP‐glucuronosyltransferase 1a1 [Ugt1a1]). However, Yap deletion substantially reduced TCPOBOP‐induced hepatocyte proliferation. TCPOBOP‐driven cell cycle activation was disrupted in Yap‐KO mice because of delayed (and decreased) induction of cyclin D1 and higher expression of p21, resulting in decreased phosphorylation of retinoblastoma protein. Furthermore, the induction of other cyclins, which are sequentially involved in progression through cell cycle (including cyclin E1, A2, and B1), and important mitotic regulators (such as Aurora B kinase and polo‐like kinase 1) was remarkably reduced in Yap‐KO mice. Microarray analysis revealed that 26% of TCPOBOP‐responsive genes that were mainly related to proliferation, but not to drug metabolism, were altered by Yap deletion. Yap regulated these proliferation genes through alerting expression of Myc and forkhead box protein M1, two critical transcriptional regulators of CAR‐mediated hepatocyte proliferation. Conclusions: Our study revealed an important role of Yap signaling in CAR‐driven hepatocyte proliferation; however, CAR‐driven induction of drug metabolism genes was independent of Yap.
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c-Myc and CAR are metabolic regulators that modulate expression of genes involved in lipid and glucose homeostasis.
“In addition to their roles in proliferation, both c-Myc and CAR are metabolic regulators that modulate the expression of genes involved in lipid and glucose homeostasis.22,48, 49, 50, 51, 52”
From Constitutive Androstane Receptor Regulates Bile Acid Homeostasis and Metabolism During Liver Regeneration (2026) · cites this paper as [22]
The paper’s opening background (indexed under Abstract > Conclusions; page unavailable) calls CAR a regulator of liver energy metabolism, and the Results section on HNF4α reports CAR–HNF4α transcriptional cross-talk. Its discussion of Myc and CAR gene regulation focuses on proliferation; our copy does not establish that both regulate genes involved in lipid and glucose homeostasis. (LLM, conf=0.84)
Experimental — accuracy not yet measuredCAR's pro-proliferative role in liver is mediated through induction of c-Myc, Foxm1, and cyclin D1, and modulation of β-catenin and YAP signaling pathways.
“Prior studies have focused primarily on the pro-proliferative role of CAR in the liver, mediated through the induction of factors such as cellular-myelocytomatosis oncogene (c-Myc),15 forkhead box m1 (Foxm1),15 and cyclin D1 (Ccnd1),16 as well as modulation of β-catenin17,18 and Yes-associated protein19, 20, 21, 22 signaling pathways.”
From Constitutive Androstane Receptor Regulates Bile Acid Homeostasis and Metabolism During Liver Regeneration (2026) · cites this paper as [22]
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References 33 · Cited by 20
Research works printed but not linked: 25
- [1]Kazantseva YA, Pustylnyak YA, Pustylnyak VO. Role of nuclear constitutive androstane receptor in regulation of hepatocyte proliferation and hepatocarcinogenesis. Biochemistry (Mosc) 2016; 81: 338‐ 347. 27293091
- [4]Tschuor C, Kachaylo E, Limani P, Raptis DA, Linecker M, Tian Y, et al. Constitutive androstane receptor (Car)‐driven regeneration protects liver from failure following tissue loss. J Hepatol 2016; 65: 66‐ 74. 26948495
- [5]Ledda‐Columbano GM, Pibiri M, Loi R, Perra A, Shinozuka H, Columbano A, et al. Early increase in cyclin‐D1 expression and accelerated entry of mouse hepatocytes into S phase after administration of the mitogen 1, 4‐Bis[2‐(3,5‐Dichloropyridyloxy)] benzene. Am J Pathol 2000; 156: 91‐ 97. 10623657 PMC1868640
- [6]Dong J, Feldmann G, Huang J, Wu S, Zhang N, Comerford SA, et al. Elucidation of a universal size‐control mechanism in Drosophila and mammals. Cell 2007; 130: 1120‐ 1133. 17889654 PMC2666353
- [9]Camargo FD, Gokhale S, Johnnidis JB, Fu D, Bell GW, Jaenisch R, et al. YAP1 increases organ size and expands undifferentiated progenitor cells. Curr Biol 2007; 17: 2054‐ 2060. 17980593
- [10]Zhou D, Conrad C, Xia F, Park JS, Payer B, Yin Y, et al. Mst1 and Mst2 maintain hepatocyte quiescence and suppress hepatocellular carcinoma development through inactivation of the Yap1 oncogene. Cancer Cell 2009; 16: 425‐ 438. 19878874 PMC3023165
- [11]Zhang N, Bai H, David KK, Dong J, Zheng Y, Cai J, et al. The Merlin/NF2 tumor suppressor functions through the YAP oncoprotein to regulate tissue homeostasis in mammals. Dev Cell 2010; 19: 27‐ 38. 20643348 PMC2925178
- [12]Benhamouche S, Curto M, Saotome I, Gladden AB, Liu CH, Giovannini M, et al. Nf2/Merlin controls progenitor homeostasis and tumorigenesis in the liver. Genes Dev 2010; 24: 1718‐ 1730. 20675406 PMC2922501
- [13]Lu L, Finegold MJ, Johnson RL. Hippo pathway coactivators Yap and Taz are required to coordinate mammalian liver regeneration. Exp Mol Med 2018; 50: e423. 29303509 PMC5992983
- [15]Abe T, Amaike Y, Shizu R, Takahashi M, Kano M, Hosaka T, et al. Role of YAP activation in nuclear receptor CAR‐mediated proliferation of mouse hepatocytes. Toxicol Sci 2018; 165: 408‐ 419. 29893953
- [16]Gibault F, Corvaisier M, Bailly F, Huet G, Melnyk P, Cotelle P. Non‐photoinduced biological properties of verteporfin. Curr Med Chem 2016; 23: 1171‐ 1184. 26980565
- [17]Ledda‐Columbano GM, Pibiri M, Concas D, Molotzu F, Simbula G, Cossu C, et al. Sex difference in the proliferative response of mouse hepatocytes to treatment with the CAR ligand. TCPOBOP. Carcinogenesis 2003; 24: 1059‐ 1065. 12807759
- [18]Tian J, Huang H, Hoffman B, Liebermann DA, Ledda‐Columbano GM, Columbano A, et al. Gadd45β is an inducible coactivator of transcription that facilitates rapid liver growth in mice. J Clin Invest 2011; 121: 4491‐ 502. 21965327 PMC3204825
- [20]Costa RH, Kalinichenko VV, Tan Y, Wang IC. The CAR nuclear receptor and hepatocyte proliferation. Hepatology 2005; 42: 1004‐ 1008. 16250031
- [22]Kazantseva YA, Yarushkin AA, Mostovich LA, Pustylnyak YA, Pustylnyak VO. Xenosensor CAR mediates down‐regulation of miR‐122 and up‐regulation of miR‐122 targets in the liver. Toxicol Appl Pharmacol 2015; 288: 26‐ 32. 26171734
- [23]Walesky C, Apte U. Role of hepatocyte nuclear factor 4α (HNF4α) in cell proliferation and cancer. Gene Expr 2015; 16: 101‐ 108. 25700366 PMC5841246
- [24]Walesky C, Edwards G, Borude P, Gunewardena S, O’Neil M, Yoo B, et al. Hepatocyte nuclear factor 4 alpha deletion promotes diethylnitrosamine‐induced hepatocellular carcinoma in rodents. Hepatology 2013; 57: 2480‐ 2490. 23315968 PMC3669646
- [25]Walesky C, Gunewardena S, Terwilliger EF, Edwards G, Borude P, Apte U. Hepatocyte‐specific deletion of hepatocyte nuclear factor‐4α in adult mice results in increased hepatocyte proliferation. Am J Physiol Gastrointest Liver Physiol 2013; 304: G26‐ G37. 23104559 PMC3543634
- [26]Miao J, Fang S, Bae Y, Kemper JK. Functional inhibitory cross‐talk between constitutive androstane receptor and hepatic nuclear factor‐4 in hepatic lipid/glucose metabolism is mediated by competition for binding to the DR1 motif and to the common coactivators, GRIP‐1 and PGC‐1alpha. J Biol Chem 2006; 281: 14537‐ 14546. 16492670
- [27]Yarushkin AA, Kachaylo EM, Pustylnyak VO. The constitutive androstane receptor activator 4‐[(4R,6R)‐4,6‐diphenyl‐1,3‐dioxan‐2‐yl]‐N, N‐dimethylaniline inhibits the gluconeogenic genes PEPCK and G6Pase through the suppression of HNF4alpha and FOXO1 transcriptional activity. Br J Pharmacol 2013; 168: 1923‐ 1932. 23231652 PMC3623062
- [28]Tian J, Marino R, Johnson C, Locker J. Binding of drug‐activated CAR/Nr1i3 alters metabolic regulation in the liver. iScience 2018; 9: 209‐ 228. 30396153 PMC6222290
- [29]Croci O, De Fazio S, Biagioni F, Donato E, Caganova M, Curti L, et al. Transcriptional integration of mitogenic and mechanical signals by Myc and YAP. Genes Dev 2017; 31: 2017‐ 2022. 29141911 PMC5733494
- [30]Cho SW, Xu J, Sun R, Mumbach MR, Carter AC, Chen YG, et al. Promoter of lncRNA gene PVT1 is a tumor‐suppressor DNA boundary element. Cell 2018; 173: 1398‐ 1412.e1322. 29731168 PMC5984165
- [31]Uslu VV, Petretich M, Ruf S, Langenfeld K, Fonesca NA, Marioni JC et al. Long‐range enhancers regulating Myc expression are required for normal facial morphogenesis. Nat Genet 2014; 46: 753‐ 758. 24859337
- [33]Grijalva JL, Huizenga M, Mueller K, Rodriguez S, Brazzo J, Camargo F, Sadri‐Vakili G, et al. Dynamic alterations in Hippo signaling pathway and YAP activation during liver regeneration. Am J Physiol Gastrointest Liver Physiol 2014; 307: G196‐ G204. 24875096
- Authors
- Bhushan, B., Molina, L., Koral, K., Stoops, J. W., Mars, W. M., Banerjee, S., Orr, A., Paranjpe, S., Monga, S. P., Locker, J., & Michalopoulos, G. K.
- Affiliations
- University of Pittsburgh (US)
- Corresponding author
- Michalopoulos, George K.
- ORCID
- Venue
- Hepatology
- Published
- 2021-04-29
- Volume
- 73
- Issue
- 5
- Pages
- 2005-2022
- ISSN
- 0270-9139 (ISSN-L)
- 1527-3350
- PMC
- PMC7885729
- OpenAlex
- W3049375741
- Cited by
- 20
- References
- 33
- Citations by year
- 2026: 1 · 2025: 5 · 2024: 1 · 2023: 4 · 2022: 4 · 2021: 5
- FWCI
- 1.62
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Hepatology (publishedVersion, cc-by-nc)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- Funders
- Cleveland Foundation
- National Cancer Institute (R01CA204586)
- National Institute of Diabetes and Digestive and Kidney Diseases (R01 DK62277, F30DK121393, P30DK120531, R01DK062277)
- National Institute of Biomedical Imaging and Bioengineering (T32EB001026)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Hippo pathway signaling and YAP/TAZ
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
- Keywords
- YAP, constitutive androstane receptor, hepatocyte proliferation, hepatomegaly, TCPOBOP, cell cycle regulation, cyclin D1, p21, retinoblastoma protein phosphorylation, c-Myc
- MeSH terms
- Constitutive Androstane Receptor, YAP-Signaling Proteins, Animals, Cell Cycle, Female, Gene Expression Regulation, Genes, Humans, Liver Regeneration, Inactivation, Metabolic, Mice, Knockout, Hepatocytes, Cell Proliferation, Mice, Transcriptome
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