Original ResearchFull Report: Basic and Translational—LiverEstrogen Activation of G-Protein–Coupled Estrogen Receptor 1 Regulates Phosphoinositide 3-Kinase and mTOR Signaling to Promote Liver Growth in Zebrafish and Proliferation of Human Hepatocytes
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Section snippets
Zebrafish Husbandry
Zebrafish were maintained according to Institutional Animal Care and Use Committee guidelines at Harvard Medical School.
Chemical Exposures
Zebrafish larvae were exposed to chemical modifiers, dissolved in 0.1% (volume/volume) dimethyl sulfoxide (DMSO) for 5 hours from 110 to 115 hours postfertilization (hpf), and analyzed at 120 hpf (unless otherwise specified).11 Adults were treated for 5–7 hours per exposure. Chemicals are listed in Supplementary Table 1.
Morpholino Injection
Morpholino oligonucleotides designed against esr1, esr2a,
Estrogen Enhances Liver Growth
To demonstrate the potential impact of endogenous estrogen levels on liver size, adult zebrafish livers were assessed in females and males. Livers of adult female zebrafish were bigger than those of male siblings, as visualized in hepatocyte reporter fish expressing green fluorescent protein (GFP) regulated by the fatty acid binding protein 10a promoter, Tg(fabp10a:GFP) (Figure 1A). Baseline liver-to-body weight ratios in 10 WT males and females at 7 months showed a significant 2.6-fold
Discussion
Liver cancer is the second most common cause of cancer death worldwide, and it predominantly affects men.2 Here, we identified a novel role for GPER1 in hepatocytes that cell-autonomously regulates larval liver growth and contributes to the sex dimorphisms observed in liver cancer incidence and growth. E2 promotes hepatocyte cell cycle progression and proliferation through GPER1 and downstream activation of PI3K-mTOR signaling. Importantly, the GPER1-mTOR pathway remains essential for liver
Acknowledgments
The authors thank Isaac Oderberg from the Goessling lab for creating the visual abstract, John Asara at the Metabolomics Core at the Beth Israel Deaconess Medical Center for assistance with the metabolomics experiments, and staff of Beth Israel Deaconess zebrafish core facility for fish husbandry.
Author contributions: Saireudee Chaturantabut and Wolfram Goessling conceived the study, designed experiments, and analyzed data. Saireudee Chaturantabut conducted all zebrafish experiments. Arkadi
References (52)
- et al.
Estrogen receptors and the metabolic network
Cell Metab
(2011) - et al.
Estrogen defines the dorsal-ventral limit of VEGF regulation to specify the location of the hemogenic endothelial niche
Dev Cell
(2014) - et al.
Genetic interaction of PGE2 and Wnt signaling regulates developmental specification of stem cells and regeneration
Cell
(2009) - et al.
APC mutant zebrafish uncover a changing temporal requirement for wnt signaling in liver development
Dev Biol
(2008) - et al.
AKT/PKB signaling: navigating downstream
Cell
(2007) - et al.
mTOR signaling in growth control and disease
Cell
(2012) The regulation of cell size
Cell
(2013)- et al.
Increased 4E-BP1 expression protects against diet-induced obesity and insulin resistance in male mice
Cell Rep
(2016) - et al.
G protein-coupled estrogen receptor deficiency accelerates liver tumorigenesis by enhancing inflammation and fibrosis
Cancer Lett
(2016) - et al.
High-dose tamoxifen in the treatment of inoperable hepatocellular carcinoma: a multicenter randomized controlled trial
Hepatology
(2002)
Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012
Int J Cancer
Sex difference in alcohol-related organ injury
Alcohol Clin Exp Res
Sex hormone imbalance in male alcoholic cirrhotic patients with and without hepatocellular carcinoma
Cancer
Local estrogen formation by nontumoral, cirrhotic, and malignant human liver tissues and cells
Cancer Res
Hormonal and enzymatic parameters of hepatic regeneration in patients undergoing major liver resections
Hepatology
Estrogen-induced activation of Erk-1 and Erk-2 requires the G protein-coupled receptor homolog, GPR30, and occurs via trans-activation of the epidermal growth factor receptor through release of HB-EGF
Mol Endocrinol
Regulatory role of G protein-coupled estrogen receptor for vascular function and obesity
Circ Res
Activation of a novel estrogen receptor, GPER, is cardioprotective in male and female rats
Am J Physiol Heart Circ Physiol
The G protein-coupled estrogen receptor 1 (GPER1/GPR30) agonist G-1 regulates vascular smooth muscle cell Ca2⁺ handling
J Vasc Res
Targeted gene disruption in somatic zebrafish cells using engineered TALENs
Nat Biotechnol
Aberrant ligand-induced activation of G protein-coupled estrogen receptor 1 (GPER) results in developmental malformations during vertebrate embryogenesis
Toxicol Sci
PGE2-regulated wnt signaling and N-acetylcysteine are synergistically hepatoprotective in zebrafish acetaminophen injury
Proc Natl Acad Sci U S A
Yap reprograms glutamine metabolism to increase nucleotide biosynthesis and enable liver growth
Nat Cell Biol
Neoplasia in zebrafish (Danio rerio) treated with 7,12-dimethylbenz[a]anthracene by two exposure routes at different developmental stages
Toxicol Pathol
A positive/negative ion-switching, targeted mass spectrometry-based metabolomics platform for bodily fluids, cells, and fresh and fixed tissue
Nat Protoc
Antagonists selective for estrogen receptor alpha
Endocrinology
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Conflicts of interest The authors disclose the following: Wolfram Goessling is a consultant and Leonard I. Zon is a founder of Camp4 Therapeutics which develops treatments for liver diseases.
Funding This study is supported by National Institutes of Health 5K08CA172288 (to Kimberley J. Evason), R24OD017870 (to Leonard I. Zon and Wolfram Goessling), R37DK048873, R01DK056626 and R01 DK103046 (to David E. Cohen) and R01DK090311 and R01DK105198 (to Wolfram Goessling), and the Claudia Adams Barr Program in Cancer Research (to Wolfram Goessling). Wolfram Goessling is a Pew Scholar in the Biomedical Sciences.
Author names in bold designate shared co-first authorship.