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Cited by 111Liver physiology and pathologyWnt/β-catenin signaling in development and cancerDrug-Induced Hepatotoxicity and Protection

Tissue Repair in the Mouse Liver Following Acute Carbon Tetrachloride Depends on Injury‐Induced Wnt/β‐Catenin Signaling

Open access
Zhao, Ludan, Jin, Yinhua, Donahue, Katie, Tsui, Margaret, Fish, Matt, Logan, Catriona Y., Wang, Bruce, Nusse, Roel·2019·Hepatology
DOI 10.1002/hep.30563PMC PMC7043939

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Abstract

In the liver, Wnt/β‐catenin signaling is involved in regulating zonation and hepatocyte proliferation during homeostasis. We examined Wnt gene expression and signaling after injury, and we show by in situ hybridization that Wnts are activated by acute carbon tetrachloride (CCl 4 ) toxicity. Following injury, peri‐injury hepatocytes become Wnt‐responsive, expressing the Wnt target gene axis inhibition protein 2 (Axin2). Lineage tracing of peri‐injury Axin2 + hepatocytes shows that during recovery the injured parenchyma becomes repopulated and repaired by Axin2 + descendants. Using single‐cell RNA sequencing, we show that endothelial cells are the major source of Wnts following acute CCl 4 toxicity. Induced loss of β‐catenin in peri‐injury hepatocytes results in delayed repair and ultimately injury‐induced lethality, while loss of Wnt production from endothelial cells leads to a delay in the proliferative response after injury. Conclusion: Our findings highlight the importance of the Wnt/β‐catenin signaling pathway in restoring tissue integrity following acute liver toxicity and establish a role of endothelial cells as an important Wnt‐producing regulator of liver tissue repair following localized liver injury.

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  • In mouse and human liver, Wnt ligands are produced by endothelial cells of central vein branches and nearby sinusoids.

    “In the mouse and human liver, Wnt ligands are produced by endothelial cells of the central vein branches and the nearby sinusoids (17–19)”

    From Wnt signaling regulates hepatocyte cell division by a transcriptional repressor cascade (2022) · cites this paper as [19]

    The Results section (“Upregulation of endogenous and exogenous Wnt ligands”; page not supplied) says that Wnt2 and Wnt9b are expressed by central-vein endothelial cells and that Wnt2 is also expressed by sinusoidal endothelial cells. The paper’s experiments concern mouse liver; our copy does not establish the same claim for human liver. (LLM, conf=0.87)

    Experimental — accuracy not yet measured

  • Following ablation of pericentral hepatocytes, neighbouring zone 2 hepatocytes upregulated WNT/β-catenin signalling and acquired zone 3 identity to restore metabolic zonation.

    “Following ablation of pericentral hepatocytes, neighbouring zone 2 hepatocytes upregulated WNT/β-catenin signalling and acquired zone 3 identity to restore metabolic zonation.85,104,105”

    From Distinct hepatocyte identities in liver homeostasis and regeneration (2023) · cites this paper as [105]

    The Results (‘Injury induces de novo Wnt response in mid-lobular hepatocytes’; page unavailable) show that hepatocytes near the injury border become Wnt-responsive after pericentral cell loss. The lineage-tracing Results show their descendants repopulate the damaged centrilobular area, and GS and Glt1 expression patterns return by day 14. The paper does not establish that the Wnt-responsive mid-lobular hepatocytes themselves acquired zone 3 identity; during repair, it describes them as distinct from pericentral hepatocytes because they do not express those markers. (LLM, conf=0.84)

    Experimental — accuracy not yet measured

  • In mouse and human liver, Wnt ligands are produced by endothelial cells of central vein branches and nearby sinusoids, creating a zonated expression pattern of Wnt target genes and key metabolic enzymes in pericentral hepatocytes.

    “In the mouse and human liver, Wnt ligands are produced by endothelial cells of the central vein branches and the nearby sinusoids (McEnerney et al. 2017;Wang et al. 2015;Zhao et al. 2019), creating a zonated expression pattern of Wnt target genes and key metabolic enzymes in pericentral hepatocytes (Halpern et al. 2017).”

    From Hepatocyte Cell Cycle Progression Depends on a Transcriptional Repressor Cascade Downstream of Wnt Signaling (2021) · cites this paper as Zhao et al. 2019

    The Results section (‘Upregulation of endogenous and exogenous Wnt ligands following injury’; page unavailable) says that Wnt2 and Wnt9b are expressed by central-vein endothelial cells in uninjured liver, and Wnt2 by sinusoidal endothelial cells. The paper’s observations concern mouse liver; our copy does not establish the same endothelial Wnt production in human liver. The subsequent zonated-expression clause is attributed to Halpern et al., not Zhao et al. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • Tbx3 is expressed in a zonated pattern during postnatal liver growth.

    “The transcriptional repressor Tbx3, a gene required for embryonic liver development (Khan et al. 2020;Ludtke et al. 2009;Suzuki et al. 2008), is also expressed in a zonated pattern during postnatal liver growth (fig.S1A) (Wang et al. 2015;Zhao et al. 2019).”

    From Hepatocyte Cell Cycle Progression Depends on a Transcriptional Repressor Cascade Downstream of Wnt Signaling (2021) · cites this paper as Zhao et al. 2019

    The Results section ('Injury induces de novo Wnt response in mid-lobular hepatocytes'; page unavailable) describes a zonated, pericentral Tbx3 expression pattern in the uninjured adult liver and its return after injury. It does not establish that this pattern occurs during postnatal liver growth, the stage specified in the claim. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

  • Hepatocyte proliferation has previously been considered a major driver of hepatic wound closure following necro-inflammatory liver injury.

    “Hepatocyte proliferation has previously been considered a major driver of hepatic wound closure following necro-inflammatory liver injury2,24,25”

    From Multimodal decoding of human liver regeneration (2024) · cites this paper as [25]

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Research works printed but not linked: 35

  1. [1]Malato Y, Naqvi S, Schurmann N, Ng R, Wang B, Zape J, Fate tracing of mature hepatocytes in mouse liver homeostasis and regeneration. J. Clin. Invest. 2011; 121: 4850– 4860. 22105172 PMC3226005
  2. [2]Yanger K, Knigin D, Zong Y, Maggs L, Gu G, Akiyama H, Adult Hepatocytes Are Generatedby Self-DuplicationRather than Stem Cell Differentiation. Stem Cell. 2014;: 1– 22. PMC4505916 25130492
  3. [3]Newsome PN, Hussain MA, Theise ND. Hepatic oval cells: helping redefine a paradigm in stem cell biology. Curr. Top. Dev. Biol. 2004; 61: 1– 28. 15350395
  4. [4]Huch M, Dorrell C, Boj SF, Van Es JH, Li VSW, van de Wetering M, In vitro expansion of single Lgr5+ liver stem cells induced by Wnt-driven regeneration. Nature. 2013; 494: 247– 250. 23354049 PMC3634804
  5. [5]Tanimizu N, Mitaka T. Re-evaluation of liver stem/progenitor cells. Organogenesis. 2014; 10: 208– 215. 24451175 PMC4154955
  6. [6]Tarlow BD, Finegold MJ, Grompe M. Clonal tracing of Sox9+ liver progenitors in mouse oval cell injury. Hepatology. 2014; 60: 278– 289. 24700457 PMC4077948
  7. [8]Micsenyi A, Tan X, Sneddon T, Luo J-H, Michalopoulos GK, Monga SPS. Beta-catenin is temporally regulated during normal liver development. Gastroenterology. 2004; 126: 1134– 1146. 15057752
  8. [10]Thompson MD, Monga SPS. WNT/β-catenin signaling in liver health and disease. Hepatology. 2007; 45: 1298– 1305. 17464972
  9. [11]Tan X, Yuan Y, Zeng G, Apte U, Thompson MD, Cieply B, Beta-catenin deletion in hepatoblasts disrupts hepatic morphogenesis and survival during mouse development. Hepatology. 2008; 47: 1667– 1679. 18393386 PMC4449338
  10. [13]Yang J, Mowry LE, Nejak-Bowen KN, Okabe H, Diegel CR, Lang RA, Beta-catenin signaling in murine liver zonation and regeneration: A Wnt-Wnt situation! Hepatology. 2014; 60: 964– 976. 24700412 PMC4139486
  11. [14]Sekine S, Gutiérrez PJA, Yu-Ang Lan B, Feng S, Hebrok M. Liver-specific loss of β-catenin results in delayed hepatocyte proliferation after partial hepatectomy. Hepatology. 2007; 45: 361– 368. 17256747
  12. [15]Nejak-Bowen KN, Thompson MD, Singh S, Bowen WC, Dar MJ, Khillan J, Accelerated liver regeneration and hepatocarcinogenesis in mice overexpressing serine-45 mutant beta-catenin. Hepatology. 2010; 51: 1603– 1613. 20432254 PMC2908905
  13. [16]Ding B-S, Nolan DJ, Butler JM, James D, Babazadeh AO, Rosenwaks Z, Inductive angiocrine signals from sinusoidal endothelium are required for liver regeneration. Nature. 2010; 468: 310– 315. 21068842 PMC3058628
  14. [18]Rocha AS, Vidal V, Mertz M, Kendall TJ, Charlet A, Okamoto H, The Angiocrine Factor Rspondin3 Is a Key Determinant of Liver Zonation. CellReports. 2015; 13: 1757– 1764. 26655896
  15. [19]Lustig B, Jerchow B, Sachs M, Weiler S, Pietsch T, Karsten U, Negative Feedback Loop of Wnt Signaling through Upregulation of Conductin/Axin2 in Colorectal and Liver Tumors. Molecular and Cellular Biology. 2002; 22: 1184– 1193. 11809809 PMC134640
  16. [20]van Amerongen R, Bowman AN, Nusse R. Developmental stage and time dictate the fate of Wnt/β-catenin-responsive stem cells in the mammary gland. Cell Stem Cell. 2012; 11: 387– 400. 22863533 PMC13155203
  17. [21]Madisen L, Zwingman TA, Sunkin SM, Oh SW, Zariwala HA, Gu H, A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat. Neurosci. 2010; 13: 133– 140. 20023653 PMC2840225
  18. [22]Brault V, Moore R, Kutsch S, Ishibashi M, Rowitch DH, McMahon AP, Inactivation of the beta-catenin gene by Wnt1-Cre-mediated deletion results in dramatic brain malformation and failure of craniofacial development. Development. 2001; 128: 1253– 1264. 11262227
  19. [23]Carpenter AC, Rao S, Wells JM, Campbell K, Lang RA. Generation of mice with a conditional null allele for Wntless. Genesis. 2010; 48: 554– 558. 20614471 PMC3689319
  20. [24]F Schwenk UBKR. A cre-transgenic mouse strain for the ubiquitous deletion of loxP-flanked gene segments including deletion in germ cells. Nucleic Acids Research. 1995; 23: 5080. 8559668 PMC307516
  21. [25]Wang Y, Nakayama M, Pitulescu ME, Schmidt TS, Bochenek ML, Sakakibara A, Ephrin-B2 controls VEGF-induced angiogenesis and lymphangiogenesis. Nature. 2010; 465: 483– 486. 20445537
  22. [26]Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001; 25: 402– 408. 11846609
  23. [28]Ghafoory S, Breitkopf-Heinlein K, Li Q, Scholl C, Dooley S, Wölfl S. Zonation of nitrogen and glucose metabolism gene expression upon acute liver damage in mouse. PLoS ONE. 2013; 8: e78262. 24147127 PMC3798318
  24. [29]Jho E-H, Zhang T, Domon C, Joo C-K, Freund J-N, Costantini F. Wnt/beta-catenin/Tcf signaling induces the transcription of Axin2, a negative regulator of the signaling pathway. Molecular and Cellular Biology. 2002; 22: 1172– 1183. 11809808 PMC134648
  25. [30]Ding B-S, Cao Z, Lis R, Nolan DJ, Guo P, Simons M, Divergent angiocrine signals from vascular niche balance liver regeneration and fibrosis. Nature. 2014; 505: 97– 102. 24256728 PMC4142699
  26. [31]Bhathal PS, Rose NR, Mackay IR, Whittingham S. Strain differences in mice in carbon tetrachloride-induced liver injury. Br J Exp Pathol. 1983; 64: 524– 533. 6639871 PMC2040819
  27. [32]Relaix F, Zammit PS. Satellite cells are essential for skeletal muscle regeneration: the cell on the edge returns centre stage. Development. 2012; 139: 2845– 2856. 22833472
  28. [33]Gregorieff A, Liu Y, Inanlou MR, Khomchuk Y, Wrana JL. Yap-dependent reprogramming of Lgr5+ stem cells drives intestinal regeneration and cancer. Nature. 2015; 526: 715– 718. 26503053
  29. [34]Barker N, Van Es JH, Kuipers J, Kujala P, van den Born M, Cozijnsen M, Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature. 2007; 449: 1003– 1007. 17934449
  30. [35]Yan KS, Chia LA, Li X, Ootani A, Su J, Lee JY, The intestinal stem cell markers Bmi1 and Lgr5 identify two functionally distinct populations. Proceedings of the National Academy of Sciences. 2012; 109: 466– 471. PMC3258636 22190486
  31. [36]Halpern KB, Shenhav R, Massalha H, Toth B, Egozi A, Massasa EE, Paired-cell sequencing enables spatial gene expression mapping of liver endothelial cells. Nat Biotechnol. 2018; 5: 279– 15. PMC6546596 30222169
  32. [37]Hu J, Srivastava K, Wieland M, Runge A, Mogler C, Besemefelder E, Endothelial Cell-Derived Angiopoietin-2Controls Liver Regeneration as aSpatiotemporal Rheostat. Science. 2014; 343: 416– 419. 24458641
  33. [38]Manavski Y, Abel T, Hu J, Kleinlutzum D, Buchholz CJ, Belz C, Endothelial transcription factor KLF2 negatively regulates liver regeneration via induction of activin A. Proceedings of the National Academy of Sciences. 2017; 114: 3993– 3998. PMC5393189 28348240
  34. [39]Gomez-Salinero JM, Rafii S. Endothelial cell adaptation in regeneration. Science. 2018; 362: 1116– 1117. 30523098
  35. [40]Tan X, Behari J, Cieply B, Michalopoulos GK, Monga SPS. Conditional Deletion of β-Catenin Reveals Its Role in Liver Growth and Regeneration. Gastroenterology. 2006; 131: 1561– 1572. 17101329

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Authors
Zhao, L., Jin, Y., Donahue, K., Tsui, M., Fish, M., Logan, C. Y., Wang, B., & Nusse, R.
Affiliations
  • Howard Hughes Medical Institute (US)
  • University of California, San Francisco (US)
  • Institute for Stem Cell Biology and Regenerative Medicine, Department of Developmental Biology,Howard Hughes Medical Institute, Stanford School of Medicine,Stanford,CA
Corresponding author
Wang, Bruce
ORCID
Venue
Hepatology
Publisher
Ovid Technologies (Wolters Kluwer Health)
Published
2019-04-09
Volume
69
Issue
6
Pages
2623-2635
ISSN
  • 0270-9139 (ISSN-L)
  • 1527-3350
DOI
10.1002/hep.30563
PMC
PMC7043939
OpenAlex
W2907359354
Cited by
111
References
5
Citations by year
2026: 8 · 2025: 19 · 2024: 16 · 2023: 16 · 2022: 20 · 2021: 12 · 2020: 15 · 2019: 5
FWCI
8.29
Type
journal article
Language
en
Open access
Licence
Locations
  • Hepatology (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
  • eScholarship (California Digital Library) (submittedVersion)
Funders
  • Howard Hughes Medical Institute
  • Burroughs Wellcome Fund
  • National Institutes of Health
  • University of California, San Francisco
  • National Institute of General Medical Sciences (T32GM007365)
  • National Institute of Diabetes and Digestive and Kidney Diseases (K08DK101603, P30DK026743)
  • Liver Center, University of California, San Francisco
Topics
  • Health Sciences › Medicine › Hepatology › Liver physiology and pathology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Wnt/β-catenin signaling in development and cancer
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
Keywords
Wnt/β-catenin signaling, carbon tetrachloride poisoning, acute liver injury, Axin2, endothelial cells, hepatocyte proliferation, hepatocyte zonation, liver regeneration
MeSH terms
Animals, Biopsy, Needle, Carbon Tetrachloride, Cells, Cultured, Disease Models, Animal, Immunohistochemistry, Liver, Liver Regeneration, Male, Mice, Inbred C57BL, Random Allocation, Reference Values, RNA, Gene Expression, Polymerase Chain Reaction, Hepatocytes, Wnt Proteins, beta Catenin, Mice, Wnt Signaling Pathway, Axin Protein

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