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Cited by 23Liver physiology and pathologyCancer-related Molecular PathwaysGene Regulatory Network Analysis

T160-phosphorylated CDK2 defines threshold for HGF-dependent proliferation in primary hepatocytes

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Mueller, Stephanie, Huard, Jérémy, Waldow, Katharina, Huang, Xiaoyun, D'Alessandro, Lorenza A, Bohl, Sebastian, Börner, Kathleen, Grimm, Dirk +3 more·2015·Molecular Systems Biology
DOI 10.15252/msb.20156032PMC PMC4380929
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Abstract

Liver regeneration is a tightly controlled process mainly achieved by proliferation of usually quiescent hepatocytes. The specific molecular mechanisms ensuring cell division only in response to proliferative signals such as hepatocyte growth factor (HGF) are not fully understood. Here, we combined quantitative time‐resolved analysis of primary mouse hepatocyte proliferation at the single cell and at the population level with mathematical modeling. We showed that numerous G1/S transition components are activated upon hepatocyte isolation whereas DNA replication only occurs upon additional HGF stimulation. In response to HGF, Cyclin:CDK complex formation was increased, p21 rather than p27 was regulated, and Rb expression was enhanced. Quantification of protein levels at the restriction point showed an excess of CDK2 over CDK4 and limiting amounts of the transcription factor E2F‐1. Analysis with our mathematical model revealed that T160 phosphorylation of CDK2 correlated best with growth factor‐dependent proliferation, which we validated experimentally on both the population and the single cell level. In conclusion, we identified CDK2 phosphorylation as a gate‐keeping mechanism to maintain hepatocyte quiescence in the absence of HGF.

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  • The ratio of cytoplasmic to nuclear volume of primary mouse hepatocytes is 12.67/0.5 pL for frequently binucleated hepatocytes.

    “The previously determined (Mueller et al., 2015) ratio of cytoplasmic to nuclear volume of primary mouse hepatocytes (12.67/0.5 pL, for frequently binucleated hepatocytes) facilitated modeling of concentration changes due to inter-compartmental transport processes.”

    From Model Based Targeting of IL-6-Induced Inflammatory Responses in Cultured Primary Hepatocytes to Improve Application of the JAK Inhibitor Ruxolitinib (2017) · cites this paper as Mueller et al., 2015

    The Results section (Analysis of G1/S phase progression and abundance of pathway proteins; page unavailable) says the authors measured a total primary mouse hepatocyte volume of 12.92 ± 0.62 pL by confocal microscopy. Our copy does not give the claimed 12.67/0.5 pL cytoplasmic-to-nuclear breakdown or tie those values to binucleated hepatocytes. (LLM, conf=0.82)

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Authors
Mueller, S., Huard, J., Waldow, K., Huang, X., D'Alessandro, L. A., Bohl, S., Börner, K., Grimm, D., Klamt, S., Klingmüller, U., & Schilling, M.
Affiliations
  • German Cancer Research Center (DE)
  • Max Planck Society (DE)
  • University Hospital Heidelberg (DE)
Corresponding author
  • Klingmüller, Ursula
  • Schilling, Marcel
ORCID
Venue
Molecular Systems Biology
Publisher
Springer Science and Business Media LLC
Published
2015-03-01
Volume
11
Issue
3
Pages
795-795
ISSN
1744-4292 (ISSN-L)
DOI
10.15252/msb.20156032
PMC
PMC4380929
OpenAlex
W2098282516
Cited by
23
References
82
Citations by year
2025: 3 · 2024: 3 · 2023: 1 · 2022: 3 · 2021: 1 · 2019: 2 · 2018: 1 · 2017: 5 · 2016: 4
FWCI
1.98
Type
journal article
Language
en
Open access
Licence
Locations
  • Molecular Systems Biology (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
  • MPG.PuRe (Max Planck Society) (submittedVersion, other-oa)
Funders
  • Virtual Liver Network initiative of the German Federal Ministry of Education and Research
  • IMI/EFPIA
  • MIP‐DILI Consortium
  • German Center for Lung Research
  • German Center for Infection Research
Topics
  • Health Sciences › Medicine › Hepatology › Liver physiology and pathology
  • Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene Regulatory Network Analysis
Keywords
hepatocyte proliferation, hepatocyte growth factor, liver regeneration, hepatocyte quiescence, G1/S transition, restriction point, cyclin-CDK complexes, p21, p27, Rb, E2F1, single-cell analysis
MeSH terms
Animals, DNA Replication, Gene Expression Regulation, Male, Models, Theoretical, Phosphorylation, Tyrosine, Hepatocyte Growth Factor, Hepatocytes, Cell Proliferation, Cyclin-Dependent Kinase 2, Mice, Single-Cell Analysis, Primary Cell Culture

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