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Journal of Molecular Medicine

, Volume 97, Issue 11, pp 1589–1600 | Cite as

The PNPLA3 I148M variant promotes lipid-induced hepatocyte secretion of CXC chemokines establishing a tumorigenic milieu

  • Hans Dieter NischalkeEmail author
  • Philipp Lutz
  • Eva Bartok
  • Benjamin Krämer
  • Bettina Langhans
  • Regina Frizler
  • Thomas Berg
  • Jochen Hampe
  • Stephan Buch
  • Christian Datz
  • Felix Stickel
  • Gunther Hartmann
  • Christian P. Strassburg
  • Jacob Nattermann
  • Ulrich Spengler
Original Article

Abstract

The I148M variant of the Patatin-like phospholipase domain-containing 3 (PNPLA3) protein is associated with an increased risk for liver inflammation and hepatocellular carcinoma (HCC), but the underlying mechanism is unknown. We hypothesized that enhanced CXC chemokine secretion mediates hepatic inflammation that accelerates development of HCC. Expandable primary human (upcyte®) hepatocytes and human PLC/PRF/5 hepatoma cells were lentivirally transduced with both PNPLA3 I148M variants and stimulated with lipids. Cytokine levels in culture supernatant and patient sera (n = 80) were analyzed by ELISA. Supernatants were assessed in transmigration experiments, tube formation, and proliferation assays. In vitro, lipid stimulation of transduced hepatocytes dose-dependently induced the production of interleukin-8 and CXCL1 in hepatocytes carrying the PNPLA3 148M variant. In line, sera from PNPLA3 148M-positive patients with alcoholic liver cirrhosis contained higher levels of interleukin-8 and CXCL1 than patients with wild-type PNPLA3. Supernatants from lipid-stimulated hepatocytes with the PNPLA3 148M variant induced enhanced migration of white blood cells, angiogenesis, and cell proliferation in comparison with supernatants from wild-type hepatocytes via CXC receptors 1 and 2. Increased production of interleukin-8 and CXCL1 by hepatocytes carrying the PNPLA3 148M variant contributes to a pro-inflammatory and tumorigenic milieu in patients with alcoholic liver disease.

Key messages

  • The PNPLA3 148M variant is associated with cirrhosis and hepatocellular carcinoma.

  • Lipid stimulation of hepatocytes with this variant induces IL-8 and CXCL1.

  • Supernatants from hepatocytes with this variant promote migration and angiogenesis.

  • Sera from patients with this variant contained enhanced levels of IL-8 and CXCL1.

  • The PNPLA3 148M variant contributes to a tumorigenic milieu via IL-8 and CXCL1.

Keywords

PNPLA3 148M IL-8 CXCL1 cirrhosis HCC alcoholic liver disease rs738409 

Notes

Acknowledgments

We thank Jennifer Söhne, Gudrun Hack, Lisa Behnke, and Saskia Schmitz for their perfect technical assistance.

Funding information

This study was funded by the Deutsche Krebshilfe (70112169) to HDN and US. FS was funded by grant SNF 310030_169196 from the Swiss National Fund. JN received funding by the German Research Foundation (DFG SFB/TRR 57). The funding organizations had no role in the design of the study, and collection, analysis, and interpretation of the data, or writing of the manuscript.

Compliance with ethical standards

The study protocol was approved by the ethics committee of the University of Bonn (number 351/15) and complied with the ethical guidelines given by the Declaration of Helsinki. Written informed consent was given from all patients.

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

109_2019_1836_MOESM1_ESM.docx (548 kb)
ESM 1 (DOCX 547 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Hans Dieter Nischalke
    • 1
    Email author
  • Philipp Lutz
    • 1
  • Eva Bartok
    • 2
  • Benjamin Krämer
    • 1
  • Bettina Langhans
    • 1
  • Regina Frizler
    • 1
  • Thomas Berg
    • 3
  • Jochen Hampe
    • 4
  • Stephan Buch
    • 4
  • Christian Datz
    • 5
  • Felix Stickel
    • 6
  • Gunther Hartmann
    • 2
  • Christian P. Strassburg
    • 1
  • Jacob Nattermann
    • 1
  • Ulrich Spengler
    • 1
  1. 1.Department of Internal Medicine I, University HospitalUniversity of BonnBonnGermany
  2. 2.Institute of Clinical Chemistry and Clinical Pharmacology, University HospitalUniversity of BonnBonnGermany
  3. 3.Department of GastroenterologyUniversity Hospital LeipzigLeipzigGermany
  4. 4.Medical Department 1, University Hospital DresdenTU DresdenDresdenGermany
  5. 5.Department of Internal Medicine, Hospital OberndorfTeaching Hospital of the Paracelsus Private University of SalzburgSalzburgAustria
  6. 6.Department of Gastroenterology and HepatologyUniversity Hospital of ZürichZürichSwitzerland

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