Pigment Epithelium-Derived Factor (PEDF) Inhibits Wnt/β-catenin Signaling in the Liver

• Published in: Cellular and molecular gastroenterology and hepatology Vol. 1; no. 5; pp. 535 - 549.e14
• Main Authors: Protiva, Petr, Gong, Jingjing, Sreekumar, Bharath, Torres, Richard, Zhang, Xuchen, Belinsky, Glenn S, Cornwell, Mona, Crawford, Susan E, Iwakiri, Yasuko, Chung, Chuhan
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.09.2015; Elsevier
• Subjects: Extracellular Matrix; Gastroenterology and Hepatology; Original Research; PEDF; Wnt/β-Catenin; conditioned medium; extracellular signal-regulated kinase; small interfering RNA; LRP6; PEDF; SHG; FDR; second harmonic generation; glyceraldehyde-3-phosphate dehydrogenase; WT; benzyl alcohol/benzyl benzoate; Gene Ontology; GSK; hepatocellular carcinoma; polymerase chain reaction; knockout; BABB; Extracellular Matrix; KO; GO; GSEA; HCC; CM; siRNA; pigment epithelium-derived factor; ECM; false-discovery rate; low-density lipoprotein receptor-related protein 6; glycogen synthase kinase; gene set enrichment analysis; wild type; GAPDH; Wnt/β-Catenin; PCR; ERK
• ISSN: 2352-345X; 2352-345X
• DOI: 10.1016/j.jcmgh.2015.06.006

Background & Aims Pigment epithelium-derived factor (PEDF) is a secretory protein that inhibits multiple tumor types. PEDF inhibits the Wnt coreceptor, low-density lipoprotein receptor-related protein 6 (LRP6), in the eye, but whether the tumor-suppressive properties of PEDF occur in organs such as the liver is unknown. Methods Wnt-dependent regulation of PEDF was assessed in the absence and presence of the Wnt coreceptor LRP6. Whole genome expression analysis was performed on PEDF knockout (KO) and control livers (7 months). Interrogation of Wnt/β-catenin signaling was performed in whole livers and human hepatocellular carcinoma (HCC) cell lines after RNA interference of PEDF and restoration of a PEDF-derived peptide. Western diet feeding for 6 to 8 months was used to evaluate whether the absence of PEDF was permissive for HCC formation (n = 12/group). Results PEDF levels increased in response to canonical Wnt3a in an LRP6-dependent manner but were suppressed by noncanonical Wnt5a protein in an LRP6-independent manner. Gene set enrichment analysis (GSEA) of PEDF KO livers revealed induction of pathways associated with experimental and human HCC and a transcriptional profile characterized by Wnt/β-catenin activation. Enhanced Wnt/β-catenin signaling occurred in KO livers, and PEDF delivery in vivo reduced LRP6 activation. In human HCC cells, RNA interference of PEDF led to increased levels of activated LRP6 and β-catenin, and a PEDF 34-mer peptide decreased LRP6 activation and β-catenin signaling, and reduced Wnt target genes. PEDF KO mice fed a Western diet developed sporadic well-differentiated HCC. Human HCC specimens demonstrated decreased PEDF staining compared with hepatocytes. Conclusions PEDF is an endogenous inhibitor of Wnt/β-catenin signaling in the liver.