Crenigacestat, a selective NOTCH1 inhibitor, reduces intrahepatic cholangiocarcinoma progression by blocking VEGFA/DLL4/MMP13 axis
Open Access
- 10 February 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Cell Death & Differentiation
- Vol. 27 (8), 2330-2343
- https://doi.org/10.1038/s41418-020-0505-4
Abstract
Intrahepatic cholangiocarcinoma (iCCA) is a deadly disease with rising incidence and few treatment options. An altered expression and/or activation of NOTCH1–3 receptors has been shown to play a role in iCCA development and progression. In this study, we established a new CCA patient-derived xenograft model, which was validated by immunohistochemistry and transcriptomic analysis. The effects of Notch pathway suppression by the Crenigacestat (LY3039478)-specific inhibitor were evaluated in human iCCA cell lines and the PDX model. In vitro, LY3039478 significantly reduced Notch pathway components, including NICD1 and HES1, but not the other Notch receptors, in a panel of five different iCCA cell lines. In the PDX model, LY3039478 significantly inhibited the Notch pathway and tumor growth to the same extent as gemcitabine. Furthermore, gene expression analysis of iCCA mouse tissues treated with LY3039478 revealed a downregulation of VEGFA, HES1, and MMP13 genes. In the same tissues, DLL4 and CD31 co-localized, and their expression was significantly inhibited in the treated mice, as it happened in the case of MMP13. In an in vitro angiogenesis model, LY3039478 inhibited vessel formation, which was restored by the addition of MMP13. Finally, RNA-sequencing expression data of iCCA patients and matched surrounding normal liver tissues downloaded from the GEO database demonstrated that NOTCH1, HES1, MMP13, DLL4, and VEGFA genes were significantly upregulated in tumors compared with adjacent nontumorous tissues. These data were confirmed by our group, using an independent cohort of iCCA specimens. Conclusion: We have developed and validated a new iCCA PDX model to test in vivo the activity of LY3039478, demonstrating its inhibitory role in Notch-dependent angiogenesis. Thus, the present data provide new knowledge on Notch signaling in iCCA, and support the inhibition of the Notch cascade as a promising strategy for the treatment of this disease.Keywords
Funding Information
- Associazione Italiana per la Ricerca sul Cancro (18737)
This publication has 42 references indexed in Scilit:
- Notch signaling: targeting cancer stem cells and epithelial-to-mesenchymal transitionOncoTargets and Therapy, 2013
- EPD and EPDnew, high-quality promoter resources in the next-generation sequencing eraNucleic Acids Research, 2012
- Intrahepatic cholangiocarcinoma can arise from Notch-mediated conversion of hepatocytesJCI Insight, 2012
- Cholangiocarcinomas can originate from hepatocytes in miceJCI Insight, 2012
- AltAnalyze and DomainGraph: analyzing and visualizing exon expression dataNucleic Acids Research, 2010
- MMP13 as a stromal mediator in controlling persistent angiogenesis in skin carcinomaCarcinogenesis: Integrative Cancer Research, 2009
- Stromal Expression of MMP-13 Is Required for Melanoma Invasion and MetastasisJournal of Investigative Dermatology, 2009
- Intrahepatic CholangiocarcinomaAnnals of Surgery, 2008
- Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profilesProceedings of the National Academy of Sciences of the United States of America, 2005
- Notch Signaling: Cell Fate Control and Signal Integration in DevelopmentScience, 1999