Crenigacestat Blocking Notch Pathway Reduces Liver Fibrosis in the Surrounding Ecosystem of Intrahepatic CCA ViaTGF-β Inhibition

Overview

Journal J Exp Clin Cancer Res

Publisher Biomed Central

Specialty Oncology

Date 2022 Nov 29

PMID 36443822

Authors

Serena Mancarella

Isabella Gigante

Grazia Serino

Elena Pizzuto

Francesco Dituri

Maria F Valentini

Jingxiao Wang

Xin Chen

Raffaele Armentano

Diego F Calvisi

Gianluigi Giannelli

Affiliations

Soon will be listed here.

Abstract

Background: Intrahepatic cholangiocarcinoma (iCCA) is a highly malignant tumor characterized by an intensive desmoplastic reaction due to the exaggerated presence of the extracellular (ECM) matrix components. Liver fibroblasts close to the tumor, activated by transforming growth factor (TGF)-β1 and expressing high levels of α-smooth muscle actin (α-SMA), become cancer-associated fibroblasts (CAFs). CAFs are deputed to produce and secrete ECM components and crosstalk with cancer cells favoring tumor progression and resistance to therapy. Overexpression of Notch signaling is implicated in CCA development and growth. The study aimed to determine the effectiveness of the Notch inhibitor, Crenigacestat, on the surrounding microenvironment of iCCA.

Methods: We investigated Crenigacestat's effectiveness in a PDX model of iCCA and human primary culture of CAFs isolated from patients with iCCA.

Results: In silico analysis of transcriptomic profiling from PDX iCCA tissues treated with Crenigacestat highlighted "liver fibrosis" as one of the most modulated pathways. In the iCCA PDX model, Crenigacestat treatment significantly (p < 0.001) reduced peritumoral liver fibrosis. Similar results were obtained in a hydrodynamic model of iCCA. Bioinformatic prediction of the upstream regulators related to liver fibrosis in the iCCA PDX treated with Crenigacestat revealed the involvement of the TGF-β1 pathway as a master regulator gene showing a robust connection between TGF-β1 and Notch pathways. Consistently, drug treatment significantly (p < 0.05) reduced TGF-β1 mRNA and protein levels in tumoral tissue. In PDX tissues, Crenigacestat remarkably inhibited TGF-β signaling and extracellular matrix protein gene expression and reduced α-SMA expression. Furthermore, Crenigacestat synergistically increased Gemcitabine effectiveness in the iCCA PDX model. In 31 iCCA patients, TGF-β1 and α-SMA were upregulated in the tumoral compared with peritumoral tissues. In freshly isolated CAFs from patients with iCCA, Crenigacestat significantly (p < 0.001) inhibited Notch signaling, TGF-β1 secretion, and Smad-2 activation. Consequently, Crenigacestat also inactivated CAFs reducing (p < 0.001) α-SMA expression. Finally, CAFs treated with Crenigacestat produced less (p < 005) ECM components such as fibronectin, collagen 1A1, and collagen 1A2.

Conclusions: Notch signaling inhibition reduces the peritumoral desmoplastic reaction in iCCA, blocking the TGF-β1 canonical pathway.

Citing Articles

Decoding tumor-fibrosis interplay: mechanisms, impact on progression, and innovative therapeutic strategies.

Chen H, Xu X, Li J, Xue Y, Li X, Zhang K Front Pharmacol. 2024; 15:1491400.

PMID: 39534084 PMC: 11555290. DOI: 10.3389/fphar.2024.1491400.

Tumor Immune Microenvironment in Intrahepatic Cholangiocarcinoma: Regulatory Mechanisms, Functions, and Therapeutic Implications.

Ricci A, Rizzo A, Schirizzi A, DAlessandro R, Frega G, Brandi G Cancers (Basel). 2024; 16(20).

PMID: 39456636 PMC: 11505966. DOI: 10.3390/cancers16203542.

Oncogenic GALNT5 confers FOLFIRINOX resistance via activating the MYH9/ NOTCH/ DDR axis in pancreatic ductal adenocarcinoma.

Jia Q, Zhu Y, Yao H, Yin Y, Duan Z, Zheng J Cell Death Dis. 2024; 15(10):767.

PMID: 39433745 PMC: 11493973. DOI: 10.1038/s41419-024-07110-w.

Targeting cancer-associated fibroblasts/tumor cells cross-talk inhibits intrahepatic cholangiocarcinoma progression via cell-cycle arrest.

Mancarella S, Gigante I, Pizzuto E, Serino G, Terzi A, Dituri F J Exp Clin Cancer Res. 2024; 43(1):286.

PMID: 39415286 PMC: 11484308. DOI: 10.1186/s13046-024-03210-9.

Interplay of Cardiometabolic Syndrome and Biliary Tract Cancer: A Comprehensive Analysis with Gender-Specific Insights.

Di Stasi V, Contaldo A, Birtolo L, Shahini E Cancers (Basel). 2024; 16(19).

PMID: 39410050 PMC: 11476000. DOI: 10.3390/cancers16193432.

References

Clements O, Eliahoo J, Kim J, Taylor-Robinson S, Khan S . Risk factors for intrahepatic and extrahepatic cholangiocarcinoma: A systematic review and meta-analysis. J Hepatol. 2019; 72(1):95-103. DOI: 10.1016/j.jhep.2019.09.007. View

Banales J, Marin J, Lamarca A, Rodrigues P, Khan S, Roberts L . Cholangiocarcinoma 2020: the next horizon in mechanisms and management. Nat Rev Gastroenterol Hepatol. 2020; 17(9):557-588. PMC: 7447603. DOI: 10.1038/s41575-020-0310-z. View

Matsumoto K, Onoyama T, Kawata S, Takeda Y, Harada K, Ikebuchi Y . Hepatitis B and C virus infection is a risk factor for the development of cholangiocarcinoma. Intern Med. 2014; 53(7):651-4. DOI: 10.2169/internalmedicine.53.1410. View

Alvaro D, Crocetti E, Ferretti S, Bragazzi M, Capocaccia R . Descriptive epidemiology of cholangiocarcinoma in Italy. Dig Liver Dis. 2009; 42(7):490-5. DOI: 10.1016/j.dld.2009.10.009. View

Kim Y, Kim M, Shin J, Park S, Kim S, Kim J . Hedgehog signaling between cancer cells and hepatic stellate cells in promoting cholangiocarcinoma. Ann Surg Oncol. 2014; 21(8):2684-98. DOI: 10.1245/s10434-014-3531-y. View

Okabe H, Beppu T, Hayashi H, Horino K, Masuda T, Komori H . Hepatic stellate cells may relate to progression of intrahepatic cholangiocarcinoma. Ann Surg Oncol. 2009; 16(9):2555-64. DOI: 10.1245/s10434-009-0568-4. View

Zhao W, Zhang L, Xu Y, Zhang Z, Ren G, Tang K . Hepatic stellate cells promote tumor progression by enhancement of immunosuppressive cells in an orthotopic liver tumor mouse model. Lab Invest. 2013; 94(2):182-91. DOI: 10.1038/labinvest.2013.139. View

Jing C, Fu Y, Zhou C, Zhang M, Yi Y, Huang J . Hepatic stellate cells promote intrahepatic cholangiocarcinoma progression via NR4A2/osteopontin/Wnt signaling axis. Oncogene. 2021; 40(16):2910-2922. DOI: 10.1038/s41388-021-01705-9. View

Vaquero J, Guedj N, Claperon A, Nguyen Ho-Bouldoires T, Paradis V, Fouassier L . Epithelial-mesenchymal transition in cholangiocarcinoma: From clinical evidence to regulatory networks. J Hepatol. 2016; 66(2):424-441. DOI: 10.1016/j.jhep.2016.09.010. View

10.

Kalluri R, Zeisberg M . Fibroblasts in cancer. Nat Rev Cancer. 2006; 6(5):392-401. DOI: 10.1038/nrc1877. View

11.

Sirica A . The role of cancer-associated myofibroblasts in intrahepatic cholangiocarcinoma. Nat Rev Gastroenterol Hepatol. 2011; 9(1):44-54. DOI: 10.1038/nrgastro.2011.222. View

12.

Sirica A, Gores G . Desmoplastic stroma and cholangiocarcinoma: clinical implications and therapeutic targeting. Hepatology. 2013; 59(6):2397-402. PMC: 3975806. DOI: 10.1002/hep.26762. View

13.

Chen L, Yang T, Lu D, Zhao H, Feng Y, Chen H . Central role of dysregulation of TGF-β/Smad in CKD progression and potential targets of its treatment. Biomed Pharmacother. 2018; 101:670-681. DOI: 10.1016/j.biopha.2018.02.090. View

14.

Eser P, Janne P . TGFβ pathway inhibition in the treatment of non-small cell lung cancer. Pharmacol Ther. 2017; 184:112-130. DOI: 10.1016/j.pharmthera.2017.11.004. View

15.

Katz L, Likhter M, Jogunoori W, Belkin M, Ohshiro K, Mishra L . TGF-β signaling in liver and gastrointestinal cancers. Cancer Lett. 2016; 379(2):166-72. PMC: 5107316. DOI: 10.1016/j.canlet.2016.03.033. View

16.

Claperon A, Mergey M, Aoudjehane L, Nguyen Ho-Bouldoires T, Wendum D, Prignon A . Hepatic myofibroblasts promote the progression of human cholangiocarcinoma through activation of epidermal growth factor receptor. Hepatology. 2013; 58(6):2001-11. DOI: 10.1002/hep.26585. View

17.

Xu F, Liu C, Zhou D, Zhang L . TGF-β/SMAD Pathway and Its Regulation in Hepatic Fibrosis. J Histochem Cytochem. 2016; 64(3):157-67. PMC: 4810800. DOI: 10.1369/0022155415627681. View

18.

Che L, Fan B, Pilo M, Xu Z, Liu Y, Cigliano A . Jagged 1 is a major Notch ligand along cholangiocarcinoma development in mice and humans. Oncogenesis. 2016; 5(12):e274. PMC: 5177771. DOI: 10.1038/oncsis.2016.73. View

19.

Geisler F, Strazzabosco M . Emerging roles of Notch signaling in liver disease. Hepatology. 2014; 61(1):382-92. PMC: 4268103. DOI: 10.1002/hep.27268. View

20.

Zender S, Nickeleit I, Wuestefeld T, Sorensen I, Dauch D, Bozko P . A critical role for notch signaling in the formation of cholangiocellular carcinomas. Cancer Cell. 2013; 23(6):784-95. DOI: 10.1016/j.ccr.2013.04.019. View