Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress 2023 EHA Congress 2023 ASCO Annual Meeting
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
Journal of Experimental & Clinical Cancer Research
Published in: Journal of Experimental & Clinical Cancer Research 1/2019

Open Access 01-12-2019 | Metastasis | Research

Attenuation of hedgehog/GLI signaling by NT1721 extends survival in pancreatic cancer

Authors: Claudia M. Kowolik, Min Lin, Jun Xie, Larry E. Overman, David A. Horne

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2019

Login to get access

Abstract

Background

Pancreatic cancer is one of the most lethal malignancies due to frequent late diagnosis, aggressive tumor growth and metastasis formation. Continuously raising incidence rates of pancreatic cancer and a lack of significant improvement in survival rates over the past 30 years highlight the need for new therapeutic agents. Thus, new therapeutic agents and strategies are urgently needed to improve the outcome for patients with pancreatic cancer. Here, we evaluated the anti-tumor activity of a new natural product-based epidithiodiketopiperazine, NT1721, against pancreatic cancer.

Methods

We characterized the anticancer efficacy of NT1721 in multiple pancreatic cancer cell lines in vitro and in two orthotopic models. We also compared the effects of NT1721 to clinically used hedgehog inhibitors and the standard-of-care drug, gemcitabine. The effect of NT1721 on hedgehog/GLI signaling was assessed by determining the expression of GLI and GLI target genes both in vitro and in vivo.

Results

NT1721 displayed IC50 values in the submicromolar range in multiple pancreatic cancer cell lines, while largely sparing normal pancreatic epithelial cells. NT1721 attenuated hedgehog/GLI signaling through downregulation of GLI1/2 transcription factors and their downstream target genes, which reduced cell proliferation and invasion in vitro and significantly decreased tumor growth and liver metastasis in two preclinical orthotopic mouse models of pancreatic cancer. Importantly, treatment with NT1721 significantly improved survival times of mice with pancreatic cancer compared to the standard-of-care drug, gemcitabine.

Conclusions

Favorable therapeutics properties, i.e. 10-fold lower IC50 values than clinically used hedgehog inhibitors (vismodegib, erismodegib), a 90% reduction in liver metastasis and significantly better survival times compared to the standard-of-care drug, gemcitabine, provide a rational for testing NT1721 in the clinic either as a single agent or possibly in combination with gemcitabine or other therapeutic agents in PDAC patients overexpressing GLI1/2. This could potentially result in promising new treatment options for patients suffering from this devastating disease.
Appendix
Available only for authorised users
Literature
1.
Siegel RL, Miller KD, Jemal A. Cancer statistics, 2019. CA Cancer J Clin. 2019;69(1):7–34 PubMed PMID: 30620402.CrossRefPubMed
2.
Polireddy K, Chen Q. Cancer of the pancreas: molecular pathways and current advancement in treatment. J Cancer. 2016;7(11):1497–514 PubMed PMID: 27471566. Pubmed Central PMCID: 4964134.PubMedPubMedCentralCrossRef
3.
4.
Garrido-Laguna I, Hidalgo M. Pancreatic cancer: from state-of-the-art treatments to promising novel therapies. Nat Rev Clin Oncol. 2015;12(6):319–34 PubMed PMID: 25824606.PubMedCrossRef
5.
Conroy T, Desseigne F, Ychou M, Bouche O, Guimbaud R, Becouarn Y, et al. FOLFIRINOX versus gemcitabine for metastatic pancreatic cancer. N Engl J Med. 2011;364(19):1817–25 PubMed PMID: 21561347.PubMedCrossRef
6.
Hermann PC, Huber SL, Herrler T, Aicher A, Ellwart JW, Guba M, et al. Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell. 2007;1(3):313–23 PubMed PMID: 18371365.PubMedCrossRef
7.
Quint K, Tonigold M, Di Fazio P, Montalbano R, Lingelbach S, Ruckert F, et al. Pancreatic cancer cells surviving gemcitabine treatment express markers of stem cell differentiation and epithelial-mesenchymal transition. Int J Oncol. 2012;41(6):2093–102 PubMed PMID: 23026911.PubMedCrossRef
8.
9.
Berman DM, Karhadkar SS, Maitra A, Montes De Oca R, Gerstenblith MR, Briggs K, et al. Widespread requirement for hedgehog ligand stimulation in growth of digestive tract tumours. Nature. 2003;425(6960):846–51 PubMed PMID: 14520411.PubMedCrossRef
10.
Ramaswamy B, Lu Y, Teng KY, Nuovo G, Li X, Shapiro CL, et al. Hedgehog signaling is a novel therapeutic target in tamoxifen-resistant breast cancer aberrantly activated by PI3K/AKT pathway. Cancer Res. 2012;72(19):5048–59 PubMed PMID: 22875023. Pubmed Central PMCID: 3837449.PubMedCrossRef
11.
Rajurkar M, De Jesus-Monge WE, Driscoll DR, Appleman VA, Huang H, Cotton JL, et al. The activity of Gli transcription factors is essential for Kras-induced pancreatic tumorigenesis. Proc Natl Acad Sci U S A. 2012;109(17):E1038–47 PubMed PMID: 22493246. Pubmed Central PMCID: 3340052.PubMedPubMedCentralCrossRef
12.
Javelaud D, Alexaki VI, Dennler S, Mohammad KS, Guise TA, Mauviel A. TGF-beta/SMAD/GLI2 signaling axis in cancer progression and metastasis. Cancer Res. 2011;71(17):5606–10 PubMed PMID: 21862631. Pubmed Central PMCID: 3165102.PubMedPubMedCentralCrossRef
13.
He S, Wang F, Yang L, Guo C, Wan R, Ke A, et al. Expression of DNMT1 and DNMT3a are regulated by GLI1 in human pancreatic cancer. PLoS One. 2011;6(11):e27684 PubMed PMID: 22110720. Pubmed Central PMCID: 3215738.PubMedPubMedCentralCrossRef
14.
Eichberger T, Sander V, Schnidar H, Regl G, Kasper M, Schmid C, et al. Overlapping and distinct transcriptional regulator properties of the GLI1 and GLI2 oncogenes. Genomics. 2006;87(5):616–32 PubMed PMID: 16434164.PubMedCrossRef
15.
Singh RR, Kunkalla K, Qu C, Schlette E, Neelapu SS, Samaniego F, et al. ABCG2 is a direct transcriptional target of hedgehog signaling and involved in stroma-induced drug tolerance in diffuse large B-cell lymphoma. Oncogene. 2011;30(49):4874–86 PubMed PMID: 21625222. Pubmed Central PMCID: 3165099.PubMedPubMedCentralCrossRef
16.
Liu S, Dontu G, Mantle ID, Patel S, Ahn NS, Jackson KW, et al. Hedgehog signaling and Bmi-1 regulate self-renewal of normal and malignant human mammary stem cells. Cancer Res. 2006;66(12):6063–71 PubMed PMID: 16778178. Pubmed Central PMCID: 4386278.PubMedPubMedCentralCrossRef
17.
Feldmann G, Dhara S, Fendrich V, Bedja D, Beaty R, Mullendore M, et al. Blockade of hedgehog signaling inhibits pancreatic cancer invasion and metastases: a new paradigm for combination therapy in solid cancers. Cancer Res. 2007;67(5):2187–96 PubMed PMID: 17332349. Pubmed Central PMCID: 3073370.PubMedPubMedCentralCrossRef
18.
Yao J, An Y, Wie JS, Ji ZL, Lu ZP, Wu JL, et al. Cyclopamine reverts acquired chemoresistance and down-regulates cancer stem cell markers in pancreatic cancer cell lines. Swiss Med Wkly. 2011;141:w13208 PubMed PMID: 21630164.PubMed
19.
Baumann M, Dieskau AP, Loertscher BM, Walton MC, Nam S, Xie J, et al. Tricyclic analogues of epidithiodioxopiperazine alkaloids with promising and antitumor activity. Chem Sci. 2015;6:4451–7 PubMed PMID: 26301062. Pubmed Central PMCID: 4540405.PubMedPubMedCentralCrossRef
20.
Kowolik CM, Lin M, Xie J, Overman LE, Horne DA. NT1721, a novel epidithiodiketopiperazine, exhibits potent in vitro and in vivo efficacy against acute myeloid leukemia. Oncotarget. 2016;7(52):86186–97 PubMed PMID: 27863389.PubMedPubMedCentralCrossRef
21.
Roessler E, Ermilov AN, Grange DK, Wang A, Grachtchouk M, Dlugosz AA, et al. A previously unidentified amino-terminal domain regulates transcriptional activity of wild-type and disease-associated human GLI2. Hum Mol Genet. 2005;14(15):2181–8 PubMed PMID: 15994174.PubMedCrossRef
22.
Wang X, Venugopal C, Manoranjan B, McFarlane N, O'Farrell E, Nolte S, et al. Sonic hedgehog regulates Bmi1 in human medulloblastoma brain tumor-initiating cells. Oncogene. 2012;31(2):187–99 PubMed PMID: 21685941.PubMedCrossRef
23.
Ikram MS, Neill GW, Regl G, Eichberger T, Frischauf AM, Aberger F, et al. GLI2 is expressed in normal human epidermis and BCC and induces GLI1 expression by binding to its promoter. J Invest Dermatol. 2004;122(6):1503–9 PubMed PMID: 15175043.PubMedCrossRef
24.
Regl G, Neill GW, Eichberger T, Kasper M, Ikram MS, Koller J, et al. Human GLI2 and GLI1 are part of a positive feedback mechanism in basal cell carcinoma. Oncogene. 2002;21(36):5529–39 PubMed PMID: 12165851.PubMedCrossRef
25.
Beachy PA, Karhadkar SS, Berman DM. Tissue repair and stem cell renewal in carcinogenesis. Nature. 2004;432(7015):324–31 PubMed PMID: 15549094.PubMedCrossRef
26.
Regl G, Kasper M, Schnidar H, Eichberger T, Neill GW, Philpott MP, et al. Activation of the BCL2 promoter in response to hedgehog/GLI signal transduction is predominantly mediated by GLI2. Cancer Res. 2004;64(21):7724–31 PubMed PMID: 15520176.PubMedCrossRef
27.
Katoh Y, Katoh M. Hedgehog target genes: mechanisms of carcinogenesis induced by aberrant hedgehog signaling activation. Curr Mol Med. 2009;9(7):873–86 PubMed PMID: 19860666.PubMedCrossRef
28.
Singh BN, Fu J, Srivastava RK, Shankar S. Hedgehog signaling antagonist GDC-0449 (Vismodegib) inhibits pancreatic cancer stem cell characteristics: molecular mechanisms. PLoS One. 2011;6(11):e27306 PubMed PMID: 22087285. Pubmed Central PMCID: 3210776.PubMedPubMedCentralCrossRef
29.
Kumar V, Chaudhary AK, Dong Y, Zhong HA, Mondal G, Lin F, et al. Design, synthesis and biological evaluation of novel hedgehog inhibitors for treating pancreatic cancer. Sci Rep. 2017;7(1):1665 PubMed PMID: 28490735. Pubmed Central PMCID: 5431907.PubMedPubMedCentralCrossRef
30.
Zhou Q, Zhou Y, Liu X, Shen Y. GDC-0449 improves the antitumor activity of nano-doxorubicin in pancreatic cancer in a fibroblast-enriched microenvironment. Sci Rep. 2017;7(1):13379 PubMed PMID: 29042665. Pubmed Central PMCID: 5645386.PubMedPubMedCentralCrossRef
31.
Jacobs JJ, Kieboom K, Marino S, DePinho RA, van Lohuizen M. The oncogene and Polycomb-group gene bmi-1 regulates cell proliferation and senescence through the ink4a locus. Nature. 1999;397(6715):164–8 PubMed PMID: 9923679.PubMedCrossRef
32.
Xu Z, Liu H, Lv X, Liu Y, Li S, Li H. Knockdown of the Bmi-1 oncogene inhibits cell proliferation and induces cell apoptosis and is involved in the decrease of Akt phosphorylation in the human breast carcinoma cell line MCF-7. Oncol Rep. 2011;25(2):409–18 PubMed PMID: 21152871.PubMed
33.
Balasubramanian S, Adhikary G, Eckert RL. The Bmi-1 polycomb protein antagonizes the (−)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival. Carcinogenesis. 2010;31(3):496–503 PubMed PMID: 20015867. Pubmed Central PMCID: 2832547.PubMedCrossRef
34.
Lee K, Adhikary G, Balasubramanian S, Gopalakrishnan R, McCormick T, Dimri GP, et al. Expression of Bmi-1 in epidermis enhances cell survival by altering cell cycle regulatory protein expression and inhibiting apoptosis. J Invest Dermatol. 2008;128(1):9–17 PubMed PMID: 17625597. Pubmed Central PMCID: 3043461.PubMedCrossRef
35.
Fu J, Rodova M, Roy SK, Sharma J, Singh KP, Srivastava RK, et al. GANT-61 inhibits pancreatic cancer stem cell growth in vitro and in NOD/SCID/IL2R gamma null mice xenograft. Cancer Lett. 2013;330(1):22–32 PubMed PMID: 23200667. Pubmed Central PMCID: 4153855.PubMedCrossRef
36.
Faiao-Flores F, Alves-Fernandes DK, Pennacchi PC, Sandri S, Vicente AL, Scapulatempo-Neto C, et al. Targeting the hedgehog transcription factors GLI1 and GLI2 restores sensitivity to vemurafenib-resistant human melanoma cells. Oncogene. 2017;36(13):1849–61 PubMed PMID: 27748762. Pubmed Central PMCID: 5378933.PubMedCrossRef
37.
Nanta R, Kumar D, Meeker D, Rodova M, Van Veldhuizen PJ, Shankar S, et al. NVP-LDE-225 (Erismodegib) inhibits epithelial-mesenchymal transition and human prostate cancer stem cell growth in NOD/SCID IL2Rgamma null mice by regulating Bmi-1 and microRNA-128. Oncogenesis. 2013;2:e42 PubMed PMID: 23567619. Pubmed Central PMCID: 3641359.PubMedPubMedCentralCrossRef
38.
Che J, Zhang FZ, Zhao CQ, Hu XD, Fan SJ. Cyclopamine is a novel hedgehog signaling inhibitor with significant anti-proliferative, anti-invasive and anti-estrogenic potency in human breast cancer cells. Oncol Lett. 2013;5(4):1417–21 PubMed PMID: 23599805. Pubmed Central PMCID: 3629107.PubMedPubMedCentralCrossRef
39.
Adams CR, Htwe HH, Marsh T, Wang AL, Montoya ML, Subbaraj L, et al. Transcriptional control of subtype switching ensures adaptation and growth of pancreatic cancer. Elife. 2019;8. https://​doi.​org/​10.​7554/​eLife.​45313 PubMed PMID: 31134896. Pubmed Central PMCID: 6538376.
40.
Gao J, Wang L, Xu J, Zheng J, Man X, Wu H, et al. Aberrant DNA methyltransferase expression in pancreatic ductal adenocarcinoma development and progression. J Exp Clin Cancer Res. 2013;32:86 PubMed PMID: 24423239. Pubmed Central PMCID: 4029463.PubMedPubMedCentralCrossRef
41.
Cook KM, Hilton ST, Mecinovic J, Motherwell WB, Figg WD, Schofield CJ. Epidithiodiketopiperazines block the interaction between hypoxia-inducible factor-1alpha (HIF-1alpha) and p300 by a zinc ejection mechanism. J Biol Chem. 2009;284(39):26831–8 PubMed PMID: 19589782. Pubmed Central PMCID: 2785371.PubMedPubMedCentralCrossRef
42.
Lakshmikuttyamma A, Scott SA, DeCoteau JF, Geyer CR. Reexpression of epigenetically silenced AML tumor suppressor genes by SUV39H1 inhibition. Oncogene. 2010;29(4):576–88 PubMed PMID: 19881540.PubMedCrossRef
43.
Li J, Zhang Y, Da Silva Sil Dos Santos B, Wang F, Ma Y, Perez C, et al. Epidithiodiketopiperazines inhibit protein degradation by targeting proteasome Deubiquitinase Rpn11. Cell Chem Biol. 2018;25(11):1350–8.e9 PubMed PMID: 30146242.PubMedCrossRefPubMedCentral
44.
Huang FT, Zhuan-Sun YX, Zhuang YY, Wei SL, Tang J, Chen WB, et al. Inhibition of hedgehog signaling depresses self-renewal of pancreatic cancer stem cells and reverses chemoresistance. Int J Oncol. 2012;41(5):1707–14 PubMed PMID: 22923052.PubMedCrossRef
45.
46.
Marechal R, Bachet JB, Calomme A, Demetter P, Delpero JR, Svrcek M, et al. Sonic hedgehog and Gli1 expression predict outcome in resected pancreatic adenocarcinoma. Clin Cancer Res. 2015;21(5):1215–24 PubMed PMID: 25552484.PubMedCrossRef
47.
Kurahara H, Takao S, Maemura K, Mataki Y, Kuwahata T, Maeda K, et al. Epithelial-mesenchymal transition and mesenchymal-epithelial transition via regulation of ZEB-1 and ZEB-2 expression in pancreatic cancer. J Surg Oncol. 2012;105(7):655–61 PubMed PMID: 22213144.PubMedCrossRef
48.
Song W, Tao K, Li H, Jin C, Song Z, Li J, et al. Bmi-1 is related to proliferation, survival and poor prognosis in pancreatic cancer. Cancer Sci. 2010;101(7):1754–60 PubMed PMID: 20426791.PubMedCrossRef
49.
Yin T, Wei H, Leng Z, Yang Z, Gou S, Wu H, et al. Bmi-1 promotes the chemoresistance, invasion and tumorigenesis of pancreatic cancer cells. Chemotherapy. 2011;57(6):488–96 PubMed PMID: 22248802.PubMedCrossRef
50.
Kumar K, Raza SS, Knab LM, Chow CR, Kwok B, Bentrem DJ, et al. GLI2-dependent c-MYC upregulation mediates resistance of pancreatic cancer cells to the BET bromodomain inhibitor JQ1. Sci Rep. 2015;5:9489 PubMed PMID: 25807524. Pubmed Central PMCID: 4452877.PubMedPubMedCentralCrossRef
51.
Steg A, Amm HM, Novak Z, Frost AR, Johnson MR. Gli3 mediates cell survival and sensitivity to cyclopamine in pancreatic cancer. Cancer Biol Ther. 2010;10(9):893–902 PubMed PMID: 20814245. Pubmed Central PMCID: 3230294.PubMedPubMedCentralCrossRef
52.
Yauch RL, Gould SE, Scales SJ, Tang T, Tian H, Ahn CP, et al. A paracrine requirement for hedgehog signalling in cancer. Nature. 2008;455(7211):406–10 PubMed PMID: 18754008.PubMedCrossRef
53.
Lauth M, Bergstrom A, Shimokawa T, Toftgard R. Inhibition of GLI-mediated transcription and tumor cell growth by small-molecule antagonists. Proc Natl Acad Sci U S A. 2007;104(20):8455–60 PubMed PMID: 17494766. Pubmed Central PMCID: 1866313.PubMedPubMedCentralCrossRef
Metadata
Title
Attenuation of hedgehog/GLI signaling by NT1721 extends survival in pancreatic cancer
Authors
Claudia M. Kowolik
Min Lin
Jun Xie
Larry E. Overman
David A. Horne
Publication date
01-12-2019
Publisher
BioMed Central
Published in
Journal of Experimental & Clinical Cancer Research / Issue 1/2019
Electronic ISSN: 1756-9966
DOI
https://doi.org/10.1186/s13046-019-1445-z

Other articles of this Issue 1/2019

Journal of Experimental & Clinical Cancer Research 1/2019 Go to the issue

Research

The anti-tumor growth effect of a novel agent DMAMCL in rhabdomyosarcoma in vitro and in vivo

Research

DHHC protein family targets different subsets of glioma stem cells in specific niches

Research

Activation of dynamin-related protein 1 - dependent mitochondria fragmentation and suppression of osteosarcoma by cryptotanshinone

Research

CircNFIX promotes progression of glioma through regulating miR-378e/RPN2 axis

Research

Inhibiting PAD2 enhances the anti-tumor effect of docetaxel in tamoxifen-resistant breast cancer cells

Research

TET1-mediated DNA hydroxymethylation activates inhibitors of the Wnt/β-catenin signaling pathway to suppress EMT in pancreatic tumor cells
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits