Novel Approach to the Hedgehog Signaling Pathway: Combined Treatment of SMO and PTCH Inhibitors

Purpose: Abnormal Sonic Hedgehog signaling Pathway (Shh) activation is crucial for development of cancer stem cells, neoplastic growth and epithelial-mesenchymal transition processes in adulthood. Activation of Hedgehog signaling pathway may induces the changes in cilia found in the cell membrane, iniciates the Gli1 transcription factor that is translocated to the cell nucleus and finally, the target genes are transcribed. In this study, invastigation of the antiproliferative, anti-invasive and antimigrative effect of the combined use of robotnikinin (Ptch1 antagonist) and vismodegib (Smo inhibitor) on the hedgehog signaling pathway was aimed. Material and Methods: After demonstarting the presence of the hedgehog signaling pathway in the glioblastoma cell line U87-MG, the effect of the combined use of the robotnikinin and the vismodegib on the hedgehog signaling pathway was investigated. In-vitro cell proliferation, migration, and invasion analysis of the combination of antagonist and inhibitor and in silico drug-likeness analysis were performed. Results: Two different combinations of robotnikinin and vismodegib were tested. In vitro studies show that the combined use of agents in combined treatments of Smo and Ptch1is more effective than their individual usage. Conclusion: Inhibition of the hedgehog signaling pathway with specific inhibitors and antagonists is considered an innovative strategy for cancer therapy.

Keywords:

glioblastoma, hedgehog signaling pathway, signaling pathway inhibitors robotnikinin, vismodegib,

PDF

___

(1) Nüsslein-volhard C, Wieschaus E. Mutations affecting segment number and polarity in drosophila. Nature. 1980; 287(5785):795-801
(2) Ingham PW, McMahon AP. Hedgehog signaling in animal development: Paradigms and principles. Genes and Development. 2001; 15(23):3059-87
(3) Petrova R, Joyner AL. Roles for Hedgehog signaling in adult organ homeostasis and repair. Development (Cambridge). 2014; 141(18):3445-57
(4) 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
(5) Clement V, Sanchez P, de Tribolet N, Radovanovic I, Ruiz i Altaba A. HEDGEHOG-GLI1 Signaling Regulates Human Glioma Growth, Cancer Stem Cell Self-Renewal, and Tumorigenicity. Curr Biol. 2007; 17(2):165-72
(6) Meng E, Hanna A, Samant RS, Shevde LA. The impact of hedgehog signaling pathway on DNA repair mechanisms in human cancer. Cancers. 2015; 7(3):1333-48
(7) Lézot F, Corre I, Morice S, Rédini F, Verrecchia F. SHH Signaling Pathway Drives Pediatric Bone Sarcoma Progression. Cells. 2020; 9(3):536
(8) Johnson RW, Nguyen MP, Padalecki SS, Grubbs BG, Merkel AR, Oyajobi BO, et al. TGF-β promotion of Gli2-induced expression of parathyroid hormone-related protein, an important osteolytic factor in bone metastasis, is independent of canonical Hedgehog signaling. Cancer Res. 2011; 71(3):822-31
(9) Ju B, Spitsbergen J, Eden CJ, Taylor MR, Chen W. Co-activation of hedgehog and AKT pathways promote tumorigenesis in zebrafish. Mol Cancer. 2009; 8:40
(10) Cheng SY, Michael Bishop J. Suppressor of Fused represses Gli-mediated transcription by recruiting the SAP18-mSin3 corepressor complex. Proc Natl Acad Sci U S A. 2002; 99(8):5442-7
(11) Hui CC, Angers S. Gli proteins in development and disease. Annu Rev Cell Dev Biol. 2011; 27:513-37
(12) Yang C, Chen W, Chen Y, Jiang J. Smoothened transduces Hedgehog signal by forming a complex with Evc/Evc2. Cell Res. 2012; 22(11):1593-604
(13) Briscoe J, Thérond PP. The mechanisms of Hedgehog signalling and its roles in development and disease. Nature Reviews Molecular Cell Biology. 2013; 14(7):416-29
(14) Varjosalo M, Taipale J. Hedgehog: Functions and mechanisms. Genes and Development. 2008; 22(18):2454-72
(15) Aberger F, Frischauf A-M. GLI Genes and Their Targets in Epidermal Development and Disease. In: Hedgehog-Gli Signaling in Human Disease. 2007
(16) Rubin LL, de Sauvage FJ. Targeting the Hedgehog pathway in cancer. Nature Reviews Drug Discovery. 2006; 5(12):1026-33
(17) "FDA approves Erivedge (vismodegib) capsule, the first medicine for adults with advanced basal cell carcinoma". Roche. 30 January 2012. Retrieved 9 August 2020
(18) Stanton BZ, Peng LF, Maloof N, Nakai K, Wang X, Duffner JL, et al. A small molecule that binds Hedgehog and blocks its signaling in human cells. Nat Chem Biol. 2009; 5(3):154-6
(19) Sharpe HJ, Pau G, Dijkgraaf GJ, Basset-Seguin N, Modrusan Z, Januario T, et al. Genomic Analysis of Smoothened Inhibitor Resistance in Basal Cell Carcinoma. Cancer Cell. 2015; 27(3):327-41
(20) Atwood SX, Sarin KY, Whitson RJ, Li JR, Kim G, Rezaee M, et al. Smoothened Variants Explain the Majority of Drug Resistance in Basal Cell Carcinoma. Cancer Cell. 2015; 27(3):342-53
(21) Dijkgraaf GJP, Alicke B, Weinmann L, Januario T, West K, Modrusan Z, et al. Small molecule inhibition of GDC-0449 refractory smoothened mutants and downstream mechanisms of drug resistance. Cancer Res. 2011; 71(2):435-44
(22) Chandra V, Das T, Gulati P, Biswas NK, Rote S, Chatterjee U, et al. Hedgehog Signaling Pathway Is Active in GBM with GLI1 mRNA Expression Showing a Single Continuous Distribution Rather than Discrete High/Low Clusters. PLoS One. 2015; 10(3):e0116390
(23) Huang D, Wang Y, Tang J, Luo S. Molecular mechanisms of suppressor of fused in regulating the hedgehog signalling pathway. Oncology Letters. 2018; 15(5):6077-6086
(24) Wishart DS, Feunang YD, Guo AC, Lo EJ, Marcu A, Grant JR, et al. DrugBank 5.0: A major update to the DrugBank database for 2018. Nucleic Acids Res. 2018; 46(D1):D1074-D1082
(25) Mateska I, Nanda K, Dye NA, Alexaki VI, Eaton S. Range of SHH signaling in adrenal gland is limited by membrane contact to cells with primary cilia. J Cell Biol. 2020; 219(12):e201910087
(26) Uchida H, Arita K, Yunoue S, Yonezawa H, Shinsato Y, Kawano H, et al. Role of sonic hedgehog signaling in migration of cell lines established from CD133-positive malignant glioma cells. J Neurooncol. 2011; 104(3):697-704