Effect of the sonic hedgehog inhibitor GDC-0449 on an in vitro isogenic cellular model simulating odontogenic keratocysts

Abstract

Using gene-edited cell cultures, Chinese researchers have gathered useful insights into the genetic origins and treatment of benign oral tumours called odontogenic keratocysts (OKCs). A total of 80% of OKCs are associated with mutations in the gene PTCH1, which are thought to activate a signalling pathway that drives OKC tumour growth. However, research is hindered by OKC cell culturing limitations. Tiejun Li, from the Peking University School and Hospital of Stomatology, and colleagues created an OKC cell culture model that uses CRISPR gene editing to introduce a PTCH1 gene mutation into human stem cells before differentiating them into epithelial cells. Using their model, the team confirm that PTCH1 mutation activates tumour-associated signalling, which the basal cell carcinoma drug vismodegib greatly reduces. Vismodegib also reduces the overproliferation of mutant cells and offers a potential treatment option for OKC patients.

References

[1] Kaczmarzyk T, Mojsa I, Stypulkowska J. A systematic review of the recurrence rate for keratocystic odontogenic tumour in relation to treatment modalities. Int. J. Oral. Maxillofac. Surg., 2012, 41: 756-767.

[2] Myoung H, . Odontogenic keratocyst: review of 256 cases for recurrence and clinicopathologic parameters. Oral. Surg. Oral. Med. Oral. Pathol. Oral. Radiol. Endod., 2001, 91: 328-333.

[3] Li TJ, Browne RM, Matthews JB. Epithelial cell proliferation in odontogenic keratocysts: a comparative immunocytochemical study of Ki67 in simple, recurrent and basal cell naevus syndrome (BCNS)-associated lesions. J. Oral. Pathol. Med., 1995, 24: 221-226.

[4] Slootweg PJ. p53 protein and Ki-67 reactivity in epithelial odontogenic lesions. An immunohistochemical study. J. Oral. Pathol. Med., 1995, 24: 393-397.

[5] Li TJ, Browne RM, Matthews JB. Quantification of PCNA+ cells within odontogenic jaw cyst epithelium. J. Oral. Pathol. Med., 1994, 23: 184-189.

[6] Li TJ. The odontogenic keratocyst: a cyst, or a cystic neoplasm?. J. Dent. Res., 2011, 90: 133-142.

[7] Gorlin RJ. Nevoid basal cell carcinoma syndrome. Dermatol. Clin., 1995, 13: 113-125.

[8] Lo Muzio L. Nevoid basal cell carcinoma syndrome (Gorlin syndrome). Orphanet. J. Rare. Dis., 2008, 3

[9] Johnson RL, . Human homolog of patched, a candidate gene for the basal cell nevus syndrome. Science, 1996, 272: 1668-1671.

[10] Hahn H, . Mutations of the human homolog of Drosophila patched in the nevoid basal cell carcinoma syndrome. Cell, 1996, 85: 841-851.

[11] Stone DM, . The tumour-suppressor gene patched encodes a candidate receptor for Sonic hedgehog. Nature, 1996, 384: 129-134.

[12] Toftgard R. Hedgehog signalling in cancer. Cell. Mol. Life Sci., 2000, 57: 1720-1731.

[13] Scales SJ, de Sauvage FJ. Mechanisms of hedgehog pathway activation in cancer and implications for therapy. Trends Pharmacol. Sci., 2009, 30: 303-312.

[14] Qu J, . Underestimated PTCH1 mutation rate in sporadic keratocystic odontogenic tumors. Oral. Oncol., 2015, 51: 40-45.

[15] Guo YY, . PTCH1 gene mutations in Keratocystic odontogenic tumors: a study of 43 Chinese patients and a systematic review. PLoS. One., 2013, 8: e77305.

[16] Sekulic A, . Efficacy and safety of Vismodegib in advanced basal-cell carcinoma. N. Engl. J. Med., 2012, 366: 2171-2179.

[17] Goldberg LH, . Resolution of odontogenic keratocysts of the jaw in basal cell nevus syndrome with GDC-0449. Arch. Dermatol., 2011, 147: 839-841.

[18] Booms P, Harth M, Sader R, Ghanaati S. Vismodegib hedgehog-signaling inhibition and treatment of basal cell carcinomas as well as keratocystic odontogenic tumors in Gorlin syndrome. Ann. Maxillofac. Surg., 2015, 5: 14-19.

[19] Ally MS, . The use of vismodegib to shrink keratocystic odontogenic tumors in patients with basal cell nevus syndrome. JAMA Dermatol., 2014, 150: 542-545.

[20] Thomson JA, . Embryonic stem cell lines derived from human blastocysts. Science, 1998, 282: 1145-1147.

[21] Movahednia MM, . Potential applications of keratinocytes derived from human embryonic stem cells. Biotechnol. J., 2016, 11: 58-70.

[22] Musunuru K. Genome editing of human pluripotent stem cells to generate human cellular disease models. Dis. Model Mech., 2013, 6: 896-904.

[23] Hendriks WT, Warren CR, Cowan CA. Genome editing in human pluripotent stem cells: approaches, pitfalls, and solutions. Cell. Stem. Cell., 2016, 18: 53-65.

[24] Selekman JA, Grundl NJ, Kolz JM, Palecek SP. Efficient generation of functional epithelial and epidermal cells from human pluripotent stem cells under defined conditions. Tissue Eng. Part C. Methods, 2013, 19: 949-960.

[25] Metallo CM, Ji L, de Pablo JJ, Palecek SP. Retinoic acid and bone morphogenetic protein signaling synergize to efficiently direct epithelial differentiation of human embryonic stem cells. Stem Cells, 2008, 26: 372-380.

[26] Yang A, . p63, a p53 homolog at 3q27-29, encodes multiple products with transactivating, death-inducing, and dominant-negative activities. Mol. Cell, 1998, 2: 305-316.

[27] Kasper M, Regl G, Frischauf AM, Aberger F. GLI transcription factors: mediators of oncogenic hedgehog signalling. Eur. J. Cancer, 2006, 42: 437-445.

[28] Taipale J, . Effects of oncogenic mutations in Smoothened and Patched can be reversed by cyclopamine. Nature, 2000, 406: 1005-1009.

[29] Wetmore C. Sonic hedgehog in normal and neoplastic proliferation: insight gained from human tumors and animal models. Curr. Opin. Genet. Dev., 2003, 13: 34-42.

[30] Wu SM, Choo AB, Yap MG, Chan KK. Role of Sonic hedgehog signaling and the expression of its components in human embryonic stem cells. Stem Cell Res., 2010, 4: 38-49.

[31] Mizuochi H, Fujii K, Shiohama T, Uchikawa H, Shimojo N. Hedgehog signaling is synergistically enhanced by nutritional deprivation and ligand stimulation in human fibroblasts of Gorlin syndrome. Biochem. Biophys. Res. Commun., 2015, 457: 318-323.

[32] Pan S, Xu LL, Sun LS, Li TJ. Identification of known and novel PTCH mutations in both syndromic and non-syndromic keratocystic odontogenic tumors. Int. J. Oral. Sci., 2009, 1: 34-38.

[33] Hong Y, . Heterozygous PTCH1 mutations impact the bone metabolism in patients with nevoid basal cell carcinoma syndrome likely by regulating SPARC expression. J. Bone Miner. Res., 2016, 31: 1413-1428.

[34] Takata M, . Homologous recombination and non-homologous end-joining pathways of DNA double-strand break repair have overlapping roles in the maintenance of chromosomal integrity in vertebrate cells. EMBO J., 1998, 17: 5497-5508.

[35] Mali P, . RNA-guided human genome engineering via Cas9. Science, 2013, 339: 823-826.

[36] Cong L, . Multiplex genome engineering using CRISPR/Cas systems. Science, 2013, 339: 819-823.

[37] Jinek M, . A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science, 2012, 337: 816-821.

[38] Hsu PD, Lander ES, Zhang F. Development and applications of CRISPR-Cas9 for genome engineering. Cell, 2014, 157: 1262-1278.

[39] Uhmann A, . A model for PTCH1/Ptch1-associated tumors comprising mutational inactivation and gene silencing. Int. J. Oncol., 2005, 27: 1567-1575.

[40] Calzada-Wack J, . Unbalanced overexpression of the mutant allele in murine Patched mutants. Carcinogenesis, 2002, 23: 727-733.

[41] Pan S, Li TJ. PTCH1 mutations in odontogenic keratocysts: are they related to epithelial cell proliferation?. Oral. Oncol., 2009, 45: 861-865.

[42] Chen G, . Chemically defined conditions for human iPSC derivation and culture. Nat. Methods, 2011, 8: 424-429.

Funding

National Natural Science Foundation of China (National Science Foundation of China)(81671006, 81300894, 81671006)

Natural Science Foundation of Beijing Municipality (Beijing Natural Science Foundation)(7172238)

the National Natural Science Foundation of China (81671006)