Objective：Gorlin-Goltz syndrome is an infrequent multisystemic disease with autosomal-dominant disorder with complete penetrance. The clinical features are basal cell carcinomas, recurrent odontogenic keratocysts, calcification of the falx cerebri, and skeletal anomalies. The genetic basis of the syndrome was identified with causative mutations in PTCH1. Ohba et al. reported that Ptch1 hetero mice increased bone mass caused by increasing Gli1 and decreasing Gli3 which suggest Hedgehog had important roles of osteoblast differentiation. To elucidate the pathological mechanism, we established induced pluripotent stem cells (iPSCs) from these patients and investigate Hedgehog related genes in our iPSc.
Material and method：This study has already been approved by the university ethics committee (approval number 533). Fibroblasts were derived from Gorlin patients diagnosed with a frameshift and splicing variant. We generated iPSc lines from fibroblasts obtained from NBCCS patient using a sendai vector SeVdp (KOSM) 302L. The protocol for osteoblast differentiation from iPS cells was based on a published method (Ochiai-shino et al. PloS one 2014 vol.9 e99534).
Result：We cloud successfully generated 4 individual derived iPSc. The iPScs also expressed higher GLI1 expression than control iPSc. Gorlin-iPSc treated with osteogenic medium and hedgehog activator, SAG, showed better ALP activity than control. These patient derived iPSc had suppressed WNT pathway and BMP while osteoinduction sharply up-regulated.
Discussion：In conclusion, iPSc generated from Gorlin syndrome patients had enhanced activation of Hh signaling which suppress both intrinsic Hh. G-OFiPS cells had high ability of osteogenic differentiation ability with enhanced expression of Hhs, WNTs and BMPs-RUNX2. These cell must be useful tool for not only for Gorlin syndrome study but also cancer research.