Gorlin syndrome-derived induced pluripotent stem cells are hypersensitive to Hedgehog-mediated osteogenic induction. Gorlin syndrome-derived induced pluripotent stem cells are hypersensitive to Hedgehog-mediated osteogenic induction

Gorlin syndrome is an autosomal dominant inherited syndrome that predisposes a patient to the formation of basal cell carcinomas, odontogenic keratocysts, and skeletal anomalies. Patched-1 (PTCH1) is a Hedgehog (Hh) receptor that acts as a negative regulator of constitutive Hh signaling by preventing the G protein-coupled receptor Smoothened from entering the cilium in the absence of Hh protein binding. We generated iPSCs from four unrelated Gorlin syndrome patients with loss-of-function mutations in PTCH1 using the Sendai virus vector (SeVdp(KOSM)302). The patient-derived iPSCs exhibited basic iPSC features, including stem cell marker expression, totipotency, and the ability to form teratomas. To check the Patient-derived iPSCs mimic patinet phenotype, We performted osteoblast differentiation and checked mRNA expression by hedgehog PCR array systems. Overall design: qPCR gene expression profiling. 2 control hiPSC (KD iPS and 201B7) and 4 disease specific hiPSC (G1-1,G1-6,G2-5,G2-8) were used and treated separately as indicated in the summary. Two disease specific iPSc (G1-1,G1-6) were established by same patient fibroblasts. Two disease specific iPSc (G2-5,G2-8) were established by other patient fibroblasts. There were two experimental group named iPS and OBM. iPS group cultured EB and then collect samples. OBM group cultured EB, performed osteoblast differentiation until day 10 and then collect samples. Equal amount total RNA from each donor was pooled prior to gene expression analysis.