RRC ID 52451
著者 Hasegawa D, Ochiai-Shino H, Onodera S, Nakamura T, Saito A, Onda T, Watanabe K, Nishimura K, Ohtaka M, Nakanishi M, Kosaki K, Yamaguchi A, Shibahara T, Azuma T.
タイトル Gorlin syndrome-derived induced pluripotent stem cells are hypersensitive to hedgehog-mediated osteogenic induction.
ジャーナル PLoS One
Abstract Gorlin syndrome is an autosomal dominant inherited syndrome that predisposes a patient to the formation of basal cell carcinomas, odontogenic keratocysts, and skeletal anomalies. Causative mutations in several genes associated with the sonic hedgehog (SHH) signaling pathway, including PTCH1, have been identified in Gorlin syndrome patients. However, no definitive genotype-phenotype correlations are evident in these patients, and their clinical presentation varies greatly, often leading to delayed diagnosis and treatment. 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. GLI1 expression levels were greater in fibroblasts and patient-derived iPSCs than in the corresponding control cells. Patient-derived iPSCs expressed lower basal levels than control iPSCs of the genes encoding the Hh ligands Indian Hedgehog (IHH) and SHH, the Hh acetyltransferase HHAT, Wnt proteins, BMP4, and BMP6. Most of these genes were upregulated in patient-derived iPSCs grown in osteoblast differentiation medium (OBM) and downregulated in control iPSCs cultured in OBM. The expression of GLI1 and GLI2 substantially decreased in both control and patient-derived iPSCs cultured in OBM, whereas GLI3, SHH, and IHH were upregulated in patient-derived iPSCs and downregulated in control iPSCs grown in OBM. Activation of Smoothened by SAG in cells grown in OBM significantly enhanced alkaline phosphatase activity in patient-derived iPSCs compared with control iPSC lines. In summary, patient-derived iPSCs expressed lower basal levels than the control iPSCs of the genes encoding Hh, Wnt, and bone morphogenetic proteins, but their expression of these genes strongly increased under osteogenic conditions. These findings indicate that patient-derived iPSCs are hypersensitive to osteogenic induction. We propose that Hh signaling is constituently active in iPSCs from Gorlin syndrome patients, enhancing their response to osteogenic induction and contributing to disease-associated abnormalities.
巻・号 12(10)
ページ e0186879
公開日 2017-10-31
DOI 10.1371/journal.pone.0186879
PII PONE-D-17-12732
PMID 29088246
PMC PMC5663396
MeSH Acyltransferases / genetics Adult Basal Cell Nevus Syndrome / genetics Basal Cell Nevus Syndrome / pathology Bone Morphogenetic Protein 4 / genetics Bone Morphogenetic Protein 6 / genetics Cell Differentiation / genetics Cells, Cultured Gene Expression Profiling / methods Hedgehog Proteins / genetics* Humans Induced Pluripotent Stem Cells / metabolism* Male Middle Aged Mutation Osteogenesis / genetics* Patched-1 Receptor / genetics Signal Transduction / genetics* Wnt Proteins / genetics Young Adult Zinc Finger Protein GLI1 / genetics
IF 2.74
引用数 4
リソース情報
ヒト・動物細胞 253G1(HPS0002) Nips-B2(HPS0223)