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.
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DOI：
10.1371/journal.pone.0186879
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Azuma T
中科院分区：
文献类型：
--
作者：
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
Gorlin综合征是一种常染色体显性遗传综合征，易使患者形成基底细胞癌、牙源性角化囊肿和骨骼异常。已经在Gorlin综合征患者中鉴定了与音刺猬（SHH）信号通路相关的几个基因（包括PTCH 1）的致病突变。然而，在这些患者中没有明确的基因型-表型相关性，并且其临床表现差异很大，通常导致诊断和治疗延迟。我们使用仙台病毒载体（SeVdp（KOSM）302）从四名PTCH 1功能缺失突变的不相关Gorlin综合征患者中产生iPSC。患者来源的iPSC表现出基本的iPSC特征，包括干细胞标志物表达、全能性和形成畸胎瘤的能力。成纤维细胞和患者来源的iPSC中的GLI 1表达水平高于相应的对照细胞。与对照iPSC相比，患者来源的iPSC表达的编码Hh配体Indian Hedgehog（IHH）和SHH、Hh乙酰转移酶HHAT、Wnt蛋白、BMP 4和BMP 6的基因的基础水平较低。这些基因中的大多数在成骨细胞分化培养基（OBM）中生长的患者来源的iPSC中上调，并且在OBM中培养的对照iPSC中下调。在OBM中培养的对照和患者来源的iPSC中，GLI 1和GLI 2的表达显著降低，而GLI 3、SHH和IHH在患者来源的iPSC中上调，在OBM中生长的对照iPSC中下调。与对照iPSC系相比，在OBM中生长的细胞中通过SAG激活Smoothened显著增强了患者来源的iPSC中的碱性磷酸酶活性。总之，患者来源的iPSC表达的编码Hh、Wnt和骨形态发生蛋白的基因的基础水平低于对照iPSC，但在成骨条件下，这些基因的表达强烈增加。这些发现表明患者来源的iPSC对成骨诱导高度敏感。我们认为，Hh信号在Gorlin综合征患者的iPSCs中是组成性活跃的，增强了它们对成骨诱导的反应，并导致了疾病相关的异常。
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.
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DOI：
10.1074/jbc.m114.614578
发表时间：
2015-02-06
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Konitsiotis AD;Jovanović B;Ciepla P;Spitaler M;Lanyon-Hogg T;Tate EW;Magee AI
通讯作者：
Magee AI
影响因子：
3.7
作者：
Kurosaka H;Trainor PA;Leroux-Berger M;Iulianella A
通讯作者：
Iulianella A
影响因子：
3.7
作者：
Ono M;Hamada Y;Horiuchi Y;Matsuo-Takasaki M;Imoto Y;Satomi K;Arinami T;Hasegawa M;Fujioka T;Nakamura Y;Noguchi E
通讯作者：
Noguchi E
影响因子：
10.3
作者：
Brellier, F.;Valin, A.;Magnaldo, T.
通讯作者：
Magnaldo, T.
DOI：
10.1038/nrc2503
发表时间：
2008-10
期刊：
Nature reviews. Cancer
影响因子：
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作者：
通讯作者：
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