Abstract
Gorlin syndrome (GS) is a hereditary disorder with tumorigenicity, caused by constitutive hyperactivity of hedgehog signaling. Smoothened (SMO) antagonists have been effectively used in the clinical treatment of hedgehog signaling-related cancer. However, these treatments have led to problematic side effects, including severe adverse reactions and drug resistance from additional somatic mutations. We profiled microRNAs in GS fibroblasts to explore a novel therapeutic target for controlling hyper-activated hedgehog signaling. To identify GS-related microRNAs, we analyzed dermal fibroblasts from five patients with GS and three normal controls. We used microarray comparative genomic hybridization to screen 632 human microRNAs in GS fibroblasts. We identified 16 down- and 19 upregulated microRNAs with over twofold change in expression. We validated the increased expression of four microRNAs, confirming hsa-miR-196a-5p downregulation and hsa-miR-4485 upregulation using real-time PCR. Moreover, hsa-miR-196a-5p is complementary to sites in the 3′ UTR of MAP3K1, which exhibits upregulated expression at mRNA and protein levels in GS fibroblasts. In addition, hedgehog signal induction with exogenous components decreased miR-196a-5p expression and increased map3k1 expression in a mouse mesenchymal cell line. Given that MAP3K1 has been reported to activate hedgehog signaling, hsa-miR-196a-5p may contribute to the positive feedback loop in this pathway.
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Acknowledgements
This study was supported by Grant-in-Aid for Young Scientists (B) (17K16241) and Grant-in-Aid for Scientific Research (C) (16K09960) of the Japan Society for the Promotion of Science (JSPS).
Funding
Contract grant sponsor: Grant-in-Aid for Young Scientists (B), the Japan Society for the Promotion of Science (JSPS); Contract grant number: 17K16241. Contract grant sponsor: Grant-in-Aid for Scientific Research (C), the Japan Society for the Promotion of Science (JSPS); Contract grant number: 16K09960.
Author contributions
TS: study conceptualization, design, experimental operation, and manuscript preparation. TM and HU: analysis and interpretation of genetic analysis data. TT: experimental operation. HI, TM and TU: analysis of clinical manifestations. KF and NS: manuscript revision and study supervision.
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This study was approved by the Ethics Committee of Chiba University Graduate School of Medicine (IRB number 792) and conformed to the provisions of the Declaration of Helsinki in 1995 (as revised in Seoul 2008).
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Shiohama, T., Fujii, K., Miyashita, T. et al. MicroRNAs profiling in fibroblasts derived from patients with Gorlin syndrome. J Hum Genet 64, 757–765 (2019). https://doi.org/10.1038/s10038-019-0607-3
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DOI: https://doi.org/10.1038/s10038-019-0607-3
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