Mutant MAPK7-Induced Idiopathic Scoliosis is Linked to Impaired Osteogenesis
Background/Aims: Three rare MAPK7 variants that predispose individuals to adolescent idiopathic scoliosis have previously been identified. However, the mechanism underlying the effects of the mutations remain unknown. Methods: Human mesenchymal stem cells (hMSCs) were isolated from both patients and...
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Cell Physiol Biochem Press GmbH & Co KG
2018-07-01
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doaj-3fd416206c7842298adb727f751040382020-11-24T21:45:45ZengCell Physiol Biochem Press GmbH & Co KGCellular Physiology and Biochemistry1015-89871421-97782018-07-0148388089010.1159/000491956491956Mutant MAPK7-Induced Idiopathic Scoliosis is Linked to Impaired OsteogenesisTaifeng ZhouChong ChenCaixia XuHang ZhouBo GaoDeying SuZhiheng LiaoYongyong LiShulan YangPeiqiang SuBackground/Aims: Three rare MAPK7 variants that predispose individuals to adolescent idiopathic scoliosis have previously been identified. However, the mechanism underlying the effects of the mutations remain unknown. Methods: Human mesenchymal stem cells (hMSCs) were isolated from both patients and healthy volunteer donors, and MAPK7 expression was detected by western blotting and real-time quantitative PCR (RT-qPCR). Zebrafish embryos were injected with mapk7 morpholinos or co-injected with morpholinos and wild-type (WT) MAPK7 messenger RNA (mRNA) at the one-cell stage, followed by calcein staining to evaluate bone formation. hMSCs were transfected with MAPK7 small interfering RNAs and osteogenesis was induced for 14 days. Alizarin red staining was performed and osteoblast markers were detected by western blotting and RT-qPCR. Since RPS6KA3 is a downstream target of MAPK7 and plays an important role in the osteogenesis, zebrafish embryos were then injected with rps6ka3 morpholinos, or co-injected with rps6ka3 or mapk7 morpholinos and WT RPS6KA3 mRNA at the one-cell stage. Results: MAPK7 expression in the patient group was much lower than in the control group. Morpholino-induced mapk7 knockdown in zebrafish embryos led to body curvature, which was significantly reversed by WT MAPK7 mRNA. Calcein staining revealed that mapk7-knockdown delayed the ossification of the vertebrae. MAPK7 silencing in hMSCs impaired osteogenesis and downregulated osteoblast marker expression. Morpholino-induced rps6ka3-knockdown in zebrafish embryos led to body curvature, which was reversed by WT RPS6KA3 mRNA. Interestingly, RPS6KA3 mRNA also partially reversed the phenotype induced by mapk7 morpholinos. Conclusion: Impaired osteogenesis is linked to mutant MAPK7-induced idiopathic scoliosis , and RPS6KA3 may play an important role in this process.https://www.karger.com/Article/FullText/491956Mapk7Idiopathic scoliosisZebrafishHMSCOsteogenesisRPS6KA3 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Taifeng Zhou Chong Chen Caixia Xu Hang Zhou Bo Gao Deying Su Zhiheng Liao Yongyong Li Shulan Yang Peiqiang Su |
spellingShingle |
Taifeng Zhou Chong Chen Caixia Xu Hang Zhou Bo Gao Deying Su Zhiheng Liao Yongyong Li Shulan Yang Peiqiang Su Mutant MAPK7-Induced Idiopathic Scoliosis is Linked to Impaired Osteogenesis Cellular Physiology and Biochemistry Mapk7 Idiopathic scoliosis Zebrafish HMSC Osteogenesis RPS6KA3 |
author_facet |
Taifeng Zhou Chong Chen Caixia Xu Hang Zhou Bo Gao Deying Su Zhiheng Liao Yongyong Li Shulan Yang Peiqiang Su |
author_sort |
Taifeng Zhou |
title |
Mutant MAPK7-Induced Idiopathic Scoliosis is Linked to Impaired Osteogenesis |
title_short |
Mutant MAPK7-Induced Idiopathic Scoliosis is Linked to Impaired Osteogenesis |
title_full |
Mutant MAPK7-Induced Idiopathic Scoliosis is Linked to Impaired Osteogenesis |
title_fullStr |
Mutant MAPK7-Induced Idiopathic Scoliosis is Linked to Impaired Osteogenesis |
title_full_unstemmed |
Mutant MAPK7-Induced Idiopathic Scoliosis is Linked to Impaired Osteogenesis |
title_sort |
mutant mapk7-induced idiopathic scoliosis is linked to impaired osteogenesis |
publisher |
Cell Physiol Biochem Press GmbH & Co KG |
series |
Cellular Physiology and Biochemistry |
issn |
1015-8987 1421-9778 |
publishDate |
2018-07-01 |
description |
Background/Aims: Three rare MAPK7 variants that predispose individuals to adolescent idiopathic scoliosis have previously been identified. However, the mechanism underlying the effects of the mutations remain unknown. Methods: Human mesenchymal stem cells (hMSCs) were isolated from both patients and healthy volunteer donors, and MAPK7 expression was detected by western blotting and real-time quantitative PCR (RT-qPCR). Zebrafish embryos were injected with mapk7 morpholinos or co-injected with morpholinos and wild-type (WT) MAPK7 messenger RNA (mRNA) at the one-cell stage, followed by calcein staining to evaluate bone formation. hMSCs were transfected with MAPK7 small interfering RNAs and osteogenesis was induced for 14 days. Alizarin red staining was performed and osteoblast markers were detected by western blotting and RT-qPCR. Since RPS6KA3 is a downstream target of MAPK7 and plays an important role in the osteogenesis, zebrafish embryos were then injected with rps6ka3 morpholinos, or co-injected with rps6ka3 or mapk7 morpholinos and WT RPS6KA3 mRNA at the one-cell stage. Results: MAPK7 expression in the patient group was much lower than in the control group. Morpholino-induced mapk7 knockdown in zebrafish embryos led to body curvature, which was significantly reversed by WT MAPK7 mRNA. Calcein staining revealed that mapk7-knockdown delayed the ossification of the vertebrae. MAPK7 silencing in hMSCs impaired osteogenesis and downregulated osteoblast marker expression. Morpholino-induced rps6ka3-knockdown in zebrafish embryos led to body curvature, which was reversed by WT RPS6KA3 mRNA. Interestingly, RPS6KA3 mRNA also partially reversed the phenotype induced by mapk7 morpholinos. Conclusion: Impaired osteogenesis is linked to mutant MAPK7-induced idiopathic scoliosis , and RPS6KA3 may play an important role in this process. |
topic |
Mapk7 Idiopathic scoliosis Zebrafish HMSC Osteogenesis RPS6KA3 |
url |
https://www.karger.com/Article/FullText/491956 |
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