Both JNK1 and JNK2 Are Indispensable for Sensitized Extracellular Matrix Mineralization in IKKβ-Deficient Osteoblasts

Extracellular matrix mineralization is critical for osteogenesis, and its dysregulation could result in osteoporosis and vascular calcification. IKK/NF-κB activation inhibits differentiation of osteoblasts, and reduces extracellular matrix mineralization, however the underlying mechanisms are poorly...

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Main Authors: Qianyun Hao, Zhuangzhuang Liu, Liaoxun Lu, Lichen Zhang, Li Zuo
Format: Article
Language:English
Published: Frontiers Media S.A. 2020-02-01
Series:Frontiers in Endocrinology
Subjects:
Online Access:https://www.frontiersin.org/article/10.3389/fendo.2020.00013/full
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spelling doaj-7af5ec9e00154ceea4bb188d6b147f7b2020-11-25T02:21:35ZengFrontiers Media S.A.Frontiers in Endocrinology1664-23922020-02-011110.3389/fendo.2020.00013505669Both JNK1 and JNK2 Are Indispensable for Sensitized Extracellular Matrix Mineralization in IKKβ-Deficient OsteoblastsQianyun Hao0Zhuangzhuang Liu1Liaoxun Lu2Lichen Zhang3Li Zuo4Department of Nephrology, Peking University People's Hospital, Beijing, ChinaLaboratory of Mouse Genetics, Institute of Psychiatry and Neuroscience, Xinxiang Medical University, Xinxiang, ChinaLaboratory of Mouse Genetics, Institute of Psychiatry and Neuroscience, Xinxiang Medical University, Xinxiang, ChinaLaboratory of Genetic Regulators in the Immune System, Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, Xinxiang Medical University, Xinxiang, ChinaDepartment of Nephrology, Peking University People's Hospital, Beijing, ChinaExtracellular matrix mineralization is critical for osteogenesis, and its dysregulation could result in osteoporosis and vascular calcification. IKK/NF-κB activation inhibits differentiation of osteoblasts, and reduces extracellular matrix mineralization, however the underlying mechanisms are poorly understood. In this study, we used CRISPR/Cas9 system to permanently inactivate IKKβ in preosteoblast cells and confirmed that such cells displayed dramatic increase in extracellular matrix mineralization associated with JNK phosphorylation. Such observation was also found in our study using IKKβ-deficient primary murine osteoblasts. Interestingly, we found that in Ikbkb−/−Mapk8−/− or Ikbkb−/−Mapk9−/− double knockout cells, the enhanced mineralization caused by IKKβ deficiency was completely abolished, and deletion of either Mapk8 or Mapk9 was sufficient to dampen c-Jun phosphorylation. In further experiments, we discovered that absence of JNK1 or JNK2 on IKKβ-deficient background resulted in highly conserved transcriptomic alteration in response to osteogenic induction. Therefore, identification of the indispensable roles of JNK1 and JNK2 in activating c-Jun and promoting osteoblast differentiation on IKKβ-deficient background provided novel insights into restoring homeostasis in extracellular matrix mineralization.https://www.frontiersin.org/article/10.3389/fendo.2020.00013/fullosteoblastsIKKβNF-κBJNK1JNK2CRISPR/Cas9
collection DOAJ
language English
format Article
sources DOAJ
author Qianyun Hao
Zhuangzhuang Liu
Liaoxun Lu
Lichen Zhang
Li Zuo
spellingShingle Qianyun Hao
Zhuangzhuang Liu
Liaoxun Lu
Lichen Zhang
Li Zuo
Both JNK1 and JNK2 Are Indispensable for Sensitized Extracellular Matrix Mineralization in IKKβ-Deficient Osteoblasts
Frontiers in Endocrinology
osteoblasts
IKKβ
NF-κB
JNK1
JNK2
CRISPR/Cas9
author_facet Qianyun Hao
Zhuangzhuang Liu
Liaoxun Lu
Lichen Zhang
Li Zuo
author_sort Qianyun Hao
title Both JNK1 and JNK2 Are Indispensable for Sensitized Extracellular Matrix Mineralization in IKKβ-Deficient Osteoblasts
title_short Both JNK1 and JNK2 Are Indispensable for Sensitized Extracellular Matrix Mineralization in IKKβ-Deficient Osteoblasts
title_full Both JNK1 and JNK2 Are Indispensable for Sensitized Extracellular Matrix Mineralization in IKKβ-Deficient Osteoblasts
title_fullStr Both JNK1 and JNK2 Are Indispensable for Sensitized Extracellular Matrix Mineralization in IKKβ-Deficient Osteoblasts
title_full_unstemmed Both JNK1 and JNK2 Are Indispensable for Sensitized Extracellular Matrix Mineralization in IKKβ-Deficient Osteoblasts
title_sort both jnk1 and jnk2 are indispensable for sensitized extracellular matrix mineralization in ikkβ-deficient osteoblasts
publisher Frontiers Media S.A.
series Frontiers in Endocrinology
issn 1664-2392
publishDate 2020-02-01
description Extracellular matrix mineralization is critical for osteogenesis, and its dysregulation could result in osteoporosis and vascular calcification. IKK/NF-κB activation inhibits differentiation of osteoblasts, and reduces extracellular matrix mineralization, however the underlying mechanisms are poorly understood. In this study, we used CRISPR/Cas9 system to permanently inactivate IKKβ in preosteoblast cells and confirmed that such cells displayed dramatic increase in extracellular matrix mineralization associated with JNK phosphorylation. Such observation was also found in our study using IKKβ-deficient primary murine osteoblasts. Interestingly, we found that in Ikbkb−/−Mapk8−/− or Ikbkb−/−Mapk9−/− double knockout cells, the enhanced mineralization caused by IKKβ deficiency was completely abolished, and deletion of either Mapk8 or Mapk9 was sufficient to dampen c-Jun phosphorylation. In further experiments, we discovered that absence of JNK1 or JNK2 on IKKβ-deficient background resulted in highly conserved transcriptomic alteration in response to osteogenic induction. Therefore, identification of the indispensable roles of JNK1 and JNK2 in activating c-Jun and promoting osteoblast differentiation on IKKβ-deficient background provided novel insights into restoring homeostasis in extracellular matrix mineralization.
topic osteoblasts
IKKβ
NF-κB
JNK1
JNK2
CRISPR/Cas9
url https://www.frontiersin.org/article/10.3389/fendo.2020.00013/full
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