title: Endoplasmic reticulum (ER) stress inducible factor cysteine-rich with EGF-like domains 2 (Creld2) is an important mediator of BMP9-regulated osteogenic differentiation of mesenchymal stem cells.
abstract: Mesenchymal stem cells (MSCs) are multipotent progenitors that can undergo osteogenic differentiation under proper stimuli. We demonstrated that BMP9 is one of the most osteogenic BMPs. However, the molecular mechanism underlying BMP9-initiated osteogenic signaling in MSCs remains unclear. Through gene expression profiling analysis we identified several candidate mediators of BMP9 osteogenic signaling. Here, we focus on one such signaling mediator and investigate the functional role of cysteine-rich with EGF-like domains 2 (Creld2) in BMP9-initiated osteogenic signaling. Creld2 was originally identified as an ER stress-inducible factor localized in the ER-Golgi apparatus. Our genomewide expression profiling analysis indicates that Creld2 is among the top up-regulated genes in BMP9-stimulated MSCs. We confirm that Creld2 is up-regulated by BMP9 in MSCs. ChIP analysis indicates that Smad1/5/8 directly binds to the Creld2 promoter in a BMP9-dependent fashion. Exogenous expression of Creld2 in MSCs potentiates BMP9-induced early and late osteogenic markers, and matrix mineralization. Conversely, silencing Creld2 expression inhibits BMP9-induced osteogenic differentiation. In vivo stem cell implantation assay reveals that exogenous Creld2 promotes BMP9-induced ectopic bone formation and matrix mineralization, whereas silencing Creld2 expression diminishes BMP9-induced bone formation and matrix mineralization. We further show that Creld2 is localized in ER and the ER stress inducers potentiate BMP9-induced osteogenic differentiation. Our results strongly suggest that Creld2 may be directly regulated by BMP9 and ER stress response may play an important role in regulating osteogenic differentiation.
authors.affiliation: Ministry of Education Key Laboratory of Diagnostic Medicine and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China.
authors.forename: Jiye
authors.initials: J
authors.lastname: Zhang
authors.forename: Yaguang
authors.initials: Y
authors.lastname: Weng
authors.forename: Xing
authors.initials: X
authors.lastname: Liu
authors.forename: Jinhua
authors.initials: J
authors.lastname: Wang
authors.forename: Wenwen
authors.initials: W
authors.lastname: Zhang
authors.forename: Stephanie H
authors.initials: SH
authors.lastname: Kim
authors.forename: Hongyu
authors.initials: H
authors.lastname: Zhang
authors.forename: Ruidong
authors.initials: R
authors.lastname: Li
authors.forename: Yuhan
authors.initials: Y
authors.lastname: Kong
authors.forename: Xiang
authors.initials: X
authors.lastname: Chen
authors.forename: Wei
authors.initials: W
authors.lastname: Shui
authors.forename: Ning
authors.initials: N
authors.lastname: Wang
authors.forename: Chen
authors.initials: C
authors.lastname: Zhao
authors.forename: Ningning
authors.initials: N
authors.lastname: Wu
authors.forename: Yunfeng
authors.initials: Y
authors.lastname: He
authors.forename: Guoxin
authors.initials: G
authors.lastname: Nan
authors.forename: Xian
authors.initials: X
authors.lastname: Chen
authors.forename: Sheng
authors.initials: S
authors.lastname: Wen
authors.forename: Hongmei
authors.initials: H
authors.lastname: Zhang
authors.forename: Fang
authors.initials: F
authors.lastname: Deng
authors.forename: Lihua
authors.initials: L
authors.lastname: Wan
authors.forename: Hue H
authors.initials: HH
authors.lastname: Luu
authors.forename: Rex C
authors.initials: RC
authors.lastname: Haydon
authors.forename: Lewis L
authors.initials: LL
authors.lastname: Shi
authors.forename: Tong-Chuan
authors.initials: TC
authors.lastname: He
authors.forename: Qiong
authors.initials: Q
authors.lastname: Shi
mesh_terms: D000818:Animals; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000502:physiology*; D020868:Gene Silencing; D055427:Growth Differentiation Factor 2 / Q000502:physiology*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: C507070:CRELD2 protein, human; C561361:CRELD2 protein, mouse; D015815:Cell Adhesion Molecules; D016326:Extracellular Matrix Proteins; D055427:Growth Differentiation Factor 2
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