BMP9-induced osteoblastic differentiation requires functional Notch signaling in mesenchymal stem cells

Cui, Jing; Zhang, Wenwen; Huang, Enyi; Wang, Jia; Liao, Junyi; Li, Ruidong; Yu, Xinyi; Zhao, Chen; Zeng, Zongyue; Shu, Yi; Zhang, Ruyi; Yan, Shujuan; Lei, Jiayan; Yang, Chao; Wu, Ke; Wu, Ying; Huang, Shifeng; Ji, Xiaojuan; Li, Alexander; Chen, Gong; Yuan, Chengfu; Zhang, Linghuan; Liu, Wei; Huang, Bo; Feng, Yixiao; An, Liping; Zhang, Bo; Dai, Zhengyu; Shen, Yi; Luo, Wei; Wang, Xi; Huang, Aiping; Luu, Hue H.; Reid, Russell R.; Wolf, Jennifer Moriatis; Wang, Shuai; Lee, Michael J.; He, Tong‐Chuan

doi:10.1038/s41374-018-0087-7

Cited by 57 publications

(63 citation statements)

References 68 publications

(78 reference statements)

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“…Our analysis showed that TGF-β, Notch, MAPK, and Ras signaling pathways mediate BMP9-induced differentiation. This result corroborates previous findings identifying the mechanism of BMP9-induced osteogenic differentiation [14,27,65,66,67,68]. Simultaneously, our results also revealed that insulin, PPAR, glutathione metabothyroid hormone, TNF, PI3K-Akt, and several cancer pathways are also involved in BMP9-mediated cellular differentiation.…”

Section: Resultssupporting

confidence: 93%

“…To verify our in vitro results that show exogenous BMP9-induced GC-1spg cell osteogenesis, we performed ectopic ossification experiments. Using our previously established stem cell implantation assay [14,17,38,53], we detected injection-site masses in both BMP9 and GFP groups. Masses in the BMP9 group appeared slightly larger than in the GFP group.…”

Section: Resultsmentioning

confidence: 99%

“…Several recent studies demonstrated that Notch signaling is required for BMP9-regulated bone formation in MSCs [14,27,48,64]. However, some studies also demonstrated that BMP9 can induce MSCs to differentiate into various cell types [22,65,66].…”

Section: Resultsmentioning

confidence: 99%

“…In other words, GC-1spg cells can differentiate into different cell types, including those of the osteogenic, chondrogenic, or adipogenic lineages [9,10,11,12]. However, several molecular events [13,14] and factors are involved in osteogenic differentiation of MSCs. Importantly, bone morphogenetic proteins (BMPs) are essential for this process [15,16,17].…”

Section: Introductionmentioning

confidence: 99%

“…BMP9, also known as growth differentiation factor 2 (GDF-2)[25], was originally isolated from the fetal mouse liver [25,26]. Bone formation involves BMP9 acting through the Notch signaling pathway [14,24,27]. Several metabolic processes are regulated by BMP9, including glucose and lipid metabolism [28,29], iron metabolism [30], endothelial function, and angiogenesis [31,32,33].…”

Section: Introductionmentioning

confidence: 99%

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Bone morphogenetic protein 9 induces osteogenic differentiation of germ cell 1 spermatogonial cells

Zhang

Xu²,

Chen³

et al. 2019

Preprint

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30Germ cell 1 spermatogonial (GC-1spg) cells are multipotent progenitor cells. We 31 previously confirmed that bone morphogenetic protein (BMP) 9 is among the most 32 osteogenic BMPs. However, whether GC-1spg cells are driven toward osteogenic 33 differentiation under proper stimuli is uncertain. Additionally, the molecular 34 mechanism of BMP9 remains unclear. In the present study, we aimed to determine 35 whether BMP9 can induce osteogenic differentiation of GC-1spg cells. Recombinant 36 adenoviruses were generated by the AdEasy system to regulate the BMP9 expression in 37 GC-1spg cells. We identified osteogenic markers by real-time PCR and staining 38 techniques in vitro. Ectopic ossification assays and histological analysis were also 39 performed to verify the in vivo activity of BMP9. Finally, potential signaling pathways 40 of BMP9 were assessed by transcriptome sequencing and KEGG enrichment analysis. 41Using recombinant adenoviruses, we demonstrate that BMP9 upregulates osteogenic 42 markers including Runx2, osteocalcin, osteopontin, and Sox9. BMP9 also activates 43 alkaline phosphatase activity and calcium deposition in GC-1spg cells. In vivo results 44show that BMP9 overexpression in GC-1spg cells promotes ectopic bone formation and 45 chondrogenesis. In addition, RNA-sequencing and KEGG pathway analysis 3 46 demonstrate that several signaling pathways are involved in BMP9-mediated 47 osteogenesis. GC-1spg cells not only maintain spermatogenesis but also retain the 48 ability to form bone tissue. Therefore, BMP9 activity in GC-1spg cells may help 49 identify signaling pathways implicated in bone formation and could be of use in 50 regenerative medicine. 51 Keywords: Bone morphogenetic protein 9, RNA-seq, KEGG pathway analysis, 52 recombinant adenovirus, chondrogenesis 53 Introduction 54 Germ cell 1 spermatogonial (GC-1spg) cells are mouse spermatogonia that are 55 immortalized by the SV40 large T antigen [1]. These cells can differentiate into diverse cell 56 types or undergo self-renewal, depending on different factors [2,3,4]. GC-1spg cells are 57 largely involved in spermatogenesis and have mesenchymal stem cell (MSC) differentiation 58 potential [5,6,7,8]. In other words, GC-1spg cells can differentiate into different cell types, 59including those of the osteogenic, chondrogenic, or adipogenic lineages [9,10,11,12]. 60However, several molecular events [13,14] and factors are involved in osteogenic 61 differentiation of MSCs. Importantly, bone morphogenetic proteins (BMPs) are essential for 62 this process [15,16,17]. 63 BMPs, members of the TGF-β superfamily, are required for bone formation, stem cell 64 differentiation, and male reproduction [15,16,18,19,20]. Thus far, more than 15 different 65 BMPs have been identified. Our previous work showed that BMP9 is the most osteogenic 66 BMP [21,22,23,24]. BMP9, also known as growth differentiation factor 2 (GDF-2)[25], was 67 originally isolated from the fetal mouse liver [25,26]. Bone formation involves BMP9 acting 68 through the Notch signaling ...

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Section: Resultssupporting

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Bone morphogenetic protein 9 induces osteogenic differentiation of germ cell 1 spermatogonial cells

Zhang

Xu²,

Chen³

et al. 2019

Preprint

Self Cite

View full text Add to dashboard Cite

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Bone morphogenetic protein 9 (BMP9) directly induces Notch effector molecule Hes1 through the SMAD signaling pathway in osteoblasts

et al. 2020

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Bone morphogenetic protein (BMP) 9 is one of the most osteogenic BMPs, but its mechanism of action has not been fully elucidated. Hes1, a transcriptional regulator with a basic helix‐loop‐helix domain, is a well‐known effector of Notch signaling. Here, we find that BMP9 induces periodic increases of Hes1 mRNA and protein expression in osteoblasts, presumably through an autocrine negative feedback mechanism. BMP9‐mediated Hes1 induction is significantly inhibited by an ALK inhibitor and overexpression of Smad7, an inhibitory Smad. Luciferase and ChIP assays revealed that two Smad‐binding sites in the 5′ upstream region of the mouse Hes1 gene are essential for transcriptional activation by BMP9. Thus, our data indicate that BMP9 induces Hes1 expression in osteoblasts via the Smad signaling pathway.

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Carboxymethyl chitosan prolongs adenovirus‐mediated expression of IL‐10 and ameliorates hepatic fibrosis in a mouse model

Gou

Weng

Chen

et al. 2022

Bioengineering & Transla Med

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Effective and safe liver‐directed gene therapy has great promise in treating a broad range of liver diseases. While adenoviral (Ad) vectors have been widely used for efficacious in vivo gene delivery, their translational utilities are severely limited due to the short duration of transgene expression and solicitation of host immune response. Used as a promising polymeric vehicle for drug release and nucleic acid delivery, carboxymethyl chitosan (CMC) is biocompatible, biodegradable, anti‐microbial, inexpensive, and easy accessible. Here, by exploiting its biocompatibility, controlled release capability and anti‐inflammatory activity, we investigated whether CMC can overcome the shortcomings of Ad‐mediated gene delivery, hence improving the prospect of Ad applications in gene therapy. We demonstrated that in the presence of optimal concentrations of CMC, Ad‐mediated transgene expression lasted up to 50 days after subcutaneous injection, and at least 7 days after intrahepatic injection. Histologic evaluation and immunohistochemical analysis revealed that CMC effectively alleviated Ad‐induced host immune response. In our proof‐of‐principle experiment using the CCl4‐induced experimental mouse model of chronic liver damage, we demonstrated that repeated intrahepatic administrations of Ad‐IL10 mixed with CMC effectively mitigated the development of hepatic fibrosis. Collectively, these results indicate that CMC can improve the prospect of Ad‐mediated gene therapy by diminishing the host immune response while allowing readministration and sustained transgene expression.

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BMP9-induced osteoblastic differentiation requires functional Notch signaling in mesenchymal stem cells

Cited by 57 publications

References 68 publications

Bone morphogenetic protein 9 induces osteogenic differentiation of germ cell 1 spermatogonial cells

Bone morphogenetic protein 9 induces osteogenic differentiation of germ cell 1 spermatogonial cells

Bone morphogenetic protein 9 (BMP9) directly induces Notch effector molecule Hes1 through the SMAD signaling pathway in osteoblasts

Carboxymethyl chitosan prolongs adenovirus‐mediated expression of IL‐10 and ameliorates hepatic fibrosis in a mouse model

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