title: Nanoparticle delivery of stable miR-199a-5p agomir improves the osteogenesis of human mesenchymal stem cells via the HIF1a pathway.
abstract: Elucidating the regulatory mechanisms of osteogenesis of human mesenchymal stem cell (hMSC) is important for the development of cell therapies for bone loss and regeneration. Here we showed that hsa-miR-199a-5p modulated osteogenic differentiation of hMSCs at both early and late stages through HIF1a pathway. hsa-miR-199a expression was up-regulated during osteogenesis for both of two mature forms, miR-199a-5p and -3p. Over-expression of miR-199a-5p but not -3p enhanced differentiation of hMSCs in vitro, whereas inhibition of miR-199a-5p reduced the expression of osteoblast-specific genes, alkaline phosphatase (ALP) activity, and mineralization. Furthermore, over-expression of miR-199a enhanced ectopic bone formation in vivo. Chitosan nanoparticles were used for delivery of stable modified hsa-miR-199a-5p (agomir) both in vitro and in vivo, as a proof-of-concept for stable agomir delivery on bone regeneration. The hsa-mir199a-5p agomir were mixed with Chitosan nanoparticles to form nanoparticle/hsa-mir199a-5p agomir plasmid (nanoparticle/agomir) complexes, and nanoparticle/agomir complexes could improve the in vivo regeneration of bone. Further mechanism studies revealed that hypoxia enhanced osteogenesis at early stage and inhibited osteogenesis maturation at late stage through HIF1a-Twist1 pathway. At early stage of differentiation, hypoxia induced HIF1a-Twist1 pathway to enhance osteogenesis by up-regulating miR-199a-5p, while at late stage of differentiation, miR-199a-5p enhanced osteogenesis maturation by inhibiting HIF1α-Twist1 pathway.
authors.affiliation: Zhejiang Provincial Key Laboratory of Tissue Engineering and Regenerative Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China; School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong Special Administrative Region.
authors.forename: Xiao
authors.initials: X
authors.lastname: Chen
authors.affiliation: School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong Special Administrative Region.
authors.forename: Shen
authors.initials: S
authors.lastname: Gu
authors.affiliation: School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong Special Administrative Region; Department of Bioscience and Biotechnology, School of Chemical Engineering and Life Sciences, Wuhan University of Technology, Wuhan, China.
authors.forename: Bi-Feng
authors.initials: BF
authors.lastname: Chen
authors.affiliation: Zhejiang Provincial Key Laboratory of Tissue Engineering and Regenerative Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.
authors.forename: Wei-Liang
authors.initials: WL
authors.lastname: Shen
authors.affiliation: Zhejiang Provincial Key Laboratory of Tissue Engineering and Regenerative Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.
authors.forename: Zi
authors.initials: Z
authors.lastname: Yin
authors.affiliation: Zhejiang Provincial Key Laboratory of Tissue Engineering and Regenerative Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.
authors.forename: Guo-Wei
authors.initials: GW
authors.lastname: Xu
authors.affiliation: Zhejiang Provincial Key Laboratory of Tissue Engineering and Regenerative Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.
authors.forename: Jia-Jie
authors.initials: JJ
authors.lastname: Hu
authors.affiliation: Zhejiang Provincial Key Laboratory of Tissue Engineering and Regenerative Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Zhu
authors.affiliation: School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong Special Administrative Region.
authors.forename: Gang
authors.initials: G
authors.lastname: Li
authors.affiliation: School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong Special Administrative Region.
authors.forename: Chao
authors.initials: C
authors.lastname: Wan
authors.affiliation: Zhejiang Provincial Key Laboratory of Tissue Engineering and Regenerative Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.
authors.forename: Hong-Wei
authors.initials: HW
authors.lastname: Ouyang
authors.affiliation: School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong Special Administrative Region. Electronic address: leetl@cuhk.edu.hk.
authors.forename: Tin-Lap
authors.initials: TL
authors.lastname: Lee
authors.affiliation: School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong Special Administrative Region. Electronic address: chanwy@cuhk.edu.hk.
authors.forename: Wai-Yee
authors.initials: WY
authors.lastname: Chan
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects*; D051379:Mice; D016688:Mice, Inbred NOD; D035683:MicroRNAs / Q000008:administration & dosage* / Q000494:pharmacology; D053758:Nanoparticles; D009687:Nuclear Proteins / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects*; D051381:Rats; D017207:Rats, Sprague-Dawley; D051879:Twist-Related Protein 1 / Q000378:metabolism
chemical_list: C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D035683:MicroRNAs; D009687:Nuclear Proteins; C103173:TWIST1 protein, human; D051879:Twist-Related Protein 1; C531408:mirn199 microRNA, human
keywords: Bone differentiation; Drug delivery; Human mesenchymal stem cells; Maturation; Regeneration; hsa-mir-199a