Document Type

Journal Article

Authors

Baosheng Guo, Institute for Advancing Translational Medicine in Bone AND Joint Diseases, School of Chinese Medicine, Hong Kong Baptist UniversityFollow
Baoting Zhang, School of Chinese Medicine, Department of Orthopaedics and Traumatology, The Chinese University of Hong Kong
Lizhen Zheng, School of Chinese Medicine, Department of Orthopaedics and Traumatology, The Chinese University of Hong Kong
Tao Tang, School of Chinese Medicine, Department of Orthopaedics and Traumatology, The Chinese University of Hong Kong
Jin Liu, Institute for Advancing Translational Medicine in Bone AND Joint Diseases, School of Chinese Medicine, Hong Kong Baptist University
Heng Wu, Institute for Advancing Translational Medicine in Bone AND Joint Diseases, School of Chinese Medicine, Hong Kong Baptist University
Zhijun Yang, Institute for Advancing Translational Medicine in Bone AND Joint Diseases, School of Chinese Medicine, Hong Kong Baptist UniversityFollow
Songlin Peng, Institute for Advancing Translational Medicine in Bone AND Joint Diseases, School of Chinese Medicine, Hong Kong Baptist University
Xiaojuan He, Institute of Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences
Hongqi Zhang, Institute for Advancing Translational Medicine in Bone AND Joint Diseases, School of Chinese Medicine, Hong Kong Baptist University
Kevin K.M. Yue, Institute for Advancing Translational Medicine in Bone AND Joint Diseases, School of Chinese Medicine, Hong Kong Baptist UniversityFollow
Fuchu He, State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine
Lingqiang Zhang, State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine
Ling Qin, School of Chinese Medicine, Department of Orthopaedics and Traumatology, The Chinese University of Hong Kong
Zhaoxiang Bian, Institute for Advancing Translational Medicine in Bone AND Joint Diseases, School of Chinese Medicine, Hong Kong Baptist UniversityFollow
Weihong Tan, Institute for Advancing Translational Medicine in Bone AND Joint Diseases, School of Chinese Medicine, Hong Kong Baptist University
Zicai Liang, Academician CHAN Sun Chi Albert Workroom for Advancing Translational Medicine in Bone, Joint Diseases, Kunshan RNAi Institute, Kunshan Industrial Technology Research Institute
Aiping Lu, Institute for Advancing Translational Medicine in Bone AND Joint Diseases, School of Chinese Medicine, Hong Kong Baptist UniversityFollow
Ge Zhang, Institute for Advancing Translational Medicine in Bone AND Joint Diseases, School of Chinese Medicine, Hong Kong Baptist UniversityFollow

Department/Unit

School of Chinese Medicine

Title

Therapeutic RNA interference targeting CKIP-1 with a cross-species sequence to stimulate bone formation

Language

English

Abstract

Objectives: Casein kinase 2 interacting protein 1 (CKIP-1) is a newly discovered intracellular negative regulator of bone formation without affecting bone resorption. In this study, we aimed to identify a cross-species siRNA sequence targeting CKIP-1 to facilitate developing a novel RNAi-based bone anabolic drug for reversing established osteoporosis. Methods: Eight specifically designed cross-species CKIP-1 siRNA sequences were screened in human, rhesus, rat and mouse osteoblast-like cells in vitro to identify the optimal sequence with the highest knockdown efficiency. The effect of this optimal siRNA sequence on osteogenic differentiation and matrix mineralization was further examined in osteoblast-like cells across different species, followed by an immunogenicity assessment in human peripheral blood mononuclear cells in vitro. The intra-osseous localization and silencing efficiency of the optimal siRNA were examined in vivo using a biophotonic system and real-time polymerase chain reaction, respectively. The RNAi-mediated cleavage of the CKIP-1 transcript was confirmed by rapid amplification of the 5' cDNA ends in vivo. Furthermore, the effect of the optimal siRNA sequence on osteogenic differentiation, bone turnover biomarkers, bone mass and micro-architecture parameters was investigated in healthy and osteoporotic rodents. Results: The CKIP-1 siRNA sequence (si-3) was identified as the optimal sequence, which consistently maintained CKIP-1 mRNA/protein expression at the lowest level across species in vitro. The si-3 significantly increased mRNA expression levels of osteoblast phenotypic genes and matrix mineralization across species without inducing an immunostimulatory activity in vitro. The intra-osseous localization and RNAi-mediated CKIP-1 silencing with high efficiency were confirmed in vivo. Periodic intravenous injections of si-3 promoted mRNA expression of osteoblast phenotypic genes, enhanced bone formation, increased bone mass and elevated serum level of bone formation marker without raising urine level of bone resorption marker in the healthy rodents. Moreover, the si-3 treatment promoted bone formation, improved trabecular micro-architecture and reversed bone loss in the osteoporotic mice. Conclusions: The identified optimal CKIP-1 siRNA sequence (si-3) could promote osteogenic differentiation across species in vitro, stimulate bone formation in the healthy rodents and reverse bone loss in the osteoporotic mice. © 2013.

Keywords

Bone formation, CKIP-1, Cross-species, SiRNA sequences

Publication Date

2014

Source Publication Title

Bone

Volume

59

Start Page

76

End Page

88

Publisher

Elsevier

DOI

10.1016/j.bone.2013.11.007

Link to Publisher's Edition

http://dx.doi.org/10.1016/j.bone.2013.11.007

ISSN (print)

87563282

ISSN (electronic)

18732763

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