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Non-virus-mediated transfer of siRNAs against Runx2 and Smad4 inhibit heterotopic ossification in rats

Gene Therapy volume 17, pages 370–379 (2010)Cite this article

Abstract

Heterotopic ossification of muscles, tendons and ligaments are a common problem affecting patient with trauma or received elective surgery. But the existing preventive or therapeutic methods all have disadvantages. Runt-related protein 2 (Runx2) and Smad4 are two regulators that have important roles in the differentiation of osteoblast. In this study, we attempted to examine the effect of Runx2 and Smad4 on the development of heterotopic ossification in vitro. We constructed non-virus-containing small interference RNAs (siRNAs) against Runx2 and Smad4 and tested it with reverse transcriptase-PCR and western blot. We then analyzed the independent effect of Runx2- and Smad4-specific siRNAs and their cooperative effect on the formation of heterotopic ossification induced by trauma in rats. The effects were measured with computed tomography scanning, hematoxylin and eosin staining and immunohistochemistry. We found that the Runx2- and Smad4-specific siRNAs inhibited the expression of Runx2 and Smad4 at the level of messenger RNA and protein. Runx2 and Smad4 independently inhibited the formation of heterotopic ossification. Moreover, their co-expression significantly enhanced the inhibition of heterotopic ossification compared with the independent effect. We suggest that gene therapy to inhibit Runx2 and Smad4 by RNAi could be a powerful approach to prevent or treat heterotopic ossification.

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Acknowledgements

This research was supported by the program of Gene Therapy on Sports Injury sponsored by National Natural Science Foundation of China (Grant Number: ZR09-1-01). The PiggyBac vectors were provided by the Institute of Development Biology and Molecular Medicine at Fudan University. The paper submitted does not contain information regarding medical device(s)/drug(s).

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Authors and Affiliations

  1. Institute of Sports Medicine, Peking University Third Hospital, Beijing, PR China

    T Xue, L Lin, Y Hou, X Fu, J Zhang & C Yu

  2. Department of Biochemistry and Molecular Biology, Peking University School of Basic Medical Science, Beijing, PR China

    Z Mao

  3. Department of Orthopedics Surgery, Tianjin Hospital, Tianjin, PR China

    X Wei

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  1. T Xue
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Correspondence to C Yu.

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Xue, T., Mao, Z., Lin, L. et al. Non-virus-mediated transfer of siRNAs against Runx2 and Smad4 inhibit heterotopic ossification in rats. Gene Ther 17, 370–379 (2010). https://doi.org/10.1038/gt.2009.154

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