Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Technical Report
  • Published:

A delivery system targeting bone formation surfaces to facilitate RNAi-based anabolic therapy

Nature Medicine volume 18, pages 307–314 (2012)Cite this article

Subjects

Abstract

Metabolic skeletal disorders associated with impaired bone formation are a major clinical challenge. One approach to treat these defects is to silence bone-formation–inhibitory genes by small interference RNAs (siRNAs) in osteogenic-lineage cells that occupy the niche surrounding the bone-formation surfaces. We developed a targeting system involving dioleoyl trimethylammonium propane (DOTAP)-based cationic liposomes attached to six repetitive sequences of aspartate, serine, serine ((AspSerSer)6) for delivering siRNAs specifically to bone-formation surfaces. Using this system, we encapsulated an osteogenic siRNA that targets casein kinase-2 interacting protein-1 (encoded by Plekho1, also known as Plekho1). In vivo systemic delivery of Plekho1 siRNA in rats using our system resulted in the selective enrichment of the siRNAs in osteogenic cells and the subsequent depletion of Plekho1. A bioimaging analysis further showed that this approach markedly promoted bone formation, enhanced the bone micro-architecture and increased the bone mass in both healthy and osteoporotic rats. These results indicate (AspSerSer)6-liposome as a promising targeted delivery system for RNA interference–based bone anabolic therapy.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Differential occupancy characteristics of (AspSerSer)6 compared to Asp8 at bone-formation or bone-resorption surfaces in nondecalcified bone sections using a confocal laser scanning microscope.
Figure 2: Organ-selective delivery and gene knockdown in vivo.
Figure 3: Cell-selective delivery and knockdown efficiency in vivo.
Figure 4: In vivo microCT examinations of the three-dimensional trabecular architecture and an ex vivo bone formation evaluation in nondecalcified bone sections in healthy rats.
Figure 5: In vivo microCT examination of the three-dimensional trabecular architecture in OVX-treated rats.

Similar content being viewed by others

References

  1. Black, D.M. et al. The effects of parathyroid hormone and alendronate alone or in combination in postmenopausal osteoporosis. N. Engl. J. Med. 349, 1207–1215 (2003).

    Article  CAS  Google Scholar 

  2. Hodsman, A.B. et al. Parathyroid hormone and teriparatide for the treatment of osteoporosis: a review of the evidence and suggested guidelines for its use. Endocr. Rev. 26, 688–703 (2005).

    Article  CAS  Google Scholar 

  3. Cosman, F. et al. Daily and cyclic parathyroid hormone in women receiving alendronate. N. Engl. J. Med. 353, 566–575 (2005).

    Article  CAS  Google Scholar 

  4. Lindsay, R. et al. Randomised controlled study of effect of parathyroid hormone on vertebral-bone mass and fracture incidence among postmenopausal women on oestrogen with osteoporosis. Lancet 350, 550–555 (1997).

    Article  CAS  Google Scholar 

  5. Neer, R.M. et al. Effect of parathyroid hormone (1–34) on fractures and bone mineral density in postmenopausal women with osteoporosis. N. Engl. J. Med. 344, 1434–1441 (2001).

    Article  CAS  Google Scholar 

  6. López-Fraga, M., Martinez, T. & Jimenez, A. RNA interference technologies and therapeutics: from basic research to products. BioDrugs 23, 305–332 (2009).

    Article  Google Scholar 

  7. Novina, C.D. & Sharp, P.A. The RNAi revolution. Nature 430, 161–164 (2004).

    Article  CAS  Google Scholar 

  8. Itaka, K. et al. Bone regeneration by regulated in vivo gene transfer using biocompatible polyplex nanomicelles. Mol. Ther. 15, 1655–1662 (2007).

    Article  CAS  Google Scholar 

  9. Wang, D., Miller, S.C., Kopeckova, P. & Kopecek, J. Bone-targeting macromolecular therapeutics. Adv. Drug Deliv. Rev. 57, 1049–1076 (2005).

    Article  CAS  Google Scholar 

  10. Wang, D. et al. Osteotropic peptide that differentiates functional domains of the skeleton. Bioconjug. Chem. 18, 1375–1378 (2007).

    Article  CAS  Google Scholar 

  11. Yarbrough, D.K. et al. Specific binding and mineralization of calcified surfaces by small peptides. Calcif. Tissue Int. 86, 58–66 (2010).

    Article  CAS  Google Scholar 

  12. Lu, K. et al. Targeting WW domains linker of HECT-type ubiquitin ligase Smurf1 for activation by Ckip-1. Nat. Cell Biol. 10, 994–1002 (2008).

    Article  CAS  Google Scholar 

  13. Zhang, L. et al. The PH domain containing protein Ckip-1 binds to IFP35 and Nmi and is involved in cytokine signaling. Cell. Signal. 19, 932–944 (2007).

    Article  CAS  Google Scholar 

  14. Stuart, A.J. & Smith, D.A. Use of the fluorochromes xylenol orange, calcein green, and tetracycline to document bone deposition and remodeling in healing fractures in chickens. Avian Dis. 36, 447–449 (1992).

    Article  CAS  Google Scholar 

  15. Aubin, J. & JNM., H. Bone cell biology: osteoblast, osteocyte and osteoclasts. in Pediatric Bone 43–47 (Academic Press, San Diego, California, USA, 2002).

  16. Gronthos, S. et al. Differential cell surface expression of the STRO-1 and alkaline phosphatase antigens on discrete developmental stages in primary cultures of human bone cells. J. Bone Miner. Res. 14, 47–56 (1999).

    Article  CAS  Google Scholar 

  17. Ishikawa, S. et al. Involvement of FcRγ in signal transduction of osteoclast-associated receptor (OSCAR). Int. Immunol. 16, 1019–1025 (2004).

    Article  CAS  Google Scholar 

  18. Kim, N., Takami, M., Rho, J., Josien, R. & Choi, Y. A novel member of the leukocyte receptor complex regulates osteoclast differentiation. J. Exp. Med. 195, 201–209 (2002).

    Article  CAS  Google Scholar 

  19. Posner, A.S. & Betts, F. Synthetic amorphous calcium-phosphate and its relation to bone-mineral structure. Acc. Chem. Res. 8, 273–281 (1975).

    Article  CAS  Google Scholar 

  20. Hoang, Q.Q., Sicheri, F., Howard, A.J. & Yang, D.S. Bone recognition mechanism of porcine osteocalcin from crystal structure. Nature 425, 977–980 (2003).

    Article  CAS  Google Scholar 

  21. Midura, R.J. et al. Bone acidic glycoprotein-75 delineates the extracellular sites of future bone sialoprotein accumulation and apatite nucleation in osteoblastic cultures. J. Biol. Chem. 279, 25464–25473 (2004).

    Article  CAS  Google Scholar 

  22. Steitz, S.A. et al. Osteopontin inhibits mineral deposition and promotes regression of ectopic calcification. Am. J. Pathol. 161, 2035–2046 (2002).

    Article  CAS  Google Scholar 

  23. Takahashi-Nishioka, T. et al. Targeted drug delivery to bone: pharmacokinetic and pharmacological properties of acidic oligopeptide-tagged drugs. Curr. Drug Discov. Technol. 5, 39–48 (2008).

    Article  CAS  Google Scholar 

  24. Wang, D., Miller, S., Sima, M., Kopeckova, P. & Kopecek, J. Synthesis and evaluation of water-soluble polymeric bone-targeted drug delivery systems. Bioconjug. Chem. 14, 853–859 (2003).

    Article  CAS  Google Scholar 

  25. Federman, N. & Denny, C.T. Targeting liposomes toward novel pediatric anticancer therapeutics. Pediatr. Res. 67, 514–519 (2010).

    Article  CAS  Google Scholar 

  26. Wang, G., Kucharski, C., Lin, X. & Uludag, H. Bisphosphonate-coated BSA nanoparticles lack bone targeting after systemic administration. J. Drug Target. 18, 611–626 (2010).

    Article  CAS  Google Scholar 

  27. Cullis, P.R., Mayer, L.D., Bally, M.B., Madden, T.D. & Hope, M.J. Generating and loading of liposomal systems for drug-delivery applications. Adv. Drug Deliv. Rev. 3, 267–282 (1989).

    Article  CAS  Google Scholar 

  28. Vegni, F.E., Corradini, C. & Privitera, G. Effects of parathyroid hormone and alendronate alone or in combination in osteoporosis. N. Engl. J. Med. 350, 189–192, author reply 189–192 (2004).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank X.-H. Yang for technical support with the confocal imaging, H. Yang for technical support with the flow cytometry and F.C. Chun-Wan for assistance with the bone histomorphometry. This study was supported by the Chinese National Basic Research Programs (2011CB910602), the Hong Kong Competitive Earmarked Research Grant (CUHK479111 and 473011), the Direct Grant of Faculty of Medicine of the Chinese University of Hong Kong (2041478 and 2041525), the Faculty Research Grant of Hong Kong Baptist University (30-08-089), the Chinese National Natural Science Foundation Project (30830029) and the National Key Technologies Research and Development Program for New Drugs (2009ZX09503-002).

Author information

Author notes

  1. Ge Zhang, Baosheng Guo, Heng Wu, Tao Tang and Bao-Ting Zhang: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Orthopaedics & Traumatology, Musculoskeletal Research Laboratory, The Chinese University of Hong Kong, Hong Kong, China

    Ge Zhang, Baosheng Guo, Heng Wu, Tao Tang, Bao-Ting Zhang, Lizhen Zheng, Yixin He, Xinluan Wang, Gang Li, Kwoksui Leung, Leungkim Hung & Ling Qin

  2. School of Chinese Medicine, The Chinese University of Hong Kong, Hong Kong, China

    Bao-Ting Zhang

  3. School of Chinese Medicine, Hong Kong Baptist University, Hong Kong, China

    Zhijun Yang

  4. Department of Orthopedics, Second Hospital of Medical College of Ji Nan University, Shenzhen People's Hospital, Shenzhen, China

    Xiaohua Pan

  5. School of Pharmacy, The Chinese University of Hong Kong, Hong Kong, China

    Heelum Chow & Kinwah To

  6. Center of Drug Delivery System, Shanghai Institute of Materia Medica, Shanghai, Chinese Academy of Sciences, Shanghai, China

    Yaping Li

  7. State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing, China

    Dahu Li, Fuchu He & Lingqiang Zhang

  8. Department of Imaging and Interventional Radiology, The Chinese University of Hong Kong, Hong Kong, China

    Yixiang Wang

  9. Lee Hysan Clinical Research Laboratory, The Chinese University of Hong Kong, Hong Kong, China

    Kwongman Lee

  10. Biomedical Research and Development Center, Guangdong Provincial Key Laboratory of Bioengineering Medicine, National Engineering Research Center of Genetic Medicine, Jinan University, Guangzhou, China

    Zhibo Hou

  11. Institute of Molecular Biology, Nan Kai University, Tianjin, China

    Nan Dong

  12. Translational Medicine Research & Development Center, Institute of Biomedical and Health Engineering, Shenzhen Institute of Advanced Technology, Shenzhen, China

    Ling Qin

Authors
  1. Ge Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Baosheng Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Heng Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tao Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Bao-Ting Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Lizhen Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yixin He
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Zhijun Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Xiaohua Pan
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Heelum Chow
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Kinwah To
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Yaping Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Dahu Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Xinluan Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Yixiang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Kwongman Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Zhibo Hou
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. Nan Dong
    View author publications

    You can also search for this author in PubMed Google Scholar

  19. Gang Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  20. Kwoksui Leung
    View author publications

    You can also search for this author in PubMed Google Scholar

  21. Leungkim Hung
    View author publications

    You can also search for this author in PubMed Google Scholar

  22. Fuchu He
    View author publications

    You can also search for this author in PubMed Google Scholar

  23. Lingqiang Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  24. Ling Qin
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

All the authors were involved in conducting, drafting or revising the manuscript. All the authors approved the final version of the manuscript for submission. L.Q. had full access to all of the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis. Study conception and supervision: G.Z., L.Q., L. Zhang, F.H. Design and preparation of delivery system: G.Z., H.W., Z.Y., H.C., Y.L., K.T. Design, synthesis and screening of siRNA sequences: T.T., G.Z., L. Zheng, Z.H., N.D. Analysis and interpretation of data from cell biology and molecular biology: B.G., T.T., B.-T.Z., G.Z., D.L., X.W., L.Q. Analysis and interpretation of data from immunohistochemistry: B.-T.Z., B.G., G.Z., K. Lee, L.Q. Analysis and interpretation of data from biophotonic imaging: B.G., G.Z., G.L., L.Q. Analysis and interpretation of data from microCT and bone histomorphometry: B.G., G.Z., Y.H., Y.W., L.Q. Analysis and interpretation of data for clinical relevance: G.Z., L.Q., X.P., L.H., K. Leung.

Corresponding authors

Correspondence to Ge Zhang, Lingqiang Zhang or Ling Qin.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4, Supplementary Tables 1–4, Supplementary Methods, Supplementary Discussion and Supplementary Results (PDF 3361 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhang, G., Guo, B., Wu, H. et al. A delivery system targeting bone formation surfaces to facilitate RNAi-based anabolic therapy. Nat Med 18, 307–314 (2012). https://doi.org/10.1038/nm.2617

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm.2617

This article is cited by

Search

Advanced search

Quick links

Nature Briefing: Translational Research

Sign up for the Nature Briefing: Translational Research newsletter — top stories in biotechnology, drug discovery and pharma.

Get what matters in translational research, free to your inbox weekly. Sign up for Nature Briefing: Translational Research