Commercial anti-obesity drugs acting in the gastrointestinal tract or the central nervous system have been shown to have limited efficacy and severe side effects. Anti-obesity drug development is thus focusing on targeting adipocytes that store excess fat. Here, we show that an adipocyte-targeting fusion-oligopeptide gene carrier consisting of an adipocyte-targeting sequence and 9-arginine (ATS–9R) selectively transfects mature adipocytes by binding to prohibitin. Injection of ATS–9R into obese mice confirmed specific binding of ATS–9R to fat vasculature, internalization and gene expression in adipocytes. We also constructed a short-hairpin RNA (shRNA) for silencing fatty-acid-binding protein 4 (shFABP4), a key lipid chaperone in fatty-acid uptake and lipid storage in adipocytes. Treatment of obese mice with ATS–9R/shFABP4 led to metabolic recovery and body-weight reduction (>20%). The ATS–9R/shFABP4 oligopeptide complex could prove to be a safe therapeutic approach to regress and treat obesity as well as obesity-induced metabolic syndromes.
Rosen, E. D. & Spiegelman, B. M. Adipocytes as regulators of energy balance and glucose homeostasis. Nature 444, 847–853 (2006).
Ouchi, N., Parker, J. L., Lugus, J. J. & Walsh, K. Adipokines in inflammation and metabolic disease. Nature Rev. Immunol. 11, 85–97 (2011).
Cinti, S. The adipose organ. Prostaglandins Leukot. Essent. Fatty Acids 73, 9–15 (2005).
Elangbam, C. S. Review paper: Current strategies in the development of anti-obesity drugs and their safety concerns. Vet. Pathol. 46, 10–24 (2009).
Schaffler, A., Scholmerich, J. & Salzberger, B. Adipose tissue as an immunological organ: Toll-like receptors, C1q/TNFs and CTRPs. Trends Immunol. 28, 393–399 (2007).
Ahima, R. S. Central actions of adipocyte hormones. Trends Endocrinol. Metab. 16, 307–313 (2005).
Cooke, D. & Bloom, S. The obesity pipeline: Current strategies in the development of anti-obesity drugs. Nature Rev. Drug Discov. 5, 919–931 (2006).
Nawrocki, A. R. & Scherer, P. E. Keynote review: The adipocyte as a drug discovery target. Drug Discov. Today 10, 1219–1230 (2005).
Stenkula, K. G. et al. Human, but not rat, IRS1 targets to the plasma membrane in both human and rat adipocytes. Biochem. Biophys. Res. Commun. 363, 840–845 (2007).
Watanabe, M. et al. Regulation of PPAR gamma transcriptional activity in 3T3-L1 adipocytes. Biochem. Biophys. Res. Commun. 300, 429–436 (2003).
Cao, L. et al. Molecular therapy of obesity and diabetes by a physiological autoregulatory approach. Nature Med. 15, 447–454 (2009).
Dhillon, H. et al. Central leptin gene therapy suppresses body weight gain, adiposity and serum insulin without affecting food consumption in normal rats: A long-term study. Regul. Pept. 99, 69–77 (2001).
Chen, G. et al. Disappearance of body fat in normal rats induced by adenovirus-mediated leptin gene therapy. Proc. Natl Acad. Sci. USA 93, 14795–14799 (1996).
Won, Y. W., Yoon, S. M., Lee, K. M. & Kim, Y. H. Poly(oligo-D-arginine) with internal disulfide linkages as a cytoplasm-sensitive carrier for siRNA delivery. Mol. Ther. 19, 372–380 (2011).
Won, Y. W., Kim, H. A., Lee, M. & Kim, Y. H. Reducible poly(oligo-D-arginine) for enhanced gene expression in mouse lung by intratracheal injection. Mol. Ther. 18, 734–742 (2010).
Wilson, D. S. et al. Orally delivered thioketal nanoparticles loaded with TNF-alpha-siRNA target inflammation and inhibit gene expression in the intestines. Nature Mater. 9, 923–928 (2010).
Mok, H., Lee, S. H., Park, J. W. & Park, T. G. Multimeric small interfering ribonucleic acid for highly efficient sequence-specific gene silencing. Nature Mater. 9, 272–278 (2010).
Jeong, J. H., Kim, S. H., Christensen, L. V., Feijen, J. & Kim, S. W. Reducible poly(amido ethylenimine)-based gene delivery system for improved nucleus trafficking of plasmid DNA. Bioconjug. Chem. 21, 296–301 (2010).
Hyun, H. et al. Therapeutic effects of a reducible poly (oligo-D-arginine) carrier with the heme oxygenase-1 gene in the treatment of hypoxic-ischemic brain injury. Biomaterials 31, 9128–9134 (2010).
Kolonin, M. G., Saha, P. K., Chan, L., Pasqualini, R. & Arap, W. Reversal of obesity by targeted ablation of adipose tissue. Nature Med. 10, 625–632 (2004).
Mishra, S., Murphy, L. C., Nyomba, B. L. & Murphy, L. J. Prohibitin: A potential target for new therapeutics. Trends Mol. Med. 11, 192–197 (2005).
Patel, N. et al. Rescue of paclitaxel sensitivity by repression of Prohibitin1 in drug-resistant cancer cells. Proc. Natl Acad. Sci. USA 107, 2503–2508 (2010).
Sharma, A. & Qadri, A. Vi polysaccharide of Salmonella typhi targets the prohibitin family of molecules in intestinal epithelial cells and suppresses early inflammatory responses. Proc. Natl Acad. Sci. USA 101, 17492–17497 (2004).
Kim, T. I., Ou, M., Lee, M. & Kim, S. W. Arginine-grafted bioreducible poly(disulfide amine) for gene delivery systems. Biomaterials 30, 658–664 (2009).
Kumar, P. et al. T cell-specific siRNA delivery suppresses HIV-1 infection in humanized mice. Cell 134, 577–586 (2008).
Furuhashi, M. & Hotamisligil, G. S. Fatty acid-binding proteins: Role in metabolic diseases and potential as drug targets. Nature Rev. Drug Discov. 7, 489–503 (2008).
Wang, P. et al. Profiling of the secreted proteins during 3T3-L1 adipocyte differentiation leads to the identification of novel adipokines. Cell. Mol. Life Sci. 61, 2405–2417 (2004).
Veiseh, O. et al. Cell transcytosing poly-arginine coated magnetic nanovector for safe and effective siRNA delivery. Biomaterials 32, 5717–5725 (2011).
Ter-Avetisyan, G. et al. Cell entry of arginine-rich peptides is independent of endocytosis. J. Biol. Chem. 284, 3370–3378 (2009).
Vazquez, E., Ferrer-Miralles, N. & Villaverde, A. Peptide-assisted traffic engineering for nonviral gene therapy. Drug Discov. Today 13, 1067–1074 (2008).
Uysal, K. T., Scheja, L., Wiesbrock, S. M., Bonner-Weir, S. & Hotamisligil, G. S. Improved glucose and lipid metabolism in genetically obese mice lacking aP2. Endocrinology 141, 3388–3396 (2000).
This work was partially supported by grants from the National Research Foundation of Korea (2013030789), the Brain Korea 21 plus program (22A20130011095), and the Korean Health Technology R&D project through the Ministry of Health & Welfare (HI13C-1938-010013). All the confocal microscopy and Cellvizio imaging experiments were carried out at Korea Basic Science Institute, Chuncheon Center, Chuncheon-city, Korea. We are sincerely grateful for their assistance in performing experiments and data analysis.
The authors declare no competing financial interests.
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Won, YW., Adhikary, P., Lim, K. et al. Oligopeptide complex for targeted non-viral gene delivery to adipocytes. Nature Mater 13, 1157–1164 (2014). https://doi.org/10.1038/nmat4092
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