Abstract
Adeno-associated virus (AAV) has become a leading gene transfer vector for striated muscles. However, the AAV vectors also exhibit broad tropisms after systemic delivery. In an attempt to improve muscle tropism, we inserted a 7-amino-acid (ASSLNIA) muscle-targeting peptide (MTP) in the capsids of AAV2 at residue 587 or 588, generating AAV587MTP and AAV588MTP. In vitro studies showed that both viruses diminished their infectivity on non-muscle cell lines as well as on un-differentiated myoblasts; however, preserved or enhanced their infectivity on differentiated myotubes. AAV587MTP, but not AAV588MTP, also abolished its heparin-binding capacity and infected myotubes in a heparin-independent manner. Furthermore, in vivo studies by intravenous vector administration in mice showed that AAV587MTP enhanced its tropism to various muscles and particularly to the heart (24.3-fold of unmodified AAV2), whereas reduced its tropism to the non-muscle tissues such as the liver, lungs, spleen and so on. This alteration of tissue tropism is not simply because of the loss of heparin-binding, as a mutant AAV2 (AAVHBSMut) containing heparin-binding site mutations lost infectivity on both non-muscle and muscle cells. Furthermore, free MTP peptide, but not the scrambled control peptide, competitively inhibited AAV587MTP infection on myotubes. These results suggest that AAV2 could be re-targeted to the striated muscles by a MTP inserted after residue 587 of the capsids. This proof of principle study showed first evidence of peptide-directed muscle targeting on systemic administration of AAV vectors.
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References
Cox GA, Cole NM, Matsumura K, Phelps SF, Hauschka SD, Campbell KP et al. Overexpression of dystrophin in transgenic mdx mice eliminates dystrophic symptoms without toxicity. Nature 1993; 364: 725–729.
Amalfitano A, McVie-Wylie AJ, Hu H, Dawson TL, Raben N, Plotz P et al. Systemic correction of the muscle disorder glycogen storage disease type II after hepatic targeting of a modified adenovirus vector encoding human acid-alpha-glucosidase. Proc Natl Acad Sci USA 1999; 96: 8861–8866.
Kessler PD, Podsakoff GM, Chen X, McQuiston SA, Colosi PC, Matelis LA et al. Gene delivery to skeletal muscle results in sustained expression and systemic delivery of a therapeutic protein. Proc Natl Acad Sci USA 1996; 93: 14082–14087.
Greelish JP, Su LT, Lankford EB, Burkman JM, Chen H, Konig SK et al. Stable restoration of the sarcoglycan complex in dystrophic muscle perfused with histamine and a recombinant adeno-associated viral vector. Nat Med 1999; 5: 439–443.
Wolff JA, Malone RW, Williams P, Chong W, Acsadi G, Jani A et al. Direct gene transfer into mouse muscle in vivo. Science 1990; 247: 1465–1468.
Wolff JA, Ludtke JJ, Acsadi G, Williams P, Jani A . Long-term persistence of plasmid DNA and foreign gene expression in mouse muscle. Hum Mol Genet 1992; 1: 363–369.
Xiao X, Li J, Samulski RJ . Efficient long-term gene transfer into muscle tissue of immunocompetent mice by adeno-associated virus vector. J Virol 1996; 70: 8098–8108.
Blankinship MJ, Gregorevic P, Allen JM, Harper SQ, Harper H, Halbert CL et al. Efficient transduction of skeletal muscle using vectors based on adeno-associated virus serotype 6. Mol Ther 2004; 10: 671–678.
Kay MA, Manno CS, Ragni MV, Larson PJ, Couto LB, McClelland A et al. Evidence for gene transfer and expression of factor IX in haemophilia B patients treated with an AAV vector. Nat Genet 2000; 24: 257–261.
Gregorevic P, Blankinship MJ, Allen JM, Crawford RW, Meuse L, Miller DG et al. Systemic delivery of genes to striated muscles using adeno-associated viral vectors. Nat Med 2004; 10: 828–834.
Wang Z, Zhu T, Qiao C, Zhou L, Wang B, Zhang J et al. Adeno-associated virus serotype 8 efficiently delivers genes to muscle and heart. Nat Biotechnol 2005; 23: 321–328.
Monahan PE, Samulski RJ . AAV vectors: is clinical success on the horizon? Gene Therapy 2000; 7: 24–30.
Summerford C, Samulski RJ . Membrane-associated heparan sulfate proteoglycan is a receptor for adeno-associated virus type 2 virions. J Virol 1998; 72: 1438–1445.
Summerford C, Bartlett JS, Samulski RJ . AlphaVbeta5 integrin: a co-receptor for adeno-associated virus type 2 infection. Nat Med 1999; 5: 78–82.
Qing K, Mah C, Hansen J, Zhou S, Dwarki V, Srivastava A . Human fibroblast growth factor receptor 1 is a co-receptor for infection by adeno-associated virus 2. Nat Med 1999; 5: 71–77.
Kashiwakura Y, Tamayose K, Iwabuchi K, Hirai Y, Shimada T, Matsumoto K et al. Hepatocyte growth factor receptor is a coreceptor for adeno-associated virus type 2 infection. J Virol 2005; 79: 609–614.
Asokan A, Hamra JB, Govindasamy L, Agbandje-McKenna M, Samulski RJ . Adeno-associated virus type 2 contains an integrin alpha5beta1 binding domain essential for viral cell entry. J Virol 2006; 80: 8961–8969.
Barry MA, Dower WJ, Johnston SA . Toward cell-targeting gene therapy vectors: selection of cell-binding peptides from random peptide-presenting phage libraries. Nat Med 1996; 2: 299–305.
Muller OJ, Kaul F, Weitzman MD, Pasqualini R, Arap W, Kleinschmidt JA et al. Random peptide libraries displayed on adeno-associated virus to select for targeted gene therapy vectors. Nat Biotechnol 2003; 21: 1040–1046.
Wu P, Xiao W, Conlon T, Hughes J, Agbandje-McKenna M, Ferkol T et al. Mutational analysis of the adeno-associated virus type 2 (AAV2) capsid gene and construction of AAV2 vectors with altered tropism. J Virol 2000; 74: 8635–8647.
Loiler SA, Conlon TJ, Song S, Tang Q, Warrington KH, Agarwal A et al. Targeting recombinant adeno-associated virus vectors to enhance gene transfer to pancreatic islets and liver. Gene Ther 2003; 10: 1551–1558.
Grifman M, Trepel M, Speece P, Gilbert LB, Arap W, Pasqualini R et al. Incorporation of tumor-targeting peptides into recombinant adeno-associated virus capsids. Mol Ther 2001; 3: 964–975.
Nicklin SA, Buening H, Dishart KL, de Alwis M, Girod A, Hacker U et al. Efficient and selective AAV2-mediated gene transfer directed to human vascular endothelial cells. Mol Ther 2001; 4: 174–181.
White SJ, Nicklin SA, Buning H, Brosnan MJ, Leike K, Papadakis ED et al. Targeted gene delivery to vascular tissue in vivo by tropism-modified adeno-associated virus vectors. Circulation 2004; 109: 513–519.
Work LM, Buning H, Hunt E, Nicklin SA, Denby L, Britton N et al. Vascular bed-targeted in vivo gene delivery using tropism-modified adeno-associated viruses. Mol Ther 2006; 13: 683–693.
Samoylova TI, Smith BF . Elucidation of muscle-binding peptides by phage display screening. Muscle Nerve 1999; 22: 460–466.
Samoylov AM, Samoylova TI, Hartell MG, Pathirana ST, Smith BF, Vodyanoy V . Recognition of cell-specific binding of phage display derived peptides using an acoustic wave sensor. Biomol Eng 2002; 18: 269–272.
Pacak CA, Mah CS, Thattaliyath BD, Conlon TJ, Lewis MA, Cloutier DE et al. Recombinant adeno-associated virus serotype 9 leads to preferential cardiac transduction in vivo. Circ Res 2006; 99: e3–e9.
Inagaki K, Fuess S, Storm TA, Gibson GA, McTiernan CF, Kay MA et al. Robust systemic transduction with AAV9 vectors in mice: efficient global cardiac gene transfer superior to that of AAV8. Mol Ther 2006; 14: 45–53.
Shi W, Arnold GS, Bartlett JS . Insertional mutagenesis of the adeno-associated virus type 2 (AAV2) capsid gene and generation of AAV2 vectors targeted to alternative cell-surface receptors. Hum Gene Ther 2001; 12: 1697–1711.
Park PW, Reizes O, Bernfield M . Cell surface heparan sulfate proteoglycans: selective regulators of ligand-receptor encounters. J Biol Chem 2000; 275: 29923–29926.
Thomas CE, Storm TA, Huang Z, Kay MA . Rapid uncoating of vector genomes is the key to efficient liver transduction with pseudotyped adeno-associated virus vectors. J Virol 2004; 78: 3110–3122.
Margalit H, Fischer N, Ben-Sasson SA . Comparative analysis of structurally defined heparin binding sequences reveals a distinct spatial distribution of basic residues. J Biol Chem 1993; 268: 19228–19231.
Huttner NA, Girod A, Perabo L, Edbauer D, Kleinschmidt JA, Buning H et al. Genetic modifications of the adeno-associated virus type 2 capsid reduce the affinity and the neutralizing effects of human serum antibodies. Gene Therapy 2003; 10: 2139–2147.
Xie Q, Bu W, Bhatia S, Hare J, Somasundaram T, Azzi A et al. The atomic structure of adeno-associated virus (AAV-2), a vector for human gene therapy. Proc Natl Acad Sci USA 2002; 99: 10405–10410.
Schwede T, Kopp J, Guex N, Peitsch MC . SWISS-MODEL: An automated protein homology-modeling server. Nucleic Acids Res 2003; 31: 3381–3385.
Guex N, Peitsch MC . SWISS-MODEL and the Swiss-PdbViewer: an environment for comparative protein modeling. Electrophoresis 1997; 18: 2714–2723.
Chirmule N, Propert K, Magosin S, Qian Y, Qian R, Wilson J . Immune responses to adenovirus and adeno-associated virus in humans. Gene Therapy 1999; 6: 1574–1583.
Scallan CD, Jiang H, Liu T, Patarroyo-White S, Sommer JM, Zhou S et al. Human immunoglobulin inhibits liver transduction by AAV vectors at low AAV2 neutralizing titers in SCID mice. Blood 2006; 107: 1810–1817.
Li J, Samulski RJ, Xiao X . Role for highly regulated rep gene expression in adeno-associated virus vector production. J Virol 1997; 71: 5236–5243.
Xiao X, Li J, Samulski RJ . Production of high-titer recombinant adeno-associated virus vectors in the absence of helper adenovirus. J Virol 1998; 72: 2224–2232.
Acknowledgements
We thank Dr Zhong Wang for helpful advice and Ms Chunlian Chen for technical assistance. This work is part of C Yu's PhD thesis at the University of Pittsburgh. It is supported by NIH grants AR45967 and AR50595 to X Xiao.
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Yu, CY., Yuan, Z., Cao, Z. et al. A muscle-targeting peptide displayed on AAV2 improves muscle tropism on systemic delivery. Gene Ther 16, 953–962 (2009). https://doi.org/10.1038/gt.2009.59
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DOI: https://doi.org/10.1038/gt.2009.59
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