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.

Distinct transduction of muscle tissue in mice after systemic delivery of AAVpo1 vectors

Abstract

The human musculature is a promising and pivotal target for human gene therapy, owing to numerous diseases that affect this tissue and that are often monogenic, making them amenable to treatment and potentially cure on the genetic level. Particularly attractive would be the possibility to deliver clinically relevant DNA to muscle tissue from a minimally invasive, intravenous vector delivery. To date, this aim has been approximated by the use of Adeno-associated viruses (AAV) of different serotypes (rh.74, 8, 9) that are effective, but unfortunately not specific to the muscle and hence not ideal for use in patients. Here, we have thus studied the muscle tropism and activity of another AAV serotype, AAVpo1, that was previously isolated from pigs and found to efficiently transduce muscle following direct intramuscular injection in mice. The new data reported here substantiate the usefulness of AAVpo1 for muscle gene therapies by showing, for the first time, its ability to robustly transduce all major muscle tissues, including heart and diaphragm, from peripheral infusion. Importantly, in stark contrast to AAV9 that forms the basis for ongoing clinical gene therapy trials in the muscle, AAVpo1 is nearly completely detargeted from the liver, making it a very attractive and potentially safer option.

Your institute does not have access to this article

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. Duan D. Systemic delivery of adeno-associated viral vectors. Curr Opin Virol. 2016;21:16–25.

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Muller OJ, Katus HA, Bekeredjian R. Targeting the heart with gene therapy-optimized gene delivery methods. Cardiovasc Res. 2007;73:453–62.

    Google Scholar 

  3. Muller OJ, Leuchs B, Pleger ST, Grimm D, Franz WM, Katus HA, et al. Improved cardiac gene transfer by transcriptional and transductional targeting of adeno-associated viral vectors. Cardiovasc Res. 2006;70:70–8.

    Google Scholar 

  4. Bostick B, Ghosh A, Yue Y, Long C, Duan D. Systemic AAV-9 transduction in mice is influenced by animal age but not by the route of administration. Gene Ther. 2007;14:1605–9.

    CAS  Google Scholar 

  5. Yue Y, Ghosh A, Long C, Bostick B, Smith BF, Kornegay JN, et al. A single intravenous injection of adeno-associated virus serotype-9 leads to whole body skeletal muscle transduction in dogs. Mol Ther. 2008;16:1944–52.

    CAS  PubMed  PubMed Central  Google Scholar 

  6. 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.

    CAS  PubMed  PubMed Central  Google Scholar 

  7. 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:e3–9.

    CAS  Google Scholar 

  8. Vandendriessche T, Thorrez L, Acosta-Sanchez A, Petrus I, Wang L, Ma L, et al. Efficacy and safety of adeno-associated viral vectors based on serotype 8 and 9 vs. lentiviral vectors for hemophilia B gene therapy. J Thromb Haemost. 2007;5:16–24.

    CAS  Google Scholar 

  9. Jones D. Duchenne muscular dystrophy awaits gene therapy. Nat Biotechnol. 2019;37:335–7.

    CAS  Google Scholar 

  10. Amoasii L, Long C, Li H, Mireault AA, Shelton JM, Sanchez-Ortiz E, et al. Single-cut genome editing restores dystrophin expression in a new mouse model of muscular dystrophy. Sci Transl Med. 2017;9:eaan8081. https://doi.org/10.1126/scitranslmed.aan8081.

  11. Duan D. Systemic AAV micro-dystrophin gene therapy for duchenne muscular dystrophy. Mol Ther. 2018;26:2337–56.

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Childers MK, Joubert R, Poulard K, Moal C, Grange RW, Doering JA, et al. Gene therapy prolongs survival and restores function in murine and canine models of myotubular myopathy. Sci Transl Med. 2014;6:220ra10.

    PubMed  PubMed Central  Google Scholar 

  13. Buj-Bello A, Fougerousse F, Schwab Y, Messaddeq N, Spehner D, Pierson CR, et al. AAV-mediated intramuscular delivery of myotubularin corrects the myotubular myopathy phenotype in targeted murine muscle and suggests a function in plasma membrane homeostasis. Hum Mol Genet. 2008;17:2132–43.

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Elverman M, Goddard MA, Mack D, Snyder JM, Lawlor MW, Meng H, et al. Long-term effects of systemic gene therapy in a canine model of myotubular myopathy. Muscle Nerve. 2017;56:943–53.

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Manno CS, Pierce GF, Arruda VR, Glader B, Ragni M, Rasko JJ, et al. Successful transduction of liver in hemophilia by AAV-Factor IX and limitations imposed by the host immune response. Nat Med. 2006;12:342–7.

    CAS  Google Scholar 

  16. Mingozzi F, High KA. Immune responses to AAV in clinical trials. Curr Gene Ther. 2007;7:316–24.

    CAS  Google Scholar 

  17. Mingozzi F, High KA. Therapeutic in vivo gene transfer for genetic disease using AAV: progress and challenges. Nat Rev Genet. 2011;12:341–55.

    CAS  Google Scholar 

  18. Mingozzi F, Meulenberg JJ, Hui DJ, Basner-Tschakarjan E, Hasbrouck NC, Edmonson SA, et al. AAV-1-mediated gene transfer to skeletal muscle in humans results in dose-dependent activation of capsid-specific T cells. Blood. 2009;114:2077–86.

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Pien GC, Basner-Tschakarjan E, Hui DJ, Mentlik AN, Finn JD, Hasbrouck NC, et al. Capsid antigen presentation flags human hepatocytes for destruction after transduction by adeno-associated viral vectors. J Clin Investig. 2009;119:1688–95.

    CAS  Google Scholar 

  20. Martino AT, Basner-Tschakarjan E, Markusic DM, Finn JD, Hinderer C, Zhou S, et al. Engineered AAV vector minimizes in vivo targeting of transduced hepatocytes by capsid-specific CD8+ T cells. Blood. 2013;121:2224–33.

    CAS  PubMed  PubMed Central  Google Scholar 

  21. Sun B, Zhang H, Franco LM, Brown T, Bird A, Schneider A, et al. Correction of glycogen storage disease type II by an adeno-associated virus vector containing a muscle-specific promoter. Mol Ther. 2005;11:889–98.

    CAS  Google Scholar 

  22. Sun B, Zhang H, Franco LM, Young SP, Schneider A, Bird A, et al. Efficacy of an adeno-associated virus 8-pseudotyped vector in glycogen storage disease type II. Mol Ther. 2005;11:57–65.

    CAS  Google Scholar 

  23. Mah C, Pacak CA, Cresawn KO, Deruisseau LR, Germain S, Lewis MA, et al. Physiological correction of Pompe disease by systemic delivery of adeno-associated virus serotype 1 vectors. Mol Ther. 2007;15:501–7.

    CAS  Google Scholar 

  24. Han SO, Li S, Brooks ED, Masat E, Leborgne C, Banugaria S, et al. Enhanced efficacy from gene therapy in Pompe disease using coreceptor blockade. Hum Gene Ther. 2015;26:26–35.

    CAS  Google Scholar 

  25. Bello A, Tran K, Chand A, Doria M, Allocca M, Hildinger M, et al. Isolation and evaluation of novel adeno-associated virus sequences from porcine tissues. Gene Ther. 2009;16:1320–8.

    CAS  Google Scholar 

  26. Bello A, Chand A, Aviles J, Soule G, Auricchio A, Kobinger GP. Novel adeno-associated viruses derived from pig tissues transduce most major organs in mice. Sci Rep. 2014;4:6644.

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Puppo A, Bello A, Manfredi A, Cesi G, Marrocco E, Della Corte M, et al. Recombinant vectors based on porcine adeno-associated viral serotypes transduce the murine and pig retina. PLoS One. 2013;8:e59025.

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Zincarelli C, Soltys S, Rengo G, Rabinowitz JE. Analysis of AAV serotypes 1-9 mediated gene expression and tropism in mice after systemic injection. Mol Ther. 2008;16:1073–80.

    CAS  Google Scholar 

  29. Li X, Eastman EM, Schwartz RJ, Draghia-Akli R. Synthetic muscle promoters: activities exceeding naturally occurring regulatory sequences. Nat Biotechnol. 1999;17:241–5.

    CAS  Google Scholar 

  30. Rincon MY, Sarcar S, Danso-Abeam D, Keyaerts M, Matrai J, Samara-Kuko E, et al. Genome-wide computational analysis reveals cardiomyocyte-specific transcriptional Cis-regulatory motifs that enable efficient cardiac gene therapy. Mol Ther. 2015;23:43–52.

    CAS  Google Scholar 

  31. Sarcar S, Tulalamba W, Rincon MY, Tipanee J, Pham HQ, Evens H, et al. Next-generation muscle-directed gene therapy by in silico vector design. Nat Commun. 2019;10:492.

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Miesbach W, Meijer K, Coppens M, Kampmann P, Klamroth R, Schutgens R, et al. Gene therapy with adeno-associated virus vector 5-human factor IX in adults with hemophilia B. Blood. 2018;131:1022–31.

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Mingozzi F, Maus MV, Hui DJ, Sabatino DE, Murphy SL, Rasko JE, et al. CD8(+) T-cell responses to adeno-associated virus capsid in humans. Nat Med. 2007;13:419–22.

    CAS  Google Scholar 

  34. George LA, Sullivan SK, Giermasz A, Rasko JEJ, Samelson-Jones BJ, Ducore J, et al. Hemophilia B gene therapy with a high-specific-activity factor IX variant. N Engl J Med. 2017;377:2215–27.

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Mendell JR, Al-Zaidy S, Shell R, Arnold WD, Rodino-Klapac LR, Prior TW, et al. Single-dose gene-replacement therapy for spinal muscular atrophy. N Engl J Med. 2017;377:1713–22.

    CAS  Google Scholar 

  36. Nathwani AC, Tuddenham EG, Rangarajan S, Rosales C, McIntosh J, Linch DC, et al. Adenovirus-associated virus vector-mediated gene transfer in hemophilia B. N Engl J Med. 2011;365:2357–65.

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Grimm D, Zolotukhin S. E pluribus unum: 50 years of research, millions of viruses, and one goal-tailored acceleration of AAV evolution. Mol Ther. 2015;23:1819–31.

    CAS  PubMed  PubMed Central  Google Scholar 

  38. Weinmann J, Grimm D. Next-generation AAV vectors for clinical use: an ever-accelerating race. Virus Genes. 2017;53:707–13.

    CAS  Google Scholar 

  39. Grimm D, Lee JS, Wang L, Desai T, Akache B, Storm TA, et al. In vitro and in vivo gene therapy vector evolution via multispecies interbreeding and retargeting of adeno-associated viruses. J Virol. 2008;82:5887–911.

    CAS  PubMed  PubMed Central  Google Scholar 

  40. Herrmann AK, Bender C, Kienle E, Grosse S, El Andari J, Botta J, et al. A robust and all-inclusive pipeline for shuffling of adeno-associated viruses. ACS Synth Biol. 2019;8:194–206.

    CAS  Google Scholar 

  41. Zinn E, Pacouret S, Khaychuk V, Turunen HT, Carvalho LS, Andres-Mateos E, et al. In silico reconstruction of the viral evolutionary lineage yields a potent gene therapy vector. Cell Rep. 2015;12:1056–68.

    CAS  PubMed  PubMed Central  Google Scholar 

  42. Fakhiri J, Schneider MA, Puschhof J, Stanifer M, Schildgen V, Holderbach S, et al. Novel chimeric gene therapy vectors based on adeno-associated virus and four different mammalian bocaviruses. Mol Ther Methods Clin Dev. 2019;12:202–22.

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

WT, QHP, JEA, TV, MKC, and DG are very grateful for funding and other support from the MYOCURE project. MYOCURE has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement no. 667751. DG is thankful for support by the German Center for Infection Research (DZIF, BMBF; TTU-HIV 04.803 and TTU-HIV 04.815). DG acknowledges additional funding by the German Research Foundation (DFG) through the Cluster of Excellence CellNetworks (EXC81) and the Collaborative Research Centers SFB1129 (Projektnummer 240245660) and TRR179 (Projektnummer 272983813). TV and MKC obtained funding from the Fonds Wetenschappelijk Onderzoek (FWO), VUB Industrieel Onderzoeksfonds (IOF), Koning Boudewijn Stichting (Creemers-Opdebeek) and Association Belge contre les Maladies Neuromusculaires (ABMM). We are grateful to Alexander Bello and Gary Kobinger for initially supplying the po1 sequence. The authors thank Julia Fakhiri for critical reading of the manuscript as well as Ermira Samara for her help with AAV vector production.

Author information

Authors and Affiliations

Authors

Contributions

TV, MKC, and DG conceived and designed the experiments. WT, JW, QHP, and JEA generated constructs and performed experiments. WT, JEA, TV, MKC, and DG wrote the manuscript. All authors read the manuscript and approved the final version.

Corresponding authors

Correspondence to Thierry VandenDriessche, Marinee K. Chuah or Dirk Grimm.

Ethics declarations

Conflict of interest

DG is a co-founder and shareholder of AaviGen GmbH. JW is currently an employee of Boehringer Ingelheim Pharma GmbH & Co. KG. All other authors declare that they have no conflict of interest.

Ethical approval

All animal procedures were approved by the institutional animal ethics committee of the Free University of Brussels (VUB) (Brussels, Belgium). Husbandry was carried out in individually ventilated Thoren cages that contained Hygienic Animal Bedding (Lignocel). Temperature was maintained at ~21 °C with 50–60% humidity. Animals were fed SsniFF laboratory animal food (ABEDD Vertriebs GmbH, Vienna, Austria) ad libitum.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Tulalamba, W., Weinmann, J., Pham, Q.H. et al. Distinct transduction of muscle tissue in mice after systemic delivery of AAVpo1 vectors. Gene Ther 27, 170–179 (2020). https://doi.org/10.1038/s41434-019-0106-3

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41434-019-0106-3

Search

Quick links