Skip to main content
SpringerLink
Log in

rAAV-Mediated Gene Delivery to Adipose Tissue

  • Protocol
  • First Online:
Adeno-Associated Virus Vectors

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1950))

Abstract

Recombinant adeno-associated virus (rAAV) vectors are attractive vehicles for gene therapy. Yet, it is challenging to genetically manipulate adipose tissue in adults due to the low transduction efficiency of naturally occurring AAV serotypes. We recently demonstrated that a novel engineered hybrid serotype Rec2 achieves high transduction of adipose tissue that is superior to naturally occurring serotypes via direct injection to adipose depots. Furthermore, the administration route influences the tropism and efficacy of Rec2 vector: oral administration transduces interscapular brown fat, while intraperitoneal injection preferentially targets visceral fat. Multiple in vivo studies by our lab and others have demonstrated that Rec2 vector provides a powerful tool to genetically manipulate adipose tissue for basic research and potential gene therapies of genetic and acquired diseases. Here we provide detailed protocols for AAV production and delivery to adipose tissue by direct injection, oral administration, and intraperitoneal injection.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Mingozzi F, High KA (2011) Therapeutic in vivo gene transfer for genetic disease using AAV: progress and challenges. Nat Rev Genet 12(5):341–355. https://doi.org/10.1038/nrg2988

    Article  CAS  PubMed  Google Scholar 

  2. Mizukami H, Mimuro J, Ogura T, Okada T, Urabe M, Kume A, Sakata Y, Ozawa K (2006) Adipose tissue as a novel target for in vivo gene transfer by adeno-associated viral vectors. Hum Gene Ther 17(9):921–928. https://doi.org/10.1089/hum.2006.17.921

    Article  CAS  PubMed  Google Scholar 

  3. Zhang FL, Jia SQ, Zheng SP, Ding W (2011) Celastrol enhances AAV1-mediated gene expression in mice adipose tissues. Gene Ther 18(2):128–134. https://doi.org/10.1038/gt.2010.120

    Article  CAS  PubMed  Google Scholar 

  4. Jimenez V, Munoz S, Casana E, Mallol C, Elias I, Jambrina C, Ribera A, Ferre T, Franckhauser S, Bosch F (2013) In vivo adeno-associated viral vector-mediated genetic engineering of white and brown adipose tissue in adult mice. Diabetes 62(12):4012–4022. https://doi.org/10.2337/db13-0311

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Liu X, Magee D, Wang C, McMurphy T, Slater A, During M, Cao L (2014) Adipose tissue insulin receptor knockdown via a new primate-derived hybrid recombinant AAV serotype. Mol Ther Methods Clin Dev 1. https://doi.org/10.1038/mtm.2013.8

    Article  Google Scholar 

  6. McMurphy TB, Huang W, Xiao R, Liu X, Dhurandhar NV, Cao L (2017) Hepatic expression of adenovirus 36 E4ORF1 improves glycemic control and promotes glucose metabolism through AKT activation. Diabetes 66(2):358–371. https://doi.org/10.2337/db16-0876

    Article  CAS  PubMed  Google Scholar 

  7. Huang W, McMurphy T, Liu X, Wang C, Cao L (2016) Genetic manipulation of brown fat via oral administration of an engineered recombinant adeno-associated viral serotype vector. Mol Ther 24(6):1062–1069. https://doi.org/10.1038/mt.2016.34

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Huang W, Liu X, Queen NJ, Cao L (2017) targeting visceral fat by intraperitoneal delivery of novel AAV serotype vector restricting off-target transduction in liver. Mol Ther Methods Clin Dev 6:68–78. https://doi.org/10.1016/j.omtm.2017.06.002

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Zhu Y, Gao Y, Tao C, Shao M, Zhao S, Huang W, Yao T, Johnson JA, Liu T, Cypess AM, Gupta O, Holland WL, Gupta RK, Spray DC, Tanowitz HB, Cao L, Lynes MD, Tseng YH, Elmquist JK, Williams KW, Lin HV, Scherer PE (2016) Connexin 43 mediates white adipose tissue beiging by facilitating the propagation of sympathetic neuronal signals. Cell Metab 24(3):420–433. https://doi.org/10.1016/j.cmet.2016.08.005

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Ng R, Hussain NA, Zhang Q, Chang C, Li H, Fu Y, Cao L, Han W, Stunkel W, Xu F (2017) miRNA-32 drives brown fat thermogenesis and trans-activates subcutaneous white fat browning in mice. Cell Rep 19(6):1229–1246. https://doi.org/10.1016/j.celrep.2017.04.035

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Zhang Y, Xie L, Gunasekar SK, Tong D, Mishra A, Gibson WJ, Wang C, Fidler T, Marthaler B, Klingelhutz A, Abel ED, Samuel I, Smith JK, Cao L, Sah R (2017) SWELL1 is a regulator of adipocyte size, insulin signalling and glucose homeostasis. Nat Cell Biol 19(5):504–517. https://doi.org/10.1038/ncb3514

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Xie L, Zhang Y, Gunasekar SK, Mishra A, Cao L, Sah R (2017) Induction of adipose and hepatic SWELL1 expression is required for maintaining systemic insulin-sensitivity in obesity. Channels (Austin) 11(6):673–677. https://doi.org/10.1080/19336950.2017.1373225

    Article  Google Scholar 

  13. de Jong JM, Larsson O, Cannon B, Nedergaard J (2015) A stringent validation of mouse adipose tissue identity markers. Am J Physiol Endocrinol Metab 308(12):E1085–E1105. https://doi.org/10.1152/ajpendo.00023.2015

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Cancer Biology and Genetics, The Ohio State University, Columbus, OH, USA

    Wei Huang, Nicholas J. Queen & Lei Cao

  2. The Comprehensive Cancer Center, The Ohio State University, Columbus, OH, USA

    Wei Huang, Nicholas J. Queen & Lei Cao

Authors
  1. Wei Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nicholas J. Queen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lei Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lei Cao .

Editor information

Editors and Affiliations

  1. Department of Neurosciences, University of California, San Diego, La Jolla, CA, USA

    Michael J. Castle

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Huang, W., Queen, N.J., Cao, L. (2019). rAAV-Mediated Gene Delivery to Adipose Tissue. In: Castle, M. (eds) Adeno-Associated Virus Vectors. Methods in Molecular Biology, vol 1950. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9139-6_23

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9139-6_23

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9138-9

  • Online ISBN: 978-1-4939-9139-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation