Gene delivery to adipose tissue using transcriptionally targeted rAAV8 vectors

PLoS One. 2014 Dec 31;9(12):e116288. doi: 10.1371/journal.pone.0116288. eCollection 2014.

Abstract

In recent years, the increasing prevalence of obesity and obesity-related co-morbidities fostered intensive research in the field of adipose tissue biology. To further unravel molecular mechanisms of adipose tissue function, genetic tools enabling functional studies in vitro and in vivo are essential. While the use of transgenic animals is well established, attempts using viral and non-viral vectors to genetically modify adipocytes in vivo are rare. Therefore, we here characterized recombinant Adeno-associated virus (rAAV) vectors regarding their potency as gene transfer vehicles for adipose tissue. Our results demonstrate that a single dose of systemically applied rAAV8-CMV-eGFP can give rise to remarkable transgene expression in murine adipose tissues. Upon transcriptional targeting of the rAAV8 vector to adipocytes using a 2.2 kb fragment of the murine adiponectin (mAP2.2) promoter, eGFP expression was significantly decreased in off-target tissues while efficient transduction was maintained in subcutaneous and visceral fat depots. Moreover, rAAV8-mAP2.2-mediated expression of perilipin A - a lipid-droplet-associated protein - resulted in significant changes in metabolic parameters only three weeks post vector administration. Taken together, our findings indicate that rAAV vector technology is applicable as a flexible tool to genetically modify adipocytes for functional proof-of-concept studies and the assessment of putative therapeutic targets in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adiponectin / genetics
  • Adipose Tissue / virology*
  • Animals
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism
  • Dependovirus / genetics
  • Gene Transfer Techniques*
  • Genetic Therapy / methods*
  • Genetic Vectors
  • Green Fluorescent Proteins / analysis
  • Mice
  • Perilipin-1
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism
  • Promoter Regions, Genetic

Substances

  • Adiponectin
  • Carrier Proteins
  • Perilipin-1
  • Phosphoproteins
  • Green Fluorescent Proteins

Grants and funding

The research described was funded by Boehringer Ingelheim (SUS, GL, GB, DM, HN, SK), by the Swiss National Fund and the FP7 DIABAT Project of the European Union (MG, CW) and by the Cluster of Excellence CellNetworks DFG EXC81 (SUS, DG). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.