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.
Highlights
Obesity is a leading preventable cause of death worldwide with increasing incidence in adults and children, and authorities view it as one of the most serious public health problems of the 21st century [1]
In an attempt to identify a viral vector platform allowing efficient and selective gene delivery to different adipose tissue depots in mice, we here investigated the potential of recombinant associated viruses (AAV) vectors
By comparing the transduction efficiency of five enhanced greenfluorescent protein (eGFP)-expressing recombinant Adeno-associated virus (rAAV) (AAV1, -2, -5, -6, -8) and an adenoviral vector (Ad) following local injection into the visceral fat pad of C57BL/6 mice, we identified rAAV8 as the most efficacious vector, yielding transgene expression levels that were up to 7-fold higher compared to the other tested AAV serotypes
Summary
Obesity is a leading preventable cause of death worldwide with increasing incidence in adults and children, and authorities view it as one of the most serious public health problems of the 21st century [1]. White adipose tissue (WAT) is highly adapted to store excess energy in the form of triglycerides inside lipid droplets, which are – besides other mechanisms – coated and regulated by members of the perilipin family of proteins. In contrast to WAT, brown adipose tissue (BAT) oxidizes chemical energy to produce heat as a defense against hypothermia and obesity. The development and presence of these brown in white (brite) cells – a process called browning – has recently been shown to be associated with improved metabolic phenotypes [5, 6]. Understanding the underlying mechanisms of browning and other processes that activate BAT is crucial for a therapeutic benefit which is the reduction of weight and plasma lipid levels in obese and hyperlipidemic patients. Genetic engineering of adipose tissue is essential but this, currently mostly relies on the time-consuming generation of transgenic animals [7]
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