title: Gene delivery to adipose tissue using transcriptionally targeted rAAV8 vectors.
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.
authors.affiliation: Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.
authors.forename: Silke
authors.initials: S
authors.lastname: Uhrig-Schmidt
authors.affiliation: Swiss Federal Institute of Technology, ETH Zurich, SLA C92, Institute of Food Nutrition and Health, Schwerzenbach, Switzerland.
authors.forename: Matthias
authors.initials: M
authors.lastname: Geiger
authors.affiliation: Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.
authors.forename: Gerd
authors.initials: G
authors.lastname: Luippold
authors.affiliation: Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.
authors.forename: Gerald
authors.initials: G
authors.lastname: Birk
authors.affiliation: Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.
authors.forename: Detlev
authors.initials: D
authors.lastname: Mennerich
authors.affiliation: Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.
authors.forename: Heike
authors.initials: H
authors.lastname: Neubauer
authors.affiliation: Centre for Infectious Diseases/Virology, Heidelberg University Hospital and Cluster of Excellence CellNetworks, Heidelberg University, Im Neuenheimer Feld 267, Heidelberg, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Grimm
authors.affiliation: Swiss Federal Institute of Technology, ETH Zurich, SLA C92, Institute of Food Nutrition and Health, Schwerzenbach, Switzerland.
authors.forename: Christian
authors.initials: C
authors.lastname: Wolfrum
authors.affiliation: Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Kreuz
mesh_terms: D052242:Adiponectin / Q000235:genetics; D000273:Adipose Tissue / Q000821:virology*; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000032:analysis; D051379:Mice; D000070778:Perilipin-1; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic
chemical_list: D052242:Adiponectin; D002352:Carrier Proteins; D000070778:Perilipin-1; D010750:Phosphoproteins; D049452:Green Fluorescent Proteins