PMID- 23535898
OWN - NLM
STAT- MEDLINE
DCOM- 20140307
LR  - 20211021
IS  - 1476-5462 (Electronic)
IS  - 0969-7128 (Print)
IS  - 0969-7128 (Linking)
VI  - 20
IP  - 9
DP  - 2013 Sep
TI  - Adeno-associated virus serotype 9 efficiently targets ischemic skeletal muscle 
      following systemic delivery.
PG  - 930-8
LID - 10.1038/gt.2013.16 [doi]
AB  - Targeting therapeutic gene expression to the skeletal muscle following 
      intravenous (IV) administration is an attractive strategy for treating peripheral 
      arterial disease (PAD), except that vector access to the ischemic limb could be a 
      limiting factor. As adeno-associated virus serotype 9 (AAV-9) transduces skeletal 
      muscle at high efficiency following systemic delivery, we employed AAV-9 vectors 
      bearing luciferase or enhanced green fluorescent protein (eGFP) reporter genes to 
      test the hypothesis that increased desialylation of cell-surface glycans 
      secondary to hindlimb ischemia (HLI) might help offset the reduction in tissue 
      perfusion that occurs in mouse models of PAD. The utility of the creatine 
      kinase-based (CK6) promoter for restricting gene expression to the skeletal 
      muscle was also examined by comparing it with the cytomegalovirus (CMV) promoter 
      after systemic administration following surgically induced HLI. Despite reduced 
      blood flow to the ischemic limbs, CK6 promoter-driven luciferase activities in 
      the ischemic gastrocnemius (GA) muscles were  approximately 34-,  approximately 28- and  approximately 150-fold higher than 
      in the fully perfused contralateral GA, heart and liver, respectively, 10 days 
      after IV administration. Furthermore, luciferase activity from the CK6 promoter 
      in the ischemic GA muscles was  approximately twofold higher than with CMV, while in the liver 
      CK6-driven activity was  approximately 42-fold lower than with CMV, demonstrating that the 
      specificity of ischemic skeletal muscle transduction can be further improved with 
      the muscle-specific promoters. Studies with Evans blue dye and fluorescently 
      labeled lectins revealed that vascular permeability and desialylation of the 
      cell-surface glycans were increased in the ischemic hindlimbs. Furthermore, 
      AAV9/CK6/Luc vector genome copy numbers were  approximately sixfold higher in the ischemic 
      muscle compared with the non-ischemic muscle in the HLI model, whereas this trend 
      was reversed when the same genome was packaged in the AAV-1 capsid (which binds 
      sialylated, as opposed to desialylated glycans), further underscoring the 
      importance of desialylation in the ischemic enhancement of transduction displayed 
      by AAV-9. Taken together, these findings suggest two complementary mechanisms 
      contributing to the preferential transduction of ischemic muscle by AAV-9: 
      increased vascular permeability and desialylation. In conclusion, ischemic muscle 
      is preferentially targeted following systemic administration of AAV-9 in a mouse 
      model of HLI. Unmasking of the primary AAV-9 receptor as a result of ischemia may 
      contribute importantly to this effect.
FAU - Katwal, A B
AU  - Katwal AB
AD  - Division of Cardiovascular Medicine, Department of Medicine, University of 
      Virginia, Charlottesville, VA 22903, USA.
FAU - Konkalmatt, P R
AU  - Konkalmatt PR
FAU - Piras, B A
AU  - Piras BA
FAU - Hazarika, S
AU  - Hazarika S
FAU - Li, S S
AU  - Li SS
FAU - John Lye, R
AU  - John Lye R
FAU - Sanders, J M
AU  - Sanders JM
FAU - Ferrante, E A
AU  - Ferrante EA
FAU - Yan, Z
AU  - Yan Z
FAU - Annex, B H
AU  - Annex BH
FAU - French, B A
AU  - French BA
LA  - eng
GR  - R01 AR050429/AR/NIAMS NIH HHS/United States
GR  - 5 T32 HL007284-35/HL/NHLBI NIH HHS/United States
GR  - R01 HL058582/HL/NHLBI NIH HHS/United States
GR  - T32 HL007284/HL/NHLBI NIH HHS/United States
GR  - R01 HL101200/HL/NHLBI NIH HHS/United States
GR  - T32 HL073555/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20130328
PL  - England
TA  - Gene Ther
JT  - Gene therapy
JID - 9421525
RN  - 0 (Polysaccharides)
SB  - IM
MH  - Animals
MH  - Dependovirus/genetics/metabolism/*physiology
MH  - Genes, Reporter
MH  - *Genetic Therapy
MH  - Genetic Vectors
MH  - Hindlimb/blood supply
MH  - Humans
MH  - Ischemia/genetics/metabolism/*therapy
MH  - Male
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Mice, Inbred C57BL
MH  - Muscle, Skeletal/*blood supply/*metabolism
MH  - Peripheral Arterial Disease/metabolism/*therapy
MH  - Polysaccharides/metabolism
MH  - Promoter Regions, Genetic
MH  - Transduction, Genetic
PMC - PMC3758463
MID - NIHMS448130
EDAT- 2013/03/29 06:00
MHDA- 2014/03/08 06:00
PMCR- 2014/03/01
CRDT- 2013/03/29 06:00
PHST- 2012/02/24 00:00 [received]
PHST- 2013/01/22 00:00 [revised]
PHST- 2013/02/20 00:00 [accepted]
PHST- 2013/03/29 06:00 [entrez]
PHST- 2013/03/29 06:00 [pubmed]
PHST- 2014/03/08 06:00 [medline]
PHST- 2014/03/01 00:00 [pmc-release]
AID - gt201316 [pii]
AID - 10.1038/gt.2013.16 [doi]
PST - ppublish
SO  - Gene Ther. 2013 Sep;20(9):930-8. doi: 10.1038/gt.2013.16. Epub 2013 Mar 28.