1. Volume 19, Issue 6, Pages 1079-1089 (June 2011)
A Potential Role of Distinctively Delayed Blood Clearance of Recombinant Adeno- associated Virus Serotype 9 in Robust Cardiac Transduction 
Nicole M Kotchey, Kei Adachi, Maliha Zahid, Katsuya Inagaki, Rakshita Charan, Robert S Parker, Hiroyuki Nakai 
Molecular Therapy 
Volume 19, Issue 6, Pages (June 2011)
DOI: /mt
Copyright © 2011 The American Society of Gene & Cell Therapy Terms and Conditions

2. Figure 1
Blood vector concentration-time curves following intravenous injection of various serotype or variant AAV-CMV-lacZ vectors in adult C57BL/6 mice (n = 3–7 each). (a,b) Recombinant adeno-associated virus serotype 1 (rAAV1), rAAV2, rAAV8 or rAAV9 vector was injected into mice via the tail vein in bolus at a dose of 1.0 × 1013 vg/kg. Concentrations of rAAV particles in the blood are plotted as a function of time after injection on (a) a linear-linear scale or (b) a linear-log scale. Panel (a) highlights the clearance rates during the first 8 hours following vector administration. (c,d) Various AAV1 and AAV9 hybrid vectors and (e) rAAV variants with a modification at the C-terminus of the capsid were injected into mice (please also see Figure 2a). Vertical bars represent standard errors.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2011 The American Society of Gene & Cell Therapy Terms and Conditions

3. Figure 2
Adeno-associated virus serotype 1 (AAV1) and AAV9 and other types of hybrid vectors. (a) Schematic representation of the hybrid capsids. Positions for five interstrand loops (loop I–V) are shown at the top. AAV2 capsid R585E mutation is indicated with a star. (b) AAV1.9-3 full capsid viewed down an icosahedral threefold symmetry axis at the center. The colored region indicates the 113-amino acid (a.a.). AAV9 capsid segment (a.a. 456–568) embedded in the AAV1.9-3 capsid. Each color represents the region from a different capsid monomer. (c) A closer view of (b). The 113-a.a. region from two different capsid monomers constitutes the middle and outer spikes of a protrusion as indicated with the red and green regions from the blue monomer and a gray monomer, respectively, in the circle. (d) AAV1.9-6 full capsid viewed down an icosahedral twofold symmetry axis at the center. The colored region indicates the 37-a.a. AAV9 capsid C-terminal segment (a.a ) embedded in the AAV1.9-6 capsid. Each color represents a region from a different capsid monomer. (e) The AAV9 capsid segments embedded in the AAV capsid viewed down from an outer spike of a capsid protrusion. Two separate AAV9 capsid regions from two different capsid monomers; i.e., a.a (the green region from a gray monomer in the circle) and a.a. 550–568 (the red region from a blue monomer in the circle) form the outer spike of the AAV capsid protrusion. UCSF Chimera45 was used to generate these figures. The inner, middle, and outer spikes that constitute the threefold capsid protrusion are indicated with black circles, triangles and crosses, respectively. Three and fivefold symmetry axes are shown with orange triangles and pentagons, respectively.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2011 The American Society of Gene & Cell Therapy Terms and Conditions

4. Figure 3
Hepatic and cardiac transduction with various serotype and variant recombinant adeno-associated virus (rAAV) vectors. All mice were injected with AAV-CMV-lacZ vector intravenously at a dose of 1.0 × 1013 vg/kg except for AAV2.9-6, which was injected at a 1/5 dose, and AAV9SC, which was injected subcutaneously at a dose of 1.0 × 1013 vg/kg. Tissues were harvested 11 days postinjection, and analyzed by X-Gal staining. Tissue sections were counterstained with light hematoxylin.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2011 The American Society of Gene & Cell Therapy Terms and Conditions

5. Figure 4
Relationship between the extent of endothelial vector exposure and transduction efficiency in the liver and heart. Mice were injected intravenously with 1.0 × 1013 vg/kg of (a–c) AAV9-CMV-lacZ or (d–f) AAV1.9-6-CMV-lacZ vector, then circulating vector particles were rapidly eliminated at designated time points by infusion of anti-AAV9 or AAV1.9-6 mouse sera (n = 2 each). The vector-injected control mice either did not receive any mouse sera, or received naive mouse sera 10 minutes postvector injection. (a,d) Blood vector concentration-time curves. Mouse sera containing anti-AAV9 or AAV1.9-6 capsid antibodies were infused 10 minutes, 1 hour or 4 hours postvector injection. (b,e) 72-hour areas under the curve (AUCs) in each group relative to those in the mice that received no mouse sera. (c,f) Liver and heart transduction efficiencies relative to those in the mice that received no mouse sera. Vertical bars represent differences between means and each value. (g,h) Cardiomyocyte transduction efficiencies with rAAV9 and rAAV1.9-6 in vitro. H9c2 cells were transduced with either AAV9-CMV-lacZ or AAV1.9-6-CMV-lacZ at a multiplicity of infection of 106, and transduction efficiencies were determined by (g) X-Gal staining and (h) β-galactosidase-specific enzyme-linked immunosorbent assay 9 days after infection. (i) Relative transduction efficiencies in the liver and heart by full or 8-hour endothelial exposure with either AAV9-CMV-lacZ or AAV1.9-6-CMV-lacZ. Anti-AAV9 or AAV1.9-6 sera were infused in mice 8 hours after rAAV9 or rAAV1.9-6 injection, respectively (n = 6 each). No sera control animals received only rAAV (n = 6 each). Vertical bars represent standard errors.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2011 The American Society of Gene & Cell Therapy Terms and Conditions

6. Figure 5
Effects of the absence of caveolin-1 (Cav-1) on recombinant adeno-associated virus serotype 9's (rAAV9) blood vector clearance and hepatic and cardiac transduction in mice. (a) Blood vector concentration-time curves following intravenous injection of AAV9-CMV-lacZ at a dose of 1.0 × 1013 vg/kg in caveolin-1–deficient mice and wild-type controls (n = 3 each). Asterisks indicate the time points showing a difference with statistical significance (Student's t-test, P < 0.05). (b) Transduction efficiency in the liver and heart of wild-type and caveolin-1–deficient mice injected with AAV9-CMV-lacZ (n = 5 each). Transduction efficiencies were determined by X-Gal staining of tissue sections. Vertical bars represent standard errors. KO, knockout; WT, wild type.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2011 The American Society of Gene & Cell Therapy Terms and Conditions