Gene Transfer in Skeletal and Cardiac Muscle Using Recombinant Adeno‐Associated Virus

Abstract

Adeno‐associated virus (AAV) is a DNA virus with a small (∼4.7 kb) single‐stranded genome. It is a naturally replication‐defective parvovirus of the dependovirus group. Recombinant AAV (rAAV), for use as a gene transfer vector, is created by replacing the viral rep and cap genes with the transgene of interest along with promoter and polyadenylation sequences. Only the viral inverted terminal repeats (ITRs) are required in cis for replication and packaging during production. The ITRs are also necessary and sufficient for vector genome processing and persistence during transduction. The tissue tropism of the rAAV vector is determined by the AAV capsid. In this unit we will discuss several methods to deliver rAAV in order to transduce cardiac and/or skeletal muscle, including intravenous delivery, intramuscular delivery, isolated limb infusion, intrapericardial injection in neonatal mice, and left ventricular wall injection in adult rats. Curr. Protoc. Microbiol. 28:14D.3.1‐14D.3.19. © 2013 by John Wiley & Sons, Inc.

Stability and Compatibility of Recombinant Adeno-Associated Virus Under Conditions Commonly Encountered in Human Gene Therapy Trials

“…Vector was delivered by intramuscular injection, 1 · 10 11 viral particles/mouse, in a volume of 50 ll to the right hind limb as described previously. 9 The vector was loaded into the syringe just before injection. The same investigator performed all injections.…”

The potential of adeno-associated viral vectors for gene delivery to muscle tissue

“…Currently, many routes of rAAV administration can achieve either localized or systemic muscle gene transfer for different therapeutic purposes [24], such as intramuscular injection, retrograde transvenous limb perfusion, intracoronary infusion, intrapericardial injection, transendocardial injection, recirculating delivery, intraperitoneal injection and intravenous delivery (Table 2). …”

“…This method of rAAV delivery has been used in clinical trials for muscle gene transfer to treat muscle diseases such as DMD [34]. Intrapericardial injection of rAAV is usually performed with neonatal animals, and can be used to transduce the heart and diaphragm simultaneously [24]. Transendocardial delivery of rAAV vector to achieve global cardiac gene transfer has been reported in canine models [35,36] and NHP [37].…”

“…rAAV vectors can be delivered into both skeletal and cardiac muscles in either localized or widespread manner. Multiple serotypes, such as rAAV1, 2, 5, 6, 7, 8 and 9 were developed for efficient muscle transduction [24–26]. Moreover, minimal immune toxicity of rAAV as compared with other viral vectors, such as adenovirus, retrovirus and herpes simplex virus, is a major advantage for rAAV gene therapy.…”

Delivery of Adeno-Associated Virus Gene Therapy by Intravascular Limb Infusion Methods

“…Recombinant adeno-associated virus (rAAV) targets skeletal muscle readily, and because myocytes are a nondividing cell type, therapeutic proteins can be expressed long-term, despite low levels of chromosomal vector integration. [1][2][3][4][5] This makes muscle a good gene therapy target for both musclespecific disease and as a sight for production of secreted proteins. 2,[6][7][8][9][10][11][12][13][14] While direct intramuscular (IM) vector injections are effective in targeting muscle cells, the area of spread from the injection site is limited, 11,15 meaning that only a relatively small area of myocytes can be targeted in this manner without having to administer multiple IM injections per large muscle group and the need to dose each muscle group separately.…”