Adenovirus Vectors Based on Human Adenovirus Type 19a Have High Potential for Human Muscle-Directed Gene Therapy

Abstract

Until recently, adenovirus-based gene therapy has been almost exclusively based on human adenovirus serotype 5 (Ad5). The aim of this study was to systematically compare the efficiency of transduction of primary muscle cells from various species by two adenoviral vectors from subgroups C and D. Transduction of a panel of myoblasts demonstrated a striking specificity of an Ad19a-based replication-defective E1-deleted vector (Ad19aEGFP) for human cells, whereas the Ad5-based vector had high affinity for nonhuman primate myoblasts. Transgene expression correlated well with cell-associated vector genomes. Up to 6.59% of the initially applied Ad19aEGFP vector particles were taken up by human myoblasts, as compared with 0.1% of the corresponding Ad5 vector. Remarkably, Ad19aEGFP but not Ad5EGFP efficiently transduced differentiated human myotubes, an in vitro model for skeletal muscle transduction. Uptake of Ad19aEGFP vector particles in human myotubes was 12-fold more efficient than that of Ad5EGFP. Moreover, both vectors demonstrated an early block at the level of vector uptake in mouse myoblasts and rat L6 cells. Investigation of the underlying mechanism for binding and uptake of the two vectors by human myoblasts showed high susceptibility for Ad19a to neuraminidase and wheat germ agglutinin (WGA) lectin, whereas Ad5-mediated transduction was dependent on binding to the coxsackie-adenovirus receptor (CAR) and sensitive to soluble RGD peptide and heparin. Our study offers insights into species-dependent factors that determine Ad tropism and, moreover, provides a basis for application of the novel Ad19a-based vector for gene transfer into human skeletal muscle.

Molecular evolution of human species D adenoviruses

“…Also, because of their broad tissue tropisms and the relative ease with which they can be genetically altered, adenoviruses have been a vector of choice for gene therapy protocols, representing almost one-fourth of all vectors used in current gene therapy clinical trials (http://www.abedia.com/wiley/vectors.php). Recently, due to their low seroprevalence, HAdV-D vectors are being tested for use in gene therapy (HAdVD19, D26, D28, D46, D48, D49) (Abbink et al, 2007; Kahl et al, 2010; Lemckert et al, 2006; Reddy et al, 2006; Ruzsics et al, 2006; Thirion et al, 2006). It is critically important to understand recombination mechanisms in HAdV-Ds if gene vectors based on HAdV-Ds are to be used in human patients who might become coincidently infected with a wild type virus.…”

Predicting the Next Eye Pathogen: Analysis of a Novel Adenovirus

“…Human adenoviruses (HAdV) are ubiquitous, cause infections in the respiratory, gastrointestinal, genitourinary, and ocular mucosas, can be opportunistic in the immune-compromised host, and have been associated with obesity (3–8). Adenoviruses also are common choices for gene therapy vectors (9, 10). Thus, while adenoviruses continue to cause significant morbidity and mortality in the human population, their existence also provides a potential benefit for the treatment of patients with an even broader range of ailments.…”

Gene silencing of myofibrillogenesis regulator-1 by adenovirus-delivered small interfering RNA suppresses cardiac hypertrophy induced by angiotensin II in mice

“…The Adxsi system that we used here was a vector based on E1/E3-deleted adenovirus serotype 5, whereas adenovirus serotype 5-mediated transduction was dependent on binding to the coxsackie adenovirus receptor (33). The lesser silencing efficiency of AdSiR-MR-1 in the skeletal muscle was in correlation with the lesser expression of coxsackie adenovirus receptors in skeletal muscle cells (34).…”