title: Biodistribution and inflammatory profiles of novel penton and hexon double-mutant serotype 5 adenoviruses.
abstract: The use of adenovirus serotype 5 (Ad5) vectors in the clinical setting is severely hampered by the profound liver tropism observed after intravascular delivery coupled with the pronounced inflammatory and innate immune response elicited by these vectors. Liver transduction by circulating Ad5 virions is mediated by a high-affinity interaction between the capsid hexon protein and blood coagulation factor X (FX), whilst penton-α(v)integrin interactions are thought to contribute to the induction of anti-Ad5 inflammatory and innate immune responses. To overcome these limitations, we sought to develop and characterise for the first time novel Ad5 vectors possessing mutations ablating both hexon:FX and penton:integrin interactions. As expected, intravascular administration of the FX binding-ablated Ad5HVR5*HVR7*E451Q vector (AdT*) resulted in significantly reduced liver transduction in vivo compared to Ad5. In macrophage-depleted mice, increased spleen uptake of AdT* was accompanied by an elevation in the levels of several inflammatory mediators. However ablation of the penton RGD motif in the AdT* vector background (AdT*RGE) resulted in a significant 5-fold reduction in spleen uptake and attenuated the antiviral inflammatory response. A reduction in spleen uptake and inflammatory activation was also observed in animals after intravascular administration of Ad5RGE compared to the parental Ad5 vector, with reduced co-localisation of the viral beta-galactosidase transgene with MAdCAM-1+ sinus-lining endothelial cells. Our detailed assessment of these novel adenoviruses indicates that penton base RGE mutation in combination with FX binding-ablation may be a viable strategy to attenuate the undesired liver uptake and pro-inflammatory responses to Ad5 vectors after intravascular delivery.
authors.affiliation: British Heart Foundation Glasgow Cardiovascular Research Centre, Institute of Cardiovascular and Medical Sciences, University of Glasgow, 126 University Place, Glasgow G12 8TA, UK.
authors.forename: Angela C
authors.initials: AC
authors.lastname: Bradshaw
authors.forename: Lynda
authors.initials: L
authors.lastname: Coughlan
authors.forename: Ashley M
authors.initials: AM
authors.lastname: Miller
authors.forename: Raul
authors.initials: R
authors.lastname: Alba
authors.forename: Nico
authors.initials: N
authors.lastname: van Rooijen
authors.forename: Stuart A
authors.initials: SA
authors.lastname: Nicklin
authors.forename: Andrew H
authors.initials: AH
authors.lastname: Baker
mesh_terms: D000260:Adenoviruses, Human / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D016207:Cytokines / Q000276:immunology; D004705:Endocytosis; D005170:Factor X / Q000737:chemistry / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D007113:Immunity, Innate; D007150:Immunohistochemistry; D008099:Liver / Q000276:immunology / Q000378:metabolism / Q000821:virology; D051379:Mice; D008815:Mice, Inbred Strains; D008856:Microscopy, Fluorescence; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009842:Oligopeptides / Q000378:metabolism; D011485:Protein Binding; D012703:Serotyping; D013154:Spleen / Q000276:immunology / Q000378:metabolism / Q000821:virology; D014018:Tissue Distribution; D014161:Transduction, Genetic; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D036022:Capsid Proteins; D016207:Cytokines; D009842:Oligopeptides; C018315:hexon capsid protein, Adenovirus; C043322:penton protein, adenovirus; C047981:arginyl-glycyl-aspartic acid; D005170:Factor X; D001616:beta-Galactosidase