title: Adenovirus-mediated delivery of CD46 attenuates the alternative complement pathway on RPE: implications for age-related macular degeneration.
abstract: Activation of the alternative pathway of the complement system has been implicated in the pathogenesis of age-related macular degeneration. Membrane attack complex (MAC) has been identified mainly on the Bruch's membrane and drusen underlying the retinal pigment epithelium (RPE). Membrane cofactor protein (CD46) preferentially regulates the alternative pathway of complement. The aim of this study was to evaluate the potential of increasing CD46 expression on RPE cells using an adenovirus as a gene therapy approach to reduce alternative pathway-mediated damage to RPE cells. We generated a recombinant adenovirus vector expressing human CD46 (hCD46) and delivered the vector to murine hepatocytes and RPE cells in vitro. After incubation in human serum in conditions in which the classical pathway of complement was blocked, we measured alternative pathway-mediated damage of these cells by quantifying lysis and MAC formation. Adenovirus expressing hCD46 was delivered to the subretinal space of adult mice, and 1 week later, ocular flat mounts were challenged with human serum and the levels of complement-mediated damage was quantified. Adenovirus-mediated delivery of hCD46 localizes to the basal and lateral surfaces of RPE cells where it offers protection from alternative pathway-mediated damage, but not classical, allowing the classical pathway to function unhindered.
authors.affiliation: Department of Ophthalmology and Neuroscience, Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, MA 02111, USA.
authors.forename: J H
authors.initials: JH
authors.lastname: Sweigard
authors.forename: S M
authors.initials: SM
authors.lastname: Cashman
authors.forename: R
authors.initials: R
authors.lastname: Kumar-Singh
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000328:Adult; D000818:Animals; D002460:Cell Line; D015938:Complement Membrane Attack Complex / Q000378:metabolism; D003170:Complement Pathway, Alternative / Q000235:genetics / Q000276:immunology*; D015316:Genetic Therapy; D005822:Genetic Vectors; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D008268:Macular Degeneration / Q000235:genetics* / Q000276:immunology*; D051916:Membrane Cofactor Protein / Q000235:genetics* / Q000494:pharmacology; D051379:Mice; D012160:Retina / Q000196:embryology / Q000378:metabolism; D055213:Retinal Pigment Epithelium / Q000276:immunology / Q000378:metabolism*
chemical_list: D015938:Complement Membrane Attack Complex; D051916:Membrane Cofactor Protein