title: Intravital imaging of embryonic and tumor neovasculature using viral nanoparticles.
abstract: Viral nanoparticles are a novel class of biomolecular agents that take advantage of the natural circulatory and targeting properties of viruses to allow the development of therapeutics, vaccines and imaging tools. We have developed a multivalent nanoparticle platform based on the cowpea mosaic virus (CPMV) that facilitates particle labeling at high density with fluorescent dyes and other functional groups. Compared with other technologies, CPMV-based viral nanoparticles are particularly suited for long-term intravital vascular imaging because of their biocompatibility and retention in the endothelium with minimal side effects. The stable, long-term labeling of the endothelium allows the identification of vasculature undergoing active remodeling in real time. In this study, we describe the synthesis, purification and fluorescent labeling of CPMV nanoparticles, along with their use for imaging of vascular structure and for intravital vascular mapping in developmental and tumor angiogenesis models. Dye-labeled viral nanoparticles can be synthesized and purified in a single day, and imaging studies can be conducted over hours, days or weeks, depending on the application.
authors.affiliation: Translational Prostate Cancer Research Group, London Regional Cancer Program, London, Ontario, Canada.
authors.forename: Hon Sing
authors.initials: HS
authors.lastname: Leong
authors.forename: Nicole F
authors.initials: NF
authors.lastname: Steinmetz
authors.forename: Amber
authors.initials: A
authors.lastname: Ablack
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Destito
authors.forename: Andries
authors.initials: A
authors.lastname: Zijlstra
authors.forename: Heidi
authors.initials: H
authors.lastname: Stuhlmann
authors.forename: Marianne
authors.initials: M
authors.lastname: Manchester
authors.forename: John D
authors.initials: JD
authors.lastname: Lewis
mesh_terms: D000818:Animals; D002645:Chickens; D017798:Comovirus / Q000302:isolation & purification*; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian / Q000098:blood supply*; D004625:Embryo, Nonmammalian / Q000098:blood supply*; D004730:Endothelium, Vascular / Q000033:anatomy & histology*; D007887:Fabaceae / Q000821:virology; D005456:Fluorescent Dyes / Q000032:analysis / Q000737:chemistry; D051379:Mice; D008845:Microinjections / Q000379:methods; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D053758:Nanoparticles; D036103:Nanotechnology / Q000379:methods*; D009369:Neoplasms / Q000098:blood supply*
chemical_list: D005456:Fluorescent Dyes