title: Infusion of imaging and therapeutic molecules into the plant virus-based carrier cowpea mosaic virus: cargo-loading and delivery.
abstract: This work is focused on the development of a plant virus-based carrier system for cargo delivery, specifically 30nm-sized cowpea mosaic virus (CPMV). Whereas previous reports described the engineering of CPMV through genetic or chemical modification, we report a non-covalent infusion technique that facilitates efficient cargo loading. Infusion and retention of 130-155 fluorescent dye molecules per CPMV using DAPI (4',6-diamidino-2-phenylindole dihydrochloride), propidium iodide (3,8-diamino-5-[3-(diethylmethylammonio)propyl]-6-phenylphenanthridinium diiodide), and acridine orange (3,6-bis(dimethylamino)acridinium chloride), as well as 140 copies of therapeutic payload proflavine (PF, acridine-3,6-diamine hydrochloride), is reported. Loading is achieved through interaction of the cargo with the CPMV's encapsidated RNA molecules. The loading mechanism is specific; empty RNA-free eCPMV nanoparticles could not be loaded. Cargo-infused CPMV nanoparticles remain chemically active, and surface lysine residues were covalent modified with dyes leading to the development of dual-functional CPMV carrier systems. We demonstrate cargo-delivery to a panel of cancer cells (cervical, breast, and colon): CPMV nanoparticles enter cells via the surface marker vimentin, the nanoparticles target the endolysosome, where the carrier is degraded and the cargo is released allowing imaging and/or cell killing. In conclusion, we demonstrate cargo-infusion and delivery to cells; the methods discussed provide a useful means for functionalization of CPMV toward its application as drug and/or contrast agent delivery vehicle.
authors.affiliation: Department of Biomedical Engineering, Case Western Reserve University, Schools of Medicine and Engineering, 10900 Euclid Avenue, Cleveland 44106, USA.
authors.forename: Ibrahim
authors.initials: I
authors.lastname: Yildiz
authors.forename: Karin L
authors.initials: KL
authors.lastname: Lee
authors.forename: Kevin
authors.initials: K
authors.lastname: Chen
authors.forename: Sourabh
authors.initials: S
authors.lastname: Shukla
authors.forename: Nicole F
authors.initials: NF
authors.lastname: Steinmetz
mesh_terms: D000165:Acridine Orange / Q000008:administration & dosage; D000891:Anti-Infective Agents, Local / Q000008:administration & dosage*; D045744:Cell Line, Tumor; D017798:Comovirus / Q000378:metabolism*; D004337:Drug Carriers / Q000378:metabolism*; D011992:Endosomes / Q000378:metabolism; D005456:Fluorescent Dyes / Q000008:administration & dosage*; D006801:Humans; D007211:Indoles / Q000008:administration & dosage; D053758:Nanoparticles / Q000378:metabolism*; D018532:Pisum sativum / Q000821:virology; D011370:Proflavine / Q000008:administration & dosage*; D011419:Propidium / Q000008:administration & dosage
chemical_list: D000891:Anti-Infective Agents, Local; D004337:Drug Carriers; D005456:Fluorescent Dyes; D007211:Indoles; D011419:Propidium; C007293:DAPI; D011370:Proflavine; D000165:Acridine Orange
keywords: Cowpea mosaic virus; Drug delivery; Infusion; Viral nanoparticle