title: Site-specific Genome Editing in PBMCs With PLGA Nanoparticle-delivered PNAs Confers HIV-1 Resistance in Humanized Mice.
abstract: Biodegradable poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) encapsulating triplex-forming peptide nucleic acids (PNAs) and donor DNAs for recombination-mediated editing of the CCR5 gene were synthesized for delivery into human peripheral blood mononuclear cells (PBMCs). NPs containing the CCR5-targeting molecules efficiently entered PBMCs with low cytotoxicity. Deep sequencing revealed that a single treatment with the formulation resulted in a targeting frequency of 0.97% in the CCR5 gene and a low off-target frequency of 0.004% in the CCR2 gene, a 216-fold difference. NP-treated PBMCs efficiently engrafted immunodeficient NOD-scid IL-2rγ(-/-) mice, and the targeted CCR5 modification was detected in splenic lymphocytes 4 weeks posttransplantation. After infection with an R5-tropic strain of HIV-1, humanized mice with CCR5-NP-treated PBMCs displayed significantly higher levels of CD4(+) T cells and significantly reduced plasma viral RNA loads compared with control mice engrafted with mock-treated PBMCs. This work demonstrates the feasibility of PLGA-NP-encapsulated PNA-based gene-editing molecules for the targeted modification of CCR5 in human PBMCs as a platform for conferring HIV-1 resistance.Molecular Therapy-Nucleic Acids (2013) 2, e135; doi:10.1038/mtna.2013.59; published online 19 November 2013.
authors.affiliation: Department of Therapeutic Radiology and Genetics, Yale University School of Medicine, New Haven, Connecticut, USA.
authors.forename: Erica B
authors.initials: EB
authors.lastname: Schleifman
authors.forename: Nicole Ali
authors.initials: NA
authors.lastname: McNeer
authors.forename: Andrew
authors.initials: A
authors.lastname: Jackson
authors.forename: Jennifer
authors.initials: J
authors.lastname: Yamtich
authors.forename: Michael A
authors.initials: MA
authors.lastname: Brehm
authors.forename: Leonard D
authors.initials: LD
authors.lastname: Shultz
authors.forename: Dale L
authors.initials: DL
authors.lastname: Greiner
authors.forename: Priti
authors.initials: P
authors.lastname: Kumar
authors.forename: W Mark
authors.initials: WM
authors.lastname: Saltzman
authors.forename: Peter M
authors.initials: PM
authors.lastname: Glazer