title: Non-invasive gene targeting to the fetal brain after intravenous administration and transplacental transfer of plasmid DNA using PEGylated immunoliposomes.
abstract: Research was undertaken to establish transplacental delivery of active genes to fetal brain by a non-viral vector, antibody-specific targeted therapeutic procedure. PEGylated immunoliposomes (PILs) containing firefly luciferase DNA under the influence of the SV40 promoter injected intravenously into near-term pregnant mice produced luminometric evidence of CNS tissue luciferase activity at 48-h post-injection in all newborn pups. In utero delivery of this pGL3 DNA was shown after a single i.v. injection in maternal and neonatal brains, spleen and lesser amounts in lungs, with only negligible background levels in negative controls exposed to unencapsulated pDNA. In addition to studies of normal wild-type mice, we similarly injected pregnant Lafora Knockout (EPM2a null-mutant) and demonstrated luciferase activity days later in the maternal and newborn pup brains of both types. Delivery of PILs containing a second reporter gene (the pSV40 beta-galactosidase transgene) transplacentally by the same procedure was also successful. Histochemical and biochemical demonstration of beta-galactosidase was documented for all mutant and non-mutant neonates. Brain areas of highest Lafora body development (such as the hippocampus and pontine nuclei) showed intraneuronal beta-glucosidase activity. We conclude that receptor-mediated transport of PIL-borne gene therapeutics across both the placental barrier as well as the fetal BBB in utero is feasible.
authors.affiliation: a Neurology and Research Services, VA Greater Los Angeles Healthcare System, West Los Angeles Medical Center , Los Angeles , CA , USA .|b Department of Neurology , David Geffen School of Medicine at UCLA , Los Angeles , CA , USA .
authors.forename: Eain M
authors.initials: EM
authors.lastname: Cornford
authors.affiliation: a Neurology and Research Services, VA Greater Los Angeles Healthcare System, West Los Angeles Medical Center , Los Angeles , CA , USA .|b Department of Neurology , David Geffen School of Medicine at UCLA , Los Angeles , CA , USA .
authors.forename: Shigeyo
authors.initials: S
authors.lastname: Hyman
authors.affiliation: c Department of Pathology , Harbor-UCLA Medical Center , Torrance , CA , USA .|d Department of Pathology , David Geffen School of Medicine at UCLA , Los Angeles , CA , USA .
authors.forename: Marcia E
authors.initials: ME
authors.lastname: Cornford
authors.affiliation: a Neurology and Research Services, VA Greater Los Angeles Healthcare System, West Los Angeles Medical Center , Los Angeles , CA , USA .|b Department of Neurology , David Geffen School of Medicine at UCLA , Los Angeles , CA , USA .
authors.forename: Gabriela
authors.initials: G
authors.lastname: Chytrova
authors.affiliation: a Neurology and Research Services, VA Greater Los Angeles Healthcare System, West Los Angeles Medical Center , Los Angeles , CA , USA .
authors.forename: Jennifer
authors.initials: J
authors.lastname: Rhee
authors.affiliation: e RIKEN Brain Science Institute , Saitama , Japan .
authors.forename: Toshimitsu
authors.initials: T
authors.lastname: Suzuki
authors.affiliation: e RIKEN Brain Science Institute , Saitama , Japan .
authors.forename: Tetsushi
authors.initials: T
authors.lastname: Yamagata
authors.affiliation: e RIKEN Brain Science Institute , Saitama , Japan .
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Yamakawa
authors.affiliation: f Division of Surgical Oncology , Department of Surgery, David Geffen School of Medicine at UCLA , Los Angeles , CA , USA .|g Department of Microbiology , Immunology, and Molecular Genetics, David Geffen School of Medicine at UCLA , Los Angeles , CA , USA .|h Jonsson Comprehensive Cancer Center , UCLA, Los Angeles , CA , USA .|i The Molecular Biology Institute , UCLA, Los Angeles , CA , USA , and.
authors.forename: Manuel L
authors.initials: ML
authors.lastname: Penichet
authors.affiliation: j Department of Medicine , UCLA, Los Angeles , CA , USA.
authors.forename: William M
authors.initials: WM
authors.lastname: Pardridge
mesh_terms: D061605:Administration, Intravenous; D000818:Animals; D001921:Brain / Q000378:metabolism; D004705:Endocytosis; D005260:Female; D005333:Fetus / Q000378:metabolism*; D018390:Gene Targeting / Q000379:methods*; D017930:Genes, Reporter; D020192:Lafora Disease / Q000235:genetics; D008081:Liposomes / Q000737:chemistry / Q000276:immunology*; D049409:Luciferases, Firefly / Q000008:administration & dosage / Q000235:genetics* / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010920:Placenta / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry; D011247:Pregnancy; D011401:Promoter Regions, Genetic / Q000187:drug effects; D011990:Receptors, Transferrin / Q000378:metabolism; D014018:Tissue Distribution; D019076:Transgenes / Q000187:drug effects; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D008081:Liposomes; D011990:Receptors, Transferrin; D011092:Polyethylene Glycols; D049409:Luciferases, Firefly; D001616:beta-Galactosidase
keywords: Adult and fetal blood–brain barrier; EPM2a null mutant mice; Lafora's disease; PEGylated immunoliposome; non-invasive gene delivery; transferrin receptor-mediated endocytosis; transplacental gene transfer