title: Ultrasound-guided percutaneous delivery of adenoviral vectors encoding the beta-galactosidase and human factor IX genes to early gestation fetal sheep in utero.
abstract: In utero gene therapy may provide treatment of genetic diseases before significant organ damage, allow permanent genetic correction by reaching stem cell populations, and provide immune tolerance against the therapeutic transgenes and vectors. We have used percutaneous ultrasound-guided injection as a minimally invasive fetal procedure. First-generation adenoviruses encoding the nuclear localizing beta-galactosidase reporter gene or the human factor IX (hFIX) gene, or colloidal carbon were delivered via the umbilical vein (UV, n = 4), heart (intracardiac [IC], n = 2), liver parenchyma (intrahepatic [HE], n = 11), peritoneal cavity (intraperitoneal [IP], n = 14), skeletal musculature ([intramuscular [IM], n = 11), or the amniotic cavity (intraamniotic [IA], n = 14) to early-gestation fetal sheep (0.3 gestation = day 33-61). Postmortem analysis was performed at 2, 9, or 28 days after injection. Although fetal survival was between 77% and 91% for IP, HE, IA, and IM routes, no fetuses survived UV or IC procedures. The hFIX levels reaching 1900 and 401 ng/ml (IP), 30 ng/ml (HE), 66.5 and 39 ng/ml (IA), and 83 and 65.5 ng/ml (IM), respectively, were determined 2 days after injection and decreased at birth to 16.5 ng/ml (IP), 7 ng/ml (HE), 4.5 ng/ml (IA), and 4 and 0 ng/ml (IM). Polymerase chain reaction (PCR) and immunohistochemistry showed broadest hFIX transgene spread and highest localised beta-galactosidase expression, respectively, after IP administration. Antibodies were observed against vector but not against hFIX.
authors.affiliation: Department of Obstetrics and Gynaecology, Royal Free and University College Medical School, 86-96 Chenies Mews, London, WC1E 6HX, United Kingdom. a.david@ucl.ac.uk
authors.forename: Anna
authors.initials: A
authors.lastname: David
authors.forename: Terry
authors.initials: T
authors.lastname: Cook
authors.forename: Simon
authors.initials: S
authors.lastname: Waddington
authors.forename: Donald
authors.initials: D
authors.lastname: Peebles
authors.forename: Megha
authors.initials: M
authors.lastname: Nivsarkar
authors.forename: Holly
authors.initials: H
authors.lastname: Knapton
authors.forename: Maznu
authors.initials: M
authors.lastname: Miah
authors.forename: Thomas
authors.initials: T
authors.lastname: Dahse
authors.forename: David
authors.initials: D
authors.lastname: Noakes
authors.forename: Holm
authors.initials: H
authors.lastname: Schneider
authors.forename: Charles
authors.initials: C
authors.lastname: Rodeck
authors.forename: Charles
authors.initials: C
authors.lastname: Coutelle
authors.forename: Mike
authors.initials: M
authors.lastname: Themis
mesh_terms: D000256:Adenoviridae / Q000276:immunology*; D000279:Administration, Cutaneous; D000650:Amnion; D000818:Animals; D000906:Antibodies / Q000097:blood; D005164:Factor IX / Q000235:genetics*; D005260:Female; D005333:Fetus / Q000033:anatomy & histology / Q000737:chemistry; D017930:Genes, Reporter; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage*; D005865:Gestational Age; D006801:Humans; D007108:Immune Tolerance; D007269:Injections, Intra-Arterial; D007273:Injections, Intramuscular; D007274:Injections, Intraperitoneal; D008099:Liver; D011247:Pregnancy; D012756:Sheep; D016216:Ultrasonography, Prenatal; D014471:Umbilical Veins; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics*
chemical_list: D000906:Antibodies; D005164:Factor IX; D001616:beta-Galactosidase