Ultrasound-Guided Percutaneous Delivery of Adenoviral Vectors Encoding theβ-Galactosidase and Human Factor IX Genes to Early Gestation Fetal SheepIn Utero
| Autor: | Thomas Dahse, Donald Peebles, Holm Schneider, M Nivsarkar, David E. Noakes, M Miah, Anna L. David, Simon N. Waddington, Charles Coutelle, Terence Cook, HJ Knapton, Michael Themis, Charles H. Rodeck |
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| Rok vydání: | 2003 |
| Předmět: |
Umbilical Veins
Genetic enhancement Genetic Vectors Gestational Age Administration Cutaneous Injections Intramuscular Antibodies Ultrasonography Prenatal Adenoviridae Immune tolerance Factor IX Andrology Fetus Genes Reporter Pregnancy Immune Tolerance Genetics Animals Humans Medicine Amnion Vector (molecular biology) Molecular Biology Sheep business.industry Genetic transfer Genetic Therapy beta-Galactosidase Transplantation Injections Intra-Arterial Liver In utero Immunology Molecular Medicine Female Stem cell business Injections Intraperitoneal |
| Zdroj: | Human Gene Therapy. 14:353-364 |
| ISSN: | 1557-7422 1043-0342 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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