Document Detail


Ultrasound-guided percutaneous delivery of adenoviral vectors encoding the beta-galactosidase and human factor IX genes to early gestation fetal sheep in utero.
MedLine Citation:
PMID:  12659676     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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:
Anna David; Terry Cook; Simon Waddington; Donald Peebles; Megha Nivsarkar; Holly Knapton; Maznu Miah; Thomas Dahse; David Noakes; Holm Schneider; Charles Rodeck; Charles Coutelle; Mike Themis
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Human gene therapy     Volume:  14     ISSN:  1043-0342     ISO Abbreviation:  Hum. Gene Ther.     Publication Date:  2003 Mar 
Date Detail:
Created Date:  2003-03-27     Completed Date:  2003-09-26     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9008950     Medline TA:  Hum Gene Ther     Country:  United States    
Other Details:
Languages:  eng     Pagination:  353-64     Citation Subset:  IM    
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
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MeSH Terms
Descriptor/Qualifier:
Adenoviridae / immunology*
Administration, Cutaneous
Amnion
Animals
Antibodies / blood
Factor IX / genetics*
Female
Fetus / anatomy & histology,  chemistry
Gene Therapy
Genes, Reporter
Genetic Vectors / administration & dosage*
Gestational Age
Humans
Immune Tolerance
Injections, Intra-Arterial
Injections, Intramuscular
Injections, Intraperitoneal
Liver
Pregnancy
Sheep
Ultrasonography, Prenatal*
Umbilical Veins
beta-Galactosidase / analysis,  genetics*
Chemical
Reg. No./Substance:
0/Antibodies; 9001-28-9/Factor IX; EC 3.2.1.23/beta-Galactosidase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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