| Poly (D, L-lactide-co-glycolide)/DNA microspheres to facilitate prolonged transgene expression in airway epithelium in vitro, ex vivo and in vivo. | |
| | |
MedLine Citation:
|
PMID: 12883524 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
Repeat administration of gene therapy for cystic fibrosis is likely to be essential for long-term clinical efficacy. This may be minimized by the use of slow-release gene transfer preparations with more prolonged expression and longer dosing intervals for the patient. Poly(D-L-lactide-co-glycolide) (PLG) is a biodegradable and biocompatible polymer that has been used to encapsulate plasmid DNA. PLG-DNA microspheres were generated and characterized with respect to morphology, size (80% of particles <5.2 microm), and encapsulation efficiency (50.7+/-2.3%, n=6). Gel electrophoresis of DNA re-extracted from the microspheres confirmed that despite a decrease in the proportion of supercoiled conformation, it had not been degraded by the preparation process. Gene transfer efficiency was tested using microspheres encapsulating the reporter gene beta-galactosidase in vitro on Cos 7 cells and a CF airway epithelial line (CFTEo approximately ) and ex vivo in a sheep tracheal (s.t.) model. In both cases, transgene expression was significantly (P<0.01) lower at the first time point tested (24 h in vitro, 48 h ex vivo) compared to lipid-#67-mediated gene transfer. However, PLG-mediated expression in vitro was sustained at 48 h, while lipid #67-mediated expression levels had dropped significantly (P<0.05) to 50.3+/-13.7 and 38.2+/-2.7% (Cos 7 and CFTEo approximately cells, respectively) of the 24-h level. This pattern was also seen in the s.t. model where at 72 h, PLG-mediated expression was 125.4+/-7.2% of the 48-h level demonstrating significantly (P<0.05) better retention of transfection efficiency than lipid #67, where levels had fallen to approximately half the 48 h level. By 96 h, expression was still retained in the PLG-transfected group (87.3+/-12.5% of 48 h expression) but was undetectable in the lipid -#67-transfected s.t. Finally, PLG microspheres, encapsulating the reporter gene chloramphenicol transferase (CAT, 80 microg) were instilled intranasally into Balb/C mice. Compared to lipid-#67-mediated delivery, where whole lung CAT expression was highest at 48 h (13.7 x 10(3)+/-0.05 CAT U/microg protein, n=6) and then not detectable at further time points, CAT expression was not detectable in PLG-transfected mice at 48 h, but was detectable at 7, 14 and 21 days after transfection. These data demonstrate that PLG-mediated gene transfer can produce prolonged gene expression in airway epithelia. However, gene transfer efficiency still requires significant improvement. |
| | |
Authors:
|
M Stern; K Ulrich; D M Geddes; E W F W Alton |
Related Documents
:
|
20055094 - Ionically crosslinked chitosan nanoparticles as gene delivery systems: effect of pegyla... 11072114 - Tissue engineering via local gene delivery: update and future prospects for enhancing t... 14970604 - Baculovirus-mediated gene delivery into mammalian cells. 10694804 - Gene transfer into inflamed glomeruli using macrophages transfected with adenovirus. 10187834 - The gain-of-function chinese hamster ovary mutant lec11b expresses one of two chinese h... 20712784 - A preclinical approach for gene therapy of beta-thalassemia. |
Publication Detail:
|
Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
|
Title: Gene therapy Volume: 10 ISSN: 0969-7128 ISO Abbreviation: Gene Ther. Publication Date: 2003 Aug |
Date Detail:
|
Created Date: 2003-07-28 Completed Date: 2003-09-17 Revised Date: 2006-11-15 |
Medline Journal Info:
|
Nlm Unique ID: 9421525 Medline TA: Gene Ther Country: England |
Other Details:
|
Languages: eng Pagination: 1282-8 Citation Subset: IM |
Affiliation:
|
Department of Gene Therapy, Imperial College Faculty of Medicine at the National Heart and Lung Institute, London, UK. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Animals Animals, Newborn Biocompatible Materials COS Cells Cystic Fibrosis / therapy* DNA / administration & dosage* Epithelium / enzymology* Gene Expression Gene Therapy / methods* Lactic Acid Mice Mice, Inbred BALB C Microscopy, Electron, Scanning Microspheres* Polyglycolic Acid Polymers Sheep Time Factors Trachea / enzymology Transfection / methods* beta-Galactosidase / genetics |
| Chemical | |
Reg. No./Substance:
|
0/Biocompatible Materials; 0/Polymers; 0/polylactic acid-polyglycolic acid copolymer; 26009-03-0/Polyglycolic Acid; 50-21-5/Lactic Acid; 9007-49-2/DNA; EC 3.2.1.23/beta-Galactosidase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Gene therapy progress and prospects: gene therapy of lysosomal storage disorders.
Next Document: Osteoinduction by ex vivo adenovirus-mediated BMP2 delivery is independent of cell type.