Document Detail


Retinal pigment epithelial cell-based gene therapy against hemoglobin toxicity.
MedLine Citation:
PMID:  9852279     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
To determine if overexpression of the human heme oxygenase (HO-1) protects retinal pigment (RPE) cells from hemoglobin toxicity, a human RPE cell line was infected by an adenoviral vector containing the HO-1 (Ad-HO-1) gene or transfected with a plasmid containing the cytomegalovirus promoter and HO-1 cDNA (pRc/CMV-HO-1) complexed to cationic liposomes. Phase contrast microscopy and acid phosphatase activity were examined to insure homogeneity of the cell line. Mitochondrial cytochrome and microsomal heme content were measured in both transduced and control cells. RPE cells were then challenged with hemoglobin and their viability estimated. We determined that cells transfected with Ad HO-1 overexpressed HO-1 compared to control cells: HO-1 mRNA levels were increased 3-fold within 3 days, decreasing in 7 days. In addition, we permanently transfected RPE cells with HO-1 gene. Transfected cell clones selected for neomycin resistance had elevated levels of HO activity 3-fold higher than control. Transfected cells exposed to hemoglobin had a survival rate of 93%; non-transfected cells had a 65-75% rate of survival. Transfected cells overexpressing HO-1 proved highly viable when challenged with hemoglobin. HO-1 appears to be an important component of the cellular anti-oxidant defense mechanisms against hemoglobin toxicity. However, the choice of transient or permanent expression of HO-1 against hemoglobin toxicity and hemorrhage needs to be further evaluated.
Authors:
N G Abraham; J L Da Silva; M W Dunn; K Kigasawa; S Shibahara
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  International journal of molecular medicine     Volume:  1     ISSN:  1107-3756     ISO Abbreviation:  Int. J. Mol. Med.     Publication Date:  1998 Apr 
Date Detail:
Created Date:  1999-03-01     Completed Date:  1999-03-01     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  9810955     Medline TA:  Int J Mol Med     Country:  GREECE    
Other Details:
Languages:  eng     Pagination:  657-63     Citation Subset:  IM    
Affiliation:
New York Medical College, Department of Pharmacology, Valhalla, NY 10595, USA.
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MeSH Terms
Descriptor/Qualifier:
Adenoviridae / genetics
Animals
Cell Line
Cytochromes / analysis
Gene Therapy / methods*
Genetic Vectors / pharmacology*
Heme / analysis
Heme Oxygenase (Decyclizing) / genetics*,  metabolism
Heme Oxygenase-1
Hemoglobins / pharmacology
Humans
Liposomes / pharmacology
Membrane Proteins
Microsomes / metabolism
Mitochondria / metabolism
Oxidative Stress
Pigment Epithelium of Eye / drug effects,  physiology*
RNA, Messenger / analysis*
Rabbits
Transcription, Genetic
Transfection
Grant Support
ID/Acronym/Agency:
R01 HL5-4138/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Cytochromes; 0/Hemoglobins; 0/Liposomes; 0/Membrane Proteins; 0/RNA, Messenger; 14875-96-8/Heme; EC 1.14.99.3/HMOX1 protein, human; EC 1.14.99.3/Heme Oxygenase (Decyclizing); EC 1.14.99.3/Heme Oxygenase-1

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


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