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Retroviral gene transfer: effects on endothelial cell phenotype
M Inaba1, E Toninelli, G Vanmeter
1Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Connecticut, USA.
The Journal of Surgical Research
|September 12, 1998
Summary
Somatic cell gene therapy using retroviral vectors can efficiently transduce endothelial cells (EC) ex vivo. This process does not alter EC phenotype, growth, or cell adhesion molecule expression, making it suitable for cardiovascular disease treatment.
Area of Science:
- Gene Therapy
- Cell Biology
- Cardiovascular Research
Background:
- Endothelial cells (EC) are a key target for gene therapy in cardiovascular disease.
- Viral transduction of EC may cause undesirable phenotypic changes.
- Assessing EC behavior post-gene transfer is crucial for therapeutic applications.
Purpose of the Study:
- To evaluate the impact of retroviral gene transfer (GT) on endothelial cell (EC) phenotype.
- To assess the proliferative capacity and cell adhesion molecule (CAM) profile of EC after ex vivo GT.
- To determine the safety and efficacy of a clinically relevant GT protocol for EC.
Main Methods:
- Human umbilical vein EC (HUVEC) were exposed to MFG.nlsLACZ retroviral vector.
- Control groups received empty virus (CRIP) or no virus (NT).
- Gene transfer efficiency, proliferation, and expression of activation molecules (MHC II, E-selectin, ICAM-1, VCAM-1) were quantified using FACS and cell counts.
Main Results:
- Retroviral GT achieved efficient transduction (mean 57%) without selection.
- Transduced and control EC showed identical expression profiles for key CAMs at rest and after cytokine stimulation.
- No significant differences in growth rates were observed between transduced and control EC.
Conclusions:
- High-efficiency ex vivo GT of EC is achievable with retroviral vectors without selection.
- Transduced EC maintain a non-activated phenotype with normal inducible CAM expression and growth.
- Vector and protocol specificity are critical; careful assessment is needed before in vivo application.