Abstract
Viral and non-viral gene delivery vectors are in development for human gene therapy, but both exhibit disadvantages such as inadequate efficiency, lack of cell-specific targeting or safety concerns. We have recently reported the design of hybrid delivery vectors combining retrovirus-like particles with synthetic polymers or lipids that are efficient, provide sustained gene expression and are more stable compared to native retroviruses. To guide further development of this promising class of gene delivery vectors, we have investigated their mechanisms of intracellular trafficking. Moloney murine leukemia virus-like particles (M-VLPs) were complexed with chitosan (Chi) or liposomes (Lip) comprising DOTAP, DOPE and cholesterol to form the hybrid vectors (Chi/M-VLPs and Lip/M-VLPs, respectively). Transfection efficiency and cellular internalization of the vectors were quantified in the presence of a panel of inhibitors of various endocytic pathways. Intracellular transport and trafficking kinetics of the hybrid vectors were dependent on the synthetic component and used a combination of clathrin- and caveolar-dependent endocytosis and macropinocytosis. Chi/M-VLPs were slower to transfect compared to Lip/M-VLPs due to the delayed detachment of the synthetic component. The synthetic component of hybrid gene delivery vectors plays a significant role in their cellular interactions and processing and is a key parameter for the design of more efficient gene delivery vehicles.
| Original language | English |
|---|---|
| Pages (from-to) | 120-130 |
| Number of pages | 11 |
| Journal | Journal of Controlled Release |
| Volume | 207 |
| DOIs | |
| State | Published - Jun 10 2015 |
Bibliographical note
Publisher Copyright:© 2015 Elsevier B.V.
Funding
This work was partially supported by the American Heart Association Predoctoral Fellowship ( 11PRE5960002 ) (ML), National Science Foundation ( BES 06–02636 ) (DWP) and National Institutes of Health GM085222 (DWP). In addition, we thank Sandy Mattick at the Cell Culture Media Facility at the University of Illinois for help with the preparation of cell media. Flow cytometry was performed at the Roy J. Carver Biotechnology Center Flow Cytometry Facility, University of Illinois. q-PCR and confocal fluorescence microscopy were performed at the Institute of Genomic Biology Core Facilities, University of Illinois with technical help from Mayandi Sivaguru.
| Funders | Funder number |
|---|---|
| National Science Foundation (NSF) | BES 06–02636 |
| National Institutes of Health (NIH) | |
| National Institute of General Medical Sciences | R01GM085222 |
| American Heart Association | 11PRE5960002 |
Keywords
- Chitosan
- Cholesterol
- DOPE
- DOTAP
- Gene therapy
- Hybrid vectors
- Liposomes
- Retrovirus
ASJC Scopus subject areas
- Pharmaceutical Science