Intracellular trafficking of hybrid gene delivery vectors
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 w...
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Published in | Journal of controlled release Vol. 207; pp. 120 - 130 |
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Main Authors | , , |
Format | Journal Article |
Language | English |
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Elsevier B.V
10.06.2015
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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.
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AbstractList | 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. 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. [Display omitted] |
Author | Pack, Daniel W. Keswani, Rahul K. Lazebnik, Mihael |
AuthorAffiliation | Department of Chemical and Materials Engineering and Department of Pharmaceutical Sciences, University of Kentucky, Lexington, KY 40506, USA Department of Chemical and Biomolecular Engineering, University of Illinois, Urbana, IL 61801, USA |
AuthorAffiliation_xml | – name: Department of Chemical and Biomolecular Engineering, University of Illinois, Urbana, IL 61801, USA – name: Department of Chemical and Materials Engineering and Department of Pharmaceutical Sciences, University of Kentucky, Lexington, KY 40506, USA |
Author_xml | – sequence: 1 givenname: Rahul K. surname: Keswani fullname: Keswani, Rahul K. organization: Department of Chemical and Biomolecular Engineering, University of Illinois, Urbana, IL 61801, USA – sequence: 2 givenname: Mihael surname: Lazebnik fullname: Lazebnik, Mihael organization: Department of Chemical and Biomolecular Engineering, University of Illinois, Urbana, IL 61801, USA – sequence: 3 givenname: Daniel W. orcidid: 0000-0003-0410-0423 surname: Pack fullname: Pack, Daniel W. email: dan.pack@uky.edu organization: Department of Chemical and Materials Engineering, University of Kentucky, Lexington, KY 40506, USA |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/25883029$$D View this record in MEDLINE/PubMed |
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CitedBy_id | crossref_primary_10_1002_mabi_201900183 crossref_primary_10_1002_macp_201500464 crossref_primary_10_1016_j_bbamem_2016_04_014 crossref_primary_10_1016_j_ijpharm_2018_10_010 crossref_primary_10_1007_s13346_024_01519_8 crossref_primary_10_1016_j_jcis_2016_06_071 crossref_primary_10_1016_j_colsurfb_2017_04_058 crossref_primary_10_1016_j_ijbiomac_2019_09_121 crossref_primary_10_1039_C6CS00177G crossref_primary_10_1021_acs_molpharmaceut_6b00709 crossref_primary_10_1016_j_colsurfa_2016_11_019 |
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Keywords | DOPE Retrovirus Hybrid vectors Chitosan Liposomes DOTAP Gene therapy Cholesterol |
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SubjectTerms | Blood Proteins - metabolism Caveolae - metabolism Chitosan Chitosan - chemistry Chitosan - metabolism Cholesterol Cholesterol - chemistry Cholesterol - metabolism Clathrin-Coated Vesicles - metabolism DOPE DOTAP Endocytosis - drug effects Fatty Acids, Monounsaturated - chemistry Fatty Acids, Monounsaturated - metabolism Gene therapy Genetic Vectors HEK293 Cells HeLa Cells Humans Hybrid vectors Kinetics Liposomes Microscopy, Confocal Moloney murine leukemia virus - genetics Moloney murine leukemia virus - metabolism Phosphatidylethanolamines - chemistry Phosphatidylethanolamines - metabolism Pinocytosis Quaternary Ammonium Compounds - chemistry Quaternary Ammonium Compounds - metabolism Retrovirus Temperature Transfection - methods Virion - genetics Virion - metabolism |
Title | Intracellular trafficking of hybrid gene delivery vectors |
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