Mitosis enhances transgene expression of plasmid delivered by cationic liposomes

Tseng, Wen‐Chi; Haselton, Frederick R.; Giorgio, Todd D.

doi:10.1016/s0167-4781(99)00039-1

Cited by 210 publications

(153 citation statements)

References 14 publications

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“…Fibroblasts arrive to contribute to either wound healing or fibrous capsule formation, and may be transfected by localized gene delivery [62]. The cell proliferation induced at the implant site may act to promote nuclear accumulation of the vector [120]. A long-term persistence of macrophages, however, can be problematic due to their production of oxygen radicals or formation of foreign body giant cells [119].…”

Section: Inflammation and Vector Activitymentioning

confidence: 99%

“…More generally, a range of materials with varying mechanics was investigated with a similar trend observed. The role of the matrix mechanics was attributed to an increased rate of cell proliferation on the more rigid matrices, as cell proliferation can enable nuclear accumulation of the delivered plasmid [120]. Though gene transfer may be enhanced on more rigid materials, delivery on softer materials must also be developed since softer materials may be necessary to promote the desired cellular responses [131].…”

Section: Microenvironment and Gene Transfermentioning

confidence: 99%

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Matrices and scaffolds for DNA delivery in tissue engineering

Laporte¹,

Shea²

2007

Advanced Drug Delivery Reviews

241

208

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Regenerative medicine aims to create functional tissue replacements, typically through creating a controlled environment that promotes and directs the differentiation of stem or progenitor cells, either endogenous or transplanted. Scaffolds serve a central role in many strategies by providing the means to control the local environment. Gene delivery from the scaffold represents a versatile approach to manipulating the local environment for directing cell function. Research at the interface of biomaterials, gene therapy, and drug delivery has identified several design parameters for the vector and the biomaterial scaffold that must be satisfied. Progress has been made towards achieving gene delivery within a tissue engineering scaffold, though the design principles for the materials and vectors that produce efficient delivery require further development. Nevertheless, these advances in obtaining transgene expression with the scaffold have created opportunities to develop greater control of either delivery or expression and to identify the best practices for promoting tissue formation. Strategies to achieve controlled localized expression within the tissue engineering scaffold will have broad application to the regeneration of many tissues, with great promise for clinical therapies.

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Section: Inflammation and Vector Activitymentioning

confidence: 99%

Section: Microenvironment and Gene Transfermentioning

confidence: 99%

Matrices and scaffolds for DNA delivery in tissue engineering

Laporte¹,

Shea²

2007

Advanced Drug Delivery Reviews

241

208

View full text Add to dashboard Cite

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“…18 Other studies have evidence supporting plasmid translocation across the nuclear membrane as a critical obstacle through characterization of the relation between transgene expression and mitosis by modulating cell cycle timing. 19 The implication is that plasmid transport into the nucleus is facilitated by barrier changes during mitosis. These and other observations are extremely useful in understanding the mechanisms of transgene expression, but lack the systematic and quantitative approach necessary to rationally design and evaluate nonviral delivery systems with improved function.…”

Section: Introductionmentioning

confidence: 99%

A model for the analysis of nonviral gene therapy

et al. 2003

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Further understanding of the mechanisms involved in cellular and intracellular delivery of transgene is needed to produce clinical applications of gene therapy. The compartmental and computational model designed in this work is integrated with data from previous experiments to quantitatively estimate rate constants of plasmid translocation across cellular barriers in transgene delivery in vitro. The experimental conditions between two cellular studies were held constant, varying only the cell type, to investigate how the rates differed between cell lines. Two rate constants were estimated per barrier for active transport and passive diffusion. Translocation rates of intact plasmid across the cytoplasmic and nuclear barriers varied between cell lines. CV1 cells were defined by slower rates (0.23 h À1 cytoplasmic, 0.08 h -1 nuclear) than those of the HeLa cells (1.87 h À1 cytoplasmic, 0.45 h À1 nuclear). The nuclear envelope was identified as a rate-limiting barrier by comparing the rate of intact plasmid translocation at each barrier. Slower intact plasmid translocation in CV1 cells was correlated with a reduced absolute capacity for transgene efficiency in comparison with HeLa cells. HeLa cells were three times more efficient than CV1 cells at producing green fluorescent protein per intact plasmid delivered to the nucleus. Mathematical modeling coordinated with experimental studies can provide detailed, quantitative understanding of nonviral gene therapy.

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“…A number of hurdles to efficient gene transfer using this approach have been identified. These include access of the plasmid DNA (pDNA) 3 to the nuclei of terminally differentiated airway epithelial cells (1).…”

mentioning

confidence: 99%

“…In dividing cells, the nuclear envelope disassembles at the M phase of division, and is therefore not a barrier to DNA nuclear entry (3). For non-dividing cells, pDNA cyto-nucleoplasmic transport is likely to be a facilitated process that is mediated by the interaction of soluble cytoplasmic factors with the minimal nuclear transport machinery.…”

mentioning

confidence: 99%

Identification and Functional Characterization of Cytoplasmic Determinants of Plasmid DNA Nuclear Import

Munkonge

Amin

Hyde

et al. 2009

Journal of Biological Chemistry

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Import of exogenous plasmid DNA (pDNA) into mammalian cell nuclei represents a key intracellular obstacle to efficient non-viral gene delivery. This includes access of the pDNA to the nuclei of non-dividing cells where the presence of an intact nuclear membrane is limiting for gene transfer. Here we identify, isolate, and characterize, cytoplasmic determinants of pDNA nuclear import into digitonin-permeabilized HeLa cells. Depletion of putative DNA-binding proteins, on the basis of their ability to bind immobilized pDNA, abolished pDNA nuclear import supporting the critical role of cytoplasmic factors in this process. Elution of pDNAbound proteins, followed by two-dimensional sodium dodecyl polyacrylamide gel electrophoresis identified several candidate DNA shuttle proteins. We show that two of these, NM23-H2, a ubiquitous c-Myc transcription-activating nucleoside diphosphate kinase, and the core histone H2B can both reconstitute pDNA nuclear import. Further, we demonstrate a significant increase in gene transfer in non-dividing HeLa cells transiently transfected with pDNA containing binding sequences from two of the DNA shuttle proteins, NM23-H2 and the homeobox transcription factor Chx10. These data support the hypothesis that exogenous pDNA binds to cytoplasmic shuttle proteins and is then translocated to the nucleus using the minimal import machinery. Importantly, increasing the binding of pDNA to shuttle proteins by re-engineering reporter plasmids with shuttle binding sequences enhances gene transfer. Increasing the potential for exogenously added pDNA to bind intracellular transport cofactors may enhance the potency of non-viral gene transfer.

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Mitosis enhances transgene expression of plasmid delivered by cationic liposomes

Cited by 210 publications

References 14 publications

Matrices and scaffolds for DNA delivery in tissue engineering

Matrices and scaffolds for DNA delivery in tissue engineering

A model for the analysis of nonviral gene therapy

Identification and Functional Characterization of Cytoplasmic Determinants of Plasmid DNA Nuclear Import

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