Abstract
PurposePrenyl ammonium iodides (Amino-Prenols, APs), semi-synthetic polyprenol derivatives were studied as prospective novel gene transfer agents.MethodsAP-7, -8, -11 and -15 (aminoprenols composed of 7, 8, 11 or 15 isoprene units, respectively) were examined for their capacity to form complexes with pDNA, for cytotoxicity and ability to transfect genes to cells.ResultsAll the carriers were able to complex DNA. The highest, comparable to commercial reagents, transfection efficiency was observed for AP-15. Simultaneously, AP-15 exhibited the lowest negative impact on cell viability and proliferation—considerably lower than that of commercial agents. AP-15/DOPE complexes were also efficient to introduce pDNA to cells, without much effect on cell viability. Transfection with AP-15/DOPE complexes influenced the expression of a very few among 44 tested genes involved in cellular lipid metabolism. Furthermore, complexes containing AP-15 and therapeutic plasmid, encoding the TIMP metallopeptidase inhibitor 2 (TIMP2), introduced the TIMP2 gene with high efficiency to B16-F10 melanoma cells but not to B16-F10 melanoma tumors in C57BL/6 mice, as confirmed by TIMP2 protein level determination.ConclusionObtained results indicate that APs have a potential as non-viral vectors for cell transfection.
Highlights
Gene therapy, which implies genetic material to be introduced to the target cells as a therapeutic substance, is nowadays considered as a promising alternative approach to pharmaceutical treatment of many diseases
The complex formation was observed in the gel at approximate nitrogen to DNA phosphate (N/P) ratio r ! 2.7 for AP-8, r ! 3.0 for AP-7 and AP-11, r ! 3.7 for AP-15, whereas for PEI:DNA complex migration was retarded below ratio r 1.0
In this study we demonstrated that Amino-Prenols, semi-synthetic derivatives of polyprenols, may serve as non-viral carriers for gene delivery
Summary
Gene therapy, which implies genetic material to be introduced to the target cells as a therapeutic substance, is nowadays considered as a promising alternative approach to pharmaceutical treatment of many diseases. This approach has been successful in a limited number of clinical applications, e.g. adeno-associated virus (AAV1) delivery of a transgene encoding lipoprotein lipase (LPL) to patients suffering from LPL-deficiency (LPLD) [1]. Numerous concerns are raised related to the safety of their employment in patient treatment—triggering of acute inflammatory response as well as delayed humoral and cellular immune response, risk of insertional mutagenesis, etc These obstacles have led to the exploration of alternative methods of transfection [3]. An increasing number of clinical protocols describe applications of nonviral gene therapy formulations, which in comparison to viral vectors, are less immunogenic, capable of introducing genes of unlimited size, and cheap to produce on a large scale [4]
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