Projects / Programmes
Uporaba elektroporacij v genski terapiji (Slovene)
| Code |
Science |
Field |
Subfield |
| 3.04.00 |
Medical sciences |
Oncology |
|
| Code |
Science |
Field |
| B200 |
Biomedical sciences |
Cytology, oncology, cancerology |
electroporation, gene therapy, electrochemotherapy, cisplatin, p53, beta-galactosidase, green fluorescent protein
Researchers (17)
Organisations (1)
| no. |
Code |
Research organisation |
City |
Registration number |
No. of publicationsNo. of publications |
| 1. |
0302 |
Institute of Oncology Ljubljana |
Ljubljana |
5055733000 |
15,455 |
Abstract
Gene therapy is a rapidly evolving approach for the treatment of cancer. One of the major obstacles of this approach is how to obtain safe and efficient transfection of target cells. Several gene delivery methods are currently employed, such as retroviruses, adenoviruses, liposomes, cationic lipids, direct injection, bombarding cells with particles coated with DNA and electroporation.
Electroporation is a technique widely used for introduction of different types of exogenous membrane-impermeable molecules into the cells in vitro or in vivo. A single electric pulse or a train of electric pulses in the range of kV/cm and of several microseconds length causes a reversible permeabilization of cell membrane which forms the basis of this technique. Electroporation is widely used in many laboratories for in vitro transfection due to its reproducibility and efficiency compared to other transfection methods.
Recently electroporation was successfully applied for electrochemotherapy, a treatment of tumors where electroporation is used as a drug delivery system for chemotherapeutic drugs to increase their antitumor effectiveness. Two chemotherapeutic drug, bleomycin and cisplatin proved to be effective in vitro, in vivo and in clinical trials. Objective response rates ranging from 72 percentage to 100 percentage were reported in the clinical trials.
Several mechanism, besides increased drug accumulation in the cells due to the electroporation, are involved in the antitumor effectiveness of electrochemotherapy, such as immune system of the organism, changes in tumor blood flow, whereas vascular targeting was only speculated.
Recent reports demonstrated that electroporation could be also used for in vivo gene delivery. The groups studying this method used different settings of electric pulses and different animal models. The results of the studies suggest that a single specific protocol for in vivo gene delivery by electroporation is not suitable for transfection of all tissue types. Therefore, further studies are needed to optimize electrically mediated gene delivery, such as route of plasmid administration and electrical parameters for tissue electroporation. This will lead to better understanding of mechanisms of DNA transport into the cells and will also form the basis for translation of this therapy into clinical practice.
Therefore, the proposed project deals with the two areas of usage of electric pulses in vivo. First, to further elucidate mechanisms of antitumor effectiveness of electrochemotherapy by studying the effects on endothelial cells and second, to determine the optimal electrical parameters for efficient gene delivery into tumor and endothelial cells in vivo. Based on these results, electroporation in combination with wild type p53 gene, which is one of the most important genes for regulating apoptosis, will be tested on different tumor models in vivo. The final goal is to combine electrochemotherapy with cisplatin with p53 gene delivery to increase its antitumor effectiveness. The rationale for combining cisplatin therapy with wild type p53 gene therapy is in potentiation of antitumor effectiveness of cisplatin by increased apoptosis due to the presence of wild type p53.
