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Uporaba elektroporacije v medicini: elektrokemoterapija in elektrogenska terapija
Published on Jun 26, 20135481 Views
Elektroporacija je fizikalna metoda, pri kateri z izpostavitvijo celic električnim pulzom ustreznih parametrov dosežemo prehodno povečano prepustnost celičnih membran. Povečano prepustnost membrane do
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Chapter list
Uporaba elektroporacije v medicini: elektrokemoterapija in elektrogenska terapija 00:00
History02:27
Institute of Oncology Ljubljana02:54
Main research areas – translational research03:56
What is translational oncology?04:58
Transplantable tumor models and human tumor xenografts05:38
Reversible electroporation - principle06:42
Reversible electroporation 08:24
Electro Chemo Therapy: ECT09:41
Electrochemotherapy:Increased cytotoxicity of chemotherapeutic drugs10:23
Preclinical data on electrochemotherapy11:27
Electroporation increasaes cytotoxicity of chemotherapeutic drugs on different tumour cell lines12:08
Protocol of electrochemotherapy 13:08
Complete responses of histologically different tumor types13:42
Antitumor Effectiveness15:01
Time dependence of electrochemotherapy with cisplatin injected intratumorally15:21
Mechanisms associated with antitumour effectiveness of electrochemotherapy15:44
Antitumor effectiveness of electrochemotherapy 16:14
Electroporation increases the amount of platinum bound to DNA after electrochemotherapy with cisplatin16:43
Vascularization of the tumours and effect of applied electric pulses17:11
Application of electric pulses to the tumours induces transient reduction in tumour blood flow17:49
Electroporation of tumours induces drug entrapment by vascular lock in the tumours18:04
Anti-vascular effect of electrochemotherapy with bleomycin and cisplatin18:37
Cytoskeletal F-actin changes in EP and ECT-treated HMEC-1 monolayers19:18
Cytoskeletal b-tubulin changes in EP and ECT-treated HMEC-1 monolayers20:47
Disruption of F-actin and VE-cadherin junctions in endothelial cells after EP21:04
HMEC-1 monolayer permeability changes occur quicker and to higher degree after ECT compared to EP 21:19
Proposed mechanism of EP-induced changes in tumor blood vessels – vascular lock and increased permeability of tumor blood vessels21:46
Vascular disrupting effect of electrochemotherapy at chemotheraputic drug doses that cause tumor destruction23:04
Intravital microscopy23:24
Application of EP to tumor causes increased permeability of tumor blood vessels in vivo24:25
Application of EP to tumor causes „vascular lock“ that lasts ~ 10 mins, followed by partial restoration of tumor blood flow24:56
Proposed mechanism of EP-induced changes in tumor blood vessels – vascular lock and increased peremability of tumor blood vessels25:29
ECT induce vascular lock and completely abrogates tumor blood flow; increased permeability of blood vessels26:16
EP and ECT do not affect normal blood vessels surrounding tumor26:48
Limb sparing treatment27:18
Vascular disrupting effect of electrochemotherapy27:56
Clinical application of electrochemotherapy in human medicine28:26
Electrochemotherapy - procedure28:31
Fifth framework research programme 28:59
Electric pulse generator29:31
Possible ways of performing ECT29:42
Electrochemotherapy proved to be effective in treatment of various cutaneous tumor nodules:30:05
Electrochemotherapy with bleomycin30:55
Malignant melanoma ECT with CDDP i.t., plate electrodes, single application31:11
Malignant melanoma: ECT with BLM i.v., hexagonal electrodes, single application31:13
Anal malignat melanoma32:20
Breast cancer32:44
Response rate of the patients treated before and in ESOPE study33:07
Perspectives33:50
Metastases in the Liver34:11
ECT centres in Europe -201234:53
Canine mast cell tumours35:27
Electrochemotherapy with cisplatin of squamous cell carcinoma in cat36:36
Cat SCC: Queeny36:52
Electrochemotherapy with cisplatin of sarcoid tumors in horses37:01
Sarcoid on the head treatedwith ECT with cisplatin. CR was obtained37:29
Gene electrotransfer37:34
Strategies of cancer gene therapy37:46
Gene electrotransfer: principle38:09
Optimization of gene electrotransfer 38:27
Effective gene transfer38:35
Sequence and time dependence38:53
Correlation39:14
Skeletal muscle39:40
Gene electrotransfer for treatment of cancer40:09
Translational gene therapy studies41:12
Interleukin-12 antitumor effect41:31
Concentration of mIL-12 and mIFN-g in serum of mice is dependent on the amount of plasmid DNA and number of intramuscular transfections41:59
Gene electrotransfer42:46
mIL-12 gene intramuscular electrotransfer has anti-metastatic effect on induced lung metastases43:08
Local and systemic antitumor effect43:38
Gene electrotransfer induces local and systemic release of IL-12 and IFN-γ and has antitumor effect on distant untreated tumors43:54
IL-12 gene therapy combined with tumor irradiation - radiosensitization44:23
Radiotherapy in murine sarcoma44:40
Intramuscular mIL-12 gene electrotransfer has radiosensitizing effect on solid subcutaneous tumors45:13
mIL-12 i.m. gene electrotransfer has radiosensitizing effect on induced lung metastases45:19
Endoglin (CD105)45:25
In vitro46:06
In vivo46:36
Construction of plasmid DNA encoding shRNA against endoglin from the sequence of the most effective siRNA46:52
Clinical research - EGT with IL-12 in dogs47:19
Comparison between ECT and EGT47:38
Treatment response: ECT + EGT48:43
Safety of gene therapy– preliminary results 49:06
Preliminary results – post-therapy evaluation of presence of plasmid DNA and horizontal transfer49:30
Electrogene therapy in treatment of cancer patients50:07
Gene electrotransfer to melanoma skin nodule50:25
Gene electrotransfer clinical trial with therapeutic plasmid AMEP 50:33
Patient 001-003; Treatment: IT Plasmid AMEP™ 2mg50:47
Electrotransfer of therapeutic molecules into tissues51:16
Perspectives51:54
Transfection efficacy of reporter plasmid DNA with promoter specific for skin cells52:11
Tumor irradiation combined with IL-12 gene therapy controlled by p21 radio-inducible promoter 52:24
Radio-inducible expression of IL-12 resulted in equal antitumor effectiveness compared to constitutively expressed IL-1252:34
Applications of electroporation-based technologies52:46
Acknowledgement53:27
Picture of group53:49