Research M. Pavlin involves development of new methods and applications in field of biomedicine and biotechnology, specifically delivery by nanoparticle vectors or by electric pulses. She was or is PI of several interdisciplinary research projects. Her papers were cited over 600 times (over 400 pure citations). She was also participating in 5.FP project where Cliniporator device (in collaboration with different European partners) was developed and also resulted in an international patent. Her papers are cited in several research areas. She was directly involved and lead implementation and development of several in vitro protocols of EGT in vitro, from develoment of 3D in vitro models of cells embedded in collagen gels (Haberl and Pavlin, J.Memb.Biol 2010) to protocols of gene silencing and in vitro and protocols for analysis of nanoparticles internalisation and cytotoxicity. In 2007 she received Luigi Glavani prize of International Bioelectrochemistry Socieity for young investigaters. In 2010 she established interdisciplinary young research group that includes researchers with diverse background (physicists, biologists and electrical engineers) as well as undergraduate and graduate students. This gives the students a chance to participate in active research and use state-of-the-art methods of biotechnology and biomedicine. Exiting network of collaborating groups ensures also close contact with clinicians from different fields. Young researcher Jasna Lojk (PhD mentor M. Pavlin) and the group received in last year two best poster awards in international conferences 7th Conference on Experimental and Translational Oncology - CETO 2013 and 9th International Conference on Nanosciences & Nanotechnologies (NN12) based on research and development of new nanoparticles for biomedical applications. Related PhD and diploma thesis HABERL MEGLIČ, Saša. Analiza vpliva različnih parametrov na učinkovitost genske elektrotransfekcije v celičnih kulturah in v in vitro modelu tkiva = Analysis of the influence of various parameters on gene electrotransfer efficiency in cell cultures and in in vitro tissue model : doktorska disertacija. Ljubljana: [S. Haberl], 2011. X, 174 str., ilustr. [COBISS.SI-ID 2987633] STRAŽIŠAR, Maruša. Analiza interakcije DNA s celično membrano ter učinkovitost vnosa DNA v celice CHO z elektrotransfekcijo pri nizkih koncentratih plamida : diplomsko delo. Ljubljana: [M. Stražišar], 2011. IX, 69 f., ilustr. [COBISS.SI-ID 35337989] KOTNIK, Nejc. Primerjava lipofekcije in elektroporacije pri vnosu plazmida pEGFP-N1 v človeške mioblaste in vitro = Comparison of lipofection and electroporation for transfer of plasmid pEGFP-N1 into human myoblasts in vitro : diplomska naloga. Ljubljana: [N. Kotnik], 2011. VIII, 45 f., ilustr. [COBISS.SI-ID 2947953] mentor: prof.dr. Zoran Grubič
D.08 Management and development of research activities
COBISS.SI-ID: 35337989ANG We have developed (PI lead the research team) a first 3D numerical model for optimization of gene electrotransfer in vivo. It is the first study that presents optimization of electric parameters and electrode positions for gene electrotransfer in skeletal muscle. Furthermore, our numerical model is generic and can be applied to other tissues. The results can be used as a guideline for researchers in selecting optimal parameters for in vivo studies of electro-gene therapy and electro-DNA vaccination and could advance translation into clinical environment. Based on this we had established a collaboration with the leading research medical center (Herlev Hospital, Copenhagen, head prof. Julie Gehl) in Europe for translation and application of electrogene transfer for cancer therapy. During our stay in this group we have adapted our models for their specifi configuration and performed parametric analysis of electrodes positions, depth of insertion and applied voltages, the parameters were based on animal pig study and clinical study of gene electrotransfer in muscle tissue of this group. Related publication: ČOROVIĆ, Selma, ŽUPANIČ, Anže, MIKLAVČIČ, Damijan, PAVLIN, Mojca. Numerical optimization of electric field and electrode configuration for gene electrotransfer into muscle tissue. V: SERŠA, Gregor (ur.), KOS, Janko (ur.), LAH TURNŠEK, Tamara (ur.), ČEMAŽAR, Maja (ur.), FILIPIČ, Metka (ur.), KRANJC, Simona (ur.), MARKELC, Boštjan (ur.). 7th Conference on Experimental and Translational Oncology, Portorož, Slovenia, April, 20-24, 2013. Book of abstracts. Ljubljana: Association of Radiology and Oncology, 2013, str. 114. [COBISS.SI-ID 9805396]
F.01 Acquisition of new practical knowledge, information and skills
COBISS.SI-ID: 10953300In collaboration with the Institute for pathophysiology (Faculty of medicine, University of Ljubljana) we developed the electroporation protocol and analyzed the mechanisms of introduction of short RNAs into cultured primary human myoblast cells. The protocols were optimized using siRNA against HIF-1α (Hypoxia Inducible factor 1α) mRNA. HIF is a transcription factor and key regulator of cellular oxygen homeostasis and has an important role in several pathologies. We optimized the electroporation medium and the parameters of electric pulses, which enabled us to achieve 80% silencing of HIF-1α mRNA. In parallel, we performed viability studies for the same electroporation conditions. Suppression of HIF-1α expression was first estimated by measuring HIF-1α mRNA with qPCR. Depletion of HIF-1α protein was subsequently confirmed with Western Blot. Due to involvement of HIF-1α in several muscle pathologies, the effect of HIF-1α silencing was confirmed also on two HIF-1α’s downstream gene targets; PGK (phosphoglycerate kinase) and VEGF (vascular endothelial growth factor). Manuscript of this research for the publication in an international journal is in preparation. The developed protocol will not only serve as a mean to study the mechanisms of introduction of gene material, but also for therapeutic purposes. HIF, PDK and VEGF (involved in angiogenesis) are involved in cellular metabolism and were identified as targets for cancer treatment. The research on primary human myoblast cells is performed in collaboration with the Laboratory for molecular neurobiology with the purpose of using electroporation as a method for introduction or silencing of therapeutic genes as a mean to treat specific disorders. Based on this work we were also invited to join the network of groups preparing COST proposal BIONECA.
F.18 Transfer of new know-how to direct users (seminars, fora, conferences)
COBISS.SI-ID: 9892180PI is based on accuired knowledge during previous project "Mechanisms of DNA delivery with electrogene transfer" and current project connected understanding of mechanisms with theoretical analysis of DNA mobility. Both project togehter connect exprimental work on standard in vitro models with experiments on ex vivo and 3D gel models and finally with 3D numerical models of gene electrotransfer of mucle tissue. This interdisciplinary approach and series of developed methodological skills enabled generation of new scientific knowledge and practical skills that can be now applied in different applications in collaborations with different groups froms Slovenia and abroad.
D.01 Chairing over/coordinating (international and national) projects
COBISS.SI-ID: 9632596Electroporation is a frequently used method for increasing permeability of the cell membrane, but can also cause damage to the cell: cytosol leakage, ROS formation, osmotic swelling, necrosis and induction of apoptosis. Cell stress and cell damage can also trigger inflammasome activation. However potential immune system was not analysed in many papers of electroporation. In this paper we analysed weather electroporation of macrophages in vitro can trigger NLRP3 inflammasome activation and subsequent secretion of IL-1β. Cells were exposed to electric pulses and quantity of secreted IL-1β was determined. Our results show that the observed IL-1β secretion was not NLRP3 inflammasome dependent but indicate that nevertheless, electroporation triggers a proinflammatory immune response through IL-1β secretion.
F.02 Acquisition of new scientific knowledge
COBISS.SI-ID: 10780500