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Slovak Research and Development Agency APVV:

Hypericin: biotechnology, signalome, photodynamic therapy
Program: APVV-0040-10
Principal Investigator: Prof. RNDr. Peter Fedoročko, PhD.
Duration: 01.05. 2011 - 31.10. 2014
Co-operation: Faculty of Science UK, Bratislava
Cancer Research Institute SAS

About the project
Project is aimed at complex investigation of the representatives of the genus Hypericum from the view point of genetic aspects of biosynthesis of bioactive compounds with anticancer effects, especially hypericin with perspective design of biotechnological alternative of production. We will concentrate on the study of the role of candidate genes in proposed biosynthetic pathway of hypericin using standard and transgenic experimental systems. Following stages will be focused on determination of possible genotoxic effect of photoactivated hypericin and its precursor emodin with use of prokaryotic and eukaryotic model systems. The second goal will focus on exploitation of hypericin as a photosensitizer in photodynamic therapy (PDT). Since a resistance of tumour cells may be related to low bio-availability of the drug as well as to suppressed sensitivity to stress or deregulated cell death programming, we will be concerned with key molecular pathways and transporting systems involved with determination of tumour cell response to PDT considering cell death and survival/resistance. We will endeavour to find complex solutions for PDT efficiency and onset of resistance problems with effort to prevention and suppression of negative effects using targeted pharmacological intervention.

Modulation of cell signal pathways in targed cancer therapy
Program: LPP-0062-09
Principal Investigator: Prof. RNDr. Peter Fedoročko, PhD.
Duration: 01.09. 2009 - 31.08. 2013

About the project
There have been several studies demonstrating that functional erythropoietin receptors are found on tumor cells and these findings provide evidence that erythropoietin can directly stimulate the survival, growth and proliferation of malignant cells. The presence of erythropoietin and erythropoietin receptor more significant in tumor region associated with hypoxia were seen. Erythropoietin up-regulation by tumor hypoxia may lead to increased angiogenesis both directly, via Epo's mitogenic effect on endothelial cells in concert with the hypoxic up-regulation of other angiogenic cytokines. The purpose of our project is to find out the role of endogenous Epo in angiogenesis of human ovarian adenocarcinoma cells in vitro. The role of caspases on antitumor effect of hypericin mediated photodynamic therapy is well known. However, caspase-independent pathways may be simultaneously involved with caspases in the apoptosis induced by photodynamic therapy. Many authors view apoptosis as a process of caspase activation. During the last years, there is a growing evidence for the therapeutic potential in cancer treatment, though the existence of a caspase-independent pathways does not fit the dominant scheme. The precise mechanism of hypericin induced apoptosis associated with caspase-independent pathways, sphingomyelinases or ceramide remains unclear. Therefore we will focus on characterization of apoptotic signaling as response of cancer cell to photodynamic stress from this aspect.

Scientific Grant Agency of the Ministry of Education, science, research and sport of the Slovak Republic
and the Slovak Academy of Sciences (VEGA):

Characterization of selected parameters of the exogenous S-phase markers in in vivo and in vitro conditions.
Program: VEGA 1/0322/11
Principal Investigator: RNDr. Juraj Ševc, PhD.
Duration: 01.01. 2011 - 31.12. 2013

About the project
5-bromo-2-deoxyuridine (BrdU) is synthetically derived analogue of thymidine and is probably the most used marker of proliferation in neurobiological, developmental and regenerative studies. However, the use of BrdU involves some critical issues, because BrdU can affect the proliferation rate of the examined cell population and the interpretation of obtained results. On the other hand, 5-ethynyl-2-deoxyuridine (EdU) is the novel thymidine analogue and was not very common in proliferative studies until now. In addition, there are serious assumptions that use of EdU is easier, faster and the processioning of the examined specimen is gentler in comparison with BrdU. The main goal of the presented project is the characterization and comparison of selected features of BrdU and EdU to provide an optimal and effective tool for identification of proliferating cells or for labeling the distinct cell populations at the time of their birth to examine their fate in the long term studies

Typification of human erythropoietin receptor in cancer and normal cells
Program: VEGA 1/0733/12
Principal Investigator: Assoc. Prof. Peter Solár, PhD.
Duration: 01.01. 2012-31.12. 2014

About the project
Many studies have demonstrated biological effects of erythropoietin in the cells of nonhematopoetic system and moreover some clinical studies have revealed unexpected adverse effects of such recombinant protein in cancer patients. Nevertheless, there are still not enough information allow us to characterize the molecular structure of erythropoietin receptor in cancer as well as in normal human cells. Therefore, the purpose of our study is to determine exact locality of erythropoietin receptor, analyze its function and describe using MALDI-TOF/MS sequencing erythropoietin receptor isotypes in human ovarian adenocarcinoma cells as well as in HUVEC endothelial cells.

Non-caspase signaling pathways in cancer cell programmed cell death and impact of ABC transporters on cell resistance
formation after hypericin´s photocytotoxic
Program:VEGA 1/0626/11
Principal Investigator: Prof. RNDr. Peter Fedoročko, PhD.
Duration: 01.01. 2011 3 - 31.12. 2014

About the project
The therapeutic potential of various antitumor modalities (chemo-, immuno-, photodynamic-therapy) is based on the induction of programmed cell death. Understanding of molecular mechanisms associated with this form of cell death and application of knowledge about the regulation of signaling pathways provide an opportunity to successful therapy and to development of new drugs.

The efficiency of antitumor therapy is negatively influenced by activation of the ABC transporters followed by multidrug resistance (MDR) formation which is a major reason for the failure of several chemotherapeutics in the treatment of cancers. Based on our recent results photodynamic therapy (PDT) may be negatively affected by the possibility that hypericin might be a substrate of some ABC transporters. The goal of this research project is to understand the molecular mechanisms of cell death induced by hypericin mediated PDT with respect to the ABC transporter systems that play a critical role in the effective therapeutic outcome.

Examination of proliferative and transformative processes of cells, their migration and reciprocal interaction in the spinal cord
of rat during perinatal period of life.
Program: VEGA 1/0967/12
Principal Investigator: Assoc. Prof. RNDr. Zuzana Daxnerová, PhD.
Duration: 01.01. 2012 - 31.12. 2015

About the project
Despite extensive mitotic activity in the VZ, which indicates the presence of postnatal gliogenesis, these processes weren’t described until now and the fate and function of newly-formed cells remains unknown. But despite the presence of numerous turbidities especially during the late embryonic and early postnatal gliogenesis in the mammalian spinal cord, is the development of dorsal root ganglia and also the whole peripheral nervous system much less described and germinal population of the Schwan or satellite glial cells was not definitely determined until now. The aim of the project is detailed examination of the distinct types of glial cells, which are produced in the lining of the central canal during perinatal period of life and exhaustive determination of the migratory potential of these cells not only in the spinal cord area, but also in the surrounding structures like pia mater, dorsal root ganglion and peripheral nervous system in general.

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