(1) Biomarkers in neurodegenerative and malignant processes (National project)

Focus is on amyotrophic lateral sclerosis and ischemia (MCAO). The project includes: (1) Measuring redox process in both in vivo (rat model) and tissues ex vivo by spin labels and spin traps; (2) Measuring the state of iron in brains and asses its role in production of ROS; (3) Measuring accumulation of USPIO particles into rat brain as a biomarker of the disease and its progression; (4) Measuring intactness of the BBB by using BBB permeable and BBB impermeable spin labels (in vivo. These measurements combine with studies already preformed (and future ones) using MRI on both animals and humans. We are investigating in vivo on ALS rats weather increased amount of free iron in ALS increases production of harmful free radicals through Fenton reaction and whether blood brain barrier (BBB) is breached in ALS.

(2) Evaluation of radioprotecting agents (Transnational project)

The project is realized in collaboration with Prof. G.Lubec, Medical University of Vienna. The project is based on extensive studies on potential radiopreotectors (survival, behavior, proteomics) aiming to determine to what extent free radical scavenging is included in radioprotection. We preform in vivo EPR to assess the ability of certain radioprotectors to scavenge radiation induced ROS.

(3) Elucidation of redox events during seed imbibitions and dependence of viability on different pretreatments (National project)

The project is relized in collaboration with Prof. Z.Giba, Institute of Biology, Univ. of Belgrade (FP7).

(4) Controlled release of drugs from implantable devices Collaboration with two groups from Univ. of Belgrade (National project)

The role of EPR Lab is to follow in vivo the release of drugs (spin labeled) from devices while MRI will be used for following the status of device.

(5) Studies of action of certain compounds (e.g. CeO2) on redox status of cultured tumor cells (National project)

Collaboration with two groups from Univ. of Belgrade.

(6) A detailed study of metal complexes and their role in redox processes in various biological systems (National project)

Focus is on V, Fe and Mn in fungus from which drugs for diabetes are developed.

(7) Scientific and technical collaboration contract with EmeraMed Ltd. (Transnational project)

Collaboration in research and development in the field in medical biophysics and pharmaceutics.

(8) The blood-barrier integrity and redox metals in the mouse model of Alzheimer’s disease (National project)

Collaboration with the Institute of University of Belgrade Institute for Biological Research “Siniša Stanković”, National Institute of Republic of Serbia.

(9) Electrochemistry and Electron Paramagnetic Resonance Spectroscopy of proteins and fatty acid nitroalkenes (Transnational project)

Collaboration with Department of Medical Chemistry and Biochemistry, The Palacký University, Czech Republic.

(10) New compounds as anticancer drugs (Transnational project)

Collaboration with University of Vienna, Institute of inorganic Chemistry, Austria

(11) Novel cardiovascular risk factors (Transnational project)

Collaboration with Professor Andreas Daiber, Universitäts-Professor für Molekulare Kardiologie, Medizinische Klinik und Poliklinik, Mainz, Germany. Interchange of several PhD and postdocs students and published a paper in Antioxidants 2020.

(12)  Novel nano-sized, biocompatible and stable free radical sensors for continuous in vivo hyperpolarization at ultra-low field MRI (Transnational HORIZON 2020 ERA Net RUS Plus Call 2017 – Science & Technology project)

Collaboration with Novosibirsk Institute of Organic Chemistry Of Siberian Branch of Russian Academy of Sciences, Laboratory of Nitrogen Compounds, Russian Federation.

The main goal of this project is preparation and detailed characterization of nano-sized and biocompatible free radicals that will be used as a contrast agent for continuous hyperpolarization of hydrogen nuclei at ultra-low field MRI. This research will be performed through four working packages (WPs) by a consortium consisting of four research groups with ample experience in diverse scientific disciplines. The initial step will be the preparation of small functionalized and long-lived free radicals or their precursors, which allow further synthetic modifications, i.e. coupling reactions with other functionalized molecules (WP1). The small-sized free radicals will be attached to different nano-sized biomolecular or biocompatible carriers such as the protein avidin, or dendrimers, polysiloxane-based nanoparticles and liposomes, respectively, which will further improve their biokinetic profiles and stability for in vivo utilization. Further modifications on the nano-sized carriers (incorporation of targeting vectors or other vehicle molecules) will be also performed in order to improve targeting properties of the resulting nano-sized free radicals and their accumulation in target tissue (WP2). Following the preparation of these bioconjugates, their thorough physicochemical and biophysical characterization will be performed (WP3-4), by investigating probe biocompatibility and potential toxicity towards the neurologically relevant cells in vitro, as well as biokinetic profiles in biological tissues ex vivo (WP3). The ability of the final nano-sized probes to induce continuous hyperopolarization will be performed in buffered media (phantom experiments) at the in-house made SQUID-based ULF MRI system; finally, the preliminary imaging experiments on living animal subjects (rodents) will be performed to validate the overall approach (WP4).

(13) Chemistry, biology and pharmacology of modulators of oxidative stress (Transnational joint research projects – project of particular relevance)

Collaboration with Sapienza University of Rome, Rome, Italy.

The proposed project is aimed at providing quantitative tools to thoroughly and comprehensively model and characterize new coumarin derivatives. Many naturally occuring, as well as synthesized compounds, influence the homeostasis, redox status, of the cell. Most studies on these compounds have been, so far, focused on their antioxidant activity, but not pro-oxidant activity. Since cancer cells produce high levels of ROS, which lead to increased basal oxidative stress, pro-oxidant properties of new coumarine derivatives will be addressed as well, along with anticancer and antimicrobial activity. Designing new coumarin ligands that will interact with free, positively charged, metal ions will also be one of the objectives. The effect of these compounds on redox-sensitive places within the cells and the effects on migratory and invasive capacity of cells will be also looked into. Tests will be carried out on different cancer cell lines (colorectal cancer, breast cancer, choriocarcinoma, skin cancer, lung cancer,…). Also, the assessment of their abilities to selectively kill cancer cells, but avoid/minimise effects on normal cells in the body (to limit toxic side-effects) will be investigated.

(14) NADPH oxidases in the progression of obesity – induced forms of diabetes (Transnational project)

MPNTR (Serbia) and Deutscher Akademischer Austauschdienst – DAAD bilateral project between BioScope Labs Consortium at Faculty of Physical Chemistry, University of Belgrade and Experimental and Molecular Pediatric Cardiology Department at the German Heart Center Munich.

Obesity is a rapidly growing threat to public health since it promotes the development of type 2 diabetes mellitus(T2DM), metabolic syndrome, cardiovascular diseases and even cancer. The increasing proportion of young obese subjects in the reproductive age poses an additional severe risk on pregnancy by promoting gestational diabetes and preeclampsia and various complications in the perinatal period. Moreover, the offspring of obese pregnancies is on a highly elevated risk not only to encounter problems in organ development during pregnancy but also to develop negative consequences for the post-natal health status. In fact, epidemiological data show a dramatically increased risk to develop metabolic and cardiovascular diseases in the offspring of obese pregnancies. However, the mechanisms imposing the adverse effects of obesity in particular on mother and offspring, but also on non-pregnant subjects are not well understood. Obesity and T2DM have been associated with increased load of reactive oxygen species (ROS). While there is limited evidence that ROS may promote obesity and T2DM complications in non-pregnant subjects, there are hardly any data related to the effects of ROS on the pregnant obese mother and their offspring. In particular, it remains unclear which organs suffer from increased ROS load under obesity/T2DM conditions and how this will be affected by pregnancy. Moreover, there are almost no data on the role of ROS in the placenta, whether there are indeed elevated ROS levels in the fetus in utero, and if so, whether increased ROS load is either transferred via maternal blood or generated by the fetus itself. The answers to these questions are, however, highly warranted to the further understanding of the role of obesity in pregnancy and the development of preventive and/or therapeutic strategies.

In this collaborative project, teams from Serbia and Germany will develop an approach to monitor ROS load in living (pregnant) animals and will for the first time determine ROS levels in the fetus in utero using models of obesity-induced T2DM and gestational diabetes. They will further analyse the role of ROS-generating NADPH oxidases in these pathologies as promising novel therapeutic targets against the detrimental consequences of the obesity epidemic.

(15) Spin probing of albumin from different mammalian species using continuous-wave and pulse EPR

Collaboration with Professor Dariush Hinderberger, Martin-Luther-University, Halle-Wittenberg, Germany. Postdoc research of Dr Aleksandra Pavićević.

(16) Graphene quantum dot antioxidant and proautophagic actions protect SH-SY5Y neuroblastoma cells from oxidative stress-mediated apoptotic death

Collaboration with Vladimir Trajković, Institute of Microbiology and Immunology, Faculty of Medicine, University of Belgrade

(17) Bioevaluation of glucose-modified liposomes as a potential drug delivery system for cancer treatment using 177-Lu radiotracking

Collaboration with Sanja Vranješ-Đurić” group at Vinča Institute of Nuclear Sciences”

(18) New EPR probes to monitor redox homeostasis deregulation

Collaboration with Dr. Fabienne Peyrot, French research unit/university: LCBPT UMR8601 CNRS/Université Paris Descartes, France

(19) Utilization of the food industry waste for improving honey bee health and protecting the environment (Waste2ProtectBees) (2024-2027) 

Project “PRIZMA”: Cooperation with the Faculty of Veterinary Medicine – University of Belgrade and the Institute of Forestry Belgrade, Serbia.

Budget: 300 kEUR.

Colony losses, high incidence of diseases, ineffective disease control and food shortage, have become part of beekeeper’s everyday life. Malnutrition leads to weakening of immunity and makes bees more susceptible to bee pathogens (especially Nosema ceranae and honey bee viruses) which often induces colony death. On the other side, food industry side products (grape pomace after wine production and remains of mushroom production) could present environmental risk when disposed in the nature (that usually happens). To solve these problems, this proposal has the aim to develop functional food (extracts made from food industry waste through environmentally friendly methodology) intended to reduce nutritional stress in bee colonies, enhance bee immunity and combat pathogens. The produced extracts will be tested by using the novel concept: “from molecule to hive”. This concept implies the analytic principle which will consist of physicochemical analysis of obtained extracts, extracts’ testing on cell cultures, cage bees, in the Comet assay and on whole bee colonies. It is important to underline that: (i) in vitro and in vivo EPR spectroscopy and imaging analyses of honey bees; (ii) extracts’ testing on continuous honey bee cell line and (iii) launching a freely available database regarding honey bee genome studies (obtained by third-generation RNA sequencing and isoform analyses) present a completely new approach which has never been used before in bee research. Beside the major impact of this Project (development of new extracts which would reduce bee losses and increase beekeeper income), as an indirect and long-term impact, this project creates a potential for bio-briquette production from biomass left behind the process of extraction. The utilization of winemaking and mushroom industry side products will contribute to decrease of environmental pollution and promote environmental sustainability.

(20) Role of macroautophagy in lipid nanoparticle mRNA delivery and adjuvanticity (REDIRECT) (2024-2026)

Project: “PROMIS”: Cooperation with the Faculty of Medicine, University of Belgrade, and the Institute of Virology, Vaccines and Sera “Torlak”.

Budget: 150 kEUR

Messenger RNA (mRNA) vaccines are a promising approach for the prevention of infectious diseases, but the mRNA delivery to the cell cytosol is very low. The most successful mRNA platform comprises non-immunogenic nucleoside-modified mRNA assembled within lipid nanoparticles (LNP-mRNA), which enter the cells via endocytosis, acting both as mRNA carriers and adjuvants. Macroautophagy, an intracellular autodigestion process controlling cell homeostasis and inflammation, is tightly interconnected with the endocytic pathway, thus possibly redirecting the cytoplasmic trafficking of endocytosed material, including LNP-mRNA. REDIRECT will be the first multidisciplinary project to systematically explore the role of autophagy in LNP-mRNA intracellular delivery and adjuvanticity. GFP-SARS-CoV-2 Spike mRNA will be synthesized in vitro from linearized DNA and encapsulated into four-lipid LNP using the thin-film hydration method. LNP-mRNA complexes will be characterized by dynamic light scattering and EPR spin labeling. Intracellular trafficking of flfluorescent/immunogold-labeled LNP-mRNA will be assessed by confocal/transmission electron microscopy. Cytosolic delivery/translation of mRNA will be analyzed by flow cytometry (GFP) or RT-qPCR/immunoblot (Spike). Adjuvanticity of LNP-mRNA will be determined by ELISA and immunoblot analysis of proinflammatory cytokines and inflammasome activation, respectively. Autophagy will be evaluated by immunoblot of autophagy markers, and its role in LNP-mRNA performance will be assessed in human monocytic cell lines with genetically impaired autophagy and cells treated with pharmacological modulators of autophagy. The results of the project could lay grounds for LNP-mRNA platform improvement through autophagy modulation. We expect to show a significant impact of autophagy on LNP-mRNA delivery and adjuvant performance.