Research – Department of Life Sciences

Mesenchymal stem cells (MSCs) are a key player in the field of regenerative medicine due to their numerous regenerative properties. For example, their ability to selectively home into injury/inflammation areas and their immunomodulatory effects allows the treatment of autoimmune and inflammatory diseases.

For this reason, we are planning to establish new, meaningful biological assay systems in this project that will allow to describe the underlying mechanisms that occur in stem cell therapies. Therefore, we will engineer highly standardized MSC cell lines using optogenetic manipulation, in which specific signaling pathways can be switched on and off by light. After characterization and validation under physiologic conditions, an automation of the models using the Oli-Up cell culture systems from LifeTaq-Analytics GmbH is planned.

The project is carried out in cooperation with BOKU Vienna and LifeTaq-Analytics GmbH and funded by the Austrian Research Promotion Agency under the BRIDGE programme.

The purpose of this project is to investigate the pathophysiological role of the endosteal niche, and in particular the role of osteoblast precursors in the development of breast cancer bone metastases. The objectives of the research include identifying new biomarkers that predict the potential of tumour cells to form bone metastases. A further aim is to identify substances which prevent metastases by modifying the endosteal niche. Based on initial work using a 2D co-culture system, investigations will now also be conducted in a 3D system. The project is being carried out in close cooperation with Karl Landsteiner University of Health Sciences.

Sepsis is a life-threatening organ dysfunction resulting from severe systemic inflammation caused by an infection. Despite many improved therapeutic measures, the mortality rate from sepsis is still 15-25%, and for septic shock 30-50% (Hotchkiss et al., 2016). Endothelial cells, which line the inner surface of blood vessels and capillary beds, play a central role in this life-threatening condition. In the early stages of sepsis, microbial factors such as LPS, which binds to Toll-like receptors, are responsible for directly activating endothelial cells. Endogenous agonists – released by activated leukocytes, endothelial cells or damaged cells for instance – subsequently promote the endothelial dysfunction. In this project we plan to develop new, highly-standardised cell models in which the specific signalling pathways of the Toll-like receptors can be switched on or off by means of light induction. From the light-activatable cell lines obtained in this way, we will establish physiologically relevant models of sepsis which can then be used to investigate intracellular signalling pathways and expression of different sepsis-inducing factors, microvascular endothelial permeability, and potential therapeutic agents. The objective is to mimic sepsis-inducing inflammation by switching the signalling pathways on or off (in the absence of bacterial factors) optogenetically. The project is being carried out in cooperation with Danube University Krems and UK St. Pölten.

As the incidence of osteoarthritis increased over the last years, development of new therapeutic approaches for osteochondral regeneration come into the focus of tissue engineering research.

The focus of this project lies in evaluating the therapeutic potential of extracellular vesicles isolated from mesenchymal stem cells derived from the Hoffa´s fat pad on processes occurring during osteoarthritis. The Hoffa´s fat pad is located intra-articularly behind and under the patellar. Due to its proximity to the cartilage, mesenchymal stem cells from this adipose tissue are more involved in the development and progression of osteoarthritis as mesenchymal stem cells derived from bone marrow which are often used for cartilage regeneration studies.

The majority of the project will be conducted at the Danube University Krems, with the IMC University of applied Sciences Krems taking on the proteomic analysis of the extracellular vesicles by mass spectrometry. Further project partners are the Medical University of Vienna and OrthoSera GmbH. The project is funded within the Life Science Call 2018 by the NÖ Forschungs- und Bildungsges.m.b.H (NFB).

Rare earths are used in electronic devices such as mobile phones, computers and energy-saving bulbs. However, they are scarce and cannot be recycled using eco-friendly methods. Complex and expensive mining, coupled with scarce supply, means that the prices of rare earths on the world market are rising steadily. Due to continuous technical advances, we can already predict that the supply situation for rare earths will become critical in future, which in turn could pose a threat to the development of innovative technologies.

The project partners aim to counter this trend using a new technology. This involves an approach that has never been used before: recycling by means of microorganisms (bacteria and algae). The goal of the international project partner consortium is to develop a practicable recycling technology in collaboration with regional industry, with a view to reclaiming rare earths from electronic waste and subsequently making the technology available to businesses. The consortium liaises regularly with its strategic partners, which guarantees that market needs and the technological limitations of business are taken into account in the development process.

The project is being funded as part of the EU’s INTERREG V-A Austria-Czech Republic programme.

www.at-cz.eu/reegain

During the course of this project, an alternative synthesis of hydroxytyrosol will be developed. This new route aims for a higher yield of the important natural product by combining the advantages of biotechnological procedures and modern chemical reactions. Additionally, in collaboration with our industrial partner, a new and highly promising application of hydroxytyrosol in textile industry will be evaluated.

Summarizing, the following goals can be defined:

• Establish whole cell oxidation (dihydroxylation of aromatics) as innovative and future-oriented methodology at the University of Applied Sciences Krems. By means of this environmentally benign protocol, fine chemicals and pharmaceuticals are accessible from organic waste.

• Synthesis of hydroxytyrosol, an important natural antioxidant with a wide range of applications in food industry, cosmetic industry, and pharmaceutical industry.

• Evaluation of a newly developed analytical instrument to measure the metabolic activity of various bacteria.

• Development of a new method for the mild and gentle selective bleaching of denim and related textiles.

The project is co-financed by the Province of Lower Austria and the European Regional Development Fund (ERDF).

Patients with an impaired immune system, such as HIV- positive patients and recipients of solid organ and particularly hematopoietic stem cell transplants, are at high risk of life-threatening infections with human adenoviruses. Among stem cell transplant recipients with systemic infections, mortality rates of almost 80% have been reported. The efficacy of commonly used drugs to treat adenovirus infections is limited and frequently associated with toxicity. Alternative drugs are still under investigation. In light of the fact that numbers of solid organ and hematopoietic stem cell transplant recipients are constantly rising, there is a pressing need for alternative treatment options.

Short interfering RNAs (siRNAs) and artificial microRNAs (amiRNAs) are a class of artificial small RNAs that can bring about the inactivation of cellular and viral genes via the RNA interference (RNAi) pathway. In a previous project led by the investigators, highly potent siRNAs and amiRNAs with activity against components of the adenoviral DNA replication machinery that can effectively inhibit the replication of human adenoviruses in cell culture experiments were developed and characterised. The project is aimed at investigating if adenovirus infections can be inhibited by these RNAi-triggering small RNAs in vivo.

The project is funded by the Austrian Science Fund (FWF).

The world-class standard of medical care in Austria means that many patients are now surviving in areas where previously treatments did not exist or were not widely available. This is thanks in no small part to developments in intensive care and transplantation. But one consequence, in particular of the advances in transplantation, has been a steep rise in the number of immune-deficient patients with a considerably increased risk of otherwise harmless infectious diseases, such as infections with adenoviruses. These types of infections can prove fatal for people in this patient group. While bacterial infections can for the most part be effectively controlled using antibiotics, treatment options for viral infections are unsatisfactory, and the persistently high mortality rate is clear evidence of the need for action in this area. It is no longer the original illnesses that are responsible for the death of immune-deficient patients, but rather the infections described above. We must therefore accelerate development in this area and find ways to identify new drug targets.

The aim of this research project is the systematic and scientific development of potential drug targets in the setting of infections in immune-deficient patients. The analysis of data obtained from in vitro models will afford new insights into the interplay between adenoviruses and human cells, which could lay the groundwork for further studies and the development of more effective therapies.

The project is funded by the Austrian Research Promotion Agency under the sixth call for the “Aufbau” line of the COIN.

Patients with an impaired immune system such as HIV-positive individuals or solid organ and particularly hematopoietic stem cell transplant recipients are at high risk of undergoing life-threatening infections with human adenoviruses. The efficacy of commonly used drugs to treat adenovirus infections is limited and frequently associated with toxicity. Alternative drugs are still under investigation. Hence, given the fact that numbers of solid organ and hematopoietic stem cell transplant recipients are constantly rising, alternative treatment options are highly needed.

Short interfering RNAs (siRNAs) and artificial microRNAs (amiRNAs) are a class of artificial small RNAs that can bring about the inactivation of cellular and viral genes via the RNA interference (RNAi) pathway. In a previous project led by Dr. Reinhard Klein highly potent siRNAs and amiRNAs with activity against components of the adenoviral DNA replication machinery that can effectively inhibit the replication of human adenoviruses in cell culture experiments were developed and characterized.

The project is aimed at investigating if adenovirus infections can be inhibited by these RNAi-triggering small RNAs in vivo, and which of the two approaches (i.e. siRNA versus amiRNA) is more effective. RNAi-based inhibition of adenoviruses is assessed in the Syrian hamster model which is able to mimic adenovirus infections in immunodeficient humans. Moreover, one of the two small RNA-based approaches is anticipated to lead to the selective amplification of the RNAi-triggering RNAs in adenovirus-infected cells and their transfer to neighbouring cells where they are supposed to inhibit the otherwise uncontrolled multiplication of spreading adenoviruses.

The project is funded by the Austrian Science Fund (FWF).

Abdominal obesity together with insulin resistance, dyslipidaemia and hypertension are hallmarks of the metabolic syndrome (MeS). Furthermore, this disease is also characterized by a low-grade-inflammatory status of adipose tissue causing changes in lipid metabolism of adipocytes by misbalancing uptake, deposition and release of lipids and free fatty acids (FFAs). Markers for abnormal lipid metabolism and inflammation are also found to be elevated in patients suffering from cancer cachexia (CaC). We suppose that pathological alterations of the lipid metabolism are main contributors in MeS and CaC. Therefore, we hypothesize that metabolic signatures comprising lipid, eicosanoid and cytokine patterns are reflecting disease states and disease progression and can help to monitor the impact of clinical interventions. In this study, a comprehensive plasma profiling will be applied to characterize the lipid, eicosanoid and cytokine patterns of patients suffering from MeS and CaC. The goal of the project is to identify metabolic signatures indicative for the onset and progression of the diseases and for determining the therapeutic efficacy of clinical interventions. The Karl Landsteiner Private University of Health Sciences holds the lead of the project. Additional partners are the University of Vienna and Medical Universities of Vienna and Graz.

The project was funded by the Niederösterreichische Forschungs- und Bildungsges.m.b.H. (NFB).

Testing the recombinant avian polyclonal antibody fragments against gluten peptides that are produced and made available. These antibodies will be investigated using in vitro test systems (cell culture, intestinal epithelial cells) for their potential to inhibit inflammatory reactions in the intestinal epithelium caused by gluten peptides.

The project was funded by the Austrian Research Promotion Agency under its basic programme.

Melanoma is one of the most frequent tumours in young adults. Even though it only accounts for 4% of all cases of skin cancer, melanoma is responsible for 79% of all skin cancer-related deaths. Despite the progress that has been made in the treatment of melanoma (e.g. with BRAF inhibitors), patients finally succumb due to resistance mechanisms acquired by the tumour. Many lines of evidence have shown that especially a metastatic melanoma exhibits a strong metabolic turnover, which is required to fuel cell proliferation and anabolic pathways. This increased cellular turnover also results in an increased demand to maintain the redox homeostasis. Here we propose analysing this high metabolic and therefore also ROS (reactive oxygen species) generating stress as a possible Achilles heel of melanoma. One of the major regulators of stress response in cancer is NRF2. It plays a central role in the protection of cells against oxidative and xenobiotic stresses.

The inhibition of NRF2 or its target genes might re-establish the sensitivity of melanoma to apoptosis driven by ROS. This mechanism could also prevent resistance mechanisms frequently observed in metastatic melanoma and may eliminate the frequently observed activation of endothelial cells, which surround tumour cells. It is highly likely that a combination of state of the art melanoma treatment with compounds that inhibit the generation of ROS scavengers potentiates the effectiveness of the current treatment regiments. Here we will use CRISPR-based methods as well as pharmacological inhibition to elucidate the mechanistic role of NRF2 in melanoma cells and on endothelial cells. We will also Transfer knowledge gained from our model by closely cooperating with clinicians who routinely care for melanoma patients. We propose that eliminating the antioxidative response by suppressing NRF2 directly, or its targets, will be an effective weapon in the battle against metastatic melanoma.

The project is funded by NÖ Forschungs- und Bildungsges.m.b.H (NFB).

Extremophilic microorganisms often have special properties and also special metabolic pathways due to the cellular mechanisms necessary for their natural environment. These metabolic pathways include, among other things, the ability to use unusual carbon sources and to produce various end products therefrom. These products are of interest for many applications, including pharmaceutical applications. Products are for example various polyunsaturated fatty acids and also pigments. The cultivation of these organisms is possible with the equipment available at the IMC FH Krems and through international contacts to the TU Bratislava or the Sultan Quaboos University in Oman, there are possibilities to preserve the organisms or to use the methods of analysis and to bring them to Krems in the long term. Identified substances can then be tested for bioactivity using the cellular test systems developed in-house.

The project is funded by the Province Lower Austria (Department Science and Research) under the technology fund programme ATHENOE.

Lung cancer is characterised by genderspecific differences in carcinogenesis, prevalence

and types of mutations as well as response to targeted therapies. The impact of a patient's

gender on the tumor's DNA methylation pattern and on the efficiency of latest epigenetic

therapies is still unknown and thus the research topic of this project.

The Project is funded by FFG Talente - 4th Call FEMtech Research Projects.

Proteins and peptides are key molecules in all biological processes. Their unique chemical properties make them particularly well suited for use as therapeutic agents. They have high biological activity and specificity with comparably few toxic side effects, and can be used to produce a range of highly diversified compounds that are not subject to intellectual property restrictions. The market for synthetic therapeutic peptides is growing steadily, making it an increasingly attractive area for pharmaceutical companies.

The research project’s specific focus was the development of peptides and antibodies that modulate the activity of receptor tyrosine kinases (RTKs, e.g. epidermal growth factor receptor (EGFR) and AXL). The work was performed in close cooperation with the Paracelsus Medical University Salzburg. In many cancers, the RTK signaling pathways are fundamental for proliferation, survival, angiogenesis and metastasis of cancer cells. Hence, inhibition of particular RTK pathways can induce apoptosis or senescence in the tumor cells. In addition, peptide or antibody mediated activation of RTKs could be beneficial for tissue engineering and regenerative medicine. This project has fostered the development of sustainable and cost-effective technologies for biopharmaceutical drug discovery, therapeutic apheresis, toxin neutralization and tissue engineering.

The project was funded by the Niederösterreichische Forschungs- und Bildungsges.m.b.H. (NFB).

Fermentation is an increasingly important area of the pharma- ceutical industry. Until now, academic research and industry have focused mainly on monocultures. However, it has been observed that many microorganisms only realise their full biochemical potential in tandem with others. As a result, co- cultivation has become a key pharmaceutical research Topic in the field of biotechnology.

Research in this area concentrates on identifying potential co-cultures and establishing a fermentation process that harnesses the products and capabilities of microbial communities for drug discovery and industrial applications.

The project represented an initial feasibility study of whether it is possible to identify conditions under which two selected microorganisms can grow separately from one another, and also of whether a stable co-culture can subsequently be established. The co-cultures were then analysed in terms of their ability to produce new substances.

The project was funded by the Province of Lower Austria and the European Regional Development Fund (ERDF).

The goal of the project is to combine a method comprising both cellular reporter assays and gene expression studies into a sensitive method. This would make it possible to show toxicological effects on human cells at significantly reduced concentrations. If established successfully, the combined method could make a contribution to risk minimisation for newly developed biotech products and could also lead to an innovative screening procedure for the detection of toxic environmental pollutants not detectable using current analytical methods. The technology developed will be made available to biotech companies in Lower Austria in partnership projects.

The project was financed by the Science and Research Department of the Province of Lower Austria.

The functioning of living organisms is to a large extent dependent on the interplay between the biomolecules they are composed of. Protein-protein interactions (PPIs) are a basic mechanism that regulates this interplay. Consequently, in the past few years the search for active compounds that have a therapeutic influence on protein-protein interactions has been intensified. In most cases these compounds are inhibitors of these interactions.

The aim of the project was to use a bacterial enzyme system in order to develop a prototypical workflow for the generation of hit structures for inhibition of protein-protein interactions (PPIs). Starting from the ACP/ACPS-system of Staphylococcus aureus peptides were identified that inhibit the ACP-ACPS-interaction. Peptide-ACP-interactions as well as ACPS-ACP-interactions were analyzed by NMR experiments. The insights obtained, together with structural information from available X-ray structures, led to the development of pharmacophore models that were used in virtual screening to identify potential small molecules PPI inhibitors.

The project was funded by the Austrian Research Promotion Agency under the 15th call for the BRIDGE 1 programme line.

Melanoma is the most aggressive form of skin cancer. If metastasis occurs, currently only about 10% of patients with the disease respond to standard treatment. This situation can be improved if patients at risk of metastasis are identified early, and if patients in which metastasis has already occurred undergo targeted treatment. In this project we describe the generation of new types of antibodies that can identify metastasis-specific antigens in a targeted manner, and which are being tested for suitability for diagnosis. The strength of our approach lies in the combined use of completed preclinical studies, established cell culture methods and in vivo models, as well as the antibody production expertise of Sciotec Diagnostic Technologies GmbH.

The Project under the consortial leadership of the Medical University of Vienna was funded by the Austrian Research Promotion Agency under the BRIDGE programme.

Intensive research into the molecular causes of cancer has led to the development of a range of innovative and targeted therapies which are used to selectively inactivate the molecular mechanisms responsible for tumour progression and the growth of cancer cells. These therapies can inhibit the proliferation of cancer cells and induce programmed cell death (apoptosis). They are not effective in all cancer patients due to the genetic heterogeneity of tumours. Personalised oncology aims to establish a direct link between tumour cell genotypes and sensitivity to bioactive substances, so that the patient first and foremost receives the targeted therapy with the maximum clinical benefit.

In the project an experimental approach is being developed to complement diagnostic biomarker studies. We plan to develop organotypic cancer models that enable the direct testing of the clinical efficacy of cancer therapies in cell and tissue cultures (in vitro). A large number of potential cancer therapies, which are often combinations of targeted drugs and conventional chemotherapeutic agents, could

be quickly tested for their clinical efficacy on a personalised Basis.

The project is funded by Niederösterreichische Forschungs- und Bildungsges.m.b.H. (NFB).

The aim of this project was the setup of a new technology platform in Lower Austria to make possible further developements and production of peptid-based adsorber in a quick, standardized and efficient way. Thus the aphereses could be established as an effective therapy against autoimmune disorders and sepsis in hospitals.

The project was funded by the Austrian Research Promotion Agency under the call of the Bridge Programme line.

Sepsis is one of the most frequent causes of death worldwide, including in Austria and Germany. Depending on the stage of the illness, between 25% and 60% of patients die despite receiving the maximum available treatment. In cases of sepsis, the human immune system produces a hyperinflammatory response to an infection that has entered the blood stream, and this overreaction can lead to cardiovascular failure. This hyperinflammation is followed by immunosuppression – an attempt initiated by the immune system itself to counter this overreaction. Due to the reduced attentiveness of the immune system, many patients die from serious secondary infections during this phase. Owing to the complex progression of the disease, available sepsis therapies focus predominantly on tackling symptoms and are unfortunately ineffective in many cases.

Peptides that modulate immune responses are currently regarded as promising new drug candidates for the treatment of sepsis. In this project, we aimed to develop and test new peptides that neutralise TRAIL/TNFSF10, one of the key immune regulators. Animal studies had suggested that inactivation of TRAIL/TNFSF10 is likely to reduce morbidity and mortality among patients suffering from sepsis. In addition, a gender- and cell-culture-based model for human sepsis was created, meaning that gender-specific differences (e. g. hormone status) could be taken into account when developing and validating potential new sepsis therapies.

The project was funded by the Austrian Research Promotion Agency under the first call for FEMtech research projects

in 2011.

For many cancer patients, conventional forms of chemotherapy are only partially successful due to their unspecific mode of action and their toxic side-effects. Innovative cell biological procedures involving the activation of tumour-specific T cells using dendritic cell therapy are possible alternatives. Dendritic cells are loaded or activated with tumour-associated antigens (RNA or proteins/peptides). The cells are then returned to the cancer patients in order to achieve in vivo a strong stimulation of tumour-specific cytotoxic and helper T cells, which attack and destroy the tumour. In a project conducted in collaboration with Life Research Technologies (LRT) GmbH, we have improved existing experimental strategies as well as developed new methods for the detection and quantification of the activity profiles of immune cells, in parallel with clinical studies.

The genomic revolution has boosted the development of novel cancer therapeutics targeting critical oncogenic signaling molecules. The therapeutic agents often inactivate protein kinases resulting in growth arrest and death of cancer cells. However, clinical benefit is limited to subpopulations of cancer patients. Personalized cancer medicine seeks to identify the genetic factors (biomarkers) that influence drug sensitivity. The genetic characterization of tumors will be instrumental for the individualization of treatments and for successful patient outcome and minimization of drug toxicity.

In the present project, we developed standardized diagnostic procedures that allow the detection of genetic biomarkers that can predict clinical drug response in cancer patients. The clinical relevance and the predictive value of the biomarkers have been partially validated in a retrospective clinical study focusing on breast cancer. The work required an interdisciplinary and multi-institutional collaboration between clinics, diagnostic centers and universities in Lower Austria, Tyrol, and Vienna. Genetic testing of cancer patients prior to therapy will increase the drug efficacy, safety, and cost-effectiveness of clinical treatments in cancer.

The project was funded by the Niederösterreichische Forschungs- und Bildungsges.m.b.H. (NFB).

Peptides are isolated using phage display against TNF alpha and TNFR2, and then tested for anti-inflammatory properties in the planned vascular models. The vascular models should be suitable for high-throughput screening (HTS) and provide a basis for the development of specialist detection modules by Beckman Coulter. Further tests on the isolated peptides were carried out by the industry partner. The data from the project will be used for the construction of prototype modules for the HTS market, and the peptides produced by the screening are expected to be further commercialised in follow-up projects.

The project was funded by the Austrian Research Promotion Agency under the BRIDGE programme.

Cancer-related morbidity and mortality represent a huge social and economic problem worldwide. Despite significant public and private investment in cancer research, there have still been no major advances in the treatment of most types of cancer. Success in the selection of anti-carcinogenic substances (agents) using high-throughput screening (HTS) has been very modest, due to the limited physiological relevance of the cancer models and cellular assays used up to now.

In this project, we developed innovative cell biological procedures which will help to create new generations of cancer models. A range of methods to generate three-dimensional (3D) cultures were developed. These 3D models included spheroids, heterotypic co-cultures and reporter cell lines. Using the models, it was possible to partially reconstruct the pathophysiological state of a tumour in vitro. In the course of several test series, it was shown that the standardized screening of agents was possible using the new 3D cell cultures.

The project was funded by the Austrian Research Promotion Agency under the “Aufbau” funding line of the COIN – Cooperation and Innovation programme.

The pharmaceutical industry is increasingly focusing its attention on the wide variety of natural substances developed in the form of secondary metabolites in microorganisms. Marine algae in particular contain a significant, untapped resource in the shape of chemical structures with the potential to play a major part in the development of innovative medications.

The project examined potential applications of constituents of blue-green algae in medical research on the treatment of chronic inflammations and cancer.

The project involved purification of secondary metabolites from cyanobacteria using state-of-the-art chromatography processes and characterisation by means of mass spectrometry. The different fractions were then examined using human cell-culture models to identify their impact on inflammation processes and on cancer.

The project was co-financed by the European Union through the European Regional Development Fund.

In the development phase of optimised cell-based test systems, new active substances were identified and one of the identified peptides was also mechanistically characterised. The substances bring about increased sodium uptake and oedema resorption in the lungs. Drugs currently available for the treatment of pulmonary oedema have many negative side effects, especially in the case of patients with heart disease. In collaboration with industry partners, the extent to which the substances maintain the integrity of endothelial and epithelium monolayers was tested, as well as the extent to which they were suitable for treatments of diseases of the lungs and sepsis. Additionally, a method of rapidly and reproducibly distinguishing between metastasising and non-metastasising cancer cells was developed by means of the optimised use of ECIS and confocal laser scanning microscopy.

The project was funded by the FH Plus initiative under the COIN programme.