The Institute of Neuroimmunology SAS remains the leader in the research of human brain and spinal cord disorders in the Slovak national scientific milieu, supported by strong international collaborations. We have actively participated in a variety of international projects (H2020, JPND, ERA-NET, COST) which have covered several aspects of basic and applied neuroscience research:

• Alzheimer´s and Parkinson´s disease – mechanisms leading to the neurodegeneration and biomarkers
• Blood-brain barrier damage in human tauopathies
• Astrocytes and microglia – modulators of neurodegenerative processes
• Traumatic brain injury – molecular pathways and biomarkers
• Stem cell therapy for spinal cord injury
• Interaction of pathogens with brain microvascular endothelial cells
• Structure of non-globular proteins
• Canine dementia – pathogenesis and biomarkers

The institute was involved in the Alzheimer’s disease drug development and the COVID-19 program in Slovakia, demonstrating a high translation potential of its research.

ALZHEIMER´S DISEASE RESEARCH

In order to preserve the functional brain microenvironment, the exchange of chemicals and biologicals between the periphery and the brain is tightly regulated by a functional barrier called the neurovascular unit (NVU). Andrej Kovac and his research team examined changes in the NVU in Alzheimer’s disease and other human tauopathies in close collaboration with William Banks (University of Washington, Seattle, USA) and Maria Deli (Hungarian Academy of Sciences Szeged, Hungary), key leaders in the field. The team demonstrated that disruption of the NVU, driven by neurofibrillary pathology and chronic neuroinflammation, was characterized by the production of pro-inflammatory signaling molecules such as cytokines, chemokines, and adhesion molecules by glial cells, neurons, and endothelial cells (Majerova et al., 2018, J Neural Transm; Michalicova et al., 2020, Front Mol Neurosci). These changes can modify the integrity of the barrier and migration of immune cells into the brain, which may finally lead to structural changes in capillaries (Deli and Kovac, 2020, Curr Pharm Des; Majerova et al., 2019, PLOS One).

On the other hand, a functional NVU poses a challenge for the delivery of therapeutics to the brain. Using state-of-the-art techniques, the group discovered novel delivery vectors that can transport large molecules into the brain (Majerova et al., 2020).

Tomas Smolek´s research team has studied the mechanism of spreading of tau pathology by using a transgenic rat model for tauopathy (H2020/ JPND – Pathway complexities of protein misfolding in neurodegenerative diseases: a novel approach to risks evaluation and model development, 2016-2019). They were first to demonstrate that human AD tau can spread in the rat brain (Smolek et al., 2019, Mol Neurobiol.). In addition, they showed that the immune response modulated by genetic variability is one of the factors that influences the propagation of tau neurofibrillary pathology (Smolek et al., 2019, Front Aging Neurosci.).

Research team of Rostislav Skrabana investigated the structural pathways leading to pathological switching of disordered tau protein molecule into toxic, disease-associated oligomers and polymers (Skrabana et al., 2017, J Alzheimer Dis.), highlighting the role of conformational rearrangement upon molecular truncation (Novak et al., 2018, J Alzheimer Dis.).

BIOMARKERS FOR TRAUMATIC BRAIN INJURY

Traumatic brain injury (TBI) is a risk factor for traumatic encephalopathy syndrome, especially chronic traumatic encephalopathy (CTE). The mechanisms and biomarkers of the trauma are investigated by Peter Filipcik and his research team. They have actively participated in two international projects on TBI: ERANET – Repetitive Subconcussive Head Impacts – Brain Alterations and Clinical Consequences, 2017-2019; ERANET – Neurovascular damage determines disease pathophysiology in pediatric mild traumatic brain injury: source of new biomarkers, 2020-2022. The group continues the research with several partners from Slovakian and foreign universities and has focused on developing a microRNA (miRNA) and long-noncoding RNA (lncRNA) based biomarker profile in the peripheral fluids which would reflect the brain deterioration. The analysis of blood samples from professional soccer players was performed at rest and after three conditions: accidental head impacts in a match, repetitive headers during training and high-intensity exercise. This showed that each of the activities led to a deregulation of a certain set of miRNAs and proteomic markers in the blood. More than 20 articles were published or accepted for publication by the research team (including the papers in collaboration).

PARKINSON’S DISEASE RESEARCH

Recently, the Institute established new research group focused on Parkinson’s disease pathology led by two internationally recognized researchers Dominika Fricova and Alzbeta Kralova Trancikova, who were both awarded a competitive prize L’OREAL-UNESCO for Women in Science (2020 Dominika Fricova, 2021 Alzbeta Kralova Trancikova). The team working under the supervision of Dominika Fricova is focused on the role of senescence in Parkinson’s disease. This project represents an innovative strategy for the identification of new potential targets in Parkinson’s disease treatment. Project was funded by Maire Sklodowska-Curie Cofund project (SASPRO 2). She also investigates the potential role of mesenchymal stem cells and extracellular vesicles in Parkinson’s disease treatment (Fricova et al., 2020, Nature Regenerative Medicine).

Alzbeta Kralova Trancikova studies the alpha-synuclein-associated pathology within the gastrointestinal tract and its spread to the CNS structures in mouse models of PD as well as the patient’s samples. Since the early diagnosis of PD is still insufficient, her work is also focused on the development and optimization of potential diagnostic methods from readily available, non-invasive biological material (advanced microscopic methods – FLIM and confocal microscopy from wholemount tissue micro biopsies or RT-QuIC analysis from body fluids) (Harsanyiova et al., 2020, Front Neurosci; Fricova etal., 2020, Int J Mol Sci; Pokusa et al., 2018, Physiol Res). They collaborate with the Centre for rare movement disorders at the Department of Neurology, University of Pavol Jozef Safarik in Košice, Slovakia.

STEM CELL THERAPY FOR SPINAL CORD INJURY

The main focus of the research team of Dasa Cizkova is on the stem cell therapy for the spinal cord injury (ERANET – Spinal cord repair: releasing the neuron-intrinsic brake on axon regeneration, 2017- 2019; V4 – Bridging the gap between science, education, and enterprise in regenerative medicine, 2020-2022). The team provided a comprehensive proteomic study of canine bone marrow-derived mesenchymal stem cells and conditioned media, they identified for the first time the dynamic release of various bioactive molecules, such as transcription and translation factors and osteogenic, growth, angiogenic, and neurotrophic factors from canine stem cell-conditioned medium (Humenik et al., 2019, Mol Cell Proteomics). The conditioned medium was used for further therapy of dogs with spinal cord injury. The treatment did not show any adverse effects or complications and in combination with comprehensive physiotherapy, it demonstrated clinical benefits (Vikartovska et al., 2019, Int J Mol Sci). The conditioned medium of mesenchymal stem cells improved motor function recovery and attenuated inflammation in a rat model for spinal cord injury (Cizkova et al., 2019, Int J Mol Sci).

THE MECHANISM OF TRANSLOCATION OF PATHOGENS ACROSS THE BLOOD-BRAIN BARRIER

Mangesh Bhide and his team have identified multiple mechanisms of how bacterial (Borrelia garinii, Neisseria meningitidis) or viral pathogens (West Nile virus) translocate across the blood-brain barrier (BBB) and cause menigo-encephalitis, and described the pathogen-endothelial cells interaction interfaces (Kanova et al., 2018, Front Microbiol; Tkacova et al., 2020, Ticks Tick Borne Dis). He used a variety of approaches to reveal the molecular basis of the interaction and combined with state-of-the-art bioinformatic tools generated a large dataset of multiple interaction partners (Bencurova et al., 2018, Mol Omics; Hortvatic et al., 2018, Methods Mol Biol; Mertinkova et al., 2020, Sci Report). He also created several peptide candidates or single domain antibodies for the development of novel antiviral therapeutics against West Nile Virus (Mertinkova et al., 2021, Sci Report; Hruskovicova et al., 2022, Front Microbiol). The developed peptides and antibodies are being used to generate novel drug delivery systems (DDs) made of either amphiphilic dendrimers or polymeric nanoparticles (EuroNanoMed2018-049 Nanosystems conjugated with antibody fragments for treating brain infections, EuroNanoMed2021 Developing novel nanopharmaceutics against bacterial infections at center nervous system).

ANTI-VIRAL PEPTIDES FOR THERAPY OF COVID-19

To contribute to global efforts of containing the pandemic caused by novel coronavirus – SARSCoV-2, the research team of Mangesh Bhide has developed single domain antibodies and anti-viral peptides targeting the receptor-binding domain of spike protein (APVV PP-COVID-2020 Development of therapeutic biomolecules to block SARS-CoV-2 infection). Both the biomolecules can neutralize SARS-CoV-2 virus-like particles carrying the spike protein of delta variant B.1.617 and the original D614 genotype.

STRUCTURE OF NON-GLOBULAR PROTEINS

The Institute has a long-standing tradition in the structural analyses of non-globular proteins (NGPs) that defy the traditional sequence-structure function paradigm. NGPs include intrinsically disordered regions, tandem repeats, aggregating domains, low-complexity sequences and transmembrane domains. Although growing evidence suggests that NGPs are central to many human diseases, their functional annotation is very limited. The research team of Rostislav Skrabana has participated in the pan-European project (COST, Non-globular proteins – from sequence to structure, function and application in molecular physiopathology, 2015 – 2019). One of the outputs of the project was the standardization of the design of aggregation experiments used in the research of NGP role in proteinopathies (Martins et al., 2020, Front Mol Neurosci.). Nuclear magnetic resonance spectroscopy (NMR) demonstrated how subtle differences in transient structural motifs of homologous tau and MAP2c proteins are linked to differences in their interactions and function (Melkova et al., 2019, Biomolecules; Kitoka et al., 2021, Front Mol Biosci.). Another focus of the team is the investigation of small local structures in non-globular proteins, which can determine their physiological and pathological fate (Cehlar et al., 2021, Gen Phys Biophys.).

CANINE DEMENTIA

Norbert Zilka´s research team has become the leading force in the study of the molecular mechanism of canine dementia. In 2017, he co-edited the book “Canine and feline dementia” (Springer, editors Gary Landsberg, Aladar Madari, Norbert Zilka), which summarized what’s known on the epidemiology, diagnostics, therapy and molecular basis of canine and feline dementia or cognitive dysfunction syndrome. The team described the risk factors for canine cognitive dysfunction syndrome in Slovakia (Katina et al., 2016, Acta Vet Scand), synaptic impairment and neuroinflammation associated with canine cognitive impairment (Smolek et al., 2016, J Comp Neurol.). Furthermore, they demonstrated that brain injury biomarkers (neurofilament light chain) and biochemical parameters (ALT, AST, Na, and Cl) in blood serum may predict canine cognitive impairment in aged dogs (Vikartovska et al., 2021, Front Vet Sci).

APPLIED RESEARCH

IMMUNOLOGY AND IMMUNOTHERAPY FOR NEURODEGENERATIVE DISEASES

The institute has participated through targeted research grants from a biotech firm AXON Neuroscience SE on the development of new, first-in-man, first-in-kind, tau biological therapeutics for Alzheimer´s disease, namely active tau vaccine AADavac1 (Kontsekova et al., Alz Res Ther, 2014a, b). The vaccine proved to be safe and stimulated high levels of antibodies, slowed down neurodegeneration in the brain and reduced the cognitive impairment (Novak et al., 2018, Front. Neurosci.; Novak et al., 2021, Nature Ageing). Within this project, the team of Eva Kontsekova investigated the molecular mechanisms of spreading of neurofibrillary pathology of tau across the brain, how a novel therapeutic monoclonal antibody DC8E8 can efficiently prevent the neuronal internalization of the pathological AD tau species (Weissova et al., 2019, Acta Neuropathol. Comm.), and how human primary microglia, isolated from the brains of deceased Alzheimer’s patients, can be promoted by DC8E8 antibody to effectively eliminate extracellular pathologic tau proteins (Zilkova et al., 2020, Acta Neuropathol. Comm.). This work is a result of a collaboration with Dr. Hoozemans from Amsterdam UMC of Vrije Universiteit Amsterdam and Department of Pathology of Amsterdam Neuroscience (Netherlands).

SLOVAK AD COHORT

The Institute has become the driving force behind the innovative approaches to improve diagnosis of Alzheimer’s disease and care for the patients with dementia and their caregivers in Slovak republic. The institute (supervised by Petr Novak) in collaboration with Centre MEMORY were engaged in an international project which aimed to bring new understanding of which aspects are most important to patients and caregivers to preserve and improve their autonomy, dignity and quality of life (H2020/JPND – Alzheimer’s disease data-driven insights on individual outcomes of importance, 2019 -2022). We created the very first Slovak database of well-defined AD cohort, and set up new diagnostic algorithm for AD diagnosis in the MEMORY Centre.

THERAPEUTIC ANTIBODIES AGAINST COVID-19

The Institute was actively involved in COVID-19 program in Slovakia, in collaboration with Biomedical centre of Slovak Academy of Sciences, we created the screening network for the whole Academy to monitor the SARS-CoV-2 positive cases. In a collaboration with COVIDAX a.s., the research team of Eva Kontsekova developed several assays for antibody and cell response which were used to monitor immune response to COVID-19 infection and vaccination.

Moreover, the Institute took part in the development of therapeutic antibodies against COVID-19. The antibodies developed by hybridoma technology displayed high affinity to all variants of concern (Kovacech et al., 2022, EBioMedicine by Lancet).