Student Researchers' Society Topics

Co-supervisor: Dr. POZSGAI, Gábor

The treatment of the chronic inflammatory diseases, like allergic dermatitis and inflammatory bowel diseases (IBD) means still a problem of modern medicine. In the progression of these diseases neurogenic factors play an important role. Neuropeptides released from the capsaicin-sensitive sensory neurons, as well as inflammatory and immune cells influence the inflammatory processes, but their complex roles have not been cleared in precise details. Some of these peptides exert proinflammatory (SP, NKA, CGRP), while others exert antiinflammatory effects (opioid peptides, somatostatin, cortistatin, PACAP). Beside the classical peptide mediators of neural origin, hemokinin-1 and endokinins, expressed mainly in inflammatory and immune cells, were discovered a few years ago. All these peptide mediators are released on response to endogenous or exogenous stimuli, through the activation of Transient Receptor Potential Vanilloid 1 (TRPV1) and Ankyrin 1 (TRPA1), or independently of that. Antagonising the effect of the proinflammatory peptides, as well as the precise identification of the mechanisms of antiinflammatory mediators may enable the development of antiinflammatory drugs acting on new target structures. Therefore, our experiments aim to investigate the interactions between the neurogenic components and the immune system in animal models of dermatitis and IBD, with the help of functional, morphological, immunological, molecular biological, as well as in vivo imaging techniques.
Key words: neuro-immune interaction, neuropeptides, skin, dermatitis, bowel, IBD, proinflammatory, antiinflammatory

Co-supervisor: Dr. POZSGAI, Gábor

Among other members of the TRP receptor family, capsaicin-sensitive sensory neurons express Transient Receptor Potential Ankyrin 1 (TRPA1) ion channels. TRPA1 can be activated by several exogenous and endogenous chemical stimuli, such as pungent agents (mustard oil, cinnamaldehyde, allicin) and toxic gases (acrolein, formaldehyde) resulting in the release of inflammatory peptides (SP, CGRP). Hydrogen-sulphide (H2S) is an important gaseous mediator of the human body. In the recent years, there have been several data published, indicating that H2S may be involved in various vascular and inflammatory processes. Moreover, it has also been suggested that H2S may act on capsaicin-sensitive peptidergic neurons. The aim of our investigation is to reveal the interaction of neural and non-neural TRPA1 receptors and H2S in microcirculatory processes, in dermal, intestinal and airway inflammations, as well as in neuropathic states, with the help of in vivo and in vitro experimental methods.

Co-supervisor: Dr. HORVÁTH, Ádám István

TRPA1 and TRPV4 receptors are expressed on capsaicin-sensitive sensory neurons, immune cells and keratinocytes. TRPA1 receptors are activated by pungent agents (mustard oil, cinnamaldehyde), exogenous and endogenous toxic chemicals, as well as inflammatory mediators and noxious cold (<17°C). The activation and sensitization mechanisms and interactions of TRPV4 are less known. Therefore, we aim to analyze their roles and cross-talks in inflammatory and neuropathic pain models with thermo- and mechanonociceptive testing methods using gene-deleted mice pharmacological interventions.

Co-supervisor: Dr. HORVÁTH, Ádám István

Amino-oxidases neutralise the pharmacological effects of exogenous and endogenous biogen amines. However, their reaction products (aldehyde, hydrogen-peroxide, ammonia) are biologically active, potentially toxic compounds. Although the substrate-specificity of these enzymes and their sensitivity to enzyme inhibitors is well-known, their physiological and pathophysiological role is not cleared yet. Therefore, as a continuation of our former experiments, we investigate their function in neuropathic, postoperative and inflammatory pain conditions.

Clinical and experimental data indicates that chronic stress increases pain sensitivity, but the underlying mechanisms are largely unknown. The pain perception of mice exposed to chronic restraint stress is assessed with functional tests in our experiments. Neuronal activation caused by stress and pain is examined with immunohistochemistry both in the central and peripheral nervous system. The role of peripheral capsaicin-sensitive sensory neurones is explored with desensitized animals. Investigation of the stress-induced hyperalgesia may contribute to the understanding of stress-related pain syndromes (e.g.: fibromyalgia) and to the identification of new drug targets.

Measurement of the noxious heat threshold is a new concept in the investigation of heat-induced pain, i.e. thermonociception in awake animals. Conventional methods of thermonociception measure the latency (i.e. time) of pain-avoiding (nocifensive) behavioral reactions evoked by a suprathreshold heat stimulus. In contrast, with the recently developed increasing-temperature water bath it is possible to measure the noxious heat threshold of the hind paw or tail in rats or mice by determining the lowest bath temperature that can evoke a nocifensive reaction. This novel equipment has been validated in several experimental models in which heat hyperalgesia manifested itself as a decrease of the noxious heat threshold. Future work aims at investigating other paradigms associated with a drop of heat threshold and revealing their mediators as well as pharmacological modulation including testing of analgesics.

Co-supervisor: Dr. BÖRZSEI, Rita Judit

Computational docking is an essential tool of drug design widely used by pharmaceutical companies. The project will focus on the prediction of structure and energy of drug-target interactions by computational docking. Both capabilities and limitations of the method will be investigated on a set of drug-target systems. The set will be assembled on the basis of common interest of the student and the supervisor.

Co-supervisor: Dr. ZSIDÓ, Balázs Zoltán

Epigenetics plays an important role in the pathomechanisms of various diseases. Modifications of histone proteins of the nucleosome, the “histone code” is fundamental in understanding of diseases with epigenetic background. In this project, we will study the effect of the modifications on structure and interactions of histones focusing on their role in disease pathomechanisms. Structural bioinformatics tools will be applied in our investigations.

Co-supervisor: Dr. BÖRZSEI, Rita Judit

Application of pharmacoinformatics tools is essential in modern drug design. The project will focus on the development of computational tools facilitating efficient discovery of potent drug candidates. The tools work with atomic resolution structures of (protein) targets and calculate the strength of drug-target interactions. Students interested in physical chemistry with programming and/or scripting skills (interest) are good candidates for participating in this project.

Co-supervisor: Dr. ZSIDÓ, Balázs Zoltán

Attacking the assembly of HIV capsid is promising strategy of fighting AIDS. Proteins of the viral capsid are good targets of drug design due to their low mutation rates. In the project, available assembly inhibitors will be collected and their mechanisms will be investigated by computational techniques. Based on the collected information and the uncovered mechanisms, new inhibitors will be also designed.

Co-supervisor: Dr. BÖRZSEI, Rita Judit

Signal transduction is a key to understanding pathomechamisms of various diseases. Signaling pathways are often based on the formation of complexes between protein partners. In many cases, experimental determination of atomic resolution structures of protein complexes (required by drug design) is difficult. In the present project, we will calculate the structure of such complexes using computational tools. We will also study their molecular dynamics to understand their role in signaling and provide starting information for drug development.

Co-supervisor: Dr. ZSIDÓ, Balázs Zoltán

GFR is an essential clinical parameter describing kidney function. The present topic involves a comparative literature survey of the methods of determination of GFR as creatinine clearance. In case we can find available literature and electronic healthcare records, we will apply statistical approaches to evaluate the significance of the state of art methods and will propose probable changes. Likewise, depending on the availability of data we will investigate the connection between physiological parameters (such as blood pressure) and GFR.

Co-supervisor: Dr. ZSIDÓ, Balázs Zoltán

Current drug repositioning possibilities are investigated via generation and evaluation of target-ligand complex structures. During the research work, up-to-date pharmacoinformatic methods are applied.

Co-supervisor: Dr. ZSIDÓ, Balázs Zoltán

Calculation of the structure, dynamics, and energy of pharmacologically important molecular complexes using computational molecular modelling tools.

Co-supervisor: SZÉL, Viktor

Computational docking is an essential tool of drug design widely used by pharmaceutical companies. The project will focus on the prediction of structure and energy of drug-target interactions by computational docking. Both capabilities and limitations of the method will be investigated on a set of drug-target systems. The set will be assembled on the basis of common interest of the student and the supervisor.

Co-supervisor: Dr. BÁTAI, István Zoárd

In this topic effects of hydrogen sulfide, sodium polysulfide and dimethyl trisulfide are investigated. Beside classical models of nociception and inflammation, animal modles of neuropathic pain, arthritis and pancreatitis are utilized. Mechanical and heat sensitivity, myeloperoxidase enzyme activity, oedema formation, rate of plasma extravasation as well as activation of corresponding parts of the CNS by immunohistochemistry are detected during the experiments. 
 

Co-supervisor: GÖNTÉR, Kitti

In this topic we will study the effects of DMTS, an organic polysulphide, in a mouse model of anxiety and depression. Regarding the underlying mechanism, we investigate the endocannabinoid system. DMTS is likely to inhibit the main enzymes that degrade endocannabinoids by forming a disulfide bond with their cysteine side chains. Inhibition of these enzymes increases the endocannabinoid level in the CNS, reducing the effects of acute and chronic stress and the risk of depression.

TRPV1 and A1 receptor expression in the central nervous system suggests the role of these ion channels in neuropsychiatric disorders. Their activation releases tachykinins (such as Substance-P, neurokinin A) acting at neurokinin 1 and 2 (NK1 and NK2) receptors and NK antagonists have been previously reported to be effective in animal models of anxiety and depressive disorders. The newest tachykinin, hemokinin-1 (HK-1), is also present in many brain areas, but its binding site, receptor activation mechanisms and signalling, which are different from those of other tachykinins. Therefore, in our experiments, tachykinins and neurokinin receptors are investigated with complex in vivo behavioural methodology in gene-deficient mice and with the help of receptor antagonists.

Insight into the functional changes of tissues and cells may by examined by visualization of mRNA molecules and their protein products. With the help of ultrasensitive in situ hybridization (RNAscope) technique, it is even possible to detect individual molecules at cellular level. The technique may also be combined with immunohistological methods, so the changes can be examined and quantified both at mRNA and protein level by confocal microscopy even within the same tissue samples or cells.

This approach may help to determine the molecular background of functional or behavioral changes observed in in vivo models for various diseases. A wide range of animal models including mutant mouse lines are available in our lab also.

Co-supervisor: Dr. ALOMARI, Ammar

Ten percent of the population suffers from this neurological disease. It is three times more common in women than in men. Migraine headaches are typically recurrent, throbbing, unilateral, and seizure-like. Nausea, vomiting, photosensitivity may be the accompanying symptoms. In sensitive individuals, certain substances and situations trigger seizures. As migraine attacks deteriorate the quality life considerably, our main goal here is to gain deeper insight into the underlying brain territories to identify new potential therapeutical targets in the brain. The exact pathomechanism of migraine is still not fully understood, but triggering factors include hormonal changes, stress exposure, disturbances of the daily rhythm affecting sleep, and certain weather conditions.

Interestingly, the recruitment of the Edinger-Westphal nucleus (EW) by stress adaptation response, circadian rhythm, and estrus cycle is known. In addition, a number of neurotransmitters are found in EW that are also involved in the pathophysiology of migraine. Gene-deficient animals and in vitro techniques are available for our research, which can be used to describe the functional and neuromorphological changes in various migraine models at both mRNA (RNAscope, PCR) and protein (IHC) level.

Co-supervisor: Dr. ALOMARI, Ammar

Alcohol abuse is responsible for 3 million deaths worldwide each year, 5.3% of all deaths. Alcohol is known to be pathogenic for more than 200 different diseases. Young active people are the main group affected, with 13.5% of deaths among 20-39 years of age due to excessive alcohol consumption.

Our most recent study has shown that dorsal raphe nucleus (DR) neurons and urocortinergic neurons of the centrally projecting Edinger-Westphal (EWcp) nucleus carry the transient receptor potential ankyrin 1 (TRPA1) ion channel, whose expression is reduced in chronic stress and suicide. Alcohol abuse/dependence and stress-related (mal)adaptation are often co-occurring psychopathological conditions. The role of urocortin 1 (UCN1) and DR serotonin (5HT) is also known to be involved in alcohol abuse. Since ethanol and its metabolites (acetaldehyde and acetic acid) are able to activate TRPA1 and efficiently cross the blood-brain barrier, the question arises whether they are able to act directly on TRPA1 receptors on EWcp/UCN1 and/or DR neurons.

We aim to investigate how TRPA1 contributes to DR/5HT function in a mouse model of chronic alcohol exposure. We plan to perform complex behavioural, neuroendocrine and functional-neuromorphological studies in Trpa1 wild-type (WT) and gene-deficient (KO) mice, with a focus on DR serotonergic neurons.

We hypothesize that ethanol and its metabolites may influence alcohol dependence and reward mechanisms via TRPA1 ion channels expressed in EWcp and DR.

For our research, we have available TRPA1 gene-deficient animals, in vivo and in vitro techniques to monitor functional and neuromorphological changes at mRNA (RNAscope, PCR) and protein (IHC) levels in a chronic alcohol exposure model.

Co-supervisor: Dr. ALOMARI, Ammar

Alcohol abuse is responsible for 3 million deaths worldwide each year, accounting for 5.3% of all deaths, and has been shown to play a pathogenic role in more than 200 diseases. It mainly affects the young active population, accounting for 13.5% of deaths in the 20-39 age group.

Based on our preliminary studies and literature data, we know that the transient receptor potential ankyrin 1 (TRPA1) cation channel is expressed in the liver. Among the many activators of the ion channel, ethanol and its metabolites acetaldehyde and acetate are thought to have the potential to act directly on liver cells and to influence alcohol-induced liver injury.

Our aim is to detect TRPA1 in liver tissue, characterize TRPA1 positive cells and elucidate the possible role of this ion channel in a mouse model of chronic alcohol exposure-induced liver injury. A complex pharmacokinetic, functional-morphological and molecular biological study of TRPA1 gene-deficient mice with a focus on liver tissue is planned. A better understanding of the mechanisms underlying alcohol-induced liver damage may bring us closer to mitigating the adverse consequences of alcohol abuse.

For our research, we have available TRPA1 gene-deficient animals, in vivo and in vitro techniques to monitor functional and morphological changes in the liver at both mRNA (RNAscope, PCR) and protein (IHC) levels in a chronic alcohol exposure model.

Co-supervisor: MILICA, Milicic

Alcohol abuse is responsible for 3 million deaths worldwide each year, 5.3% of all deaths. Alcohol is known to be pathogenic for more than 200 different diseases.

Our most recent study has shown that the urocortinergic neurons of the centrally projecting Edinger-Westphal (EWcp) nucleus carry the transient receptor potential ankyrin 1 (TRPA1) ion channel, whose expression is reduced in chronic stress and suicide. Alcohol abuse/dependence and stress-related (mal)adaptation are often co-occurring psychopathological conditions. Urocortin 1 (UCN1) is also known to have a role in alcohol abuse, besides the stress adaptation. Since ethanol and its metabolites (acetaldehyde and acetic acid) can activate TRPA1 and efficiently cross the blood-brain barrier, the question arises as to whether they are able to act directly on TRPA1 receptors on EWcp/UCN1 neurons and contribute to alcohol abuse. There is a direct connection between the urocortinergic EWcp and the dopaminergic ventral tegmental area (VTA). VTA has a key role in rewarding and addiction, as it belongs to the mesolimbic dopaminergic system.

Our aim is to investigate how TRPA1 contributes to the function of dopaminergic system in a mouse model of chronic alcohol exposure. We plan to perform complex behavioral, neuroendocrine and functional-neuromorphological studies with Trpa1 wild-type (WT) and gene-deficient (KO) mice.

We hypothesize that ethanol and its metabolites may influence alcohol dependence and reward mechanisms via TRPA1 ion channels expressed in EWcp.

TRPA1 gene-deficient animals, in vivo and in vitro techniques are available to monitor functional and neuromorphological changes at mRNA (RNAscope, PCR) and protein (IHC) levels in a model of chronic alcohol exposure.

Co-supervisor: MILICA, Milicic

Post-traumatic stress disorder (PTSD) is a chronic anxiety disorder that develops in response to intense emotional/physical stress. Many factors are unknown about its complex pathophysiological mechanisms. It is known that the overactive bladder is more common among patients with PTSD, the exact pathomechanism of which is unknown. The target of the proposed research is the Barrington's nucleus (pontine micturition center), which plays a very important role in micturition, mainly through its glutamatergic neurons.

The aim of our research is to elucidate the pathomechanism of overactive bladder in a mouse model of PTSD using complex behavioural, neuroendocrine, functional neuromorphological and molecular biological methods, with a special focus on Barrington’s nucleus.

The TDK topic will provide the opportunity to learn in vivo, neuromorphological and molecular biological methods.

TRPA1 is a non-selective cation channel activated by a wide range of irritants, nonxious cold and pro-inflammatory cytokines. Since it is permeable for Ca²⁺, Na⁺ and K⁺, its activation results in membrane depolarization, action potential discharges and neurotransmitter release both centrally and peripherally. It is predominantly expressed in primary sensory neurons and epithelial cells. Trpa1 mRNA expression in adult and developing mouse brain has been poorly investigated. Therefore, the main goals of this study are (i) to map Trpa1 mRNA expression both spatially and temporally on sagittal and coronal sections of developing mouse brain by using the ultrasensitive RNAscope in situ hybridization technology, since currently no specific α-TRPA1 antibody is commercially available; (ii) to co-localize Trpa1-positive regions and stages with specific neuronal markers either performing multiplex RNAscope or RNAscope combined immunostaining procedure depending on the availability of the antibody specific to the neuronal marker of interest.

It is well known that the transient receptor potential vanilloid-1 (TRPV1) ion channel is expressed primarily in primary sensory neurons located in the dorsal root and trigeminal sensory ganglia, and plays a significant role in the mechanism of pain and neurogenic inflammation. However, much less is known about its central nervous system distribution and role of the capsaicin receptor researched by Nobel Laureate, David Julius in 2021. In our work, we plan to (i) identify the TRPV1-expressing neuronal groups in coronal sections of mouse brain using RNAscope in situ hybridization technology, as a specific anti-TRPV1 antibody is not commercially available; (ii) further characterize the TRPV1-positive regions with specific neuronal markers, either by multiple RNAscope or RNAscope combined with immunostaining technology.

Co-supervisor: Prof. Dr. HELYES, Zsuzsanna

Endometriosis is a complex estrogen-dependent an inflammatory condition defined as the presence of endometrium-like tissue at ectopic sites. The disease affects around 10-12 % of women of reproductive age, causing chronic pain and infertility. Complex sensory-vascular and immune reactions play a role in its development and progression, but the underlying mechanism and therapy remains to be clarified. Capsaicin-sensitive nerve ending-related cytokines and neuropeptides as well as Transient Receptor Potential Ankyrin 1 (TRPA1) and Vanilloid 1 (TRPV1) receptors are assumed to be involved in endometriosis and consequently chronic pain, but their role is yet unclear. During this thesis, we investigate inflammatory cytokines, chemokines, growth factors, neuropeptides, and nociceptive receptors in human serum and tissue samples as well as in human endometriosis and stromal cell lines.

Co-supervisor: TÓTH, Norbert

Transient Receptor Potential Vanilloid 1 (TRPV1) and Ankyrin 1 (TRPA1) are non-selective peptidergic cation channels, which can be activated by various exogenous and endogenous irritants and inflammatory mediators. They are proven to play a role in a number of inflammatory and pain-related pathologies, and therefore have the potential for drug development. Cell lines and primary cells are important in vitro model systems in drug development, with cells expressing TRPV1 and TRPA1 contributing to preclinical findings of receptorial mechanisms. Examination of the expression and in vitro activation of these ion channels will provide data for further experiments.

Complex regional pain syndrome (CRPS) and fibromyalgia (FM) are chronic pain conditions with different origin. 2% of the populations suffer from FM, while a prevalence of CRPS is about 1:2,000, is a chronic pain condition experienced by humans. However CRPS and FM have distinct clinical phenotypes, they do share many other features. Pain, allodynia and abnormal sensation are develop in each syndrome and might to exist on a similar spectrum. Both CRPS and FM can be developed after specific traumatic events, although in FM is mostly associated with psychological, in CRPS it is mainly triggered by minor physical trauma. Their pathophysiological mechanisms are poorly understood, the most likely mechanisms involve the classic and neurogenic paths of inflammation mediated by cytokines and neuropeptides, intertwined with changes of the autonomic and central nervous system, psychological mechanisms and possible role of autoantibodies may also suggested.

In this work we explore the mechanisms, pathways and potential therapeutic targets in CRPS-related extensive, sustained pain, and examine the role of neuroinflammation and cytokine signaling.

Co-supervisor: Dr. BORBÉLY, Éva

There is clinical and animal evidence that chronic stress increases pain, but many of the underlying mechanisms are unknown. In our experiments, we investigate changes in pain perception in mice exposed to acute or chronic cold environment stress using functional tests. Central and peripheral nervous system neuronal activation in response to pain and stress is investigated by immunohistochemistry. The role of Transient Receptor Potential Ankyrin 1 (TRPA1) mainly expressed on peripheral capsaicin-sensitive sensory nerve terminals will be investigated using gene-deleted mice and pharmacological intervention. The study of stress-induced hyperalgesia may contribute to understanding the pathomechanism of stress-related pain syndromes (e.g. fibromyalgia) and to identify new therapeutic targets.

Co-supervisor: Dr. TÉKUS, Valéria

Chronic pain affects a remarkable proportion of the popoluation worldwide, however, its treatment remains a constant problem in the medical practice. Among the pain states with various etiology, the treatment of neuropathic pain means the greatest challenge, since these diseases cannot be treated sufficiently with either form of the currently available drugs (NSAIDs, opioid compounds, adjuvant analgesics). Previous results of our workgroup showed that sensory neuropeptides released from the capsaicin-sensitive nerve endings and/or acting locally (SP, NKA, HK-1) play an important role in the development of neuropathic pain. Besides, somatostatin, released from primary sensory neurons into the systemic circulation exerts remarkable analgesic and anti-inflammatory effects via the sst4 receptors. Therefore, we aim to develop new analgesic drugs that are able to act on the neuropathic component serving as a novel mechanism of action, in animal models of traumatic neuropathic pain.

Chronic pain affects about 20% of the population worldwide. Regarding its etiology, it can be inflammatory, degenerative or neuropathic in origin. The therapeutic options for the relief of chronic pain are often inadequate, therefore, it is essential to develop new analgesic agents, as well as to improve new opportunities for their routes of administration.
The activation of capsaicin-sensitive sensory nerve endings via the Transient Receptor Potential Vanilloid 1 (TRPV1) receptor plays an important role in the pathogenesis of chronic pain. When the receptor is continuously activated with low concentrations of capsaicin, somatostatin released from the nerve endings produces a significant analgesic and anti-inflammatory effect.
Sincet transdermal therapeutic systems (TTS) could provide a good opportunity for local administation of these compounds, the aim of the present project is to perform in vivo preclinical testing of a newly developed transdermal therapeutic system (TTS) containing capsaicin and/or other active compounds, as well as to demonstrate their antinociceptive effect and other beneficial properties in rat models of plantar incision-induced thermal allodynia, and carrageenan-induced acute inflammatory mechanical hyperalgesia.