Development of brain organoid models to study the role of cellular senescence in the pathogenesis of glioblastoma
Provider:
Czech Science Foundation
Investigator:
Zdenek Hodny (AS CR), Dasa Bohaciakova (MUNI), Marie Vajrychova (University hospital Hradec Kralove)
Abstract:
N-methyl-D-aspartate receptors (NMDARs) are essential for developing many central nervous system diseases. Although approved blockers acting at NMDARs are available, including memantine and ketamine, a high demand exists for developing compounds acting by a different mechanism. In this project, we will investigate novel potent blockers of GluN1/GluN2 NMDARs based on the structure of K2060 that will (i) exhibit enhanced "membrane-to-channel" inhibition and (ii) exert different inhibition kinetics even in the presence of Mg2+. We will evaluate the inhibitory effect of K2060 derivatives by electrophysiology in HEK293 cells expressing normal (WT) and mutant NMDAR, as well as in hippocampal neurons from WT and "knock-in" (KI; carrying the GluN2A-N615S mutation) mice, in combination with computational approaches (QSAR, docking, and modeling). Finally, using in vivo approaches, we will determine how the observed in vitro parameters affect therapeutic efficacy in modulating audiogenic seizures in the KI mouse model.
Project duration:
1-January 2024–31-December 2026
Novel open-channel blockers of NMDA receptors with enhanced membrane-to-channel inhibition
Provider:
Czech Science Foundation
Investigator:
Ondrej Soukup (University hospital Hradec Kralove), Martin Horak (AS CR)
Abstract:
N-methyl-D-aspartate receptors (NMDARs) are essential for developing many central nervous system diseases. Although approved blockers acting at NMDARs are available, including memantine and ketamine, a high demand exists for developing compounds acting by a different mechanism. In this project, we will investigate novel potent blockers of GluN1/GluN2 NMDARs based on the structure of K2060 that will (i) exhibit enhanced "membrane-to-channel" inhibition and (ii) exert different inhibition kinetics even in the presence of Mg2+. We will evaluate the
inhibitory effect of K2060 derivatives by electrophysiology in HEK293 cells expressing normal (WT) and mutant NMDAR, as well as in hippocampal neurons from WT and "knock-in" (KI; carrying the GluN2A-N615S mutation) mice, in combination with computational approaches (QSAR, docking, and modeling). Finally, using in vivo approaches, we will determine how the observed in vitro parameters affect therapeutic efficacy in modulating audiogenic seizures in the KI mouse model.
Project duration:
1-January 2024–31-December 2026
Biotransformation of tacrine implicating its hepatotoxicity
Provider:
Czech Science Foundation
Investigator:
Ondrej Soukup (University hospital Hradec Kralove)
Abstract:
In this project we would like to explore the toxicity of tacrine which led to its withdrawal from the
market. First, we need to reveal the toxic mechanism of tacrine, i.e., to confirm the current
hypothesis of the toxic pathway 7-OH tacrine- quinone methide – hepatotoxicity. Secondly, we
aim to prove that by a suitable substitution on the tacrine core, we may prevent triggering of the
toxic cascade and develop a compound possessing favorable pharmacological properties while
lacking the toxic ones. We will used human primary spheroides to evaluate the biotransformation of tacrine, and by use of different -species orignating material, we will select the most “man-like” animal model, where our hypothesis will be confirmed. We also believe that by confirming our hypotesis we open a new avenue in the drug design of potential drugs based on tacrine scaffold
avoiding toxic biotransformation and subsequent hepatotoxicity.
Project duration:
1-January 2023–31-December 2025
Structure-activity guided design of novel long-acting antagonists of muscarinic receptors
Provider:
Czech Science Foundation
Investigator:
Jan Jakubik, Jan Korabecny (University hospital Hradec Kralove)
Abstract:
Given the broad range of functions that muscarinic receptors subserve, it is of fundamental importance to find subtype-selective ligands for therapeutic use in specific disorders. Antagonists with long-residence time at receptors are acting longer which allows for a lower daily dose to reach a maximal therapeutic effect. A fundamentally novel class of antagonists are PAM-antagonists that exert positive cooperativity with endogenous neurotransmitter. PAMantagonists preferentially block activated receptors. PAM-antagonists of muscarinic receptors have a therapeutic potential in a selective reversal of persistent excessive agonism under certain pathological conditions, like bronchospasm in asthma. We aim to delineate the molecular mechanisms underlying long-residence time and PAM-antagonism. Based on this knowledge we will design novel long-acting PAM-antagonists.
Project duration:
1-January 2023–31-December 2025
Pnpt1 as a regulator of metabolic reprogramming in phagocytes
Provider:
Czech Science Foundation
Investigator:
Tomas Rousar (University of Pardubice), Ivo Fabrik (University hospital Hradec Kralove)
Abstract:
Excessive activation of tissue resident phagocytes can lead to tissue damage and
autoimmunity. It is known that the pathology is driven mainly by specific phagocyte subtypes
which reprogram their immunometabolism to fuel inflammation. However, the underlying
mechanisms controlling phagocyte metabolic rewiring remain poorly understood. Our data
suggest that the oxidative metabolism in activated phagocytes is regulated by mitochondrial
exoribonuclease Pnpt1. In this project we aim to describe molecular mechanisms and functional
impacts of Pnpt1-dependent regulation of immunometabolism in activated phagocytes in the
context of immune response. To achieve this, we will use modern omics methods, high
resolution respirometry, and functional assays applied to Pnpt1-deficient cells. The obtained
data will provide new insights into the dynamics and control of mitochondrial metabolism in
activated phagocytes.
Project duration:
1-January 2023–31-December 2025
Minimization and potential utilization of negative effects of high temperature of column and mobile phase in proteomic analyzes
Provider:
Czech Science Foundation
Investigator:
Juraj Lenco (Faculty of Pharmacy in Hradec Kralove), Ivo Fabrik (University hospital Hradec Kralove)
Abstract:
High-temperature liquid chromatography (HTLC) significantly improves the separation of peptides. Thanks to the progress in stationary phase development, it is currently possible to separate peptides at column temperatures of 80-90 °C. At such high temperatures, maximum separation efficiency can be achieved at the flow rates commonly used in peptide analysis by liquid chromatography with mass detection (LC-MS). Nevertheless, our recent study revealed that the combination of elevated column temperature and acidic pH of the mobile phase induces cleavage and artificial modification of peptides. Therefore, we aimed this project to develop HTLC methods in which the risk of cleavage and modification of peptides will be eliminated or at least significantly reduced while maintaining the advantages of peptide separation at high column temperatures. Contrariwise, in the second research direction, we will exploit thermally accelerated acid hydrolysis of peptide bonds to develop a straightforward online method for protein cleavage for fast proteomic LC-MS analyses.
The project aims to develop high-temperature liquid chromatography methods for highly efficient separation of peptides without significant risks of their thermal degradation and, contrariwise, methods that will use thermally accelerated acid peptide bonds hydrolysis for fast online protein cleavage.
Project duration:
1-April 2022–31-December 2024
Novichok nerve agents - toxicity and countermeasures
Provider:
Czech Science Foundation
Investigator:
Daniel Jun (University of Defence), Ondřej Soukup (University hospital Hradec Kralove)
Abstract:
Despite all the chemical weapons eradication efforts made by the Chemical Weapons Convention, nerve agents still represent a permanent threat both in the military and civilian sector. A novichok is a group of nerve agents of the fourth generation, which differ in their scaffold from known nerve agents and there is a lack of information about their toxicity and efficacy of therapeutically used oxime reactivators in the treatment of intoxication. Therefore, there is a necessity to thoroughly evaluate the toxic effects of novichoks and to find the effective treatment of the poisoning with these nerve agents. In this project, we would like to systematically study and uncover all the important aspects of novichoks toxicity and available countermeasures.
Project duration:
1-January 2022–31-December 2024
The concept of rationally designed triple agents against Alzheimer's disease
Provider:
Czech Science Foundation
Investigator:
Jan Korabecny (Uhospital Hradec Kralove); Martin Horak (Czech Academy of Science)
Abstract:
Alzheimer's disease is a severe neurologic disorder with an enormous economic burden. Current treatments only temporarily delay the disease progression. The main limiting factor for developing new therapeutic compounds is the complexity of the disease. Thus, a new group of potential drugs called multi-target directed ligands (MTDLs) emerged as an alternative option to combat the disease. To this date, dozens of MTDLs have been published; however, their clinical potential is minimal, probably due to random selection of biological targets. In the proposed project, we will validate the concept of MTDLs with a rationally designed series of small molecules based on quinoline and quinazoline derivates, which simultaneously target impaired cholinergic, glutamatergic, and monoaminergic neurotransmission. The putative targets of these molecules are cholinesterases, monoamine oxidases, and NMDA receptors. To confirm the neuroprotective and procognitive efficacy of the novel molecules, we will employ a battery of in vitro, in silico, and in vivo experiments.
Project duration:
1-January 2022–31-December 2024
Non-specific and versatile cholinesterase reactivators to countermeasure organophosphorus intoxication
Provider:
Czech Science Foundation
Investigator:
Jana Zdarova Karasova, Lukas Gorecki
Abstract:
Even nowadays, there is no sufficient protection against the most toxic chemical weapons of mass destruction. Current causal antidotes are based on reactivation of enzyme acetylcholinesterase (AChE). The most of the clinical and experimental reactivators are composed of two parts, one with oxime group responsible for reactivation and the other serves as an anchor to enzyme. This molecule act as “bipolar” and could have productive or nonproductive orientation. This situation could be reversed by addition of another oxime group into molecule. We will prepare series of hybrids with two reactivating moieties. These compounds will be designed to be effective against both cholinesterases AChE and butyrylcholinesterase (BChE) inhibited by various organophosphates. We will use specific fragments that are known for their different selectivity and activity. Structure‑activity relationship will be observed in vitro on both enzymes AChE and BChE. Two most versatile compounds will be investigated in vivo for toxicity assay, their pharmacokinetics, and in vivo efficacy will be determined.
Aim of the project is preparation of non‑selective reactivators of cholinesterase. We will find candidate with wide range of action against inhibited cholinesterases with various organophosphates. Highlighted compounds will be inspected in vivo as a proof of concept.
Project duration:
1-January 2022–31-December 2024
