National projects

At the national level, within the submission of project applications, we most often participate in the project calls of the following grant agencies: GAČR, NAZV, TAČR, AZV, MPO, MŽP, MV, IGA Mendelu

LPR

Ferritin nanocages as advanced platforms for targeted drug delivery to neuroblastoma
Program:
Number:
Provider: LPR
Investigator: Ing. Simona Rex, Ph.D.
Project solution period: 2021

Abstract:

The project will use ferritin nanocells prepared by molecular cloning and protein engineering techniques. To achieve active targeting of selected molecules overexpressed on neuroblastoma cell membranes, the surface of ferritin cages will be modified with short biomimetic peptides derived from ligands with natural affinity for these molecules, such as conotoxins, neurotrophins, neurotrophic factors and the target molecules themselves. The sequence of these peptides will be designed using in silico molecular dynamics and docking techniques. Clinically used cytostatics with high toxicity to non-target cells (such as doxorubicin) and experimental cytotoxic molecules (such as ellipticine) will be delivered. Modern biophysical and biochemical methods will be used for complex characterization of prepared nanocarriers. Much attention will be paid to specific toxicity to neuroblastoma target cells comparable to that of the free drug, while ensuring the protection of non-target cells. Ideally, the results obtained could serve as a basis for the design of selective drug nanocarriers with high translational potential for oncological nanotherapy.

MV

An ultrafast portable system to detect SARS-CoV-2
Program: BV – VS. 4
Number: VI04000057
Provider: MV
Investigator:

Co-investigator:

prof. Pavel Neužil, Ph.D. (VUT)

doc. RNDr. Ondřej Zítka, Ph.D.

Project solution period: 2018 – 2021

Abstract:

The goal of this project is to develop a unique portable system to diagnose the presence of SARS-CoV-2 virus within 20 min based on its specific RNA sequence. It will be achieved by using a microfluidic system where we load the sample potentially containing the virus of interest. The sample will be automatically processed following amplification and detection of nucleic acids, assuming the ones from the virus will be presented in the sample. Key system parameters are high sensitivity, small size and low manufacturing cost thus this proposed system would become a common tool used by mobile health workers and other security personnel of the Czech Republic. During future pandemic similar to current COVID-19 the system will be modified to quickly identify the pathogen source to be quarantine

AZV ČR

Ferritin as a tool for enzymes-directed prodrug activation
Program: VES 2021
Number: NU21J-08-00043
Provider: AZV ČR
Investigator: Mgr. Zbyněk Heger, Ph.D.
Project solution period: 2021 – 2024

Abstract:

Targeted therapy, delivering biologically active substances to tumors without interacting with the healthy cells, is the golden grail of cancer chemotherapy. Despite some therapeutics benefiting from targeted therapy have been employed in anticancer treatment, their widespread use is complicated by severe drawbacks. The proposed project aims to establish a novel concept of smart therapy, based on active transport of enzymes for a catalytic activation of nontoxic prodrugs. In the project, advanced bio-nano delivery vehicles – ferritins for active transport of enzymes will be developed. In the project, broad spectrum of prodrugs inactivated by enzyme- sensitive protective masking moieties will be investigated. Activation kinetics of enzyme-prodrug pairs will be examined in vitro and in vivo, while a particular attention will be paid towards elimination of side-effects with a simultaneous enhancement of therapeutic efficiency.

Smart biocompatible nanotools for a selective delivery of drug-siRNA cocktails for combination therapy of breast cancer
Program: VES 2020
Number: NU20-03-00477
Investigator: Prof. Ing. Kamil Kuča, Ph.D. – Fakultní nemocnice Hradec Králové
Co-investigator: prof. RNDr. Vojtěch Adam, Ph.D.
Project solution period: 2020 – 2023

Abstract:

Advances in understanding of cancer biology have only slowly been translated into improvements in cancer care. Among the main reasons belongs the lack of selective transport of anticancer drugs towards tumor tissue. With these limitations in mind, and in attempts to apply Paul Ehrlich´s “magic bullet” concept, liposomes and polymeric drugs were developed in 1960s. Recent developments in nanotechnologies are expected to improve drug delivery, primarily through enabling the drug solubilization, protection from degradation and decreasing the side effects. Nanomaterials can be functionalized with biomolecules, enabling to target specific cells within certain tissues or even specific organelles. However, only little improvements have been achieved in case of chemoresistant diseases, which is most likely due to inefficiency of cargo. Therefore, we propose to combine cutting-edge smart bionanotechnologies based on biocompatible protein nanocages – ferritins, with a heterogeneous cargo of short interfering RNA (to make cells vulnerable) and cytostatic drugs (a killing ingredient).

Non-invasive biomarker analyses for early detection of urinary tract malignancies
Program: VES 2019
Number: NV19-03-00208
Investigator: prof.MUDr. Dalibor Pacík, CSc.
Co-investigator: prof. RNDr. Vojtěch Adam, Ph.D.
Project solution period: 2019 – 2022

Abstract:

Urinary tract malignancies (UTM), involving bladder and kidney carcinoma, are one of the most frequently diagnosed cancers causing death. There is an urgent need for search of suitable, noninvasive biomarkers that would allow for early diagnosis of UTM. Metabolomic profiles of the cancer cells represent unique phenotypes, suitable for identification of biomarkers specific for development stages and progression of the cancer. Therefore the present clinical study is focused on identification of potential volatile biomarkers in urine samples of bladder and kidney carcinoma using the approach of solid phase microextraction (SPME) coupled to gas chromatography with mass spectrometry detection. Different coatings of the SPME fibers, applying nanomaterials, will be used in order to select the optimal conditions for the trapping of volatile biomarkers. Subsequently, the method will be applied in the clinical cohort of one hundred samples for each type of carcinoma. The novelty of this project relay on the application of the nanomaterials for the identification of noninvasive biomarkers, which may considerably improve the timely diagnosis of UTM.

Role of monocyte metabolism and transcription factor networks in progression of human sepsis.
Program: VES 2018
Number: NV18-06-00529
Provider: AZV ČR
Investigator:

Co-investigator:

Mgr. Jan Frič, Ph.D. FNUSA-ICRS

Ing. Lucie Vaníčková, Ph.D.

Project solution period: 2018 – 2021

Abstract:

The host immune system plays a critical role in the development and
progression of sepsis, but the mechanisms that lead to cellular dysfunction, organ failure and death remain poorly defined. Peripheral blood monocytes are a major source of the effector molecules that are closely correlated with sepsis development and clinical outcomes. We aim to dissect the signalling pathways and transcription factor networks that control monocyte function in human sepsis. We will also use a newly-developed 3D organoid model to assess how patient monocyte signalling is influenced by lung tissue, which is the most
common site of infection in sepsis and serves as a reservoir of monocytes for release into the blood. Given that sepsis-linked changes in monocyte function alter the metabolite content of blood, we will also use a metabolomic approach to identify novel biomarkers within the volatile fraction of patient serum. This project will better define the molecular determinants of monocyte activity in human sepsis and assist the identification of new prognostic biomarkers for use in the clinic.

GAČR

Encapsulation of cholinesterase reactivators using apoferritin for enhanced bioavailability in central nervous systém
Program: Standardní projekty
Number: 19-13628S
Provider: GAČR
Investigator:

Co-investigator:

Doc. PharmDr. Kamil Musílek, Ph.D.

Mgr. Zbyněk Heger, Ph.D.

Project solution period: 2019 – 2021

Abstract:

Organophosphorus compounds are produced as chemical warfare agents or used as insecticides and cause life endangering intoxications. For the treatment of intoxication, acetylcholinesterase reactivators (oximes) are used as causal antidotes. Currently, clinically used or promising experimental reactivators (so-called quaternary reactivators) have major limitation in a very limited penetration into the central nervous system, where intoxication and irreversible changes in nerve tissue also occur. For this reason, biocompatible delivery systems are developed to encapsulate the reactivator´s molecules, transfer them to the central nervous system, and then release for reactivation of organophosphorus intoxication. This project focuses on acetylcholinesterase reactivators encapsulated with apoferritin and preclinical research on their bioavailability in the central nervous system.

Paperfluidic-based rapid low cost portable analytical devices with instrumentation-free readout
Program: Standardní projekty
Number: 19-02108S
Provider: GAČR
Investigator:

Co-investigator:

prof. RNDr. Miroslav Macka, Ph.D.

doc. Markéta Vaculovičová, Ph.D.

Project solution period: 2019 – 2021

Abstract:

Paper-based microfluidic analytical devices (μ-PADs) significantly advance the options of rapid and low-cost portable analysis. Instrumentation-free readout based on distance-based detection enhances the value of μ-PADs due to their extremely low cost, and simplicity of use without any instrumentation required. Selective reagents have to be immobilised on the μ-PAD to achieve formation of an accurate reaction boundary allowing distance-based detection, but so far only very limited range of immobilised reagent have been used. Herein we propose new ways of reagent immobilisation, thus broadening the range of reagents that can be used. We propose to utilise electrostatic interaction on μ-PADs with paper substrate modified to carry ionic charges, and explore other approaches including paper-embedded nano- or microparticles. Finally, integrated sample preparation compatible with distance-based detection μ-PADs will be studied for the determination of selected (bio)analytes in samples with various types of matrices.

Diffusive gradient in thin film technique (DGT): an effective tool for predicting mercury
bioavailability
Program: Standardní projekty
Number: 19-11528S
Provider: GAČR
Investigator: doc. Mgr. Pavlína Pelcová, Ph.D.
Project solution period: 2019 – 2021

Abstract:

The proposed project deals with the optimisation and utilisation of the diffusive gradient in thin film technique (DGT) for the determination of bioavailable mercury species (Hg2+, CH3Hg+,C2H5Hg+, C6H5Hg+) in contaminated soil and aquatic environment (in the locality of Jedová
hora – Brdy, Czech Republic), and with the assessing the capability of the diffusive gradient in thin film (DGT) technique to predict the bioavailability of mercury for agricultural crops and for aquatic plants. Furthermore, the influence of acid rain and soil parameters on the transport of mobile mercury forms from soil to plants will be also observed, and the contamination of the Záskalská aquatic ecosystem, which is located at the foothills of Jedová hora, will be evaluated.
The obtained results will serve to better understand the mercury bioaccumulation in similarly
mercury-contaminated areas.

MŠMT

Multidisciplinary research to increase the application potential of nanomaterials in agricultural practice
Program: OP VVV – PAV
Number: CZ.02.1.01/0.0/16_025/0007314
Provider: MŠMT
Investigator: prof. RNDr. Vojtěch Adam, Ph.D.
Project solution period: 2018 – 2022

Abstract:

The project uses a multidisciplinary approach for the design, development and testing of advanced nanomaterials, mainly based on selenium, but also other metals, semi-metals and biomacromolecular materials, as alternatives to antibiotics. The developed nanomaterials will be applied to three areas in order to accelerate their use in agricultural practice. They will be tested to validate their application potential, primarily in veterinary and phytosanitary medicine, where bacterial resistance presents major socio-economic problems. The project itself contains four interlinked research projects:

• VZ1 Advanced nanomaterials and the use of protein cages for their targeted transport
• VZ2 Use of advanced nanomaterials in the treatment of mastitis in livestock
• VZ3 Use of advanced nanomaterials to ensure sterility and antimicrobiality of coating materials in agricultural practice
• VZ4 Use of advanced nanomaterials for the treatment of plants against bacterial strains of Xanthomonas campestris pv. Campestris

The main output of the project will be, on the one hand, publications in highly impacted ISI indexed journals, to which all presentations of the research activity of the VZ1 research plan will be directed, and, on the other hand, three international patent applications, which will be the output of VZ2, VZ3 and VZ4. Specifically, it will be research and development of intramammary injection as protection against mastitis (VZ2), sterile covering material with antimicrobial properties (VZ3) and plant protection product (VZ4). All three outputs are directed to the field of agricultural practice, and their use will be a significant socio-economic benefit in this sector.

TAČR

Nanocomposite materials based on sulfur in crop nutrition
Program: ZÉTA
Number: TJ04000519
Provider: TAČR
Investigator: Ing. Dalibor Húska Ph.D.
Project solution period: 2020 – 2022
Gastro-processing waste into a solid carbon product for material use
Program: ZÉTA
Number: TJ02000262
Provider: TAČR
Investigator: Ing. Martina Vršanská, Ph.D.
Project solution period: 2019 – 2021

Abstract:

The aim of this project is treatment of gastro waste (GW) into a solid carbonaceous product for material utilization by drying, pretreatment with the addition of additives for the pyrolysis process. The first is focused on defining energy in dried GW, as well as pyrolysis products: biochar, pyrolysis oil and syngas. The second direction focuses on the use of biochar in agriculture. The third direction focuses on biochar as a filter medium for removing pollutants from WW. Part of the project is design, production and testing of a functional sample: drying equipment, pyrolysis and filtration units. The project will determine which direction will be acceptable from the point of view of legislation, technical solution and economy. This goal will be achieved at the end of this project.

IGRÁČEK

An Invisible Threat: Nanoplastics VS. microalgae, bacteria and plants
Program: SP
Number: SGC-2021-012
Provider: IGRÁČEK
Investigator: Mgr. Ondřej Pěnčík
Project solution period: 2021 – 2023

Abstract:

350-400 tonnes of plastic waste are produced worldwide. This amount increases every year – especially in the current pandemic situation with the usage of disposable items. PET plastics are important pollutants in the aquatic environment. It has long been known that they can form microparticles and nanoparticles that affect the health of the environment and humans. At the same time, information on the effects is incomplete, so further research is required. This project is trying to fix it, because main aim of the project will be to map the behavior of PET nanoplastics in aquatic and terrestrial environments.

An Invisible Threat: Nanoplastics VS. microalgae, bacteria and plants
Program: SP
Number: SGC-2021-005
Provider: IGRÁČEK
Investigator: Ing. Martin Řiháček
Project solution period: 2021 – 2023

Abstract:

Zinc oxide (ZnO) and zinc oxide nanoparticles (ZnO NPs) have been used in agriculture as fertilizers and feed supplements. In the European Union (EU), from June 2022, due to contamination of the environment, zinc will be allowed as a feed additive only in quantities that meet daily animal requirements. Application of zinc for extended periods of time promotes induction and spread of antibiotic resistance and virulence traits among bacteria in the digestive tract of domestic animals and the environment.

Our research is focused on biology of Escherichia coli exposed to sub-inhibitory concentrations of ZnO/ZnO NPs for 40 subcultivations. For 20 sub-culturings, E. coli will be treated with ZnO/ZnO NPs. From 20th to 40th sub-culturing, bacteria will be separated into 2 aliquots: the first aliquot will remain exposed to ZnO/ZnO NPs and the second aliquot will be without treatment. Selected sub-cultures (0th, 5th, 10th, 20th and 40th) will be exposed to antibiotics: aminoglycosides, carbapenems, cephalosporins, penicillins, sulfonamides, fluoroquinolones, and tetracyclines and minimum inhibitory concentration (MIC) will be measured to determine the effect of zinc on development of antibiotic resistance. Furthermore, the effect of zinc treatments on biology of E. coli after 40 sub-culturings will be studied by: A) genome sequencing by MiniSeq, Illumina, B) transcriptome sequencing by NextSeq 500 Illumina, C) proteome analysis by nanoflow reverse-phase liquid chromatography-mass spectrometry, and D) metabolome analysis by Agilent Technologies 6460 Triple Quad LC/MS with focus on metabolites in the Krebs cycle. Selected differentially expressed genes will be confirmed by the RT-qPCR. The obtained multiple omics data will be combined and analyzed with the goal to understand how E. coli adapts to exposure of ZnO/ZnO on genomic, transcriptomic, proteomic and metabolomic levels. PhD students will learn how to prepare, process, and analyze samples on these four omics levels in collaboration with different research groups with specific expertise at Mendelu. In addition, the principal investigator will attend an internship at the University of Koln, Germany to learn new skills in transcriptomics. The output of this project is also to publish two articles in a top peer reviewed journals.