Katarzyna Anna Radaszkiewicz

Masaryk University

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Jiřı́ Pachernı́k

Masaryk University

Petr Humpolíček

Tomas Bata University in Zlín

Tomasz Radaszkiewicz

Masaryk University

Věra Kašpárková

Tomas Bata University in Zlín

Zdenka Capáková

Tomas Bata University in Zlín

Marián Lehocký

Tomas Bata University in Zlín

Tomáš Gybeľ

Masaryk University

Jaroslav Stejskal

Tomas Bata University in Zlín

Lukáš Kubala

Czech Academy of Sciences, Institute of Biophysics

Antonín Minařík

Tomas Bata University in Zlín

Cooperative Institutions

Masaryk University

Czech Academy of Sciences

Tomas Bata University in Zlín

Czech Academy of Sciences, Institute of Biophysics

University Hospital Brno

St. Anne's University Hospital Brno

Charles University

Central European Institute of Technology

Central European Institute of Technology – Masaryk University

International Clinical Research Center, St. Anne's University Hospital Brno

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Cell Differentiation and Aging Lead To Up-Regulation of FTO, While the ALKBH5 Protein Level Was Stable During Aging but Up-Regulated During in vitro-Induced Cardiomyogenesis

Physiological Research (2023)

Jana Krejčí OA Arcidiacono Radim Čegan Katarzyna Anna Radaszkiewicz Jiřı́ Pachernı́k

FTO and ALKBH5 proteins are essential erasers of N6-adenosine methylation in RNA. We studied how levels of FTO and ALKBH5 proteins changed during mouse embryonic development, aging, cardiomyogenesis, and neuroectodermal differentiation. We observed that aging in male and female mice was associated with FTO up-regulation in mouse hearts, brains, lungs, and kidneys, while the ALKBH5 level remained stable. FTO and ALKBH5 proteins were up-regulated during experimentally induced cardiomyogenesis, but the level of ALKBH5 protein was not changed when neuroectodermal differentiation was induced. HDAC1 depletion in mouse ES cells caused FTO down-regulation. In these cells, mRNA, carrying information from genes that regulate histone signature, RNA processing, and cell differentiation, was characterized by a reduced level of N6-adenosine methylation in specific gene loci, primarily regulating cell differentiation into neuroectoderm. Together, when we compared both RNA demethylating proteins, the FTO protein level undergoes the most significant changes during cell differentiation and aging. Thus, we conclude that during aging and neuronal differentiation, m6A RNA demethylation is likely regulated by the FTO protein but not via the function of ALKBH5.

RNA methylation

10.33549/physiolres.935078

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Citations (1)

Orthotopic model for the analysis of melanoma circulating tumor cells

Scientific Reports (2024)

Markéta Pícková Zuzana Kahounová Tomasz Radaszkiewicz Jiřina Procházková Radek Fedr

Abstract Metastatic melanoma, a highly lethal form of skin cancer, presents significant clinical challenges due to limited therapeutic options and high metastatic capacity. Recent studies have demonstrated that cancer dissemination can occur earlier, before the diagnosis of the primary tumor. The progress in understanding the kinetics of cancer dissemination is limited by the lack of animal models that accurately mimic disease progression. We have established a xenograft model of human melanoma that spontaneously metastasizes to lymph nodes and lungs. This model allows precise monitoring of melanoma progression and is suitable for the quantitative and qualitative analysis of circulating tumor cells (CTCs). We have validated a flow cytometry-based protocol for CTCs enumeration and isolation. We could demonstrate that (i) CTCs were detectable in the bloodstream from the fourth week after tumor initiation, coinciding with the lymph node metastases appearance, (ii) excision of the primary tumor accelerated the formation of metastases in lymph nodes and lungs as early as one-week post-surgery, accompanied by the increased numbers of CTCs, and (iii) CTCs change their surface protein signature. In summary, we present a model of human melanoma that can be effectively utilized for future drug efficacy studies.

Circulating tumor cell

Primary tumor

10.1038/s41598-024-58236-y

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Stimuli-responsive thin film composites of conducting polymers and cellulose nanocrystals for tissue engineering

International Journal of Biological Macromolecules (2024)

Eva Korábková Zhanna A. Boeva Katarzyna Anna Radaszkiewicz Kateřina Skopalová Věra Kašpárková

Polypyrrole

10.1016/j.ijbiomac.2024.131036

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The role of prickle proteins in vertebrate development and pathology

Molecular and Cellular Biochemistry (2023)

Katarzyna Anna Radaszkiewicz Marie Šulcová E. Kohoutkova Jakub Harnoš

Abstract Prickle is an evolutionarily conserved family of proteins exclusively associated with planar cell polarity (PCP) signalling. This signalling pathway provides directional and positional cues to eukaryotic cells along the plane of an epithelial sheet, orthogonal to both apicobasal and left–right axes. Through studies in the fruit fly Drosophila , we have learned that PCP signalling is manifested by the spatial segregation of two protein complexes, namely Prickle/Vangl and Frizzled/Dishevelled. While Vangl, Frizzled, and Dishevelled proteins have been extensively studied, Prickle has been largely neglected. This is likely because its role in vertebrate development and pathologies is still being explored and is not yet fully understood. The current review aims to address this gap by summarizing our current knowledge on vertebrate Prickle proteins and to cover their broad versatility. Accumulating evidence suggests that Prickle is involved in many developmental events, contributes to homeostasis, and can cause diseases when its expression and signalling properties are deregulated. This review highlights the importance of Prickle in vertebrate development, discusses the implications of Prickle-dependent signalling in pathology, and points out the blind spots or potential links regarding Prickle, which could be studied further.

Dishevelled

Signalling

10.1007/s11010-023-04787-z

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Conducting composite films based on chitosan or sodium hyaluronate. Properties and cytocompatibility with human induced pluripotent stem cells

Carbohydrate Polymers (2020)

Daniela Jasenská Věra Kašpárková Katarzyna Anna Radaszkiewicz Zdenka Capáková Jiřı́ Pachernı́k

10.1016/j.carbpol.2020.117244

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Citations (19)

Hierarchically Structured Polystyrene-Based Surfaces Amplifying Fluorescence Signals: Cytocompatibility with Human Induced Pluripotent Stem Cell

International Journal of Molecular Sciences (2021)

Kateřina Skopalová Katarzyna Anna Radaszkiewicz Markéta Kadlečková Jiřı́ Pachernı́k Antonín Minařík

An innovative multi-step phase separation process was used to prepare tissue culture for the polystyrene-based, hierarchically structured substrates, which mimicked in vivo microenvironment and architecture. Macro- (pore area from 3000 to 18,000 µm2; roughness (Ra) 7.2 ± 0.1 µm) and meso- (pore area from 50 to 300 µm2; Ra 1.1 ± 0.1 µm) structured substrates covered with micro-pores (area around 3 µm2) were prepared and characterised. Both types of substrate were suitable for human-induced pluripotent stem cell (hiPSC) cultivation and were found to be beneficial for the induction of cardiomyogenesis in hiPSC. This was confirmed both by the number of promoted proliferated cells and the expressions of specific markers (Nkx2.5, MYH6, MYL2, and MYL7). Moreover, the substrates amplified the fluorescence signal when Ca2+ flow was monitored. This property, together with cytocompatibility, make this material especially suitable for in vitro studies of cell/material interactions within tissue-mimicking environments.

Polystyrene

10.3390/ijms222111943

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Distribution of formins in cardiac muscle: FHOD1 is a component of intercalated discs and costameres

European Journal of Cell Biology (2014)

Abdulatif Al Haj Antonina Joanna Mazur Katarzyna Anna Radaszkiewicz Tomasz Radaszkiewicz Aleksandra Makowiecka

Immunostaining

Polyclonal antibodies

Intercalated disc

10.1016/j.ejcb.2014.11.003

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Citations (18)

Linking planar polarity signalling to actomyosin contractility during vertebrate neurulation

Open Biology (2024)

Sarka Novotna Lorena A. Maia Katarzyna Anna Radaszkiewicz Pavel Roudnický Jakub Harnoš

Actomyosin contractility represents an ancient feature of eukaryotic cells participating in many developmental and homeostasis events, including tissue morphogenesis, muscle contraction and cell migration, with dysregulation implicated in various pathological conditions, such as cancer. At the molecular level, actomyosin comprises actin bundles and myosin motor proteins that are sensitive to posttranslational modifications like phosphorylation. While the molecular components of actomyosin are well understood, the coordination of contractility by extracellular and intracellular signals, particularly from cellular signalling pathways, remains incompletely elucidated. This study focuses on WNT/planar cell polarity (PCP) signalling, previously associated with actomyosin contractility during vertebrate neurulation. Our investigation reveals that the main cytoplasmic PCP proteins, Prickle and Dishevelled, interact with key actomyosin components such as myosin light chain 9 (MLC9), leading to its phosphorylation and localized activation. Using proteomics and microscopy approaches, we demonstrate that both PCP proteins actively control actomyosin contractility through Rap1 small GTPases in relevant in vitro and in vivo models. These findings unveil a novel mechanism of how PCP signalling regulates actomyosin contractility through MLC9 and Rap1 that is relevant to vertebrate neurulation.

Contractility

Dishevelled

10.1098/rsob.240251

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The acceleration of cardiomyogenesis in embryonic stem cells in vitro by serum depletion does not increase the number of developed cardiomyocytes

PLoS ONE (2017)

Katarzyna Anna Radaszkiewicz Dominika Sýkorová Lucia Binó Jana Kudová Markéta Bébarová

The differentiation of pluripotent embryonic stem (ES) cells into various lineages in vitro represents an important tool for studying the mechanisms underlying mammalian embryogenesis. It is a key technique in studies evaluating the molecular mechanisms of cardiomyogenesis and heart development and also in embryotoxicology. Herein, modest modifications of the basic protocol for ES cell differentiation into cardiomyocytes were evaluated in order to increase the yield and differentiation status of developed cardiomyocytes. Primarily, the data show that ES cell cultivation in the form of non-adherent embryoid bodies (EBs) for 5 days compared to 8 days significantly improved cardiomyogenic differentiation. This is illustrated by the appearance of beating foci in the adherent EBs layer at earlier phases of differentiation from day 10 up to day 16 and by the significantly higher expression of genes characteristic of cardiomyogenic differentiation (sarcomeric alpha actinin, myosin heavy chain alpha and beta, myosin light chain 2 and 7, and transcriptional factor Nkx2.5) in EBs cultivated under non-adherent conditions for 5 days. The ratio of cardiomyocytes per other cells was also potentiated in EBs cultivated in non-adherent conditions for only 5 days followed by cultivation in adherent serum-free culture conditions. Nevertheless, the alteration in the percentage of beating foci among these two tested cultivation conditions vanished at later phases and also did not affect the total number of cardiomyocytes determined as myosin heavy chain positive cells at the end of the differentiation process on day 20. Thus, although these modifications of the conditions of ES cells differentiation may intensify cardiomyocyte differentiation, the final count of cardiomyocytes might not change. Thus, serum depletion was identified as a key factor that intensified cardiomyogenesis. Further, the treatment of EBs with N-acetylcysteine, a reactive oxygen species scavenger, did not affect the observed increase in cardiomyogenesis under serum depleted conditions. Interestingly, a mild induction of the ventricular-like phenotype of cardiomyocytes was observed in 5-day-old EBs compared to 8-day-old EBs. Overall, these findings bring crucial information on the mechanisms of ES cells differentiation into cardiomyocytes and on the establishment of efficient protocols for the cardiomyogenic differentiation of ES cells. Further, the importance of determining the absolute number of formed cardiomyocyte-like cells per seeded pluripotent cells in contrast to the simple quantification of the ratios of cells is highlighted.

Embryoid body

10.1371/journal.pone.0173140

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Citations (10)

RNF43 inhibits WNT5A driven signaling and suppresses melanoma invasion

bioRxiv (Cold Spring Harbor Laboratory) (2021)

Tomasz Radaszkiewicz Michaela Nosková Kristína Gömöryová Olga Vondálová Blanářová Katarzyna Anna Radaszkiewicz

Abstract RNF43 is a E3 ubiquitin ligase and known negative regulator of WNT/β-catenin signaling. We demonstrate that RNF43 is also regulator of noncanonical WNT5A-induced signaling in human cells. Analysis of the RNF43 interactome using BioID and immunoprecipitation showed that RNF43 can interact with the core receptor complex components dedicated to the noncanonical Wnt pathway such as ROR1, ROR2, VANGL1 and VANGL2. RNF43 triggers VANGL2 ubiquitination and proteasomal degradation and clathrin-dependent internalization of ROR1 receptor. This activity of RNF43 is physiologically relevant and blocks pro-metastatic WNT5A signaling in melanoma. RNF43 inhibits responses to WNT5A, which results in the suppression of invasive properties of melanoma cells. Furthermore, RNF43 prevented WNT5A-assisted development of resistance to BRAF V600E inhibitor. In line with these findings, RNF43 expression decreases during melanoma progression and RNF43-low patients have worse prognosis. We conclude that RNF43 is a newly discovered negative regulator of WNT5A-mediated biological responses that desensitizes cells to WNT5A.

ROR1

Internalization

WNT5A

10.1101/2021.02.08.430210

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Citations (2)