Latest Publications

@article{ondrisova_foxo1rictor_2024,

title = {FoxO1/Rictor axis induces a non-genetic adaptation to Ibrutinib via Akt activation in chronic lymphocytic leukemia.},

author = {Laura Ondrisova and Vaclav Seda and Krystof Hlavac and Petra Pavelkova and Eva Hoferkova and Giorgia Chiodin and Lenka Kostalova and Gabriela Mladonicka Pavlasova and Daniel Filip and Josef Vecera and Pedro Faria Zeni and Jan Oppelt and Zuzana Kahounova and Rachel Vichova and Karel Soucek and Anna Panovska and Karla Plevova and Sarka Pospisilova and Martin Simkovic and Filip Vrbacky and Daniel Lysak and Stacey M. Fernandes and Matthew S. Davids and Alba Maiques-Diaz and Stella Charalampopoulou and Jose I. Martin-Subero and Jennifer R. Brown and Michael Doubek and Francesco Forconi and Jiri Mayer and Marek Mraz},

doi = {10.1172/JCI173770},

issn = {1558-8238 0021-9738},

year  = {2024},

date = {2024-10-01},

journal = {The Journal of clinical investigation},

pages = {e173770},

abstract = {BTK inhibitor therapy induces peripheral blood lymphocytosis in chronic lymphocytic leukemia (CLL) lasting for several months. It remains unclear whether non-genetic adaptation mechanisms exist, allowing CLL cells' survival during BTK inhibitor-induced lymphocytosis and/or playing a role in therapy resistance. We show that in approximately 70 % of CLL cases, ibrutinib treatment in vivo increases Akt activity above pre-therapy levels within several weeks, leading to compensatory CLL cell survival and a more prominent lymphocytosis on therapy. Ibrutinib-induced Akt phosphorylation (pAktS473) is caused by the upregulation of FoxO1 transcription factor, which induces expression of Rictor, an assembly protein for mTORC2 protein complex that directly phosphorylates Akt at serine 473 (S473). Knock-out or inhibition of FoxO1 or Rictor led to a dramatic decrease in Akt phosphorylation and growth disadvantage for malignant B cells in the presence of ibrutinib (or PI3K inhibitor idelalisib) in vitro and in vivo. FoxO1/Rictor/pAktS473 axis represents an early non-genetic adaptation to BCR inhibitor therapy not requiring PI3Kδ or BTK kinase activity. We further demonstrate that FoxO1 can be targeted therapeutically, and its inhibition induces CLL cells' apoptosis alone or in combination with BTK inhibitors (ibrutinib, acalabrutinib, pirtobrutinib) and blocks their proliferation triggered by T-cell factors (CD40L, IL-4, and IL-21).},

note = {Place: United States},

keywords = {Drug therapy, Hematology, Leukemias, Oncology, Signal Transduction},

pubstate = {published},

tppubtype = {article}

}

@article{kvokackova_single-cell_2023,

title = {Single-cell protein profiling defines cell populations associated with triple-negative breast cancer aggressiveness.},

author = {Barbora Kvokačková and Radek Fedr and Daniela Kužílková and Jan Stuchlý and Adéla Vávrová and Jiří Navrátil and Pavel Fabian and Róbert Ondruššek and Petra Ovesná and Ján Remšík and Jan Bouchal and Tomáš Kalina and Karel Souček},

doi = {10.1002/1878-0261.13365},

issn = {1878-0261 1574-7891},

year  = {2023},

date = {2023-06-01},

journal = {Molecular oncology},

volume = {17},

number = {6},

pages = {1024–1040},

abstract = {Triple-negative breast cancer (TNBC) is an aggressive and complex subtype of breast cancer that lacks targeted therapy. TNBC manifests characteristic, extensive intratumoral heterogeneity that promotes disease progression and influences drug response. Single-cell techniques in combination with next-generation computation provide an unprecedented opportunity to identify molecular events with therapeutic potential. Here, we describe the generation of a comprehensive mass cytometry panel for multiparametric detection of 23 phenotypic markers and 13 signaling molecules. This single-cell proteomic approach allowed us to explore the landscape of TNBC heterogeneity, with particular emphasis on the tumor microenvironment. We prospectively profiled freshly resected tumors from 26 TNBC patients. These tumors contained phenotypically distinct subpopulations of cancer and stromal cells that were associated with the patient's clinical status at the time of surgery. We further classified the epithelial-mesenchymal plasticity of tumor cells, and molecularly defined phenotypically diverse populations of tumor-associated stroma. Furthermore, in a retrospective tissue-microarray TNBC cohort, we showed that the level of CD97 at the time of surgery has prognostic potential.},

note = {Place: United States},

keywords = {*Triple Negative Breast Neoplasms/metabolism, Cell Line, Humans, mass cytometry, phenotypic plasticity, Proteomics, Retrospective Studies, Signal Transduction, single-cell profiles, Stromal Cells/metabolism, triple-negative breast cancer, Tumor, tumor heterogeneity, Tumor microenvironment, unsupervised machine learning algorithm},

pubstate = {published},

tppubtype = {article}

}

@article{simeckova_vitro_2022,

title = {In vitro profiling of toxic effects of environmental polycyclic aromatic hydrocarbons on nuclear receptor signaling, disruption of endogenous metabolism and induction of cellular stress.},

author = {Pavlína Šimečková and Kateřina Pěnčíková and Ondrej Kováč and Josef Slavík and Martina Pařenicová and Jan Vondráček and Miroslav Machala},

doi = {10.1016/j.scitotenv.2021.151967},

issn = {1879-1026 0048-9697},

year  = {2022},

date = {2022-04-01},

journal = {The Science of the total environment},

volume = {815},

pages = {151967},

abstract = {Polycyclic aromatic hydrocarbons (PAHs) may interact with multiple intracellular receptors and related signaling pathways. We comprehensively evaluated the toxicity profiles of six environmentally relevant PAHs differing in structure, genotoxicity and their ability to activate the aryl hydrocarbon receptor (AhR). We focused particularly on their impact on intracellular hormone-, xenobiotic- and lipid-sensing receptors, as well as on cellular stress markers, combining a battery of human reporter gene assays and qRT-PCR evaluation of endogenous gene expression in human hepatocyte-like HepaRG cells, with LC/MS-MS analysis of cellular sphingolipids. The effects of PAHs included: activation of estrogen receptor α (in case of fluoranthene (Fla), pyrene (Pyr), benz[a]anthracene (BaA), benzo[a]pyrene (BaP)), suppression of androgen receptor activity (Fla, BaA, BaP and benzo[k]fluoranthene (BkF)), enhancement of dexamethasone-induced glucocorticoid receptor activity (chrysene (Chry), BaA, and BaP), and potentiation of triiodothyronine-induced thyroid receptor α activity (all tested PAHs). PAHs also induced transcription of endogenous gene targets of constitutive androstane receptor (Fla, Pyr), or repression of target genes of pregnane X receptor and peroxisome proliferator-activated receptor α (in case of the AhR-activating PAHs - Chry, BaA, BaP, and BkF) in HepaRG cells. In the same cell model, the AhR agonists reduced the expression of glucose metabolism genes (PCK1, G6PC and PDK4), and they up-regulated levels of glucosylceramides, together with a concomitant induction of expression of UGCG, glucosylceramide synthesis enzyme. Finally, both BaP and BkF were found to induce expression of early stress and genotoxicity markers: ATF3, EGR1, GDF15, CDKN1A/p21, and GADD45A mRNAs, while BaP alone increased levels of IL-6 mRNA. Overall, whereas low-molecular-weight PAHs exerted significant effects on nuclear receptors (with CYP2B6 induction observed already at nanomolar concentrations), the AhR activation by 4-ring and 5-ring PAHs appeared to be a key mechanism underlying their impact on nuclear receptor signaling, endogenous metabolism and induction of early stress and genotoxicity markers.},

note = {Place: Netherlands},

keywords = {*Polycyclic Aromatic Hydrocarbons/toxicity, Aryl Hydrocarbon/genetics/metabolism, Benzo(a)pyrene, Cellular stress response, Cytoplasmic and Nuclear/genetics, Energy Metabolism, Humans, Nuclear receptors, Polycyclic aromatic hydrocarbons, Receptors, Signal Transduction, Sphingolipids, Xenobiotics},

pubstate = {published},

tppubtype = {article}

}

@article{vondracek_environmental_2020,

title = {Environmental six-ring polycyclic aromatic hydrocarbons are potent inducers of the AhR-dependent signaling in human cells.},

author = {Jan Vondráček and Kateřina Pěnčíková and Miroslav Ciganek and Jakub Pivnička and Martina Karasová and Martina Hýžďalová and Simona Strapáčová and Lenka Pálková and Jiří Neča and Jason Matthews and Michal Vojtíšek Lom and Jan Topinka and Alena Milcová and Miroslav Machala},

doi = {10.1016/j.envpol.2020.115125},

issn = {1873-6424 0269-7491},

year  = {2020},

date = {2020-11-01},

journal = {Environmental pollution (Barking, Essex : 1987)},

volume = {266},

number = {Pt 2},

pages = {115125},

abstract = {The toxicities of many environmental polycyclic aromatic hydrocarbons (PAHs), in particular those of high-molecular-weight PAHs (with MW higher than 300), remain poorly characterized. The objective of this study was to evaluate the ability of selected environmentally relevant PAHs with MW 302 (MW302 PAHs) to activate the aryl hydrocarbon receptor (AhR), since this represents a major toxic mode of action of PAHs. A large number of the evaluated compounds exhibited strong AhR-mediated activities, in particular in human models. The studied MW302 PAHs also significantly contributed to the overall calculated AhR activities of complex environmental mixtures, including both defined standard reference materials and collected diesel exhaust particles. The high AhR-mediated activities of representative MW302 PAHs, e.g. naphtho[1,2-k]fluoranthene, corresponded with the modulation of expression of relevant AhR target genes in a human lung cell model, or with the AhR-dependent suppression of cell cycle progression/proliferation in estrogen-sensitive cells. This was in a marked contrast with the limited genotoxicity of the same compound(s). Given the substantial levels of the AhR-activating MW302 PAHs in combustion particles, it seems important to continue to investigate the toxic modes of action of this large group of PAHs associated with airborne particulate matter.},

note = {Place: England},

keywords = {*Polycyclic Aromatic Hydrocarbons, *Receptors, AhR, Anti-estrogenicity, Aryl Hydrocarbon, Carcinogenic PAHs, Genotoxicity, Humans, Lung cell toxicity, Particulate Matter, Signal Transduction, Vehicle Emissions},

pubstate = {published},

tppubtype = {article}

}

@article{svobodova_2378-tetrachlorodibenzo-p-dioxin_2019,

title = {2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) Disrupts Control of Cell Proliferation and Apoptosis in a Human Model of Adult Liver Progenitors.},

author = {Jana Svobodová and Jiřina Procházková and Markéta Kabátková and Martin Krkoška and Lenka Šmerdová and Helena Líbalová and Jan Topinka and Jiří Kléma and Alois Kozubík and Miroslav Machala and Jan Vondráček},

doi = {10.1093/toxsci/kfz202},

issn = {1096-0929},

year  = {2019},

date = {2019-12-01},

journal = {Toxicological sciences : an official journal of the Society of Toxicology},

volume = {172},

number = {2},

pages = {368–384},

abstract = {The aryl hydrocarbon receptor (AhR) activation has been shown to alter proliferation, apoptosis, or differentiation of adult rat liver progenitors. Here, we investigated the impact of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-mediated AhR activation on a human model of bipotent liver progenitors, undifferentiated HepaRG cells. We used both intact undifferentiated HepaRG cells, and the cells with silenced Hippo pathway effectors, yes-associated protein 1 (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ), which play key role(s) in tissue-specific progenitor cell self-renewal and expansion, such as in liver, cardiac, or respiratory progenitors. TCDD induced cell proliferation in confluent undifferentiated HepaRG cells; however, following YAP, and, in particular, double YAP/TAZ knockdown, TCDD promoted induction of apoptosis. These results suggested that, unlike in mature hepatocytes, or hepatocyte-like cells, activation of the AhR may sensitize undifferentiated HepaRG cells to apoptotic stimuli. Induction of apoptosis in cells with silenced YAP/TAZ was associated with upregulation of death ligand TRAIL, and seemed to involve both extrinsic and mitochondrial apoptosis pathways. Global gene expression analysis further suggested that TCDD significantly altered expression of constituents and/or transcriptional targets of signaling pathways participating in control of expansion or differentiation of liver progenitors, including EGFR, Wnt/β-catenin, or tumor growth factor-β signaling pathways. TCDD significantly upregulated cytosolic proapoptotic protein BMF (Bcl-2 modifying factor) in HepaRG cells, which could be linked with an enhanced sensitivity of TCDD-treated cells to apoptosis. Our results suggest that, in addition to promotion of cell proliferation and alteration of signaling pathways controlling expansion of human adult liver progenitors, AhR ligands may also sensitize human liver progenitor cells to apoptosis.},

note = {Place: United States},

keywords = {*Models, Adaptor Proteins, Apoptosis, Apoptosis/*drug effects/genetics, Aryl hydrocarbon receptor, Aryl Hydrocarbon/metabolism, Biological, Cell Line, Cell Proliferation, Cell Proliferation/*drug effects/genetics, Gene Expression/drug effects, HepaRG cells, Hippo signaling, Humans, Liver/*drug effects/pathology, Polychlorinated Dibenzodioxins/*toxicity, Receptors, RNA, Signal Transducing/genetics, Signal Transduction, Small Interfering/genetics, Stem Cells/*drug effects/pathology, Trans-Activators/genetics, Transcription Factors/genetics, Transcriptional Coactivator with PDZ-Binding Motif Proteins, Transfection, YAP-Signaling Proteins},

pubstate = {published},

tppubtype = {article}

}

@article{benes_inhibition_2011,

title = {Inhibition of topoisomerase IIα: novel function of wedelolactone.},

author = {Petr Benes and Lucia Knopfova and Filip Trcka and Alice Nemajerova and Diana Pinheiro and Karel Soucek and Miroslav Fojta and Jan Smarda},

doi = {10.1016/j.canlet.2011.01.002},

issn = {1872-7980 0304-3835},

year  = {2011},

date = {2011-04-01},

journal = {Cancer letters},

volume = {303},

number = {1},

pages = {29–38},

abstract = {The naturally occurring coumestan wedelolactone has been previously shown to reduce growth of various cancer cells. So far, the growth-suppressing effect of wedelolactone has been attributed to the inhibition of the NFκB transcription factor and/or androgen receptors. We found that wedelolactone suppressed growth and induced apoptosis of androgen receptor-negative MDA-MB-231 breast cancer cells at concentrations that did not inhibit the NFκB activity. The cells responded to wedelolactone by the S and G2/M phase cell cycle arrest and induction of the DNA damage signaling. Wedelolactone interacted with dsDNA and inhibited the activity of DNA topoisomerase IIα. We conclude that wedelolactone can act as growth suppressor independently of NFκB and androgen receptors.},

note = {Place: Ireland},

keywords = {Antigens, Antineoplastic Agents/pharmacology, Apoptosis/drug effects, Breast Neoplasms/*drug therapy/enzymology/pathology, Cell Cycle/drug effects, Cell Growth Processes/drug effects, Cell Line, Cell Survival/drug effects, Coumarins/*pharmacology, DNA Damage, DNA Topoisomerases, DNA-Binding Proteins/*antagonists & inhibitors/metabolism, Enzyme-Linked Immunosorbent Assay, Female, Humans, Immunoblotting, Neoplasm/metabolism, Signal Transduction, Topoisomerase Inhibitors/*pharmacology, Tumor, Type II/metabolism},

pubstate = {published},

tppubtype = {article}

}

@article{starsichova_tgf-beta1_2010,

title = {TGF-beta1 suppresses IL-6-induced STAT3 activation through regulation of Jak2 expression in prostate epithelial cells.},

author = {Andrea Starsíchová and Eva Lincová and Zuzana Pernicová and Alois Kozubík and Karel Soucek},

doi = {10.1016/j.cellsig.2010.06.014},

issn = {1873-3913 0898-6568},

year  = {2010},

date = {2010-11-01},

journal = {Cellular signalling},

volume = {22},

number = {11},

pages = {1734–1744},

abstract = {Chronic inflammation plays an important role in the initiation and progression of various human diseases including benign prostatic hyperplasia or prostate cancer. Here we show that the proinflammatory cytokine interleukin-6 (IL-6) has prosurvival effects and chronically activates the Jak2/STAT3 signalling pathway in a model of benign prostatic hyperplasia (BPH-1). We demonstrate that the antiinflammatory cytokine transforming growth factor-beta1 (TGF-beta1), which also permanently activates its canonical signalling pathway through SMAD proteins in BPH-1 cells, modifies the effects of IL-6 on cell proliferation. Importantly, TGF-beta1 inhibits IL-6 signal transduction by decreasing the phosphorylation levels of STAT3. This effect is associated with decreased expression of Jak2 at both mRNA and protein levels. Moreover, we showed that TGF-beta1 inhibits IL-6-induced expression of the cancer-associated gene MUC1. These observations demonstrated a novel interaction between TGF-beta1 and IL-6 signalling and suggested another mechanism of how defects in TGF-beta signalling, frequently associated with prostate pathologies, can contribute to the disruption of tissue homeostasis.},

note = {Place: England},

keywords = {Cell Line, Cell Proliferation, Epithelial Cells/*metabolism, Humans, Interleukin-6/*antagonists & inhibitors/pharmacology, Janus Kinase 2/genetics/*metabolism, Male, Mucin-1/metabolism, Phosphorylation, Prostate/cytology/enzymology/*metabolism, Prostatic Hyperplasia/enzymology/*metabolism, RNA, RNA Interference, Signal Transduction, Smad Proteins/metabolism, Small Interfering/metabolism, STAT3 Transcription Factor/*metabolism, Transforming Growth Factor beta1/*pharmacology},

pubstate = {published},

tppubtype = {article}

}

@article{soucek_fetal_2010,

title = {Fetal colon cell line FHC exhibits tumorigenic phenotype, complex karyotype, and TP53 gene mutation.},

author = {Karel Soucek and Pavla Gajdusková and Marie Brázdová and Martina Hýzd'alová and Lenka Kocí and David Vydra and Radek Trojanec and Zuzana Pernicová and Lenka Lentvorská and Marián Hajdúch and Jirina Hofmanová and Alois Kozubík},

doi = {10.1016/j.cancergencyto.2009.11.009},

issn = {1873-4456 0165-4608},

year  = {2010},

date = {2010-03-01},

journal = {Cancer genetics and cytogenetics},

volume = {197},

number = {2},

pages = {107–116},

abstract = {Stable cell lines obtained by spontaneous immortalization might represent early stages of malignant transformation and be useful experimental models for studies of mechanisms of cancer development. The FHC (fetal human cells) cell line has been established from normal fetal colonic mucosa. Detailed characterization of this cell line and mechanism of spontaneously acquired immortality have not been described yet. Therefore, we characterized the FHC cell line in terms of its tumorigenicity, cytogenetics, and TP53 gene mutation analysis. FHC cells displayed capability for anchorage-independent growth in semisolid media in vitro and formed solid tumors after transplantation into SCID (severe combined immunodeficiency) mice. This tumorigenic phenotype was associated with hypotriploidy and chromosome number ranging from 66 to 69. Results of comparative genetic hybridization arrays showed that most chromosomes included regions of copy number gains or losses. Region 8q23 approximately 8q24.3 (containing, e.g., MYC proto-oncogene) was present in more than 20 copies per nucleus. Moreover, we identified mutation of TP53 gene in codon 273; triplet CGT coding Arg was changed to CAG coding His. Expression of Pro codon 72 polymorphic variant of p53 was also detected. Mutation of TP53 gene was associated with abolished induction of p21(Waf1/Cip1) and MDM-2 proteins and resistance to apoptosis after genotoxic treatment. Because of their origin from normal fetal colon and their relative resistance to the induction of apoptosis, FHC cells can be considered a valuable experimental model for various studies.},

note = {Place: United States},

keywords = {*Genes, Animals, Apoptosis/physiology, Carcinoembryonic Antigen/metabolism, Cell Adhesion/physiology, Cell Growth Processes/physiology, Cell Line, Cell Transformation, Colon/cytology/metabolism/*physiology, Colonic Neoplasms/*genetics/*pathology, Comparative Genomic Hybridization, Cytogenetic Analysis/methods, DNA Damage, DNA Mutational Analysis/methods, Female, Fetus/cytology, Fluorescence, HCT116 Cells, Humans, In Situ Hybridization, Karyotyping, Keratins/metabolism, Mice, Neoplasm Transplantation, Neoplastic/genetics/pathology, p53, Phenotype, Proto-Oncogene Mas, SCID, Signal Transduction, Transformed},

pubstate = {published},

tppubtype = {article}

}

@article{stika_mk-886_2006,

title = {MK-886 enhances tumour necrosis factor-alpha-induced differentiation and apoptosis.},

author = {Jirí Stika and Jan Vondrácek and Jirina Hofmanová and Vladimír Simek and Alois Kozubík},

doi = {10.1016/j.canlet.2005.06.012},

issn = {0304-3835},

year  = {2006},

date = {2006-06-01},

journal = {Cancer letters},

volume = {237},

number = {2},

pages = {263–271},

abstract = {We investigated the role of the 5-lipoxygenase (5-LOX) pathway of arachidonic acid metabolism in tumour necrosis factor-alpha (TNF-alpha)-induced differentiation of human leukemic HL-60 cells using MK-886, an inhibitor of 5-LOX activating protein. MK-886 augmented cell cycle arrest and differentiation induced by TNF-alpha; however, both effects were probably 5-LOX-independent, because a general LOX inhibitor, NDGA, had no effect. Apoptosis was significantly elevated after combined TNF-alpha and MK-886 treatment, which could be partially associated with changes of Mcl-1 protein expression. NF-kappaB signalling or activation of JNKs were not modulated by MK-886. Thus, in addition to apoptosis, MK-886 can enhance TNF-alpha-induced differentiation.},

note = {Place: Ireland},

keywords = {*Apoptosis, Arachidonate 5-Lipoxygenase/metabolism, Cell Cycle, Cell Differentiation, Cell Line, Cell Survival, HL-60 Cells, Humans, Indoles/*pharmacology, Lipoxygenase Inhibitors/*pharmacology, Signal Transduction, Time Factors, Tumor, Tumor Necrosis Factor-alpha/*metabolism},

pubstate = {published},

tppubtype = {article}

}