1st Faculty of Medicine Charles University 1st Faculty of Medicine Charles University Hematology laboratories
lvysa 05.03.2024

Hematopoietic stem cells use chromatin-remodeling factors for specific processes: the story of Smarca5

Turkova T, Kokavec J, Zikmund T, Dibus N, Pimkova K, Nemec D, Holeckova M, Ruskova L, Sedlacek R, Cermak L, Stopka T. Differential requirements for Smarca5 expression during hematopoietic stem cell commitment. Commun Biol. 2024 Feb 29;7(1):244. doi: 10.1038/s42003-024-05917-z.

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Mouse transgenic research suggests that lymphocyte development requires a robust chromatin remodeling. Turkova et al. found that although the chromatin-remodeling enzyme known as Smarca5 was thought to be a general factor, lymphocyte development in stem cells is most sensitive to its decrease.

By Tereza Turkova & Tomas Stopka


Figure legend: Hematopoietic stem cells in bone marrow regulate the expression of the epigenetic regulator Smarca5, which influences the development of blood elements through ATP-dependent effects on nucleosome architecture. Our data show that the development of lymphoid progenitors of both the B and T lineages is most dependent on Smarca5 levels, erythropoiesis is less dependent, while a very low level of the studied ISWI ATPase is sufficient for myeloid cell development. Created with BioRender.com.

The production of blood cells relies on coordinated regulation of gene expression, timing of the cell cycle, and suppression of inappropriate processes by apoptosis. Like any stem cell, the hematopoietic stem cell deals with the balance between stemness and differentiation, but also with the quantitative requirements for the production of different elements at specific stages of development and during everyday situations that might include bleeding or infections. The different decisions in the cell nucleus of stem cells are related to the structure of the DNA, which is wrapped in a bun of histone proteins. Different critical sequences have to be made available, and this is done by an enzymatic motor that releases DNA from the histone octamer, called the nucleosome, with the expenditure of energy, creating a region of freely accessible DNA, while other unnecessary DNA sequences are twisted like a thread around a spool. The unwinding of the DNA strand from the nucleosome is called the process of chromatin remodeling. It is generally accepted that stem cells have very little gene expression and replication, and thus we were interested in how dependent the various blood subtypes are on the function of the chromatin remodeling enzymatic engine in stem cells during their early differentiation. Specifically, we tested how the ISWI ATPase SMARCA5 (SNF2H), which represents an absolutely essential and unique enzyme, effects the production of lymphoid, erythroid and myeloid cells.

This type of project involves mouse transgenic models that we have gradually developed over the past 20 years. My lab group took up the project around 2005 in Prague, Czech Republic, but the roots go back to 1998 when we cloned the mouse mRNA for Smarca5 and found that Smarca5 is very important for blood production. In 2003, we published with Art Skoultchi (Albert Einstein College of Medicine, Bronx, NY) a mouse knockout for Smarca5 that showed its indispensability for early embryonic development. Further research led to the development of a mouse model of Smarca5 conditional inactivation that demonstrated the importance of Smarca5 for hematopoietic stem cell, lymphoid and erythroid development.

When we used up the possibilities of the knockout mouse and decided to create a conditional transgenic model for Smarca5. Although we created models for human as well as murine Smarca5, a publication now coming out in Communications Biology covers the first results with the transgene for human SMARCA5 entitled: Differential requirements for Smarca5 expression during hematopoietic stem cell commitment. We observed that the transgenic allele only provides about 10% of the expression of the normal allele. Therefore, we crossed the transgenic model with the previous model with a knockout allele to generate a set of supermodels in which the different Smarca5 expression levels exist. This allowed us to show that the hypomorphic (producing lower protein levels) SMARCA5 allele (S5tg) is fully functional and can rescue, in a dose-dependent manner, the gene inactivation phenotypes from previous k.o. projects, i.e. defects in lymphoid and erythroid development. In addition, transgenic Smarca5 binds all canonical complexes that have been incrementally discovered. Once we express low level of Smarca5 in stem cells, we see that progenitors entering lymphoid T and B development are the most sensitive to Smarca5 reduction, while other progenitors are less or not affected (see figure & legend).

To support our hypothesis that SMARCA5 expression level has a direct effect on development at stem cell level, we employed competitive transplantation experiment of transgenic and normal stem cells into normal but also transgenic recipients. Both of these experiments confirmed our hypothesis that it is primarily lymphocyte development that is strongly dependent on Smarca5 levels and that the defect originates in stem cells. Low SMARCA5 levels lead to significant permissiveness of the bone marrow niche for wild type stem cells. Transgenic hematopoietic stem cells were not able to produce lymphocytes, indicating that these lymphoid early progenitors are most sensitive to SMARCA5 levels. You think that's the end of the story? But no. Our current results show that Smarca5 is indeed a very important factor that affects the life of cancer stem cells. Therefore, we are also focusing on the question of how to effectively get rid of these cancer stem cells and one of the mechanisms is to focus on the biology of Smarca5 in this regard.

Article link HERE
lvysa 13.02.2024

Poslechněte si videokomentáře z letošní konference HEMATOLOGIE 2024

Co se událo na poli české a světové hematologie? ZDE  294

lvysa 07.03.2024

Cena kolegia děkana 1.LF UK za excelentní publikaci 2022

Stopka T, Minařík L, Dusilková N, Pešta M, Kulvait V, Špaček M, Zemanová Z, Kalousová M, Jonášová A. G-CSF plus azacitidine versus azacitidine alone for patients with high-risk myelodysplastic syndrome: academic, open label, randomized trial. Blood Cancer J. 2022 Jul 7;12(7):105. doi: 10.1038/s41408-022-00698-2. (PubMed)

Pimkova K, Jassinskaja M, Munita R, Ciesla M, Guzzi N, Cao Thi Ngoc P, Vajrychova M, Johansson E, Bellodi C, Hansson J. Quantitative analysis of redox proteome reveals oxidation-sensitive protein thiols acting in fundamental processes of developmental hematopoiesis. Redox Biol. 2022 Jul;53:102343. doi: 10.1016/j.redox.2022.102343. (PubMed)

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lvysa 21.06.2023

EHA 2023 Frankfurt

Comments from EHA2023 in Frankfurt - prof. Stopka and Dr. Pimková (in Czech).   285

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lvysa 29.03.2023

Czech physicians and researchers to prolong the lives of people with leukaemia

A team of researchers and physicians from the BIOCEV Centre, 1st Department of Medicine - Hematology (the First Faculty of Medicine of Charles University) and the General University Hospital in Prague has designed a new approach for the treatment of acute leukaemia. This innovative method involves the combination of two medicines that, when administered together, have significantly prolonged the survival of patients. The clinical trial lasted five years and included 76 patients from the Czech Republic. Information about the new type of treatment was published by the prestigious Blood Cancer Journal from the Nature Publishing Group. At the moment, negotiations are underway to expand the trial to the European-wide level.

Doc. Anna Jonášová and Prof. Tomáš Stopka  

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Myelodysplastic syndrome (MDS) is a cancer of the bone marrow causing a low production of non-functioning blood cells. As a result, patients are deficient in both red and white blood cells, which are crucial for the body’s immunity, as well as platelets for blood coagulation. Patients suffering from this anaemia suffer from bruising, frequent internal bleeding and are less resistant to bacterial and viral infections, for example pneumonia.

Depending on the severity of the disease, antibiotics or repeated blood transfusions are given as treatment. In exceptional cases, patients can be treated with a bone marrow transplant. In Europe, four MDS patients per 100,000 people are diagnosed annually. In the Czech Republic, the General University Hospital registers approximately 80 patients each year. The critical phase of MDS is the transition to acute myeloid leukaemia (AML), characterised with a very short survival time.

"The current globally applied treatment for MDS patients during their transition to AML is based on 5-azacytidine, a medicine which was invented in the 1970s by the Czechoslovak Academy of Sciences. Azacytidine is an inhibitor of nuclear enzymes, referred to as DNA methylases, which are genetically and functionally disrupted in leukemic cells, and therefore, their activity helps to eliminate leukemic cells. Based on previous research, much of which has been published in reputable journals, our MDS/AML group has come up with the concept of “differentiation therapy for MDS”. We propose combining azacytidine with another substance called G-CSF, which promotes the formation and development of white blood cells in the bone marrow. We have been able to demonstrate the therapeutic effects of the new combination, and, above all, its safe effect on patients, but we have also been able to prolong their survival by months,” says Prof. Tomáš Stopka, head of research from the First Faculty of Medicine, Charles University, and Biocev. “At the same time, we have discovered that the combination of these agents induced significantly more treatment responses, which is ground-breaking and essential for further research and clinical testing in patients.”

A clinical trial demonstrating the benefit of the new treatment method in high-risk MDS was approved by the State Institute for Drug Control in 2017. The coordinator of the clinical trial was the Head of MDS and AML Treatment and Diagnosis, Doc. Anna Jonášová from the 1st Internal Clinic – Haematology, First Faculty of Medicine, Charles University in Prague: “We had to make a huge effort even before the trial itself was launched as we had to secure an application with the State Institute for Drug Control and have the trial pass through the ethics committees. The proposed therapeutic combination advances the use of the cytokine G-CSF in AML therapy, while its synergistic activity with azacytidine makes this treatment more effective and safer for patients, representing a proven strategy that is immediately applicable in practice. We would not have achieved the results if it had not been for the great willingness of the patients who participated in the trial."

At the moment, intensive negotiations are underway for a new European-wide trial in which another medicine, Venetoclax, will be investigated alongside 5-azacytidine and G-CSF. "I am happy that we are successfully linking genetic and clinical data in the Czech Republic and thanks to this, we are able to determine how acute leukaemia will be treated in the near future," adds Prof. Tomáš Stopka.

Link to the publication: HERE

Rozhovor pro UK FORUM: Nová kombinace léčiv prodlužuje život pacientů s leukémií (ukforum.cz)

lvysa 17.04.2023

Medical Tribune - Ročník XIX, číslo 6

 Article about our research in Medical Tribune (in czech)         

lvysa 29.03.2023

Tomáš Stopka: G-CSF plus azacitidine versus azacitidine alone

G-CSF plus azacitidine versus azacitidine alone for patients with high-risk myelodysplastic syndrome: academic, open label, randomized trial.Stopka T, Minařík L, Dusilková N, Pešta M, Kulvait V, Špaček M, Zemanová Z, Kalousová M, Jonášová A. Blood Cancer J. 2022 Jul 7;12(7):105. doi: 10.1038/s41408-022-00698-2. (IF 11.98) [link]

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Rozhovor pro UK FORUM: Nová kombinace léčiv prodlužuje život pacientů s leukémií (ukforum.cz)

TZ BIOCEV: Čeští lékaři a vědci prodlouží život lidem s leukémií | Biocev

lvysa 29.03.2023

Kristýna Pimková: Quantitative analysis of redox proteome

Quantitative analysis of redox proteome reveals oxidation-sensitive protein thiols acting in fundamental processes of developmental hematopoiesis. Pimkova K, Jassinskaja M, Munita R, Ciesla M, Guzzi N, Cao Thi Ngoc P, Vajrychova M, Johansson E, Bellodi C, Hansson J. Redox Biology. Volume 53, July 2022, 102343. ISSN 2213-2317. https://doi.org/10.1016/j.redox.2022.102343. (IF 11.79) [PubMed]

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lvysa 28.03.2023

Tomáš Stopka: Circulating microRNAs in Cerebrospinal Fluid and Plasma

Circulating microRNAs in Cerebrospinal Fluid and Plasma: Sensitive Tool for Detection of Secondary CNS Involvement, Monitoring of Therapy and Prediction of CNS Relapse in Aggressive B-NHL Lymphomas. Krsmanovic P, Mocikova H, Chramostova K, Klanova M, Trnkova M, Pesta M, Laslo P, Pytlik R, Stopka T, Trneny M, Pospisil V. Cancers. 2022; 14(9):2305. https://doi.org/10.3390/cancers14092305 (IF 6.6) [link]

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lvysa 29.03.2023

Petra Bašová: Combined Approach to Leukemic Differentiation

Combined Approach to Leukemic Differentiation Using Transcription Factor PU.1-Enhancing Agents. Bašová P, Paszeková H, Minařík L, Dluhošová M, Burda P, Stopka T. Int J Mol Sci. 2022 Jun 16;23(12):6729. doi: 10.3390/ijms23126729. (IF 5.9) [PubMed]

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lvysa 29.03.2023

Shefali Thakur: Chromatin remodeler Smarca5

Chromatin Remodeler Smarca5 Is Required for Cancer-Related Processes of Primary Cell Fitness and Immortalization. Thakur S, Cahais V, Turkova T, Zikmund T, Renard C, Stopka T, Korenjak M, Zavadil J. Cells 2022, 11,808. https://doi.org/10.3390/cells11050808 (IF 6.6) [PDF

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lvysa 29.03.2023

Kristýna Pimková: Peroxiredoxin 6

Peroxiredoxin 6 protects irradiated cells from oxidative stress and shapes their senescence-associated cytokine landscape. Salovska B, Kondelova A, Pimkova K, Liblova Z, Pribyl M, Fabrik I, Bartek J, Vajrychova M, Hodny Z. Redox Biol. 2022 Feb;49:102212. doi: 10.1016/j.redox.2021.102212. (IF 10.79) [PubMed]

 

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lvysa 29.03.2023

Ľubomír Minařík: Analysis of 5-Azacytidine Resistance

Analysis of 5-Azacytidine Resistance Models Reveals a Set of Targetable Pathways. Minařík L, Pimková K, Kokavec J, Schaffartziková A, Vellieux F, Kulvait V, Daumová L, Dusilková N, Jonášová A, Vargová KS, Králová Viziová P, Sedláček R, Zemanová Z, Stopka T. Cells. 2022; 11(2):223. https://doi.org/10.3390/cells11020223. (IF 6.6) [PDF]    209

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lvysa 29.03.2023

Pavel Čabart: Universal Promoter Scanning

Universal promoter scanning by Pol II during transcription initiation in Saccharomyces cerevisiae. Qiu C, Jin H, Vvedenskaya I, Llenas JA, Zhao T, Malik I, Visbisky AM, Schwartz SL, Cui P, Čabart P, Han KH, Lai W, Metz RP, Johnson CD, Sze SH, Pugh BF, Nickels BE & Kaplan, CD (2020). Genome biology, 21(1), 132. https://doi.org/10.1186/s13059-020-02040-0. (IF 14.028) [PubMed] (131)

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lvysa 29.03.2023

Tomáš Zikmund - Helena Paszeková: Loss of ISWI ATPase SMARCA5 (SNF2H)

Loss of ISWI ATPase SMARCA5 (SNF2H) in Acute Myeloid Leukemia Cells Inhibits Proliferation and Chromatid Cohesion. Zikmund T, Paszekova H, Kokavec J, Kerbs P, Thakur S, Turkova T, Tauchmanova P, Greif PA, Stopka T. Int J Mol Sci. 2020;21(6):E2073. Published 2020 Mar 18.doi:10.3390/ijms21062073. (IF 4.183) [PubMed]


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lvysa 29.03.2023

Tomáš Stopka: Aberrantly elevated suprabasin

Aberrantly elevated suprabasin in the bone marrow as a candidate biomarker of advanced disease state in myelodysplastic syndromes. Pribyl M, Hubackova S, Moudra A, Vancurova M, Polackova H, Stopka T, Jonasova A, Bokorova R, Fuchs O, Stritesky J, Salovska B, Bartek J, Hodny Z. [published online ahead of print, 2020 Jul 21]. Mol Oncol. 2020;10.1002/1878-0261.12768. doi:10.1002/1878-0261.12768. (IF 6.574) [PubMed] 202

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lvysa 29.03.2023

Tomáš Stopka: Chromatin remodeler Snf2h

The chromatin remodeler Snf2h is essential for oocyte meiotic cell cycle progression. Zhang C, Chen Z, Yin Q, Fu X, Li Y, Stopka T, Skoultchi AI, Zhang Y. Genes Dev. 2020 Jan 9. doi: 10.1101/gad.331157.119. (IF 8.998) [Pubmed


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lvysa 29.03.2023

Ľubomír Minařík: Azacitidine Switch to Lenalidomide

Azacitidine Switch to Lenalidomide Eradicated the TP53/ CDKN2A Co-Mutated Clone and Induced Long-Term Erythroid Response in Del(5q) MDS. Minarik L, Zemanova Z, Kulvait V, Dluhosova M, Jonasova A and Stopka T. Ann Hematol Oncol. 2019; 6(1): 0000. [PDF] (IF 2.219

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lvysa 29.03.2023

Tomáš Stopka: MDS News

Randomizovaná otevřená akademická studie: srovnání standardního podání azacytidinu oproti azacytidinu s preinkubací G-CSF u myelodysplastického syndromu vyššího rizika – interim analýza. Stopka T, Minařík Ĺ, Kulvait V, Pešta M, Schaffartziková A, Kislik G, Dusílková N, Zemanová Z, Jonášová A. Myelodysplastic Syndrome News. Listopad 2019. Ročník 7 / číslo 2. [PDF]


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lvysa 29.03.2023

Tomáš Zikmund: ISWI ATPase Smarca5

ISWI ATPase Smarca5 Regulates Differentiation of Thymocytes Undergoing b-selection. Zikmund T, Kokavec J, Turkova T, Savvulidi F, Paszekova H, Vodenkova S, Sedlacek R, Skoultchi AI, Stopka T.. J Immunol. 2019 Jun 15;202(12):3434-3446. doi: 10.4049/jimmunol.1801684. Epub 2019 May 8. [PubMed] (IF 4.539) 185

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lvysa 29.03.2023

Reputable publication in Haematologica July 2018

BCR-ABL1 mediated miR-150 downregulation through MYC contributed to myeloid differentiation block and drug resistance in chronic myeloid leukemia. Srutova K, Curik N, Burda P, Savvulidi F, Silvestri G, Trotta R, Klamova H, Pecherkova P, Sovova Z, Koblihova J, Stopka T, Perrotti D, Machova Polakova K. Haematologica. 2018 Jul 26. pii: haematol.2018.193086. doi: 10.3324/haematol.2018.193086. [PubMed] (IF 9.09)

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lvysa 29.03.2023

Anna Jonášová: Lenalidomide treatment in lower risk myelodysplastic syndromes

Lenalidomide treatment in lower risk myelodysplastic syndromes—The experience of a Czech hematology center. (Positive effect of erythropoietin±prednisone addition to lenalidomide in refractory or relapsed patients). Jonasova A, Neuwirtova R, Polackova H, Siskova M, Stopka T, Cmunt E, Belickova M, Moudra A, Minarik L, Fuchs O, Michalova K, Zemanova Z. Leukemia Research 69 (2018) 12-17. 2018 March 27. https://doi.org/10.1016/j.leukres.2018.03.015 . (IF 2.501) [PDF]

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lvysa 29.03.2023

Kamila Polgárová: Somatic mutation dynamics in MDS patients

Somatic mutation dynamics in MDS patients treated with azacitidine indicate clonal selection in patients-responders. Polgarova K, Vargova K, Kulvait V, Dusilkova N, Minarik L, Zemanova Z, Pesta M, Jonasova A and Stopka T. Oncotarget. 2017 Dec 6. (IF 5.168) [PDF] (B, N, H, 131)

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lvysa 29.03.2023

Nina Dusílková & Petra Bašová: Plasma microRNAs in Cutaneous Lymphoma, in press IJMS

Plasma miR-155, miR-203, and miR-205 are Biomarkers for Monitoring of Primary Cutaneous T-cell Lymphomas. Dusilkova N, Basova P, Polivka J, Kodet O, Kulvait V, Pesta M, Trneny M, Stopka T. Int J Mol Sci. 2017 Oct 15;18(10). pii: E2136. doi: 10.3390/ijms18102136. [PDF] (IF: 3.226)

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lvysa 29.03.2023

Petra Bašová: Serum microRNAs in Breast Cancer Relapse prediction, in press in IJMS

Prediction Potential of Serum miR-155 and miR-24 for Relapsing Early Breast Cancer. Basova P, Pesta M, Sochor M, Stopka T. Int J Mol Sci. 2017 Oct 10;18(10). pii: E2116. doi: 10.3390/ijms18102116[PDF] (IF: 3.226) 121

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lvysa 29.03.2023

Juraj Kokavec: mouse model of Smarca5 in HSCs

Kokavec J, Zikmund T, Savvulidi F, Kulvait V, Edelmann W, Skoultchi AI, Stopka T. The ISWI ATPase Smarca5 (Snf2h) Is Required for Proliferation and Differentiation of Hematopoietic Stem and Progenitor Cells. Stem Cells. 2017 Jun;35(6):1614-1623. doi: 10.1002/stem.2604. [https] (IF: 5.599)

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lvysa 29.03.2023

Hana Hušková - MS on cancer driver NOW in Oncogene

Huskova H, Ardin M, Weninger A, Vargova K, Barrin S, Villar S, Olivier M, Stopka T, Herceg Z, Hollstein M, Zavadil J, Korenjak M. Modeling cancer driver events in vitro using barrier bypass-clonal expansion assays and massively parallel sequencing. Oncogene. 2017 Jul 10. doi: 10.1038/onc.2017.215. [PDF] (IF: 7.519)
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lvysa 29.03.2023

Karina Vargová - MS on miR-155/miR-150 network in CLL now in press in BCJ

Vargova K, Pesta M, Obrtlíkova P, Dusilkova N, Minarik L, Vargova J, Berkova A, Zemanova Z, Michalova K, Spacek M, Trneny M and Stopka T. MiR-155/miR-150 network regulates progression through the disease phases of chronic lymphocytic leukemia. Blood Cancer J. 2017 Jul 21;7(7):e585. doi: 10.1038/bcj.2017.63. [PDF] (IF: 6.126)

 

 

 

lvysa 29.03.2023

Jarmila Vargová: MARCKS: new therapeutic target in MCL

Vargova J, Vargova K, Dusilkova N, Kulvait V, Pospisil V, Zavadil J, Trneny M, Klener P, Stopka T. Differential expression, localization and activity of MARCKS between mantle cell lymphoma and chronic lymphocytic leukemia. Blood Cancer J. 2016 Sep 23;6(9):e475. doi: 10.1038/bcj.2016.80. [PDF] (IF: 4.502)

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ohlasy v tisku:

http://www.ctidoma.cz/zpravodajstvi-zdravi/2016-10-05-cesti-odbornici-zjistili-jak-vylecit-rakovinu-krve-26743

http://sciencemag.cz/cesti-vedci-na-stope-novym-lekum-proti-lymfomum/

lvysa 29.03.2023

Pavel Burda: addition to BBA, collaborative project with John Strouboulis

Papageorgiou DN, Karkoulia E, Amaral-Psarris A, Burda P, Kolodziej K, Demmers J, Bungert J, Stopka T, Strouboulis J. Distinct and overlapping DNMT1 interactions with multiple transcription factors in erythroid cells: Evidence for co-repressor functions. Biochim Biophys Acta. 2016 Dec;1859(12):1515-1526. doi: 10.1016/j.bbagrm.2016.09.007. [https] (IF: 5.373) 132