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Leukemic stem cells
Vobořil, Matouš ; Stöckbauer, Petr (advisor) ; Šídová, Monika (referee)
The biology of stem cells came to the foreground not only due to scientific but also due to clinical interest and it is one of the most developing fields of current biomedical research. The idea, that all tumor cells contain population of cells like stem cells, leads to the "cancer stem cells hypothesis". It says, that each tumor cell contains small population of cells capable to initiate and maintain the tumor growth. Tumors of the hematopoietic tissue were the first, where cancer or leukemic stem cells were isolated. Therefore, leukemic stem cells are so far the best understood cancer stem cells. However, despite the huge advances in the biology of leukemic stem cells, there are many properties still unknown. This thesis initially presents basic knowledges in the stem cell biology, including their origin and identification. Later it focuses on the stem cell hypothesis and describes of the main properties of stem cells in solid tumors. The main part of this thesis also shows in details the origin and properties of the leukemic stem cells and decribes some new directions in the targeted therapy of hematological malignancies.
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Histone deacetylase inhibitors in plasma cell leukemia treatment: effect of the bone marrow microenvironment
Burianová, Ilona ; Stöckbauer, Petr (advisor) ; Pour, Luděk (referee) ; Benson, Veronika (referee)
Multiple myeloma and its aggressive variant, plasma cell leukemia, are still considered to be incurable diseases despite the progressive treatment approaches comprising novel drugs. This can be attributed to the presence of the bone marrow microenvironment which plays an important role in drug resistance of myeloma cells. Hematopoietic cell lines derived from hematologic malignancies are suitable models for the study of etiopathogenesis of these malignant diseases and for testing new potential drugs. Establishment of these cell lines is still considered to be coincidental and rare event. The first part of the thesis is focused on establishment and characterization of the cell line UHKT-944 derived from a patient with primary plasma cell leukemia, and on completion of characterization of the cell line UHKT-893 derived from a patient with multiple myeloma. Additional analysis of UHKT-893 cell line were performed including sequence analysis of IgVH gene rearrangements and cytogenetic analysis which contributed to more detailed characterization of this cell line. During cultivation of UHKT-944 cells, we monitored the cell growth and confirmed dependence on interleukin-6 (IL-6). Immunophenotype analysis revealed the presence of surface markers characteristic of malignant plasma cells. UHKT-944 cells...
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Expression of adaptor protein PAG/Cbp in non-neoplastic and neoplastic lymphoid tissue
Švec, Alexandr ; Mandys, Václav (advisor) ; Plank, Lukáš (referee) ; Kodet, Roman (referee) ; Stöckbauer, Petr (referee)
New notions about cell development, cell cycle, differentiation and cell signalling are now being used for refinement of classification of malignant lymphomas (ML) and for development of new drugs specifically targeting deregulated proteins some of which play a role in immune signalling. The first step in most of the signalling pathways is transduction of the signal through the plasma membrane mediated by a plasma cell receptor. Certain proximal signalling molecules such as a family of Src-kinases (SFK) are segregated in particular plasma membrane compartments called lipid rafts or glycosphingolipid-enriched microdomains (GEM). I studied expression of PAG in non-neoplastic and neoplastic lymphoid tissue in order to determine its expression pattem and to investigate its utility in routine diagnostic haematopathology.
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Immunologic and epigenic modulation of gene expression of human leukemia cell lines
Elknerová, Klára ; Stöckbauer, Petr (advisor) ; Špíšek, Radek (referee) ; Škvor, Jiří (referee)
Therapy by monoclonal antibody to CD34 molecule Growth-inhibitory and proapoptotic effect of the monoclonal antibody to CD34 molecule, clone 4H11, were tested on CD34+ leukemic cell lines (MOLM-9, JURL-MK1) and CD34- cell line (PS-1). We have found that the monoclonal antibody to CD34 inhibited the proliferation and induced apoptosis of all CD34+ cell lines tested, however it has similar effect on CD34- leukemic cell line (PS-1) in concentration higher than 32 µg/ml. We did not observe induction of differentiation by anti-CD34 antibody, but a growth arrest of cells in the G0/G1 phase of the cell cycle was detected in CD34+ cell lines. Combinations of anti-CD34 antibody with both type I (α, β) or type II ( ) interferons did not enhance the effects on the cell growth or inhibition of cellular proliferation of the antibody alone. Combinations of anti-CD34 antibody with hematopoietic cytokines or INF-γ did not induced diferentiation. Our data suggest that the monoclonal antibody to CD34 molecule prepared from clone 4H11, after sufficient experimental and preclinical testing on laboratory animals, may provide a new basis for possible targeted antibody therapy of acute or chronic myeloid leukemia. Therapy by histone deacetylase inhibitors Histone deacetylase inhibitors (HDACi) are emerging new class of...
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Histone deacetylase inhibitors in plasma cell leukemia treatment: effect of the bone marrow microenvironment
Burianová, Ilona ; Stöckbauer, Petr (advisor) ; Pour, Luděk (referee) ; Benson, Veronika (referee)
Multiple myeloma and its aggressive variant, plasma cell leukemia, are still considered to be incurable diseases despite the progressive treatment approaches comprising novel drugs. This can be attributed to the presence of the bone marrow microenvironment which plays an important role in drug resistance of myeloma cells. Hematopoietic cell lines derived from hematologic malignancies are suitable models for the study of etiopathogenesis of these malignant diseases and for testing new potential drugs. Establishment of these cell lines is still considered to be coincidental and rare event. The first part of the thesis is focused on establishment and characterization of the cell line UHKT-944 derived from a patient with primary plasma cell leukemia, and on completion of characterization of the cell line UHKT-893 derived from a patient with multiple myeloma. Additional analysis of UHKT-893 cell line were performed including sequence analysis of IgVH gene rearrangements and cytogenetic analysis which contributed to more detailed characterization of this cell line. During cultivation of UHKT-944 cells, we monitored the cell growth and confirmed dependence on interleukin-6 (IL-6). Immunophenotype analysis revealed the presence of surface markers characteristic of malignant plasma cells. UHKT-944 cells...
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Establishment and characterization of novel IL-6-dependent myeloma cell line ÚHKT-893
Vančurová, Irena ; Stöckbauer, Petr (advisor) ; Škvor, Jiří (referee)
Multiple myeloma is an incurable fatal neoplasm of plasma cells affecting mainly elderly people. There are many research laboratories in the world where multiple myeloma is studied. Permanent cell lines are indispensable tools for both basic and applied research. However, the establishment of new cell lines is difficult with poor success. We established 96 primary cultures of bone marrow samples from myeloma patients. Only one culture succeeded in permanet myeloma cell line. The novel plasmacytic cell line ÚHKT-893 was established from bone marrow sample of 57 years old female relapsed with multiple myeloma of IgGκ isotype. The cell line is however dependent on the continuous presence of interleukin-6 in culture media. ÚHKT-893 cells grow continuously already more than 1 year. The growth of cells was regularly monitored according to growth curves and by measurement of cell viability. Surface antigenic profile was repeatedly determined by flow cytometry. The simultaneous expression of both CD138 and CD38 surface molecules confirmed the plasma cell origin of cells. The establishment of the ÚHKT-893 myeloma cell line will extend the panel of existing myeloma cell lines as a new ex vivo model for the study of the etiology and pathogenesis of multiple myeloma. It will also provide the source of new...
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Pre-clinical model of acute promyelocytic leukemia:/study of the anti-leukemic effect induced by ATRA and DNA vaccination
Pokorná, Kateřina ; Holáň, Vladimír (advisor) ; Stöckbauer, Petr (referee) ; Degos, Laurent (referee)
DOCTORAL THESIS 2012 POKORNA Abstract We have used a well characterized transplantable transgenic mouse model which mimics human acute promyelocytic leukemia (APL), both in its biological characteristics and its response to conventional therapeutic drugs. The aim of our study was to better characterize the efficacy of the combined treatment and to determine molecular markers of clinical outcome. We established a minimal residual disease monitoring based on the high sensitivity of detection of PML-RAR transcripts by polymerase chain reaction (PCR) technology in APL mice. We showed that oncogene-specific PCR-based assays allow, like in patients, the diagnosis, follow-up and prediction of disease evolution. Furthermore, PCR assay was used to assess various tissues and organs for the presence of PML-RAR-positive cells in minimal residual disease free long-term survivors. As expected, majority of mice had no measurable tissue level of PML-RAR demonstrating the efficacy of immunotherapy. However, tracking the oncogene-positive cells reveals for the first time that extramedullary PML-RAR-positive cell reservoirs such as the brain may persist and be involved in the leukemia relapse. We aimed at investigating the immune responses involved in the anti-leukemic effect of the combined immutherapy. To evaluate the...
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Leukemic stem cells
Vobořil, Matouš ; Stöckbauer, Petr (advisor) ; Šídová, Monika (referee)
The biology of stem cells came to the foreground not only due to scientific but also due to clinical interest and it is one of the most developing fields of current biomedical research. The idea, that all tumor cells contain population of cells like stem cells, leads to the "cancer stem cells hypothesis". It says, that each tumor cell contains small population of cells capable to initiate and maintain the tumor growth. Tumors of the hematopoietic tissue were the first, where cancer or leukemic stem cells were isolated. Therefore, leukemic stem cells are so far the best understood cancer stem cells. However, despite the huge advances in the biology of leukemic stem cells, there are many properties still unknown. This thesis initially presents basic knowledges in the stem cell biology, including their origin and identification. Later it focuses on the stem cell hypothesis and describes of the main properties of stem cells in solid tumors. The main part of this thesis also shows in details the origin and properties of the leukemic stem cells and decribes some new directions in the targeted therapy of hematological malignancies.
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Immunologic and epigenic modulation of gene expression of human leukemia cell lines
Elknerová, Klára ; Stöckbauer, Petr (advisor) ; Špíšek, Radek (referee) ; Škvor, Jiří (referee)
Therapy by monoclonal antibody to CD34 molecule Growth-inhibitory and proapoptotic effect of the monoclonal antibody to CD34 molecule, clone 4H11, were tested on CD34+ leukemic cell lines (MOLM-9, JURL-MK1) and CD34- cell line (PS-1). We have found that the monoclonal antibody to CD34 inhibited the proliferation and induced apoptosis of all CD34+ cell lines tested, however it has similar effect on CD34- leukemic cell line (PS-1) in concentration higher than 32 µg/ml. We did not observe induction of differentiation by anti-CD34 antibody, but a growth arrest of cells in the G0/G1 phase of the cell cycle was detected in CD34+ cell lines. Combinations of anti-CD34 antibody with both type I (α, β) or type II ( ) interferons did not enhance the effects on the cell growth or inhibition of cellular proliferation of the antibody alone. Combinations of anti-CD34 antibody with hematopoietic cytokines or INF-γ did not induced diferentiation. Our data suggest that the monoclonal antibody to CD34 molecule prepared from clone 4H11, after sufficient experimental and preclinical testing on laboratory animals, may provide a new basis for possible targeted antibody therapy of acute or chronic myeloid leukemia. Therapy by histone deacetylase inhibitors Histone deacetylase inhibitors (HDACi) are emerging new class of...
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