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Science and research

Grant Projects (ongoing in the year 2012)

View ongoing grants in the year: 2009, 2010, 2011, 2012, 2013, 2014, 2015, 2016, 2017, 2018, 2019, 2020, 2021, 2022, 2023, 2024.

Quality of life measurement and its influence to overall survival in hematopoietic stem cell transplantation in CZ

IGA MZd NT 11299 [2010 – 2015]

prof. MUDr. Marek Trněný, CSc.
MUDr. Veronika Válková, CSc.

The co-recipient from the transplant center of Institute of Hematology and Blood Transfusion is responsible for anonymous data of patients after autologous and allogeneic hematopoietic stem cells transplant to the database of EBMT (The European Group for Blood and Marrow Transplantation). This data will be used for characterization of the particular diagnostic groups within the project frame. The main aim of the presenting grant project is to analyze the quality of life (QOL) of the transplanted patients using standardized QOL surveys. The co-recipient´s team consists from a physician and datamanager, who will be responsible for the data collecting using the QOL surveys and their subsequent processing to the Principal investigators team. The main benefit is an expansion of the co-investigators view to the psycho-social perspective of the patients.

The next generation sequencing as a tool of personal medicine in patients with myelodysplastic syndrome and chronic myeloid leukemia

IGA MZd NT 13899-4 [2012 – 2015]

prof. MUDr. Jaroslav Čermák, CSc.

The proposed project applies Next-generation sequencing for ultra-deep mutation detection in genes involved in genome methylation processes (TET2, ASXL1, IDH1 / 2, TP53, EZH2) in patients with myelodysplastic syndrome (MDS). We will determine the relationship between the success of therapy and the gene mutation status in patients receiving the demethylation treatment. In addition, we will study the relationship between quantity of TP53 mutations and treatment failure in patients treated with Lenalidomide. In chronic myeloid leukemia (CML) we would like to determine the predictive value of ultra-deep sequencing of BCR-ABL mutations for resistance development on tyrosine kinase inhibitors. We will also study the association between polymorphisms found in promoter regions of genes belonging to the superfamilies of membrane transporters (ABC and SLC) and response to treatment. This project introduces the possibility of using molecular high-technology as a tool for individualizing therapy of MDS and CML.

Diagnostic and prognostic relevance of selected microRNAs in myelodysplastic syndromes

IGA MZd NT 13847-4 [2012 – 2015]

Ing. Michaela Dostálová Merkerová, Ph.D.

Recently, analyses of gene expression identified several miRNAs (miR-34a, miR-224, miR-10a, miR-10b, miR-126, miR-181 family, miRNAs from commonly deleted region, miRNA cluster at 14q32, miR-29b) whose expression changes in patients with myelodysplastic syndromes (MDS) during the course of the disease or in response to its treatment. We suppose that these molecules may be implicated in disease development or progression. In the project, we want to prove applicability of these miRNAs as molecular markers for monitoring of MDS patients (by quantification of miRNA transcription by qRT-PCR) and to study them using an in vitro system.

CMV specific mRNA vaccine for ex vivo stimulation of T cells in patiens after hematopoietic stem cell transplantation

IGA MZd NT 13898-4 [2012 – 2015]

RNDr. Šárka Němečková, DrSc.

Here, we propose to prepare mRNA vaccine against a number of candidate immunogenic CMV proteins. The vaccine will include antigen presenting cells electroporated mRNA coding for viral protective antigens and immunostimulatory molecules known to enhance activation of T-cells directed to Th1. To evaluate the protective potential of each CMV vaccine component the magnitude of CMV T-cell response of seropositive donors or patients after HSCT to each individual protein will be monitored and analyzed in the context of CMV DNA levels of HSCT patient. Tcell response will be characterized phenotypicaly and functionally by multiparametric flow cytometry. The result of the study will enable to elaborate immunotherapy protocol under good manufacturing practise conditions. mRNA vaccine will be also used for monitoring in CMV-specific T cell response of patients after HSCT.

Utilization of centrosomal proteins for prognostication and vaccinotherapy of chronic myeloid leukemia

IGA MZd NT 13862-4 [2012 – 2015]

RNDr. Michal Šmahel, Ph.D.

Search for prognostic markers of chronic myelogenous leukemia (CML) and development of DNA vaccines against this disease based on increased production of centrosomal proteins. Detection of HMMR/RHAMM, AURKA and ESPL1 mRNA level in blood cells and antibodies against the corresponding proteins in CML patients at disease diagnosis and during subsequent monitoring. Utilization of the obtained findings for optimization of therapy. Development of DNA vaccines against HMMR and AURKA that should be efficient against CML stem cells resistant to imatinib. Enhancement of vaccine immunogenicity by the addition of cell localization signals and sequences encoding strong helper epitopes. Analysis of the effect of xenogeneic origin of the genes on vaccine immunogenicity. Reduction of HMMR and AURKA potencial oncogenicity by the mutagenesis of functional domains and by the removal of their initiation codons in fusion genes. Observation of immune responses induced with the developed DNA vaccines in mouse models.

The study of the myelodysplastic syndrome pathogenesis in patients with isolated del(5q) abnormality and the analysis of the effect of lenalidomide

IGA MZd NT 13836-4 [2012 – 2015]

Ing. Ota Fuchs, CSc.
MUDr. Anna Jonášová, M.D., RNDr. Ludmila Doležalová, CSc.

We want to contribute to further understanding of the 5q- syndrome pathogenesis mechanism by the study of the important transcription factors, EKLF (erythroid Krüppel like factor, also called KLF1), Fli1 (Friend leukemia virus integration 1) and GATA1, which take part together with other factors on the differentiation of common MEP progenitor cell into erythroid or megakaryocytic line. The expression of these and other important genes in this mechanism will be analysed in separated bone marrow and blood cells of patients MDS with 5q- syndrom in comparison with low-risk MDS with normal chromosome 5 and with healthy controls by TaqMan real-time PCR. Our second aim is to contribute to the understanding of the mechanism of the effective agent lenalidomide in patients with 5q- syndrome in comparison with low-risk MDS with normal chromosome 5. Mutations of EKLF, which can significantly affect lenalidomide treatment, will be also detected.

The role of microRNA expression in HPV-associated and HPV-independent head and neck tumors

GA ČR P304/12/2244 [2012 – 2015]

RNDr. Ruth Tachezy, Ph.D.

MicroRNAs (miRNAs) participate in the regulation of gene expression. miRNA genes are frequently located in cancer-associated genomic regions, suggesting their role in the development of malignant tumors. Levels of expression of some miRNAs are diferent in tumors and in normal tissues and miRNA expression profiles are specific for different tumors. Studies on head and neck tumors (HNC) confirmed the difference in the level of expression of some miRNAs in the tumor and normal tissue but the results have not been conclusive. Sources of the variability of the data have been the heterogeneity of the studied cohorts in terms of the anatomic localization and etiology of tumors. The study of miRNA expression in a matched set of HPV- dependent and independent HNCs will allow us to characterize the di_erences in miRNA expression in tumors of di_erent etiology. The comparison of miRNA expression profiles in tumor tissues and in isogenic primary human keratinocyte clones immortalized by HPV will provide information about pathways involved in the development of tumors of different etiology.

Genital warts, premalignant lesions and vulvar carcinoma diagnostic and therapeutical management optimalisation

IGA MZd NT 13167-4 [2012 – 2015]

Doc. MUDr. Helena Robová, Ph.D., 2. LF UK Praha
RNDr. Ruth Tachezy, Ph.D.

Current knowledge about pathogenesis of vulvar lesions including genital warts, precancerous lesions, vulvar cancer has led to an update of classification of precancerous lesions. The aim is to find optimal algorithms for each entity and to enhance our knowledge on predictive factors which can be used for clinical praxis. In this project we plan to correlate HPV profiles of lesions with histopathological characteristics especially morphological diagnostic sings of HPV positive and HPV negative lesions. HPV profiles will be correlated with expression of immunohistochemical markers, mainly p161NK4a(p16). Another predictive factors will be assessed including minichromosome maintenance protein 2 (MCM2), DNA topoisomerase II Alfa (TOP IIA), cycline E and ProExC (mixture of antibodies against MCM2 and TOP IIA). When combined the predictive factors could improve sensitivity and specificity of histopathological examination. Diagnostic and therapeutic algorithms should be clinical outcome of the project.

Development of a Novel Recombinogenic Technique for Chloroplast Transformation and its Use for Production of human papillomavirus E7 Protein in Plants

IAA500960903 [2009 – 2012]

Doc. RNDr. Josef Vlasák, CSc., Biologické centrum AV ČR, v.v.i.
RNDr. Viera Ludvíková

Our aim is to develop more efficient method for plant chloroplast transformation using biolistic bombardment with linear DNA vectors together with chloroplast cassettes expressing bacteriophage lambda Red proteins. The idea stems from experiments with targeted insertion mutagenesis in yeast, cyanobacteria, and E. coli, where DNA ends promote recombination, and lambda Red proteins Exo, Beta, and Gam are very efficient in short recombinogenic ends protection and processing. Linear vectors can be generated conveniently as PCR products with synthetic primer ends that are homologous to sequence targets in the chloroplast genome. Chloroplast vectors will carry marker genes and also human papillomavirus oncogene E7 in the form of highly immunogenic but nononcogenic fusion E7GGG/GUS. It is our goal to obtain transplastomic plants producing this antigen with potential use as therapeutic vaccine. Viral antigens produced in plants will be assayed with antibodies and tested in mice.

Development of a Novel Recombinogenic Technique for Chloroplast Transformation and its Use for Production of human papillomavirus E7 Protein in Plants

COST Action BM0801 [2010 – 2012]

haskovec

Molecular analysis of epigenetic changes in AML and MDS patients for prognosis, progression and monitoring of treatment of the patients. Comparison of methylation in promotor parts of selected genes at diagnosis and during treatment of AML and MDS patients. Methods: DNA methylation arrays, bisulfite sequencing, methylation-specific PCR in real-time.

Nanobiophotonics for future health care (2012-2018, GA0/GB)

GA ČR P205/12/G118 [2012 – 2018]

doc. Ing. Jiří Homola, CSc., DSc., Ústav fotoniky a elektroniky AV ČR
prof. Ing. Jan E. Dyr, DrSc.

As modern medicine evolves towards quantitative and molecular based science, biophotonics is envisioned to play an increasingly important role in multiple areas of medicine, contributing to quality of health care, reduction of health care costs, and sustainability of the medical care in the ageing society. The proposed project aims to advance research in selected areas of nanobiophotonics with focus on photonic molecular biosensors based on plasmonic nanostructures. The main areas of research in this project include research into plasmonic phenomena on metallic nanostructures, development of novel tools for analysis and design of plasmonic nanostructures, fabrication and experimental characterization of plasmonic nanostructures with potential for surface plasmon resonance (SPR) and surface-enhanced Raman scattering (SERS) sensing, interfacing biomolecules with inorganic nanostructures and investigation of interactions between such biophotonic structures and biological samples, and realization of SPR and SERS biosensors for detection of biomarkers of onco-hematological diseases.

Effect of vaccination in patients with recurrent respirátory papillomatosis – can we improve the quality of life of these patients?

Merc & Dohme s.r.o. IIS ID 37651 [2011 – 2016]

RNDr. Ruth Tachezy, Ph.D.
MUDr. Jitka Vydrová – Medical Healthcom., s.r.o., Hlasové centrum, Praha

Phase III b clinical study. The main objective of the study will be to evaluate the impact of the tetravalent HPV vaccine on the occurrence of recurrent papillomatous lesions and remission duration in patients with recurrent respiratory papillomatosis (RRP). Secondary objectives will be to determine the HPV type involved in a RRP lesion and to monitor the levels of antibodies against the vaccine antigens or immune system components in the blood serum.

Origin of childhood leukaemias

IGA MZd NT 12328-5 [2011 – 2015]

doc. MUDr. Jan Zuna, Ph.D., 2. LF UK Praha
MUDr. Ivan Fales

Dr Ivan Fales and Dr Šárka Rahmatová are working in the Cord Blood Bank Czech Republic (CBB) as medical doctors and coordinators. They are responsible for related and unrelated part of CBB, for selection of mother-donors, for processing, cryopreservation and long-term storage of the cord blood grafts. Further more, they are responsible for the control of the processed grafts and for submission of the completely tested grafts to the Czech Stem Cells Registry (CSCR). The cord blood grafts are offered through CSCR for searches and transplantations. CBB Czech Republic closely cooperates with CSCR on selection and shipment of cord blood grafts for transplantation and also for elimination of the grafts in case of the child's (donor's) illness. In this project CBB will be responsible for the collection of cord blood samples of donors (mothers) with signed informed consent and for elimination of the cord blood grafts from the CSCR including the cases of haematological malignancy of the donor child. In these cases the cord blood samples and the grafts will be handed over to the CLIP laboratories for research purposes.

Risk factors for development of CMV virostatic resistance in the patients after allogeneic stem cell transplantation

IGA MZd NT 13691-4 [2012 – 2015]

MUDr. Petr Hubáček, PhD., 2. LF UK Praha
prof. MUDr. Petr Cetkovský, Ph.D., MBA

CMV resistance of virostatic treatment is severe complication of the CMV therapy in patients after allogeneic haematopoietic stem cell transplantation (HSCT). Therefore, we plan to clarify risk factors for development of CMV resistance associated with clinical symptoms, immunosuppressive therapy etc., genetic CMV variability of glycoprotein B, glycoprotein H and UL144. Efficacy of therapy will be monitored by measuring of the blood levels of ganciclovir/valganciclovir and length of the virostatic treatment too. Retrospectively, CMV genotypes and presence of CMV resistance strains in the patients after HSCT in the Institute of Haematology and Blood Transfusion and Dept. of Paediatric Haematology and Oncology of Motol University Hospital between the years 2002-2011. For statistical analysis of the risk factors, we'll collect the clinical data from tested patients. Prospective testing will be performed also in the centres with different historical frequency of this complication.

Human mesenchymal stromal cells for therapeutic purposes. Preclinical and pharmacological testing and approval of the cellular product for clinical use.

IGA MZd NT 13531-4 [2012 – 2015]

MUDr. Robert Pytlík 1. LF UK Praha
MUDr. Ivan Fales

Human mesenchymal stromal cells (hMSC) will be prepared form bone blood of patients undergoing bone marrow examination for diagnostic purposes and will be expanded by patented method developed by submitters of the grant project proposal. Previously obtained knowledge of pharmacodynamics will be completed with studies performed to proof the concept, to characterize the cells and to test safety of the cellular product in vitro and in vivo. Patented method of cultivation will be transferred to conditions of good manufacturing practice (GMP). Preclinical documentation concerning components used for preparation of the cellular product will be compiled and request for approval of the product for clinical use will be submitted to the Czech State Institute for Drug Control (SÚKL) to obtain approval for clinical use. If necessary, documentation will be completed to conform with SUKL requirements.


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Ústav hematologie a krevní transfuze
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