Technologie indukované pluripotence, která umožňuje přípravu prakticky jakéhokoliv buněčného modelu, potřebného pro výzkum vybraného onemocnění, se stává významným nástrojem v boji proti pandemii koronaviru SARS‐CoV-2 a případně i jiným formám pandemií, které se mohou s velkou pravděpodobností objevit v blízké, či vzdálené budoucnosti. Je výhodné, pokud lidstvo v předstihu disponuje arzenálem vhodných modelových systémů, které je možné v případě potřeby nasadit do boje a využít i pro vývoj nových terapeutických a diagnostických strategií. S ohledem na aktuální epidemickou situaci se tak významným způsobem může zkrátit čas, nezbytný pro nalezení vhodného léčebného postupu či zavedení spolehlivého diagnostického setu. Indukované pluripotentní kmenové buňky mají zároveň potenciál použití při regeneraci tkáně, poškozené samotnou infekcí. Jejich značnou výhodou na poli regenerativní medicíny je imunologicky atraktivní možnost využití tělu vlastních, tzv. autologních buněk, bez nutnosti nasazení imunosupresivní terapie po transplantaci.
Technology of induced pluripotency allows the preparation of any cell model for research of any selected disease and becomes the important tool also in the field of SARS‐CoV-2 coronavirus pandemic and other forms of pandemics that can appear in near or far future. It is beneficial for humankind to have an arsenal of useful model systems that can be immediately used in development of novel therapeutic or diagnostic strategies whenever needed. The possible use of newly developed therapy or diagnostic set can be greatly accelerated during pandemic situation. Induced pluripotent stem cells that can be prepared by technology of induced pluripotency also have great potential in tissue regeneration for tissues damaged by coronavirus infection. Immunologically very attractive is the fact that induced pluripotent stem cells can be applied (transplanted) without the use of aggressive immunosuppressive treatment.
- MeSH
- biologické modely MeSH
- COVID-19 terapie MeSH
- indukované pluripotentní kmenové buňky * MeSH
- lidé MeSH
- organoidy MeSH
- techniky in vitro * metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
Here, we present newly derived in vitro model for modeling Duchenne muscular dystrophy. Our new cell line was derived by reprogramming of peripheral blood mononuclear cells (isolated from blood from pediatric patient) with Sendai virus encoding Yamanaka factors. Derived iPS cells are capable to differentiate in vitro into three germ layers as verified by immunocytochemistry. When differentiated in special medium, our iPSc formed spontaneously beating cardiomyocytes. As cardiomyopathy is the main clinical complication in patients with Duchenne muscular dystrophy, the cell line bearing the dystrophin gene mutation might be of interest to the research community.
We present here a new iPS cell line for modeling sporadic form of ALS. Cell line was generated by reprogramming skin fibroblasts isolated with explant culture technology from skin biopsy, donated by ALS patient. For reprogramming, polycistronic self-replicating RNA vector was used and derived iPS cells were characterized by immunocytochemistry and FACS (pluripotent factors expression), karyotyping, STR fingerprinting analysis and in vitro differentiation assay. New cell line showed normal (46, XY) karyotype and differentiated in vitro into cells from three germ layers. STR analysis proved the origin and originality of the cell line.
- MeSH
- amyotrofická laterální skleróza * patologie MeSH
- buněčná diferenciace MeSH
- buněčné linie MeSH
- fibroblasty metabolismus MeSH
- indukované pluripotentní kmenové buňky * metabolismus MeSH
- lidé MeSH
- technologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The number of individuals diagnosed with colorectal cancer (CRC) has been on an alarming upward trajectory over the past decade. In some countries, this cancer represents one of the most frequently diagnosed types of neoplasia. Therefore, it is an important demand to study the pathology underlying this disease to gain insights into the mechanism of resistance to treatment. Resistance of tumors to chemotherapy and tumor aggressiveness have been associated with a minor population of neoplastic cells, which are considered to be responsible for tumor recurrence. These types of neoplastic cells are known as cancer stem cells, which have been previously reported to serve an important role in pathogenesis of this malignant disease. Slovakia has one of the highest incidence rates of CRC worldwide. In the present study, the aim was to classify the abundance of selected stem cell markers (CD133, CD166 and Lgr5) in CRC tumors using flow cytometry. In addition, the methylation status of selected genomic regions of CRC biomarkers (ADAMTS16, MGMT, PROM1 (CD133), LGR5 and ALCAM) was investigated by pyrosequencing in a cohort of patients from Martin University Hospital, Martin, Slovakia. Samples from both primary tumors and metastatic tumors were tested. Analysis of DNA methylation in the genomic regions of indicated five CRC biomarkers was also performed, which revealed the highest levels of methylation in the A disintegrin and metalloproteinase with thrombospondin motifs 16 and O6-methyguanine-DNA methyl transferase genes, whereas the lowest levels of methylation were found in genes expressing prominin-1, leucine-rich repeat-containing G-protein-coupled receptor 5 and activated leukocyte cell adhesion molecule. Furthermore, tumor tissues from metastases showed significantly higher levels of CD133+ cells compared with that in primary tumors. Higher levels of CD133+ cells correlated with TNM stage and the invasiveness of CRC into the lymphatic system. Although relatively small number of samples was processed, CD133 marker was consider to be important marker in pathology of CRC.
- Publikační typ
- časopisecké články MeSH
Objev indukované pluripotence v roce 2006 umožnil revoluční způsob získávaní autologních terapeuticky aplikovatelných buněk, a mož‐ nost modelovat jakékoliv onemocnění v in vitro podmínkách. Možnost vrátit libovolnou, finálně diferencovanou buňku „v čase“ zpátky do stádia pluripotence je zajímavé i pro oblast onkologického výzkumu. Tato technologie umožnila studium procesů spojených s roz‐ vojem nádorového fenotypu buňky a taky s přechodem nádorové buňky do stádia s nižší mírou diferenciace. Reprogramování buněk do indukovaných pluripotentních kmenových buněk také pomáhá mnohem lépe studovat raritní populaci buněk, přítomných v nádo‐ rech – tzv. nádorové kmenové buňky. Indukovaná pluripotence některých typů nádorových buněk, spojená s jejich následnou řízenou diferenciací by se zároveň mohla stát jednou z možných terapeutických aplikací v onkologii.
Discovery of technology of induced pluripotency that allows the generation of autologous therapeutically applicable cells and generati‐ on of in vitro cell models for diseases with limited (or highly invasive) access to tested cells has also opened new horizons in the field of oncology research. The unique ability to reprogram the cancer cell into pluripotency with subsequent directed differentiation into cell with no malignant phenotype should be considered as a challenge in the field of new oncotherapy development. Although still conside‐ red to be realistic only on the level of experimental approach, the recent progress in the field of induced pluripotency gives the hope that dedifferentiation‐based therapies connected with the erase of malignant phenotype of original cancer cell will be more realistic in near future. By then, the most important role of induced pluripotency in oncology remains in the field of regenerative therapy as a source of autologous cells for regeneration of tissues or organs damaged by tumor growth or aggressive therapy
Colorectal cancer (CRC) is one of the leading cancers in both genders. TNM staging system is still the most commonly used tumor classification and prognostic system. The disadvantage of TNM is that the prognostic information it provides is incomplete, and patients with the same histological tumor stages may differ significantly in the clinical outcome. Therefore, the identification of new prognostic parameters is crucial. The carcinogenic process that gives rise to an individual tumor is unique and tumor microenviroment should be taken into consideration. In CRC, T-cell infiltration is not homogenous, and recent studies are mostly focusing on memory T-cells and CD8 cells in predicting disease-free survival (DFS) and overall survival (OS). It seems that DFS and OS are not only dependent on microsatellite instable or stable status but mostly on the levels of expression of the immune signatures. Also, patients with high infiltration of cytotoxic and memory cells have significantly better outcome. This review consolidates current knowledge and recent research about importance of immune-cell-associated proteins, specific gene profiles of immune cells and immunotherapy in CRC. We also discussed cell-specific signatures in cancer treatment.
- MeSH
- imunoterapie MeSH
- inhibitory kontrolních bodů terapeutické užití MeSH
- kolorektální nádory genetika imunologie patologie MeSH
- lidé MeSH
- nádorové mikroprostředí MeSH
- prognóza MeSH
- sekvenční analýza RNA MeSH
- staging nádorů MeSH
- výpočetní biologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Pankreatický duktální adenokarcinom (PDAC) představuje jeden z nejagresivnějších typů lidských malignit. V současnosti je toto zhoubné onemocnění čtvrtou nejčastější příčinou úmrtí na rakovinu. Pětileté přežití pacientů s duktálním adenokarcinomem je méně než 8 %. Nové in vitro a in vivo modely jsou proto nutně potřebné pro vývoj nových terapií. S vlastní technologií pro derivaci nových, unikátních 3D nádorových buněčných linií izolovaných z lidských nádorů a ve spolupráci s Ústavem živočišné fyziologie a genetiky AV ČR v Liběchově představujeme plán vývoje velkého zvířecího modelu pro modelování lidského PDAC. S využitím tohoto modelu a nejmodernějších laboratorních technik provedeme profilovací analýzy (detekce a fenotypování cirkulujících rakovinných buněk, izolace a sekvenování cirkulující DNA, metabolomické profilování a analýzu onkoproteinů, a detekce cytokinů pomocí multiplexních protilátkových čipů za účelem nalezení biomarkerů nádoru pankreatu. Nově vyvinutý model zároveň poskytneme pro potřeby výzkumu spolupracujícím vědeckým pracovištím a farmaceutickým společnostem.
Pancreatic ductal adenocarcinoma represents one of the most aggressive type of human malignancy. Currently, this malignancy is the fourth most frequent cause of dead. 5-year survival of patients with ductal adenocarcinoma is less than 8 %. New in vitro and in vivo models are therefore desperately needed for new therapy development. With our own technology for derivation of new, unique 3D cancer cell lines from human tumors and in collaboration with Institute of Animal Physiology and Genetics, AS CR, in Libechov, we present here the plan for the development of large xenograft animal model (pig) bearing human pancreatic tumor. With the use of this model and state -of -the -art lab.techniques, we will perform multiple profiling analyses (circulating cancer cell detection and phenotyping, circulating DNA isolation and sequencing, metabolomic profiling and cancer -related proteins and cytokines detection with multiplex antibody array chips in order to find the hallmarks of pancreatic tumor.
Melanoma-bearing Libechov minipig (MeLiM) represents a large animal model for melanoma research. This model shows a high incidence of complete spontaneous regression of melanoma - a phenomenon uncommon in humans. Here, we present the first metabolomic characterisation of the MeLiM model comparing animals with progressing and spontaneously regressing melanomas. Plasma samples of 19 minipigs with progression and 27 minipigs with evidence of regression were analysed by a targeted metabolomic assay based on mass spectrometry detection. Differences in plasma metabolomics patterns were investigated by univariate and multivariate statistical analyses. Overall, 185 metabolites were quantified in each plasma sample. Significantly altered metabolomic profile was found, and 42 features were differentially regulated in plasma. Besides, the machine learning approach was used to create a predictive model utilising Arg/Orn and Arg/ADMA ratios to discriminate minipigs with progressive disease development from minipigs with regression evidence. Our results suggest that progression of melanoma in the MeLiM model is associated with alteration of arginine, glycerophospholipid and acylcarnitines metabolism. Moreover, this study provides targeted metabolomics characterisation of an animal model of melanoma with progression and spontaneous regression of tumours.
- MeSH
- metabolomika metody MeSH
- miniaturní prasata MeSH
- modely nemocí na zvířatech MeSH
- prasata MeSH
- progrese nemoci MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Tumor hypoxia is described as an oxygen deprivation in malignant tissue. The hypoxic condition is a consequence of an imbalance between rapidly proliferating cells and a vascularization that leads to lower oxygen levels in tumors. Hypoxia-inducible factor 1 (HIF-1) is an essential transcription factor contributing to the regulation of hypoxia-associated genes. Some of these genes modulate molecular cascades associated with the Warburg effect and its accompanying pathways and, therefore, represent promising targets for cancer treatment. Current progress in the development of therapeutic approaches brings several promising inhibitors of HIF-1. Flavonoids, widely occurring in various plants, exert a broad spectrum of beneficial effects on human health, and are potentially powerful therapeutic tools against cancer. Recent evidences identified numerous natural flavonoids and their derivatives as inhibitors of HIF-1, associated with the regulation of critical glycolytic components in cancer cells, including pyruvate kinase M2(PKM2), lactate dehydrogenase (LDHA), glucose transporters (GLUTs), hexokinase II (HKII), phosphofructokinase-1 (PFK-1), and pyruvate dehydrogenase kinase (PDK). Here, we discuss the results of most recent studies evaluating the impact of flavonoids on HIF-1 accompanied by the regulation of critical enzymes contributing to the Warburg phenotype. Besides, flavonoid effects on glucose metabolism via regulation of HIF-1 activity represent a promising avenue in cancer-related research. At the same time, only more-in depth investigations can further elucidate the mechanistic and clinical connections between HIF-1 and cancer metabolism.
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
We generated new in vitro model for sporadic form of amyotrophic lateral sclerosis by reprogramming isolated skin fibroblasts into iPSCs. Fibroblasts were reprogrammed with commercially available synthetic polycistronic, self-replicating RNA vector. As verified by FISH, an early passages of a new iPSC line showed mosaic karyotype (cells with normal and abnormal karyotype 46,XY,t(2;14)(q13;p12) were present), while late passages contained only cells with abnormal karyotype. New iPSCs differentiated into all three germ layers and formed a teratoma in nude mice. Our iPSC line represents a new model for therapy testing and drug development in the field of ALS research.
- MeSH
- amyotrofická laterální skleróza * genetika MeSH
- buněčná diferenciace MeSH
- fibroblasty MeSH
- indukované pluripotentní kmenové buňky * MeSH
- myši nahé MeSH
- myši MeSH
- přeprogramování buněk MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
