Selection of the proper target is crucial for clinically relevant monitoring of minimal residual disease (MRD) in patients with acute lymphoblastic leukemia using the quantitation of clonal-specific immunoreceptor (immunoglobulin/T cell receptor) gene rearrangements. Consequently, correct interpretation of the results of the entire analysis is of utmost importance. Here we present an overview of the quality control measures that need to be implemented into the process of marker identification, selection, and subsequent quantitation of the MRD level.
- MeSH
- Precursor Cell Lymphoblastic Leukemia-Lymphoma * diagnosis genetics MeSH
- Biomarkers MeSH
- Immunoglobulins genetics MeSH
- Humans MeSH
- Neoplasm, Residual diagnosis genetics MeSH
- Quality Control MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Review MeSH
Práce shrnuje dosud známé poznatky o vybraných imunologických ukazatelích klinického stavu u pacientů v sepsi a prezentuje výsledky pilotní studie 24 pacientů hospitalizovaných na Klinice anesteziologie a resuscitace (KAR) Fakultní nemocnice Královské Vinohrady s diagnózou sepse. 1., 3. a 5. den hospitalizace na KAR byly měřeny koncentrace IL-6, IL-10, HLA-DR exprese monocyty a počítáno APACHE II skóre. Koncentrace IL-10 měřené 3. (p=0,02) a 5. (p=0,05) den hospitalizace na KAR a maximální naměřené koncentrace IL-6 (p=0,04) a IL-10 (p=0,05) byly statisticky významně vyšší a HLA-DR exprese 5. den (p=0,01) nižší u později zemřelých pacientů (oproti těm, kteří se ze sepse vyléčili). Skóre APACHE II korelovalo 1. den statisticky významně (p=0,03) negativně s HLA-DR expresí monocyty.Význam IL-6 jako ukazatele souvisejícího s rozvojem septického šoku není z výsledků naší studie zřejmý. Z výsledků práce vyplývá, že hodnoty měřené při příjmu jako jediné nestačí k vytipování pacientů se závažnější prognózou, ideální by bylo dlouhodobé monitorování imunologických znaků umožňující sledovat vývoj imunologického stavu pacientů.
The paper summarises current knowledge on markers of the immune status in patients with sepsis and presents the results of a pilot study of 24 patients hospitalised at the Intensive Care Unit (ICU) of the University hospital Královské Vinohrady. IL-6 and IL-10 concentrations and the HLA-DR expression on monocytes were measured and APACHE II score counted on day 1, 3 and 5 following admission to the ICU. IL-10 concentrations measured on day 3 (p=0.02) and 5 (p=0.05) and the maximal measured IL-6 and IL-10 concentrations (p=0.04 and p= 0.05 respectively) were significantly higher and HLA-DR expression on monocytes on day 5 (p=0.01) was significantly lower in patients who had later died as a consequence of the sepsis episode (compared to those who had survived). APACHE II score negatively correlated with HLA-DR expression on monocytes on day 1 (p=0.03). The value of IL-6 as a marker related to the presence of septic shock is not obvious from the results of our study. Based on the results of the study, the values measured on admission to the hospital are insufficient to sort out the patients with a more serious prognosis; the ideal would be long-term monitoring of the immune status markers, which would enable us study the dynamics of the patient’s immune status.
- MeSH
- Biomarkers blood MeSH
- Adult MeSH
- HLA-DR Antigens blood MeSH
- Immunologic Techniques methods MeSH
- Interleukin-10 blood MeSH
- Interleukin-6 blood MeSH
- Middle Aged MeSH
- Humans MeSH
- Monocytes MeSH
- Aged MeSH
- Sepsis immunology mortality physiopathology MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Review MeSH
- Comparative Study MeSH
Východiská: AML je agresívne, fenotypovo a geneticky heterogénne klonálne ochorenie krvotvorných progenitorových buniek s veľkou molekulárnou variabilitou. Nová klasifikácia WHO 2008 delí de novo AML podľa cytogenetických a molekulárnych prognostických a prediktívnych markerov. V poslednej dobe je stále väčšia možnosť určiť podskupinu rizikových pacientov so zlou prognózou medzi tými s normálnym karyotypom. Cieľom našej štúdie bolo zistiť prognosticky významné molekulárne markery u detí s AML na stratifikáciu pacientov s normálnym karyotypom a na sledovanie ochorenia podľa genetického nálezu. Súbor pacientov a metódy: V rokoch 2008–2010 sme analyzovali vzorky kostnej drene a periférnej krvi u 20 detí s de novo AML konvenčnou cytogenetickou analýzou, fluorescenčnou in situ hybridizáciou a molekulovou analýzou. Molekulárna analýza bola vykonaná na úrovni cDNA restrikčnou analýzou PCR produktov (FLT3-TKD), konvenčnou PCR (MLL-PTD, NPM1mut, FLT3-ITD) a kvantifikačnou RT-PCR (expresia fúznych génov, génov BAALC a WT1). Výsledky: Vzorky 20 detí s AML boli analyzované využitím konvenčnej cytogenetiky, FISH a molekulovo-genetickými metódami. Abnormálny karyotyp bol zistený u 13 pacientov (65 %). Ďalšia analýza ukázala FLT3-ITD v 5/20 (25 %), FLT3-TKD v 3/20 (15 %), NPM1mut 2/20 (10 %) a MLL-PTD v 1/20 (5 %), nadmernej expresie WT1 génu v 15/20 (75 %) a nadmernej expresie BAALC v 13/20 (65 %) pacientov. Záver: Široký cytogenetický a molekulárny skríning pomohol nájsť aspoň jeden genetický marker u všetkých 20 pacientov pre ďalšie sledovanie a stratifikáciu rizika detí s AML. Na progresiu ochorenia umreli 4/20 (20 %) detí.
Backgrounds: AML is an aggressive, phenotypically and genetically heterogenous clonal disease of hematopoietic progenitor cells with a great molecular variability. New WHO classification 2008 divides de novo AML according to cytogenetic and molecular prognostic and predictive markers. Recently, it is increasingly possible to identify a subgroup of poorer prognosis patients among those with normal karyotype AML. The aim of our study was to identify prognostically important molecular markers in children with AML, to stratify patients with normal karyotype and to monitor the disease according the genetic findings. Material and Methods: In 2008–2010, we analyzed bone marrow and peripheral blood samples of 20 children with de novo AML by conventional cytogenetic analysis, fluorescence in situ hybridisation and molecular diagnostics. The molecular analysis was performed on the cDNA level, with the restriction analysis of PCR products (FLT3-TKD), conventional PCR (MLL-PTD, NPM1mut, FLT3- ITD) and quantification RT-PCR method (expression of fusion transcripts, BAALC, WT1). Results: Samples from 20 children with AML were analyzed using the conventional cytogenetics, FISH and molecular methods. Abnormal karyotype was identified in 13 patients (65%). Further analysis revealed FLT3-ITD in 5/20 (25%), FLT3-TKD in 3/20 (15%), NPM1mut in 2/20 (10%) and MLL-PTD in 1/20 (5%), overexpression of WT1 gene in 15/20 (75%) and overexpression of BAALC in 13/20 (65%) patients. Conclusion: Wide cytogenetic and molecular screening helped to find at least one genetic marker in all 20 patients for later follow-up and risk stratification. 4/20 (20%) patients died of the disease progression.
- MeSH
- Leukemia, Myeloid, Acute diagnosis genetics blood MeSH
- Cytogenetic Analysis methods utilization MeSH
- Child MeSH
- Gene Expression genetics MeSH
- Genetic Markers genetics MeSH
- In Situ Hybridization, Fluorescence methods utilization MeSH
- Nuclear Proteins genetics isolation & purification MeSH
- Humans MeSH
- Mutation genetics MeSH
- Neoplasm Proteins genetics isolation & purification MeSH
- Polymerase Chain Reaction methods utilization MeSH
- WT1 Proteins genetics isolation & purification MeSH
- Myeloid-Lymphoid Leukemia Protein genetics isolation & purification MeSH
- Statistics as Topic MeSH
- fms-Like Tyrosine Kinase 3 genetics isolation & purification MeSH
- Check Tag
- Child MeSH
- Humans MeSH
Contemporary issues in genetics and evolution ; vol. 4
213 s. : il.
- MeSH
- DNA Fingerprinting * MeSH
- Conspectus
- Kriminalistika
- NML Fields
- soudní lékařství
- genetika, lékařská genetika
- NML Publication type
- kolektivní monografie
BACKGROUND: Melanoma is the most aggressive skin cancer with ability to recur also after early-stage tumor surgery. The aim was to identify early-stage melanoma patients at high risk of recurrence using liquid biopsy, estimating of mutated BRAF ctDNA and the level of tumor marker S100B in plasma. METHODS: Eighty patients were enrolled in the study. BRAF V600E mutation was determined in FFPE tissue and plasma samples using ultrasensitive ddPCR with pre-amplification. The level of S100B was determined in plasma by immunoassay chemiluminescent method. RESULTS: The best prediction of melanoma recurrence after surgery was observed in patients with combined high level of S100B (S100Bhigh) and ctDNA BRAFV600E (BRAFmut) in preoperative (57.1% vs. 12.5%, p = 0.025) as well as postoperative blood samples (83.3% vs. 14.3%, resp., p = 0.001) in comparison with low S100B and BRAF wild-type. Similarly, patients with preoperative and postoperative S100Bhigh and BRAFmut experienced worse prognosis (DFI p = 0.05, OS p = 0.131 and DFI p = 0.001, OS = 0.001, resp.). CONCLUSION: We observed the benefit of the estimation of combination of S100B and ctDNA BRAFmut in peripheral blood for identification of patients at high risk of recurrence and unfavorable prognosis. SIGNIFICANCE: There is still no general consensus on molecular markers for deciding the appropriateness of adjuvant treatment of early-stage melanoma. We have shown for the first time that the combined determination of the ctDNA BRAFmut oncogene (liquid biopsy) and the high level of tumor marker S100B in pre- and postoperative plasma samples can identify patients with the worst prognosis and the highest risk of tumor recurrence. Therefore, modern adjuvant therapy would be appropriate for these patients with resectable melanoma, regardless of disease stage.
- MeSH
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Neoplasm Recurrence, Local * genetics blood MeSH
- Melanoma * genetics blood pathology diagnosis surgery MeSH
- Mutation * MeSH
- Biomarkers, Tumor * blood genetics MeSH
- Skin Neoplasms * blood genetics pathology diagnosis surgery MeSH
- Predictive Value of Tests MeSH
- Prognosis MeSH
- Proto-Oncogene Proteins B-raf * genetics blood MeSH
- S100 Calcium Binding Protein beta Subunit * blood genetics MeSH
- Aged, 80 and over MeSH
- Aged MeSH
- Neoplasm Staging MeSH
- Liquid Biopsy methods MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged, 80 and over MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
A key requirement for precision medicine is the accurate identification of patients that would respond to a specific treatment or those that represent a high-risk group, and a plethora of molecular biomarkers have been proposed for this purpose during the last decade. Their application in clinical settings, however, is not always straightforward due to relatively high costs of some tests, limited availability of the biological material and time, and procedural constraints. Hence, there is an increasing interest in constructing tissue-based surrogate biomarkers that could be applied with minimal overhead directly to histopathology images and which could be used for guiding the selection of eventual further molecular tests. In the context of colorectal cancer, we present a method for constructing a surrogate biomarker that is able to predict with high accuracy whether a sample belongs to the "BRAF-positive" group, a high-risk group comprising V600E BRAF mutants and BRAF-mutant-like tumors. Our model is trained to mimic the predictions of a 64-gene signature, the current definition of BRAF-positive group, thus effectively identifying histopathology image features that can be linked to a molecular score. Since the only required input is the routine histopathology image, the model can easily be integrated in the diagnostic workflow.
Kolorektální karcinom je jedním z nejčastěji se vyskytujících nádorových onemocnění. Bohužel, významná část pacientů v klinických stádiích II a III umírá do pěti let po radikálním chirurgickém zákroku následkem progrese onemocnění. Racionální přístup k indikaci adjuvantní léčby pro tyto pacienty nabízí molekulární charakterizace jejich rizika napfříč oběma klinickými stádii pomocí technologie DNA čipů. Do studie byly zařazeny bioptické vzorky dvanácti pacientů s histologicky potvrzeným kolorektálním karcinomem (KRK) v klinickém stádiu II a III tvořené minimálnu ze 70% maligními buňkami. Šest pacientů mělo dobrou prognózu s délkou bezpříznakového přežití (DFS) delší než 36 měsíců, Šest mělo špatnou prognózu s DFS kratším než 36 měsíců. Pomocí nízkohustotních oligonukleotidových makročipů společnosti SuperArray určených k relativní kvantifikaci exprese 128 genů potenciálně zapojených do procesu metastazování, byly identifikovány profily genové exprese primárních KRK všech 12 pacientů. Analýzou expresních profilů pomocí t-testu (á = 0,01) a metody SAM jsme identifikovali 10 genů rozdílnu exprimovaných (9 upregulovaných, 1 down-regulovaný) v primárních nádorech pacientů se špatnou prognózou. Naše výsledky nasvědčují, že technologie nízkohustotních oligonukleotidových makročipů je užitečnou pomůckou pro lep- ší pochopení molekulární podstaty progrese nádorového onemocnění a zlepšení predikce metastatického potenciálu primárních nádorů u pacientů s lokoregionálně pokročilým kolorektálním karcinomem.
Colorectal cancer (CRC) is one of the most common malignancies. Unfortunately, a significant proportion of surgically cured patients in the early stage of the disease develop progression and die from the disease. Twelve patients who had histologically confirmed left-sided colon adenocarcinoma with a volume fraction showing at least 70% of malignant tumor cells were included. Only stage II-III patients according to IUCC with no prior chemotherapy or radiotherapy were eligible for the study. Six patients were poor prognosis cases with disease free survival (DFS) lower then 36 month and six were good prognosis cases with DFS>36 month. Relative gene expression levels of 128 genes potentially involved in cancer progression and dissemination were obtained by low-density oligonucleotide microarrays (SuperArray Bioscience Corp., Bethesda, MD) from 12 primary colon cancer samples. Gene expression data analysis based on the SAM and ttest (á = 0, 01) methods identified 10 genes with significantly different expression in primary tumors of patients with poor prognosis. Our preliminary data suggest that oligonucleotide microarray technology should contribute to a better understanding of the progression of colorectal cancer, and facilitate prediction of their metastatic potential.
- MeSH
- DNA Fingerprinting methods utilization MeSH
- Gene Expression genetics MeSH
- Financing, Organized MeSH
- Genetic Markers genetics immunology MeSH
- Colorectal Neoplasms diagnosis etiology genetics MeSH
- Medical Oncology methods trends MeSH
- Humans MeSH
- Pilot Projects MeSH
- Reverse Transcriptase Polymerase Chain Reaction methods utilization MeSH
- Disease-Free Survival MeSH
- Prognosis MeSH
- Oligonucleotide Array Sequence Analysis methods utilization MeSH
- Neoplasm Staging methods utilization MeSH
- Check Tag
- Humans MeSH
