Východiská: V managemente pacientov s kolorektálnym karcinómom (colorectal cancer – CRC) stále existuje priestor pre zlepšenie stratifikácie rizika a tým presnejšie „ušitie“ liečby na mieru. Veľmi sľubným sa v tomto ohľade javia biomarkery získavané prostredníctvom takzvanej tekutej biopsie, čo je neinvazívna metóda odberu telesných tekutín pacienta, najčastejšie periférnej krvi. Analyzujú sa rozličné biomarkery súvisiace s nádorom, ktoré môžu mať ako prognostickú, tak aj prediktívnu hodnotu. Jedným z najviac prebádaných nádorových biomarkerov je práve nádorová cirkulujúca DNA, ktorej spektrum využitia bolo spočiatku len u pokročilých a metastatických karcinómov a spočívalo v molekulárnom profilovaní alebo zisťovaní získanej rezistencie k liečbe. V súčasnosti sa využitie cirkulujúcej nádorovej DNA (ctDNA) posunulo už k skorým štádiám karcinómov, kde okrem iného slúži k identifikácii minimálnej reziduálnej choroby alebo k skorej diagnostike CRC. Doterajšie štúdie ukazujú veľmi sľubný potenciál týchto biomarkerov, ale k využitiu v klinickej praxi bude potrebné získať viac informácií a počkať na výsledky prebiehajúcich výskumov. Cieľ: V tomto prehľadovom článku sa budeme venovať ctDNA, jej aspektom, možnostiam diagnostiky a súčasnému využitiu v rámci CRC.
Background: Space still exists in the management of patients with colorectal cancer (CRC) for improving risk stratification and thus the precision of treatment tailoring. Quite promising in this regard are biomarkers acquired via liquid biopsy, which is a non-invasive method of body fluid draw, most commonly peripheral blood. A variety of biomarkers associated with the tumor are analyzed, which can have either prognostic or predictive value. Circulating tumor DNA (ctDNA) is one of the most explored tumor biomarkers. Initially, its utility spectrum was only in advanced or metastatic cancers and consisted of molecular profiling and detecting acquired resistance to treatment. Nowadays, the use of circulating tumor DNA has shifted to earlier cancer stages, where it can identify minimal residual disease or diagnose colorectal cancer early. Existing studies show promising potential of these biomarkers, but more information needs to be gathered and information from ongoing studies needs to be obtained in order to use them in everyday practice. Aim: In this review article, we will discuss ctDNA, its aspects, diag- nostic possibilities and current use in CRC.
BACKGROUND: To validate the clinical utility of a previously identified circulating tumor DNA methylation marker (meth-ctDNA) panel for disease detection and survival outcomes, meth-ctDNA markers were compared to PSA levels and PSMA PET/CT findings in men with different stages of prostate cancer (PCa). METHODS: 122 PCa patients who underwent [68Ga]Ga-PSMA-11 PET/CT and plasma sampling (03/2019-08/2021) were analyzed. cfDNA was extracted, and a panel of 8 individual meth-ctDNA markers was queried. PET scans were qualitatively and quantitatively assessed. PSA and meth-ctDNA markers were compared to PET findings, and their relative prognostic value was evaluated. RESULTS: PSA discriminated best between negative and tumor-indicative PET scans in all (AUC 0.77) and hormone-sensitive (hsPC) patients (0.737). In castration-resistant PCa (CRPC), the meth-ctDNA marker KLF8 performed best (AUC 0.824). CHST11 differentiated best between non- and metastatic scans (AUC 0.705) overall, KLF8 best in hsPC and CRPC (AUC 0.662, 0.85). Several meth-ctDNA markers correlated low to moderate with the tumor volume in all (5/8) and CRPC patients (6/8), while PSA levels correlated moderately to strongly with the tumor volume in all groups (all p < 0.001). CRPC overall survival was independently associated with LDAH and PSA (p = 0.0168, p < 0.001). CONCLUSION: The studied meth-ctDNA markers are promising for the minimally-invasive detection and prognostication of CRPC but do not allow for clinical characterization of hsPC. Prospective studies are warranted for their use in therapy response and outcome prediction in CRPC and potential incremental value for PCa monitoring in PSA-low settings.
- MeSH
- cirkulující nádorová DNA genetika krev MeSH
- EDTA analogy a deriváty MeSH
- izotopy gallia * MeSH
- lidé středního věku MeSH
- lidé MeSH
- metylace DNA * genetika MeSH
- nádorové biomarkery * genetika krev MeSH
- nádory prostaty rezistentní na kastraci genetika krev diagnostické zobrazování MeSH
- nádory prostaty * genetika krev diagnostické zobrazování MeSH
- PET/CT * metody MeSH
- prognóza MeSH
- prostatický specifický antigen * krev genetika MeSH
- průřezové studie MeSH
- radioizotopy galia * MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
High-grade B-cell lymphomas (HGBCLs) are aggressive blood cancers with a severe disease course, especially when the central nervous system (CNS) is involved. Standard histological examination depends on tissue availability and is currently supplemented with molecular tests, as the status of MYC, BCL2, or BCL6 gene rearrangements is required for proper lymphoma classification. This case report demonstrates the relevance of cerebrospinal fluid (CSF) cell-free DNA testing by integrative next-generation sequencing (NGS) panel. The benefit of this approach resided in tumor genotyping alongside the proof of CNS progression despite MRI negativity, revealing a clonal relationship with the primary tumor lesion. In addition, our strategy allowed us to classify the tumor as DLBCL/HGBL-MYC/BCL2 entity. In clinical practice, such a minimally invasive approach provides a more sensitive tool than standard imaging and cell analyzing techniques, enabling more accurate disease monitoring and relapse prediction in particular cases.
- MeSH
- B-buněčný lymfom genetika patologie diagnóza diagnostické zobrazování MeSH
- cirkulující nádorová DNA genetika MeSH
- difúzní velkobuněčný B-lymfom genetika patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- lokální recidiva nádoru patologie genetika MeSH
- magnetická rezonanční tomografie * MeSH
- nádorové biomarkery genetika MeSH
- nádory centrálního nervového systému genetika patologie diagnostické zobrazování MeSH
- vysoce účinné nukleotidové sekvenování * MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
Gliomas are the most common brain tumor type in children and adolescents. To date, diagnosis and therapy monitoring for these tumors rely on magnetic resonance imaging (MRI) and histopathological as well as molecular analyses of tumor tissue. Recently, liquid biopsies (LB) have emerged as promising tool for diagnosis and longitudinal tumor assessment potentially allowing for a more precise therapeutic management. However, the optimal strategy for monitoring gliomas by LB remains to be determined. In this study, we analyzed circulating tumor DNA (ctDNA) from 78 liquid biopsies (plasma n = 44, cerebrospinal fluid n = 34 (CSF)) of 35 glioma patients, determining H3F3A K28M (K27M) and BRAF V600E mutation allele frequency using droplet digital PCR (ddPCR). All results were correlated to clinically relevant parameters including diagnostic imaging and CSF aspiration site (ventricular vs lumbar) with respect to tumor localization. Regarding diagnostic accuracy, the calculated sensitivity score in the H3F3A K27M cohort was 84.61% for CSF and 73.68% for plasma. In the BRAF V600E cohort, we determined a sensitivity of 83.3% in plasma and 80% in CSF. The overall specificity was 100%. With respect to the CSF aspiration, the intra-operatively obtained CSF demonstrated 100% detection rate, followed by ventricular CSF obtained via Ommaya Reservoir/shunt puncture (93%) and CSF obtained via lumbar puncture (66%). Notably, this further correlated with the proximity of the CSF site to tumor localization. Longitudinal CSF monitoring demonstrated a good correlation to clinical and radiological disease evolution. Importantly, we show for the first time that monitoring BRAF V600E by ddPCR could serve as treatment response assessment in gliomas. In summary, our observation may inform recommendations with regard to location of CSF aspiration when incorporating LB into future treatment protocols.
- MeSH
- cirkulující nádorová DNA mozkomíšní mok genetika MeSH
- dítě MeSH
- dospělí MeSH
- gliom * genetika patologie diagnóza MeSH
- histony * genetika MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mutace MeSH
- nádorové biomarkery * genetika mozkomíšní mok MeSH
- nádory mozku * genetika diagnóza patologie MeSH
- předškolní dítě MeSH
- protoonkogenní proteiny B-Raf * genetika MeSH
- tekutá biopsie metody MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- předškolní dítě MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Obesity is considered an important factor contributing to the development of atherosclerosis. Inflammation plays a key role in endothelial dysfunction (ED), an initial stage of the atherosclerotic process. Several microRNAs (miRNAs) may play an important role in the inflammatory process, but there is a lack of information about their participation in the early stages of atherosclerosis development in patients with obesity. We aimed to assess the relations between plasma concentration of selected miRNAs, ED evaluated by reactive hyperemia index (RHI), inflammatory markers and other factors involved in the pathogenesis of atherosclerosis in adolescents and young adults with obesity. Participants (30 males, 30 females; aged 15 25 years) were divided into two groups: those with overweight/obesity (OW/O) (20 males, 20 females) and controls (C) (10 males, 10 females). The plasma concentrations of inflammatory markers, cytokines, adipocytokines, markers of lipid profile and glucose metabolism and selected miRNAs (miR 92, 126, -146a, -155) were analyzed. No significant differences in any of the miRNAs were found between the groups. MiR-146a correlated positively with RHI. Dividing the group by sex showed more significant associations between miRNA and analyzed parameters (IL-6, fasting glycemia) in men. Several observed correlations indicate a potential role of miRNAs in inflammation, the atherosclerotic process and glycemic control, primarily in male subjects with obesity. The relatively low number of observed associations between assessed parameters related to obesity and the pathogenesis of its complications could be attributed to the early stage of the atherosclerotic process in young subjects with obesity, where only subtle abnormalities are expectedly found. Key words Endothelial dysfunction, Atherosclerosis, Obesity, MicroRNA, Reactive hyperemia index.
- MeSH
- ateroskleróza * krev genetika MeSH
- biologické markery krev MeSH
- cirkulující mikroRNA * krev genetika MeSH
- dospělí MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- obezita * krev komplikace genetika MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Tekutá biopsie představuje novou nadějnou možnost v diagnostice, monitoraci průběhu onemocnění a časné detekci relapsu. Velkou výhodou je její miniinvazivita, možnost častého opakovaní, a tím monitorace onemocnění v reálném čase. V tomto článku budou shrnuty dosavadní výsledky jejího použití u urologických nádorů.
The liquid biopsy is a new promising option in diagnosis, monitoring during the treatment and early detection of relapse. A big advantage of liquid biopsy is its mini-invasivity, the possibility of frequent repetition and thus real-time monitoring of the disease. This article summarizes the current results of liquid biopsy in urological malignancies.
- MeSH
- cirkulující nádorová DNA MeSH
- lidé MeSH
- nádorové biomarkery MeSH
- nádorové cirkulující buňky MeSH
- nádory ledvin diagnóza MeSH
- nádory močového měchýře diagnóza MeSH
- nádory prostaty diagnóza MeSH
- tekutá biopsie * metody MeSH
- testikulární nádory diagnóza MeSH
- urologické nádory diagnóza patologie terapie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- komentáře MeSH
Výskyt krátkých fragmentů volných DNA (cell-free DNA – cfDNA) jaderného či mitochondriálního původu v tělesných tekutinách je fyziologický. Jejich koncentrace násobně narůstá následkem zranění, zánětu či intenzivní fyzické námahy. Zvýšené koncentrace se rychle vracejí na původní hladiny díky masivní degradaci restrikčními endonukleázami a extenzivnímu katabolizmu v játrech, slezině a částečně i díky vylučování ledvinami. Vyšší koncentrace cfDNA byly popsány v souvislosti s přítomností většiny rizikových faktorů aterosklerózy. Existuje silná korelace mezi cfDNA a koncentracemi kreatinkinázy a troponinu. cfDNA může být použita jako marker cévní mozkové příhody či k rozlišení STEMI a nonSTEMI infarktu myokardu. Mitochondriální cfDNA pak stimuluje zánětlivé procesy. Přestože řada nálezů potřebuje ověřit na rozsáhlejších souborech, cfDNA se mohou v budoucnu stát novými slibnými markery onemocnění a zdravotních prognóz i v kardiologii.
The presence of short fragments of cell-free DNA (cfDNA), of nuclear or mitochondrial origin, in body fluids is physiological. Their concentration increases manifold as a result of injury, inflammation or intensive physical exercise. Increased concentrations quickly return to their baseline levels due to intense degradation by restriction endonucleases and extensive catabolism in the liver, spleen and partly excretion by the kidneys. Higher cfDNA concentrations have been described in association with the presence of majority atherosclerosis risk factors. There is a strong correlation between cfDNA and creatine kinase and troponin concentrations. CfDNA can be used as a marker of stroke or to differentiate between STEMI and non-STEMI myocardial infarction. Mitochondrial cfDNA then stimulates inflammatory processes. Although many of the findings need validation on larger datasets, cfDNAs may become new promising markers of disease and health prognosis in the future also in cardiology.
Background/Objectives: Although the overall survival prognosis of patients in advanced stages of pancreatic ductal adenocarcinoma (PDAC) is poor, typically ranging from days to months from diagnosis, there are rare cases of patients remaining in therapy for longer periods of time. Early estimations of survival prognosis would allow rational decisions on complex therapy interventions, including radical surgery and robust systemic therapy regimens. Understandably, there is great interest in finding prognostic markers that can be used for patient stratification. We determined the role of various KRAS mutations in the prognosis of PDAC patients using biopsy samples and circulating tumor DNA. Methods: A total of 118 patients with PDAC, clinically confirmed by endoscopic ultrasound-guided fine-needle aspiration biopsy (EUS-FNB), were included in the study. DNA was extracted from cytological slides following a standard cytology evaluation to ensure adequacy (viability and quantity) and to mark the tumor cell fraction. Circulating tumor DNA (ctDNA) was extracted from plasma samples of 45 patients in stage IV of the disease. KRAS mutations in exons 12 and 13 were detected by denaturing capillary electrophoresis (DCE), revealing a minute presence of mutation-specific heteroduplexes. Kaplan-Meier survival curves were calculated for individual KRAS mutation types. Results:KRAS mutations were detected in 90% of tissue (106/118) and 44% of plasma (20/45) samples. All mutations were localized at exon 2, codon 12, with G12D (GGT > GAT) being the most frequent at 44% (47/106) and 65% (13/20), followed by other types including G12V (GGT > GTT) at 31% (33/106) and 10% (2/20), G12R (GGT > CGT) at 17% (18/106) and 10% (2/20), G12C (GGT/TGT) at 5% (5/106) and 0% (0/20) and G12S (GGT/AGT) at 1% (1/106) and 5% (1/20) in tissue and plasma samples, respectively. Two patients had two mutations simultaneously (G12V + G12S and G12D + G12S) in both types of samples (2%, 2/106 and 10%, 2/20 in tissue and plasma samples, respectively). The median survival of patients with the G12D mutation in tissues was less than half that of other patients (median survival 101 days, 95% CI: 80-600 vs. 228 days, 95% CI: 184-602), with a statistically significant overall difference in survival (p = 0.0080, log-rank test), and furthermore it was less than that of all combined patients with other mutation types (101 days, 95% CI: 80-600 vs. 210 days, 95% CI: 161-602, p = 0.0166). For plasma samples, the survival of patients with this mutation was six times shorter than that of patients without the G12D mutation (27 days, 95% CI: 8-334 vs. 161 days, 95% CI: 107-536, p = 0.0200). In contrast, patients with detected KRAS G12R in the tissue survived nearly twice as long as other patients in the aggregate (286 days, 95% CI: 70-602 vs. 162 days, 95% CI: 122-600, p = 0.0374) or patients with other KRAS mutations (286 days, 95% CI: 70-602 vs. 137 days, 95% CI: 107-600, p = 0.0257). Conclusions: Differentiation of specific KRAS mutations in EUS-FNB and ctDNA (above all, the crucial G12D and G12R) is feasible in routine management of PDAC patients and imperative for assessment of prognosis.
- MeSH
- biopsie tenkou jehlou pod endosonografickou kontrolou * MeSH
- cirkulující nádorová DNA genetika krev MeSH
- dospělí MeSH
- duktální karcinom slinivky břišní * genetika patologie krev MeSH
- lidé středního věku MeSH
- lidé MeSH
- mutace * MeSH
- nádorové biomarkery genetika MeSH
- nádory slinivky břišní * genetika patologie mortalita MeSH
- prognóza MeSH
- protoonkogenní proteiny p21(ras) * genetika MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- tekutá biopsie metody MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Plasma donor-derived cell-free DNA (dd-cfDNA) is used to screen for rejection in heart transplants. We launched the Trifecta-Heart study ( ClinicalTrials.gov No. NCT04707872), an investigator-initiated, prospective trial, to examine the correlations between genome-wide molecular changes in endomyocardial biopsies (EMBs) and plasma dd-cfDNA. The present report analyzes the correlation of plasma dd-cfDNA with gene expression in EMBs from 4 vanguard centers and compared these correlations with those in 604 kidney transplant biopsies in the Trifecta-Kidney study ( ClinicalTrials.gov No. NCT04239703). METHODS: We analyzed 137 consecutive dd-cfDNA-EMB pairs from 70 patients. Plasma %dd-cfDNA was measured by the Prospera test (Natera Inc), and gene expression in EMBs was assessed by Molecular Microscope Diagnostic System using machine-learning algorithms to interpret rejection and injury states. RESULTS: Top transcripts correlating with dd-cfDNA were related to genes increased in rejection such as interferon gamma-inducible genes (eg, HLA-DMA ) but also with genes induced by injury and expressed in macrophages (eg, SERPINA1 and HMOX1 ). In gene enrichment analysis, the top dd-cfDNA-correlated genes reflected inflammation and rejection pathways. Dd-cfDNA correlations with rejection genes in EMB were similar to those seen in kidney transplant biopsies, with somewhat stronger correlations for TCMR genes in hearts and ABMR genes in kidneys. However, the correlations with parenchymal injury-induced genes and macrophage genes were much stronger in hearts. CONCLUSIONS: In this first analysis of Trifecta-Heart study, dd-cfDNA correlates significantly with molecular rejection but also with injury and macrophage infiltration, reflecting the proinflammatory properties of injured cardiomyocytes. The relationship supports the utility of dd-cfDNA in clinical management of heart transplant recipients.
- MeSH
- biologické markery krev MeSH
- biopsie MeSH
- dárci tkání * MeSH
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- myokard * patologie metabolismus MeSH
- prediktivní hodnota testů MeSH
- prospektivní studie MeSH
- rejekce štěpu * genetika imunologie patologie krev diagnóza MeSH
- senioři MeSH
- stanovení celkové genové exprese MeSH
- transplantace ledvin škodlivé účinky MeSH
- transplantace srdce * škodlivé účinky MeSH
- volné cirkulující nukleové kyseliny * krev genetika MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- multicentrická studie MeSH
- srovnávací studie MeSH
