BACKGROUND: Glioblastoma is the commonest malignant brain tumor and has a very poor prognosis. Reduced expression of the MGMT gene (10q26.3), influenced primarily by the methylation of two differentially methylated regions (DMR1 and DMR2), is associated with a good response to temozolomide treatment. However, suitable methods for detecting the methylation of the MGMT gene promoter and setting appropriate cutoff values are debated. RESULTS: A cohort of 108 patients with histologically and genetically defined glioblastoma was retrospectively examined with methylation-specific Sanger sequencing (sSeq) and methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) methods. The DMR2 region was methylated in 29% of samples, whereas DMR1 was methylated in 12% of samples. Methylation detected with the MS-MLPA method using probes MGMT_215, MGMT_190, and MGMT_124 from the ME012-A1 kit (located in DMR1 and DMR2) correlated with the methylation of the corresponding CpG dinucleotides detected with sSeq (p = 0.005 for probe MGMT_215; p < 0.001 for probe MGMT_190; p = 0.016 for probe MGMT_124). The threshold for methylation detection with the MS-MLPA method was calculated with a ROC curve analysis and principal components analysis of the data obtained with the MS-MLPA and sSeq methods, yielding a weighted value of 0.362. Thus, methylation of the MGMT gene promoter was confirmed in 36% of samples. These patients had statistically significantly better overall survival (p = 0.003). CONCLUSIONS: Our results show that the threshold for methylation detection with the MS-MLPA method determined here is useful from a diagnostic perspective because it allows the stratification of patients who will benefit from specific treatment protocols, including temozolomide. Detailed analysis of the MGMT gene promoter enables the more-precise and personalized treatment of patients with glioblastoma.

• MeSH
• CpG ostrůvky genetika   MeSH
• DNA modifikační methylasy * genetika   MeSH
• dospělí MeSH
• enzymy opravy DNA * genetika   MeSH
• glioblastom * genetika   farmakoterapie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• metylace DNA * genetika   MeSH
• nádorové supresorové proteiny * genetika   MeSH
• nádory mozku * genetika   MeSH
• promotorové oblasti (genetika) * genetika   MeSH
• retrospektivní studie MeSH
• sekvenční analýza DNA metody   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• temozolomid terapeutické užití   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
• srovnávací studie MeSH
• validační studie MeSH

BACKGROUND AND PURPOSE: Diffuse gliomas, a heterogeneous group of primary brain tumors, have traditionally been stratified by histology, but recent insights into their molecular features, especially the IDH mutation status, have fundamentally changed their classification and prognosis. Current diagnostic methods, still predominantly relying on invasive biopsy, necessitate the exploration of noninvasive imaging alternatives for glioma characterization. MATERIALS AND METHODS: In this prospective study, we investigated the utility of the spherical mean technique (SMT) in predicting the IDH status and histologic grade of adult-type diffuse gliomas. Patients with histologically confirmed adult-type diffuse glioma underwent a multiparametric MRI examination using a 3T system, which included a multishell diffusion sequence. Advanced diffusion parameters were obtained using SMT, diffusional kurtosis imaging, and ADC modeling. The diagnostic performance of studied parameters was evaluated by plotting receiver operating characteristic curves with associated area under curve, specificity, and sensitivity values. RESULTS: A total of 80 patients with a mean age of 48 (SD, 16) years were included in the study. SMT metrics, particularly microscopic fractional anisotropy (μFA), intraneurite voxel fraction, and μFA to the third power (μFA3), demonstrated strong diagnostic performance (all AUC = 0.905, 95% CI, 0.835-0.976; P < .001) in determining IDH status and compared favorably with diffusional kurtosis imaging and ADC models. These parameters also showed a strong predictive capability for tumor grade, with intraneurite voxel fraction and μFA achieving the highest diagnostic accuracy (AUC = 0.937, 95% CI, 0.880-0.993; P < .001). Control analyses on normal-appearing brain tissue confirmed the specificity of these metrics for tumor tissue. CONCLUSIONS: Our study highlights the potential of SMT for noninvasive characterization of adult-type diffuse gliomas, with a potential to predict IDH status and tumor grade more accurately than traditional ADC metrics. SMT offers a promising addition to the current diagnostic toolkit, enabling more precise preoperative assessments and contributing to personalized treatment planning.

• MeSH
• difuzní magnetická rezonance metody   MeSH
• dospělí MeSH
• gliom * diagnostické zobrazování   patologie   MeSH
• isocitrátdehydrogenasa * genetika   MeSH
• lidé středního věku MeSH
• lidé MeSH
• magnetická rezonanční tomografie metody   MeSH
• mutace MeSH
• nádory mozku * diagnostické zobrazování   patologie   MeSH
• prospektivní studie MeSH
• senioři MeSH
• senzitivita a specificita MeSH
• stupeň nádoru MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Glioblastoma is the commonest primary malignant brain tumor, with a very poor prognosis and short overall survival. It is characterized by its high intra- and intertumoral heterogeneity, in terms of both the level of single-nucleotide variants, copy number alterations, and aneuploidy. Therefore, routine diagnosis can be challenging in some cases. We present a complicated case of glioblastoma, which was characterized with five cytogenomic methods: interphase fluorescence in situ hybridization, multiplex ligation-dependent probe amplification, comparative genomic hybridization array and single-nucleotide polymorphism, targeted gene panel, and whole-genome sequencing. These cytogenomic methods revealed classical findings associated with glioblastoma, such as a lack of IDH and TERT mutations, gain of chromosome 7, and loss of chromosome 10. At least three pathological clones were identified, including one with whole-genome duplication, and one with loss of 1p and suspected loss of 19q. Deletion and mutation of the TP53 gene were detected with numerous breakends on 17p and 20q. Based on these findings, we recommend a combined approach to the diagnosis of glioblastoma involving the detection of copy number alterations, mutations, and aneuploidy. The choice of the best combination of methods is based on cost, time required, staff expertise, and laboratory equipment. This integrated strategy could contribute directly to tangible improvements in the diagnosis, prognosis, and prediction of the therapeutic responses of patients with brain tumors.

• MeSH
• glioblastom * genetika   patologie   diagnóza   MeSH
• hybridizace in situ fluorescenční metody   MeSH
• jednonukleotidový polymorfismus MeSH
• lidé středního věku MeSH
• lidé MeSH
• mutace MeSH
• nádorové biomarkery genetika   MeSH
• nádory mozku * genetika   patologie   diagnóza   MeSH
• prognóza MeSH
• srovnávací genomová hybridizace metody   MeSH
• variabilita počtu kopií segmentů DNA MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH

OBJECTIVE: The highly infiltrative growth of glioblastoma (GBM) makes distinction between the tumor and normal brain tissue challenging. Therefore, fluorescence-guided surgery is often used to improve visual identification of radiological tumor margins. The aim of this study was to evaluate the ability of recently developed molecularly targeted near-infrared (NIR) protease-activated probes to visualize GBM tissue and to compare the most promising candidate with the gold standard, 5-aminolevulinic acid (5-ALA). METHODS: Single-substrate probes 6QC-ICG and 6QC-Cy5 (cysteine cathepsin cleavable), double-substrate probes AG2-FNIR and AG2-Cy5 (cysteine cathepsin and caspase 3 cleavable), and 5-ALA were administered intravenously to mice with orthotopic tumors. Activation of the probes was also evaluated in cell cultures in vitro and in biopsy material from patients with GBM ex vivo. The tumor to normal brain tissue fluorescence ratio (TNR) was quantified in brain sections using preclinical and clinical visualization platforms, and in tissue homogenates and cell suspensions using spectrofluorimetry. Subcellular localization of the fluorophores was visualized by confocal microscopy. RESULTS: In vitro, the single-substrate probe 6QC-ICG was cleaved in glioma cells and macrophages, and the resulting fluorophore accumulated intracellularly. In experimental GBMs, both single- and double-substrate probes visualized tumor tissue, while in healthy brain tissue the signal was minimal. TNR was highest for 6QC-ICG and AG2-FNIR, but the signal intensity was higher for 6QC-ICG. Using xenograft and syngeneic mouse models, as well as human GBM biopsy material ex vivo, the authors confirmed the ability of 6QC-ICG to specifically visualize the glioma tissue using preclinical and clinical visualization platforms. Finally, a comparison with 5-ALA in animals coadministered with both compounds revealed a higher TNR for 6QC-ICG in experimental GBMs. CONCLUSIONS: The cysteine cathepsin-cleavable probe 6QC-ICG is activated by glioma cells and tumor-associated macrophages, leading to a high contrast between tumor and nontumorous brain tissue that is superior to that of the current standard, 5-ALA. In addition to a well-defined mechanism of action, protease-activated probes that use NIR fluorophores (e.g., indocyanine green) have the advantage of low absorption and scattering of the NIR light and lower tissue autofluorescence. These results suggest that 6QC-ICG has the potential to become the targeted agent in intraoperative detection of GBM tissue using fluorescence imaging.

• MeSH
• fluorescenční barviva MeSH
• glioblastom * diagnostické zobrazování   patologie   MeSH
• kyselina aminolevulová * MeSH
• lidé MeSH
• molekulární sondy MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory mozku * diagnostické zobrazování   patologie   MeSH
• optické zobrazování metody   MeSH
• proteasy metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

Neurochirugie je zásadní pro získání histologických vzorků, provádění správných chirurgických výkonů, pro indikace k výkonům a následné pooperační sledování. V posledních letech došlo jak ke zpřesnění indikací neurochirurgických intervencí, tak ke snížení komplikací. Zásadní je použití moderních zobrazovacích, navigačních a elektrofyziologických technik. Péče o pacienty s nádory je však multioborová, proto je nezbytná kooperace s dalšími odbornostmi.

Neurosurgery plays a crutial role in obtaining histological samples, in performance of adequate surgical procedures, in decision about surgical procedures and postoperative follow-up. Indications of neurosurgical procedures has been refined as well as the rate of complications has been reduced. Modern imaging, navigation and eletrophysiological techniques are of utmost importance. Care has to be multidisciplinar, thus cooperation between key care givers is vital.

• MeSH
• astrocytom chirurgie   diagnóza   patologie   MeSH
• chemoradioterapie metody   MeSH
• ependymom chirurgie   diagnóza   patologie   MeSH
• glioblastom diagnóza   patologie   terapie   MeSH
• gliom diagnóza   klasifikace   terapie   MeSH
• kraniofaryngeom chirurgie   diagnóza   MeSH
• lidé MeSH
• lymfom diagnóza   patologie   terapie   MeSH
• meningeom chirurgie   diagnóza   patologie   MeSH
• metastázy nádorů diagnóza   terapie   MeSH
• nádory centrálního nervového systému * diagnóza   klasifikace   terapie   MeSH
• nádory hypofýzy chirurgie   diagnóza   patologie   MeSH
• nádory míchy diagnóza   terapie   MeSH
• neurochirurgické výkony metody   MeSH
• stupeň nádoru MeSH
• vestibulární schwannom chirurgie   diagnóza   MeSH
• Check Tag
• lidé MeSH

Brain metastases are a very common and serious complication of oncological diseases. Despite the vast progress in multimodality treatment, brain metastases significantly decrease the quality of life and prognosis of patients. Therefore, identifying new targets in the microenvironment of brain metastases is desirable. Fibroblast activation protein (FAP) is a transmembrane serine protease typically expressed in tumour-associated stromal cells. Due to its characteristic presence in the tumour microenvironment, FAP represents an attractive theranostic target in oncology. However, there is little information on FAP expression in brain metastases. In this study, we quantified FAP expression in samples of brain metastases of various primary origin and characterised FAP-expressing cells. We have shown that FAP expression is significantly higher in brain metastases in comparison to non-tumorous brain tissues, both at the protein and enzymatic activity levels. FAP immunopositivity was localised in regions rich in collagen and containing blood vessels. We have further shown that FAP is predominantly confined to stromal cells expressing markers typical of cancer-associated fibroblasts (CAFs). We have also observed FAP immunopositivity on tumour cells in a portion of brain metastases, mainly originating from melanoma, lung, breast, and renal cancer, and sarcoma. There were no significant differences in the quantity of FAP protein, enzymatic activity, and FAP+ stromal cells among brain metastasis samples of various origins, suggesting that there is no association of FAP expression and/or presence of FAP+ stromal cells with the histological type of brain metastases. In summary, we are the first to establish the expression of FAP and characterise FAP-expressing cells in the microenvironment of brain metastases. The frequent upregulation of FAP and its presence on both stromal and tumour cells support the use of FAP as a promising theranostic target in brain metastases.

• MeSH
• fibroblasty patologie   MeSH
• individualizovaná medicína MeSH
• karcinom z renálních buněk * patologie   MeSH
• kvalita života MeSH
• lidé MeSH
• membránové proteiny metabolismus   MeSH
• nádorové mikroprostředí MeSH
• nádory ledvin * patologie   MeSH
• nádory mozku * patologie   MeSH
• serinové endopeptidasy metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

• Publikační typ
• abstrakt z konference MeSH

• Publikační typ
• komentáře MeSH

Difuzní nízkostupňové gliomy (stupeň II) jsou heterogenní neuroepiteliální tumory, které se vyznačují infiltrativním růstem a histologicky nesplňují kritéria pro dia­gnózu vysokostupňového gliomu (stupeň III, IV). I přes signifikantně nižší incidenci ve skupině primárních mozkových tumorů si zasluhují plnou pozornost z důvodu častého výskytu u mladších pacientů a závažných důsledků plynoucích z růstu tumoru i onkologické léčby na jejich další život. Rozvoj na poli histopatologie, cytogenetiky a zobrazovacích metod postupem času vede ke zpřesňování dia­gnostiky a individualizaci léčebného postupu. Hlavní léčebnou metodou je v současnosti radikální chirurgické odstranění tumoru s důrazem na minimalizaci rizika vzniku nového neurologického deficitu. I přes zdánlivě dostatečnou šíři resekce bývají do léčebného protokolu zahrnuty další metody, jako jsou radioterapie a/nebo chemoterapie. Načasování a modifikace adjuvantní terapie závisí na specifických molekulárních vlastnostech gliomů v kombinaci s dalšími prognostickými faktory. Dosáhnutí plného vyléčení difuzních nízkostupňových gliomů je však nemožné, a to především z důvodu jejich infiltrativního růstu. Hlavním cílem neurochirurgického výkonu a onkologické léčby je tak oddálit čas do progrese a následné dediferenciace a tím prodloužit život těchto pacientů se snahou co nejvíce zachovat kvalitu jejich života. Článek si dává za cíl shrnout dosavadní poznatky o difuzních nízkostupňových gliomech.

Diffuse low grade gliomas (grade II) are heterogenous neuroepithelial tumours, which are known for their infiltrative growth and their distinct histological profile does not match the criteria of the dia­gnosis of high grade gliomas (grade III, IV). The incidence of diffuse low grade gliomas is significantly lower in comparison to other primary brain tumours. Nevertheless these tumours deserve attention because of their frequent occurence in younger patients and the negative consequences of their bio­logical behaviour in combination with mostly long-term complications associated with oncological treatment. Dia­gnostics and the individualization of treatment are constantly improving thanks to the ongoing development in the fields of histopathology, cytogenetics and neuroimaging methods. Currently the main treatment option is a radical surgical removal of the tumour with the effort to minimalize the risk of new neurological deficit. Despite the apparently sufficient extent of the resection, further means, such as radiotherapy and/or chemotherapy, are included in the treatment regimen. The timing and modifications of this adjuvant treatment depend on the specific molecular features of the tumour in a combination with other prognostic factors. However, the curability of diffuse low grade gliomas cannot be achieved, mainly due to their infiltrative growth. The main goal of the neurosurgical procedure and oncological treatment is to delay the time to progression and subsequent dediferenciation and thus to prolong the survival of the glioma patients with emphasis on preserving their quality of life. The article aims to summarize current knowledge about diffuse low grade gliomas.

• MeSH
• astrocytom * diagnóza   terapie   MeSH
• chirurgie operační metody   MeSH
• gliom * diagnóza   terapie   MeSH
• kvalita života MeSH
• lidé MeSH
• oligodendrogliom * diagnóza   terapie   MeSH
• protokoly protinádorové léčby MeSH
• radioterapie metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH