Patients below 55 years were genetically studied because the prevalence of isocitrate dehydrogenase 1 (IDH1) decreases in older patients and on grounds of cost-effectiveness, as suggested by the World Health Organization (WHO) in 2016. The aim of our study was to use novel massively parallel sequencing (MPS) approaches to examine rare variants of IDH1/2 in Czech diffuse astrocytic and oligodendroglial tumors (gliomas) patients below 55 years of age who had been immunohistochemically (IHC) diagnosed as IDH1 R132H negative. The IHC IDH1 status (wild type or mutant) of 275 tissue samples was analyzed using antibodies against the IDH1 R132H protein. Sixty-three samples of 55 years old patients with IHC IDH1 WT status were genotyped using two different MPS technologies to detect rare IDH1 and IDH2 variants. The tiered IHC (60 positive) and molecular (10 positive) approach thus revealed that 70 of the 275 samples (25%) bore IDH1/IDH2 mutations. The combined molecular and IHC approach thus revealed that 70 of the 275 samples (25%) considered in the study bore IDH1/IDH2 mutations. IHC detection of the IDH1 R132H variant should be routinely complemented with MPS to detect rare IDH1/2 variants in glioma patients below 55 years of age with negative IHC result of IDH R132H variant.
- MeSH
- gliom * patologie MeSH
- isocitrátdehydrogenasa genetika metabolismus MeSH
- lidé středního věku MeSH
- lidé MeSH
- mutace MeSH
- nádory mozku * diagnóza MeSH
- retrospektivní studie MeSH
- senioři MeSH
- vysoce účinné nukleotidové sekvenování MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- IDH1 protein, human MeSH Prohlížeč
- IDH2 protein, human MeSH Prohlížeč
- isocitrátdehydrogenasa MeSH
Carney triad is a multitumor syndrome affecting almost exclusively young women in a nonfamilial setting, which manifests by multifocal gastric gastrointestinal stromal tumors, paragangliomas, and pulmonary chondroma. The Carney triad-associated tumors are characterized by a deficiency of the mitochondrial succinate dehydrogenase enzymatic complex. Recently, it has been observed that the deficiency results from epigenetic silencing of the SDHC gene by its promoter hypermethylation. To elucidate anatomic distribution of SDHC promoter methylation in Carney triad patients and thus to shed some light on the possible natural development of this epigenetic change, both neoplastic and available non-neoplastic tissues of 3 patients with Carney triad were tested for hypermethylation at the SDHC promoter site. SDHC promoter hypermethylation was proven in all tumors studied. Lack of SDHC epigenetic silencing in the non-neoplastic lymphoid and duodenal tissue (ie, tissues not involved in the development of Carney triad-associated tumors) together with the finding of SDHC promoter hypermethylation in the non-neoplastic gastric wall favors the hypothesis of postzygotic somatic mosaicism as the biological background of Carney triad; it also offers an explanation of the multifocality of gastrointestinal stromal tumors of the stomach occurring in this scenario as well. However, the precise mechanism responsible for the peculiar organ-specific distribution of Carney triad-associated tumors is still unknown.
- MeSH
- chondrom * genetika metabolismus patologie MeSH
- DNA nádorová * genetika metabolismus MeSH
- leiomyosarkom * genetika metabolismus patologie MeSH
- lidé MeSH
- membránové proteiny * genetika metabolismus MeSH
- metylace DNA * MeSH
- mozaicismus * MeSH
- nádorové proteiny * genetika metabolismus MeSH
- nádory plic * genetika metabolismus patologie MeSH
- nádory žaludku * genetika metabolismus patologie MeSH
- nechromafinní paragangliom * genetika metabolismus patologie MeSH
- promotorové oblasti (genetika) * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- DNA nádorová * MeSH
- membránové proteiny * MeSH
- nádorové proteiny * MeSH
- SDHC protein, human MeSH Prohlížeč
- MeSH
- imunohistochemie MeSH
- lidé MeSH
- mucinózní adenokarcinom genetika metabolismus patologie MeSH
- nádory slinných žláz genetika metabolismus patologie MeSH
- protein ETS, translokační varianta 6 MeSH
- protoonkogenní proteiny c-ets genetika MeSH
- protoonkogenní proteiny c-ret genetika MeSH
- represorové proteiny genetika MeSH
- sekreční karcinom mamárního typu genetika metabolismus patologie MeSH
- translokace genetická MeSH
- vysoce účinné nukleotidové sekvenování MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- dopisy MeSH
- Názvy látek
- protoonkogenní proteiny c-ets MeSH
- protoonkogenní proteiny c-ret MeSH
- represorové proteiny MeSH
- RET protein, human MeSH Prohlížeč
Immunohistochemistry and immunocytochemistry (ICC) play an irreplaceable role in research and diagnostics. It is well known that antigen retrieval (AR) can, as a technique, have beneficial outcomes on immunohistochemistry results when using formalin-fixed, paraffin-embedded tissue samples. The main purpose of AR is to break protein crosslinks which are formed during formalin fixation. Although AR was originally designed for formalin-fixed, paraffin-embedded samples, the usefulness of AR in ICC has been described in previous studies. Cytologic samples are often fixed in alcohol-based fixatives which does not lead to the formation of crosslinks. Therefore, alcohol-fixed samples can be successfully immunostained without AR. We investigated the effect of heat-induced antigen retrieval (HIAR) on alcohol-fixed HEK293 cell line samples and patient cytologic samples from thyroid gland obtained by fine needle aspiration technique. We compared indirect 2-step ICC staining results performed according to the protocol with or without HIAR in citrate buffer pH 6 for several antibodies. Utilizing HIAR against intracellular antigens has beneficial effects. Therefore, more diluted antibodies can be used for satisfactory results. However, surface antigens were probably damaged by HIAR treatment. We demonstrated evident changes in cell surface topography after HIAR treatment by atomic force microscopy. Staining specificity of patient samples improves and background staining is reduced, allowing higher dilutions of primary antibody. Improving staining specificity is necessary for accurate diagnostics. Although we have shown the beneficial effect of HIAR for immunostaining intracellular antigens, proper staining protocol should be tested on appropriate controls for individual antibodies.
- MeSH
- antigeny chemie MeSH
- dospělí MeSH
- ethanol chemie MeSH
- fixace tkání * MeSH
- HEK293 buňky MeSH
- imunohistochemie * MeSH
- lidé středního věku MeSH
- lidé MeSH
- retrospektivní studie MeSH
- senioři MeSH
- štítná žláza chemie MeSH
- vysoká teplota * MeSH
- zalévání tkání do parafínu * MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antigeny MeSH
- ethanol MeSH
The 2016 World Health Organization Renal Tumor Classification defines renal oncocytoma (RO) as a benign epithelial tumor; however, malignant histopathologic features have been documented. Rare cases with metastases have been reported. We describe the case of a 62-year-old woman who was referred to the Urology Clinic for a routine work-up. Magnetic resonance imaging and computerized tomography showed a 7-cm mass in the middle and lower portions of the left kidney and 2 suspected liver metastases. The patient underwent surgery. Microscopically both renal and liver lesions presented solid, solid-nested, and microcystic architecture, composed predominantly of large eosinophilic cells without any worrisome pattern except the vascular extension. The cells were positive for S100A1, CD117, and PAX-8 and negative for CAIX, CK7, and AMACR. Fluorescence in situ hybridization showed a disomic profile for the chromosomes 1, 2, 6, 7, 10, 17. No mutation of coding sequence of the SDHB, SDHC, SDHD, VHL, and BHD genes and no loss of heterozygosity at 3p were found. The final diagnosis was "RO" according to the 2016 World Health Organization Renal Tumor Classification with "liver metastases." This report provides a wide clinical-pathologic, immunophenotypical and molecular documentation of a RO with liver metastases.
- Publikační typ
- časopisecké články MeSH
High-grade prostatic adenocarcinoma mimicking urothelial carcinoma (UC) is a rare and unusual variant, which can present a difficult diagnostic challenge. The aim of this study was to examine telomerase reverse transcriptase (TERT) mutations in order to improve differential diagnostic process in this scenario. Ten prostatic adenocarcinomas mimicking UC were retrieved by searching in-house and consultation files of Charles University Hospital, Plzen, Czech Republic, Tenon Hospital Paris, France, and University of Calgary, Canada. We performed microscopic slide review and immunohistochemical and molecular-genetic analyses using the available paraffin tissue. Patient age at diagnosis ranged from 44 to 86 years (mean, 71.8 y). All cases were transurethral resections, except one which was a prostate biopsy. Gleason score 5+5 was observed in 6 patients, whereas the remaining 4 had a Gleason score of 4+5. The tumors showed pseudopapillary, solid, nested, and cribriform architectural growth patterns. All cases were positive for prostatic markers including PSA, PAP, and NKX3.1. Immunohistochemical staining for urothelial marker, GATA3, was negative in 6 cases and only weakly positive in the remaining 4. All 10 cases showed no evidence of TERT mutations. We describe 10 high-grade prostatic adenocarcinomas that on morphology mimicked UC, but all demonstrated negative TERT mutations. A finding of negative TERT mutations in high-grade prostatic adenocarcinoma which mimics UC supports the notion that TERT promoter mutations are absent in prostate carcinoma, which may also aid the diagnostic work-up in difficult cases.
- MeSH
- adenokarcinom * diagnóza enzymologie genetika patologie MeSH
- biopsie MeSH
- diferenciální diagnóza MeSH
- dospělí MeSH
- karcinom z přechodných buněk diagnóza enzymologie genetika patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mutace * MeSH
- nádorové biomarkery * genetika metabolismus MeSH
- nádorové proteiny * genetika metabolismus MeSH
- nádory prostaty * diagnóza enzymologie genetika patologie MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- telomerasa * genetika metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
- klinické zkoušky MeSH
- multicentrická studie MeSH
- práce podpořená grantem MeSH
- Názvy látek
- nádorové biomarkery * MeSH
- nádorové proteiny * MeSH
- telomerasa * MeSH
- TERT protein, human MeSH Prohlížeč
We have studied a cohort of renal cell carcinomas (RCCs) with smooth-muscle stroma (N=6), which lacked any of following genetic aberrations: mutations in the VHL-gene-coding sequence, loss of heterozygosity of chromosome 3p, or hypermethylation of VHL. Using targeted next-generation sequencing, no intronic VHL mutations or mutations in selected genes involved in angiogenesis and genes frequently mutated in clear cell RCC were identified. Tumors were also tested for the presence of hotspot mutations in the TCEB1 gene with negative results in all cases. We conclude that there exists a group of RCCs with abundant leiomyomatous stroma, where the epithelial component is indistinguishable from conventional clear cell RCC and distinct from clear cell (tubulo-) papillary RCC and that these tumors lack aberrations related to the function of the VHL gene, mutations in genes involved in angiogenesis, and hotspot mutations in the TCEB1 gene.
- MeSH
- genetické testování MeSH
- karcinom z renálních buněk genetika MeSH
- leiomyosarkom genetika patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mutace genetika MeSH
- nádorový supresorový protein VHL genetika MeSH
- senioři MeSH
- vysoce účinné nukleotidové sekvenování trendy MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- nádorový supresorový protein VHL MeSH
BACKGROUND: Intestinal-type adenocarcinoma (ITAC) of the nasal cavity and paranasal sinuses is an uncommon tumor associated with exposure to wood and leather dust, nickel, and possibly smoking. ITAC shares phenotypical features with colorectal carcinoma. In contrast to most non-intestinal-type sinonasal adenocarcinomas, ITAC is an aggressive adenocarcinoma with poor clinical outcome; therefore, its reliable separation from non-ITAC is very important. AIM: The use of a combination of immunohistochemical markers of intestinal differentiation was tested in a cohort of sinonasal carcinomas of different types. The aim of this study was to explore a new intestinal marker, SATB2, in conjunction with CDX2 and CK20 in differential diagnosis of sinonasal adenocarcinomas. MATERIALS AND METHODS: Seven ITACs, 66 non-ITACs, and 1 case of extensive intestinal metaplasia (IM) of the nasal mucosa were included in the study and stained with SATB2, CK20, CDX2, and CK7 antibodies. Detection of mismatch repair proteins was performed in all cases of ITAC. All 7 sinonasal ITACs have been tested for KRAS, NRAS, and BRAF gene mutations. RESULTS: All ITACs showed positive expression for SATB2, whereas all non-ITAC cases were negative. The only 1 case of IM was found to be positive for SATB2, whereas the same case showed negative expression of CK20 and only focal immunostaining for CDX2. The genetic analysis showed that only 1 sinonasal ITAC (1/7) showed KRAS c.35G>C, p.(Gly12Ala) mutation, whereas BRAF and NRAS genes were wild type. Four ITACs revealed wild-type KRAS, NRAS, and BRAF, and 2 remaining cases were not analyzable. All ITACs showed preserved nuclear expression of mismatch repair proteins. CONCLUSIONS: SATB2 in combination with CDX2 and CK20 differentiates sinonasal ITAC from non-ITAC with increased diagnostic sensitivity and specificity and detects IM in the sinonasal tract more easily.
- MeSH
- adenokarcinom diagnóza metabolismus patologie MeSH
- diferenciální diagnóza MeSH
- dřevo MeSH
- imunohistochemie MeSH
- keratin-20 metabolismus MeSH
- kolorektální nádory diagnóza metabolismus patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- metaplazie MeSH
- nádorové biomarkery metabolismus MeSH
- nádory nosu diagnóza metabolismus patologie MeSH
- nádory tračníku diagnóza metabolismus patologie MeSH
- nosní dutina patologie MeSH
- paranazální dutiny patologie MeSH
- prognóza MeSH
- senioři MeSH
- střeva patologie MeSH
- transkripční faktor CDX2 metabolismus MeSH
- transkripční faktory metabolismus MeSH
- vazebné proteiny DNA v oblastech připojení k matrix metabolismus MeSH
- vystavení vlivu životního prostředí škodlivé účinky MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- CDX2 protein, human MeSH Prohlížeč
- keratin-20 MeSH
- KRT20 protein, human MeSH Prohlížeč
- nádorové biomarkery MeSH
- SATB2 protein, human MeSH Prohlížeč
- transkripční faktor CDX2 MeSH
- transkripční faktory MeSH
- vazebné proteiny DNA v oblastech připojení k matrix MeSH
Multifocal medulloblastomas (MMBs) in adults are exceedingly rare with only 5 reported cases to date. Medulloblastoma in adult differ from its childhood counterpart by being more often lateral in location, desmoplastic in morphology, and better in clinical prognosis. Little is known, however, about the characteristic features of MMB. This is particularly true for their molecular profiles. To date, molecular characteristics of multifocal medulloblastoma have been reported only once. Here, we present the second case of multifocal medulloblastoma along with its detailed morphology, imaging features, and molecular profiles with a critical review of the literature. We believe that MMB should be reported in detail to better understand their behavior, characterize their molecular profiles, and establish therapeutic protocols.
- MeSH
- infratentoriální nádory diagnóza genetika patofyziologie MeSH
- lidé MeSH
- meduloblastom genetika patofyziologie MeSH
- stanovení celkové genové exprese MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- přehledy MeSH
The purpose of this study was to test the hypothesis that intestinal lipomas occurring in patients devoid of signs of PDGFRA-mutant syndrome might represent sporadic counterparts of familial lipomatous tumors occurring in the spectrum of tumors associated with PDGFRA mutations. PDGFRA-mutant syndrome may manifest with gastrointestinal stromal tumors, Vanek tumors, fibrous tumors, and lipomatous tumors. Until now there has been no molecular genetic study of PDGFRA mutations in intestinal lipomas published in the world literature. A series of 20 intestinal lipomas were obtained from 17 patients, and mutational analysis of exons 12, 14, and 18 of the PDGFRA gene was performed. None of the 16 analyzable tumors showed mutations in PDGFRA. Thus, PDGFRA mutations probably do not play an important role in the development of sporadic lipomas of the intestines.
