OBJECTIVE: Transgenic mice with fluorescent protein (FP) reporters take full advantage of new in vivo imaging technologies. Therefore, we generated a TRPC5- and a TRPA1-reporter mouse based on FP C-terminal fusion, providing us with better alternatives for studying the physiology, interaction and coeffectors of these two TRP channels at the cellular and tissue level. METHODS: We generated transgenic constructs of the murine TRPC5- and TRPA1-gene with a 3*GGGGS linker and C-terminal fusion to mCherry and mTagBFP, respectively. We microinjected zygotes to generate reporter mice. Reporter mice were examined for visible fluorescence in trigeminal ganglia with two-photon microscopy, immunohistochemistry and calcium imaging. RESULTS: Both TRPC5-mCherry and TRPA1-mTagBFP knock-in mouse models were successful at the DNA and RNA level. However, at the protein level, TRPC5 resulted in no mCherry fluorescence. In contrast, sensory neurons derived from the TRPA1-reporter mice exhibited visible mTag-BFP fluorescence, although TRPA1 had apparently lost its ion channel function. CONCLUSIONS: Creating transgenic mice with a TRP channel tagged at the C-terminus with a FP requires detailed investigation of the structural and functional consequences in a given cellular context and fine-tuning the design of specific constructs for a given TRP channel subtype. Different degrees of functional impairment of TRPA1 and TRPC5 constructs suggest a specific importance of the distal C-terminus for the regulation of these two channels in trigeminal neurons.

• MeSH
• červený fluorescenční protein MeSH
• ganglion trigeminale metabolismus   MeSH
• genový knockin * MeSH
• kationtové kanály TRPC * genetika   metabolismus   MeSH
• kationtový kanál TRPA1 * genetika   metabolismus   MeSH
• luminescentní proteiny * genetika   metabolismus   MeSH
• myši transgenní * MeSH
• myši MeSH
• rekombinantní fúzní proteiny metabolismus   genetika   MeSH
• vápník metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Karcinom endometria patří mezi nejčastější gynekologické malignity ve vyspělých zemích a jeho incidence dlouhodobě roste. S rozvojem molekulárního porozumění biologii nádoru se otevřela cesta k cílené a imunoterapeutické léčbě, zejména u podtypů s defektním mechanismem oprav chybného párování DNA (deficient mismatch repair, dMMR) nebo s mikrosatelitovou instabilitou (microsateUite instability-high, MSI-H). Článek uvádí přehled současných možností imunoterapie v léčbě pokročilého nebo recidivujícího karcinomu endometria, včetně kombinací s chemoterapií a cílenými léky, na základě aktuálních dat z klinických studií. Imunoterapie se stala novým standardem v léčbě určité části pacientek s karcinomem endometria. Klíčovou roli hraje molekulární stratifikace, bez níž nelze imunoterapii účinně a bezpečně nasadit.

Endometrial cancer is one of the most common gynecological malignancies in developed countries, and its incidence has been increasing for a long time. With the development of molecular understanding of tumor biology, the way to targeted and immunotherapeutic treatment has been opened, especially for subtypes with deficient mismatch repair (dMMR) or microsateUite instability-high (MSI-H). The article provides an overview of current immunotherapy options for the treatment of advanced or recurrent endometrial cancer, including combinations with chemotherapy and targeted drugs, based on current clinical trial data. Immunotherapy has become the new standard of care for some patients with endometrial cancer. Molecular stratification plays a key role, without which immunotherapy cannot be used effectively and safely.

The intertumoral and intratumoral heterogeneity of colorectal adenocarcinoma (CRC) at the morphologic level is poorly understood. Previously, we identified morphological patterns associated with CRC molecular subtypes and their distinct molecular motifs. Here we aimed to evaluate the heterogeneity of these patterns across CRC. Three pathologists evaluated dominant, secondary, and tertiary morphology on four sections from four different FFPE blocks per tumor in a pilot set of 22 CRCs. An AI-based image analysis tool was trained on these tumors to evaluate the morphologic heterogeneity on an extended set of 161 stage I-IV primary CRCs (n = 644 H&E sections). We found that most tumors had two or three different dominant morphotypes and the complex tubular (CT) morphotype was the most common. The CT morphotype showed no combinatorial preferences. Desmoplastic (DE) morphotype was rarely dominant and rarely combined with other dominant morphotypes. Mucinous (MU) morphotype was mostly combined with solid/trabecular (TB) and papillary (PP) morphotypes. Most tumors showed medium or high heterogeneity, but no associations were found between heterogeneity and clinical parameters. A higher proportion of DE morphotype was associated with higher T-stage, N-stage, distant metastases, AJCC stage, and shorter overall survival (OS) and relapse-free survival (RFS). A higher proportion of MU morphotype was associated with higher grade, right side, and microsatellite instability (MSI). PP morphotype was associated with earlier T- and N-stage, absence of metastases, and improved OS and RFS. CT was linked to left side, lower grade, and better survival in stage I-III patients. MSI tumors showed higher proportions of MU and TB, and lower CT and PP morphotypes. These findings suggest that morphological shifts accompany tumor progression and highlight the need for extensive sampling and AI-based analysis. In conclusion, we observed unexpectedly high intratumoral morphological heterogeneity of CRC and found that it is not heterogeneity per se, but the proportions of morphologies that are associated with clinical outcomes.

• MeSH
• adenokarcinom * patologie   genetika   mortalita   MeSH
• dospělí MeSH
• kolorektální nádory * patologie   genetika   mortalita   MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• staging nádorů 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

I po maximální standardní léčbě, která zahrnuje maximální chirurgickou resekci, adjuvantní radioterapii a souběžnou a následně adjuvantní chemoterapii alkylační látkou temozolomidem (TMZ), což je režim ověřený klinickými studiemi fáze III, zůstává celkové přežití u glioblastomu (GB), nejčastějšího a nejmalignějšího podtypu gliomu vysokého stupně, omezené. Pětileté přežití je nižší než 10 %, přičemž obvyklou příčinou úmrtí je lokální recidiva. Nejčastěji navrhovanými alternativami čistě paliativní léčby jsou reoperace, opětovné podání alkylačních látek včetně TMZ nebo léčba relapsu lomustinem a opakované ozařování. Opakované ozařování se jeví být možností pro GB v pečlivě vybraných případech a volba ozařovací metody mezi standardní frakcionací, stereotaktickou hypofrakcionovanou radioterapií a stereotaktickou radiochirurgií musí zohledňovat technické a s pacientem související faktory a průběh relapsu. Použití alkylačních látek TMZ a lomustinu v kombinaci s opakovaným ozařováním se zdá být strategií s přínosem zejména po výběru terapie na základě metylačního stavu promotoru O6-metylguanin-DNA-metyltransferázy. Budoucími směry výzkumu jsou neoadjuvantní imunoterapie a identifikace molekulárních biomarkerů, přesnější vymezení cílového objemu na základě pozitronové emisní tomografie/výpočetní tomografie s aminokyselinami, ale také použití inhibitorů glykolýzy ve spojení s látkami proti cévnímu endoteliálnímu růstovému faktoru. Omezení kumulativních ekvivalentních dávek po 2 Gy (EQD2) na mozkovou tkáň parenchym do výše 100 Gy v případě ultrahypofrakcionovaných režimů a do výše < 120 Gy EQD2 v případě jiných frakcionačních schémat by mohlo omezit riziko mozkové radionekrózy pod 10 %.

Even after administration of a maximal standard treatment, including gross surgical resection, adjuvant radiotherapy plus concurrent and subsequently adjuvant chemotherapy with an alkylating agent, temozolomide (TMZ), a regimen validated by phase III clinical trials, the overall survival in glioblastoma (GB), which is the most frequent and malignant subtype of high-grade glioma, remains limited. With a 5-year survival rate below 10%, local recurrence is the usual cause of death. Re-operation, re-challenge with alkylating agents including TMZ or treatment of relapse with lomustine, and re-irradiation are the most frequently proposed alternatives to purely palliative treatment. Re-irradiation seems to be an option for GB in carefully selected cases, and the choice of irradiation method between standard fractionation, stereotactic hypo-fractionated radiotherapy, and stereotactic radiosurgery must take into account technical and patient-related factors and relapse pattern. The use of alkylating agents TMZ and lomustine, in combination with re-irradiation, seems to be a strategy with benefits especially after a selection of therapy based on the methylation status of the O6-methylguanine-DNA-methyltransferase promoter. Neo-adjuvant immunotherapy and identification of molecular biomarkers, more precise delineation of the target volume, based on amino-acid positron emission tomography/CT (PET-CT), but also the use of glycolytic inhibitors in association with anti-vascular endothelial growth factor agents are future research directions. Limiting the cumulative equivalent dose in 2 Gy (EQD2) received by the brain tissue to 100 Gy in the case of ultra-hypo-fractionated regimens and to < 120 Gy EQD2 in the case of other fractionation schemes could limit the risk of cerebral radio-necrosis below 10%.

• Klíčová slova
• radionekróza,
• MeSH
• glioblastom * radioterapie   MeSH
• lidé MeSH
• opakovaná terapie MeSH
• radiochirurgie metody   MeSH
• radioterapie metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

BACKGROUND: Liver glycogen storage disorders (GSDs) are rare inherited disorders of carbohydrate metabolism that are clinically characterized by hepatomegaly and fasting intolerance. This group of disorders comprises GSD Ia and Ib as well as the so-called ketotic GSDs including GSD III, VI, IX, XI and 0a. Although clinical practice guidelines exist for most GSD subtypes, diagnostics, treatment and monitoring differ significantly among metabolic centres. The aim of this study was to gain insight into current clinical practice for liver GSDs. METHODS: An international web-based survey was performed among health care professionals involved in the care of individuals with liver GSDs. RESULTS: Sixty-seven respondents from 28 different countries caring for approximately 2650 liver GSD patients completed the survey. While the diagnostic approach was generally consistent, significant differences among metabolic centres are still observed with respect to monitoring parameters and treatment approaches. Reasons for these differences are local availability of management tools and treatment options, the rarity of the different GSD subtypes, the experiences of health care professionals, and the existence of extreme phenotypes. CONCLUSION: The development of a standard set of outcomes for patients with liver GSDs is warranted as a reference for both daily care and the evaluation of safety and efficacy of future therapies. For various parameters that serve as valuable outcome measures, tools and target values should be better defined.

• MeSH
• glykogenóza * terapie   diagnóza   MeSH
• játra metabolismus   patologie   MeSH
• lidé MeSH
• management nemoci MeSH
• nemoci jater * terapie   diagnóza   MeSH
• průzkumy a dotazníky MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Alzheimer's disease (AD) is a multifaceted neurodegenerative disorder for which current treatments provide only symptomatic relief, primarily through cholinesterase (ChE) inhibition and N-methyl-d-aspartate receptor (NMDAR) antagonism. To improve therapeutic efficacy and safety, we designed and synthesized 16 novel tacrine derivatives modified at position 7 with various (hetero)aryl groups or deuterium substitution. Initially, in silico screening predicted favorable CNS permeability and oral bioavailability. Subsequent in vitro evaluations demonstrated significant inhibitory potency against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), with derivatives 5i and 5m displaying particularly promising profiles. Metabolic stability assessed using human liver microsomes revealed enhanced stability for compound 5e, whereas 5i and 5m underwent rapid metabolism. Notably, compound 7 showed improved metabolic stability attributed to deuterium incorporation. The newly synthesized compounds were further tested for antagonistic activity on the GluN1/GluN2B subtype of NMDAR, with compound 5m exhibiting the most potent and voltage-independent inhibition. The ability of these compounds to permeate the blood-brain barrier (BBB) was confirmed through in vitro PAMPA assays. In preliminary hepatotoxicity screening (HepG2 cells), most derivatives exhibited higher cytotoxicity than tacrine, emphasizing the ongoing challenge in hepatotoxicity management. Based on its overall favorable profile, compound 5m advanced to in vivo pharmacokinetic studies in mice, demonstrating efficient CNS penetration, with brain concentrations exceeding plasma levels (brain-to-plasma ratio 2.36), indicating active transport across the BBB. These findings highlight compound 5m as a promising tacrine-based multi-target-directed ligand, supporting further preclinical development as a potential therapeutic candidate for AD.

• MeSH
• acetylcholinesterasa metabolismus   MeSH
• Alzheimerova nemoc * farmakoterapie   metabolismus   MeSH
• biologická dostupnost MeSH
• butyrylcholinesterasa metabolismus   MeSH
• cholinesterasové inhibitory * farmakologie   chemie   chemická syntéza   MeSH
• hematoencefalická bariéra metabolismus   MeSH
• jaterní mikrozomy metabolismus   MeSH
• lidé MeSH
• ligandy MeSH
• molekulární struktura MeSH
• myši MeSH
• receptory N-methyl-D-aspartátu * antagonisté a inhibitory   metabolismus   MeSH
• takrin * farmakologie   chemie   chemická syntéza   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• vztahy mezi strukturou a aktivitou MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

In addition to "classic" and eosinophilic subtype, chromophobe renal cell carcinoma (RCC) is well-known to demonstrate various morphological patterns including adenomatoid, microcystic, pigmented, multicystic, papillary, neuroendocrine-like, and small cell-like, all of which are important to appreciate for accurate diagnosis. Herein, we expand on a unique chromophobe RCC morphology not previously described consisting of tumor cells with extensive stromal retraction, mimicking upper urothelial tract micropapillary carcinoma (MPC). Twelve MPC-like chromophobe RCC nephrectomies were reviewed with clinicopathological features recorded; molecular testing was performed on 7 of 12 tumors. Patients were mostly men (n=10) with a mean age of 65 years. Mean tumor size was 6.4 cm with pathological stage distribution as follows: 4 (33%) T1a, 2 (17%) T1b, 1 (8%) T2b, and 3 (25%) T3a. The extent of MPC-like chromophobe RCC foci ranged from 10% to 40% (mean=26%; there was no correlation between the extent of MPC-like chromophobe RCC foci and tumor stage). Other chromophobe RCC morphological patterns were not identified. When performed, all (100%) tumors depicted prototypic chromophobe RCC staining pattern of KIT positivity/KRT7 positivity. Molecular showed 6 of 7 (86%) with multiple chromosomal losses. Clinically significant mutations were identified in NF1, TP53, FLCN (likely somatic), CHEK2, and ZFHX3 genes. Follow up available in 9 patients showed no evidence of disease (mean=23 months). Although the etiology behind the extensive stromal retraction in our tumors is unknown, this may likely be artifactual in nature. Nonetheless, it is important to include MPC-like chromophobe RCC in the spectrum of "variant" morphologies to avoid diagnostic pitfalls from micropapillary carcinoma.

• MeSH
• artefakty * MeSH
• diferenciální diagnóza MeSH
• karcinom z přechodných buněk diagnóza   patologie   chirurgie   MeSH
• karcinom z renálních buněk * diagnóza   patologie   chirurgie   genetika   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorové biomarkery analýza   genetika   MeSH
• nádory ledvin * patologie   diagnóza   chirurgie   genetika   MeSH
• papilární karcinom patologie   diagnóza   chirurgie   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
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

The disease currently known as frontotemporal dementia (FTD) has undergone a complex evolution from its first description by Arnold Pick and later by Alois Alzheimer, through the first clinicopathological criteria introduced by David Neary and David Mann, to its current nomenclatural perception as a complex clinicopathological entity. Currently, Frontotemporal lobar degeneration is viewed as a heterogeneous syndrome caused by progressive degeneration of the frontal and temporal lobes of the brain. Clinically, it can manifest as three syndromes of frontotemporal dementia (behavioral variant of FTD, progressive non-fluent aphasia and semantic dementia) but also as so-called "overlap" syndromes involving corticobasal degeneration and progressive supranuclear palsy. Its prevalence is about 10 % among all dementias and 40 % among dementias with onset between 45 and 65 years of age. The clinical manifestation of the different subtypes varies, the common denominator being behavioral disturbances and impairment of fatic, gnostic and executive functions. Mnestic and visuo-spatial functions, although preserved for a relatively long time, are superimposed by personality disintegration, fatic, gnostic and executive dysfunction. Compared with Alzheimer's disease, it generally has an earlier age of onset, a more rapid course and more devastating impairment of individual cognitive domains. FTD has a heritability of more than 30 % according to current knowledge. The main genes involved are MAPT, C9orf72 and GRN. More rarely affected genes are VCP, TDP-43, FUS and CHMP2B. In our article, we focus on the genetics of FTD and the clinic-genetic-pathological correlations. We also aim to provide a plastic picture of how individual mutations affect the molecular mechanisms of neurodegeneration.

• MeSH
• epigeneze genetická genetika   MeSH
• frontotemporální demence * diagnóza   genetika   klasifikace   MeSH
• genetické testování metody   MeSH
• lidé MeSH
• primární progresivní nonfluentní afázie diagnóza   genetika   MeSH
• progranuliny genetika   MeSH
• proteiny tau genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

BACKGROUND: Profiling studies in small-cell lung cancer (SCLC) have mainly focused on primary tumors, omitting the potential molecular changes that might occur during lymphatic metastasis formation. Here, we assessed the molecular discordance between primary SCLCs and corresponding lymph node (LN) metastases in the light of subtype distribution and expression of clinically relevant proteins. METHODS: Comparative profiling of 32 surgically resected primary SCLCs and their LN metastases was achieved by RNA expression analysis and immunohistochemistry (IHC). In addition to subtype markers (ASCL1, NEUROD1, POU2F3, and YAP1), the expression of nine cancer-specific proteins was evaluated. RESULTS: The selected clinically relevant molecules showed no significant differences in their RNA expression profile when assessing the primary tumors and their corresponding LN metastases. Nevertheless, IHC analyses revealed significantly higher DLL3 expression in the primary tumors than in the LN metastases (P = 0.008). In contrast, NEUROD1 expression was significantly lower in the primary tumors (versus LN metastases, P < 0.001). No statistically significant difference was found by IHC analysis in the case of other clinically relevant proteins. Concerning SCLC molecular subtypes, a change in subtype distribution was detected in 21 cases. Phenotype switching from neuroendocrine (NE) subtypes toward non-NE lesions and from non-NE landscape toward NE subtypes were both detected. CONCLUSIONS: Although the molecular landscape of SCLC LN metastases largely resembles that of the tumor of origin, key differences exist in terms of DLL3 and NEUROD1 expression, and in subtype distribution. These diagnostic pitfalls should be considered when establishing the tumors' molecular profile for future clinical trials solely based on LN biopsies.

• MeSH
• imunohistochemie MeSH
• lidé středního věku MeSH
• lidé MeSH
• lymfatické metastázy * patologie   MeSH
• malobuněčný karcinom plic * chirurgie   patologie   genetika   MeSH
• nádorové biomarkery genetika   MeSH
• nádory plic * patologie   chirurgie   genetika   MeSH
• senioři MeSH
• transkripční faktory bHLH genetika   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

The diagnostic performance of molecular markers in surrogate tissues like stool may be affected by colorectal cancer (CRC) morphological heterogeneity. The mucinous histotype represents a subgroup of CRC with a peculiar molecular program and unfavorable disease progression. However, the percentage of mucinous morphology necessary to define this subtype is still a matter of debate. In this study, we investigated whether stool miRNA profiles of CRC patients differ in patients with mucinous histopathological subtypes compared to non-mucinous cancers. In this respect, we also explored how the stool miRNA signature reported in our previous multicentric study behaves in this histotype. Small-RNA sequencing was performed in fecal and tissue samples of an Italian cohort (n = 172), including 27 CRC with mucinous morphology (mucinous cancers with ≥ 50% mucinous morphology and those with mucinous component ≥ 5% but < 50%), 58 non-mucinous CRC, and 87 colonoscopy-negative controls. Results were compared with fecal miRNA profiles of a cohort from the Czech Republic (n = 98). Most of the differentially expressed (DE) stool miRNAs (n = 324) were in common between CRC with mucinous morphology and non-mucinous histopathological subtypes in comparison with healthy controls. Interestingly, the altered levels of 25 fecal miRNAs previously identified distinguishing CRC cases from controls in both cohorts were also confirmed after stratification for mucinous morphology. Forty-nine miRNAs were DE exclusively in CRC with mucinous morphology and 61 in non-mucinous CRC. Mucinous cancers and those with mucinous component showed fairly similar profiles that were comparable in the Czech cohort. Among the stool DE miRNAs observed in CRC with mucinous morphology, 20 were also altered in the comparison between tumor and adjacent mucosa tissue. This study highlights miRNAs specifically altered in CRC with mucinous morphology. Nevertheless, the performance of our stool miRNA signature in accurately distinguishing CRC cases from controls was not significantly affected by this histological subtype. This aspect further supports the use of stool miRNAs for noninvasive diagnosis and screening strategies.

• MeSH
• feces * chemie   MeSH
• kohortové studie MeSH
• kolorektální nádory * genetika   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mikro RNA * genetika   metabolismus   MeSH
• mucinózní adenokarcinom * genetika   patologie   MeSH
• nádorové biomarkery genetika   MeSH
• regulace genové exprese u nádorů MeSH
• senioři MeSH
• stanovení celkové genové exprese metody   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH