Alport syndrome (AS) is a hereditary kidney disease caused by pathogenic variants in COL4A3 and COL4A4 genes with autosomal recessive or autosomal dominant transmission or in the COL4A5 gene with X-linked inheritance. Digenic inheritance was also described. Clinically it is associated with microscopic hematuria, followed by proteinuria and chronic renal insufficiency with end-stage renal disease in young adults. Nowadays, there is no curative treatment available. The inhibitors of RAS (renin-angiotensin system) since childhood slow the progression of the disease. Sodium-glucose cotransporter-2 inhibitors seem to be promising drugs from DAPA-CKD (dapagliflozin-chronic kidney disease) study, but only a limited number of patients with Alport syndrome was included. Endothelin type A receptor and angiotensin II type 1 receptor combined inhibitors, and lipid-lowering agents are used in ongoing studies in patients with AS and focal segmental glomerulosclerosis (FSGS). Hydroxychloroquine in AS is studied in a clinical trial in China. Molecular genetic diagnosis of AS is crucial not only for prognosis prediction but also for future therapeutic options. Different types of mutations will require various types of gene, RNA, or protein therapy to improve the function, the of final protein product.
- MeSH
- autoantigeny genetika MeSH
- chronická renální insuficience * komplikace MeSH
- dědičná nefritida * farmakoterapie genetika MeSH
- diabetes mellitus 2. typu * komplikace MeSH
- dítě MeSH
- glifloziny * MeSH
- hematurie MeSH
- kolagen typu IV genetika MeSH
- lidé MeSH
- mladý dospělý MeSH
- mutace MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mladý dospělý MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Genetic testing for pathogenic COL4A3-5 variants is usually undertaken to investigate the cause of persistent hematuria, especially with a family history of hematuria or kidney function impairment. Alport syndrome experts now advocate genetic testing for persistent hematuria, even when a heterozygous pathogenic COL4A3 or COL4A4 is suspected, and cascade testing of their first-degree family members because of their risk of impaired kidney function. The experts recommend too that COL4A3 or COL4A4 heterozygotes do not act as kidney donors. Testing for variants in the COL4A3-COL4A5 genes should also be performed for persistent proteinuria and steroid-resistant nephrotic syndrome due to suspected inherited FSGS and for familial IgA glomerulonephritis and kidney failure of unknown cause.
The recent Chandos House meeting of the Alport Variant Collaborative extended the indications for screening for pathogenic variants in the COL4A5, COL4A3 and COL4A4 genes beyond the classical Alport phenotype (haematuria, renal failure; family history of haematuria or renal failure) to include persistent proteinuria, steroid-resistant nephrotic syndrome, focal and segmental glomerulosclerosis (FSGS), familial IgA glomerulonephritis and end-stage kidney failure without an obvious cause. The meeting refined the ACMG criteria for variant assessment for the Alport genes (COL4A3-5). It identified 'mutational hotspots' (PM1) in the collagen IV α5, α3 and α4 chains including position 1 Glycine residues in the Gly-X-Y repeats in the intermediate collagenous domains; and Cysteine residues in the carboxy non-collagenous domain (PP3). It considered that 'well-established' functional assays (PS3, BS3) were still mainly research tools but sequencing and minigene assays were commonly used to confirm splicing variants. It was not possible to define the Minor Allele Frequency (MAF) threshold above which variants were considered Benign (BA1, BS1), because of the different modes of inheritances of Alport syndrome, and the occurrence of hypomorphic variants (often Glycine adjacent to a non-collagenous interruption) and local founder effects. Heterozygous COL4A3 and COL4A4 variants were common 'incidental' findings also present in normal reference databases. The recognition and interpretation of hypomorphic variants in the COL4A3-COL4A5 genes remains a challenge.
The differential diagnosis of well-differentiated tumors of follicular cell origin remains the most problematic task in thyroid pathology. Specific morphologic criteria (capsular and/or vascular invasion, nuclear characteristics) are crucial in the diagnosis of these neoplasms. However, the assessment of malignant features is inconclusive in some cases. Moreover, oncocytic thyroid tumors remain controversial in a respect to their pathobiology, behavior and management. Therefore, the useful diagnostic/prognostic thyroid markers are awaited. The aim of our study was to evaluate the expression of galectin-3 and thyroid peroxidase (TPO) in benign and malignant thyroid tumors of follicular cell origin. A total of 186 archival thyroid samples including 38 non-oncocytic follicular adenomas, 53 oncocytic (Hürthle cell) adenomas, 6 non-oncocytic follicular carcinomas, 23 oncocytic (Hürthle cell) carcinomas, 43 non-oncocytic papillary carcinomas, and 23 oncocytic papillary carcinomas were analyzed for galectin-3 and TPO expression by immunohistochemistry. Both types of papillary carcinomas showed significant upregulation of galectin-3 in comparison with the other tumor types, likewise, significant differences in galectin-3 expression were discovered between non-oncocytic and oncocytic variants of studied tumors excluding follicular carcinoma. Significant lowering of TPO was revealed in oncocytic adenomas and papillary carcinomas. In conclusion, the combined use of galectin-3 and TPO markers could help to improve the differential diagnosis of thyroid tumors. Differences in the galectin-3 and TPO expression between some oncocytic and non-oncocytic tumors support their separation in the latest WHO classification of thyroid tumors.
- MeSH
- autoantigeny genetika MeSH
- diferenciální diagnóza MeSH
- galektin 3 genetika MeSH
- jodidperoxidasa genetika MeSH
- lidé MeSH
- nádorové biomarkery genetika MeSH
- nádory štítné žlázy klasifikace diagnóza MeSH
- proteiny vázající železo genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: CIP2A has been proved to play a role as an oncogene in various types of malignancies while its functionality in renal clear cell carcinoma has not been investigated. Our study aimed to investigate the role of CIP2A in renal clear cell carcinoma and to explore the possible mechanisms. METHODS: A total of 80 patients with renal clear cell carcinoma and 32 healthy people were included in the study. Expression of CIP2A was detected by qRT-PCR. CIP2A silencing renal clear cell carcinoma cell line was established. Its effects on cell proliferation and migration were verified by CCK-8 assay and Transwell cell assay, respectively. The effects of CIP2A overexpression on AKT and VEGF were investigated. RESULTS: CIP2A expression level was increased in tumor tissues compared to adjacent healthy tissues. Serum levels of CIP2A protein were higher in cancer patients than in healthy controls, and serum levels of CIP2A protein were increased with increased stage of primary tumor. Serum CIP2A protein can be used to accurately predict renal clear cell carcinoma and its prognosis. CIP2A siRNA silencing inhibited tumor cell proliferation, and treatment with Akt activator reduced this inhibitory effect. CIP2A siRNA silencing decreased the expression level of VEGF and phosphorylation levels of AKT in renal clear cell carcinoma cells, while AKT activator treatment showed no significant effects on CIP2A expression. CONCLUSION: Downregulation of CIP2A can inhibit cancer cell proliferation and vascularization in renal clear cell carcinoma through inactivation of the Akt pathway and its downstream VEGF.
- MeSH
- autoantigeny genetika MeSH
- dospělí MeSH
- genový knockdown MeSH
- intracelulární signální peptidy a proteiny genetika MeSH
- karcinom z renálních buněk krevní zásobení genetika patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- membránové proteiny genetika MeSH
- mladý dospělý MeSH
- nádorové buněčné linie MeSH
- nádory ledvin krevní zásobení genetika patologie MeSH
- patologická angiogeneze genetika MeSH
- pohyb buněk genetika MeSH
- proliferace buněk genetika MeSH
- protoonkogenní proteiny c-akt metabolismus MeSH
- senioři MeSH
- studie případů a kontrol MeSH
- vaskulární endoteliální růstový faktor A metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
AIMS: To develop an in vitro tool for assessing the efficacy and toxicity of anticancer drugs using mixed culture containing both tumor and non-tumor cells. Such in vitro tool should have high application potential in drug-screening and personalized cancer care. METHODS: Fibroblasts were spiked as non-tumor cells into tumor cells of an established line. The mixed culture was treated with a test drug at various concentrations. After the treatment, DNA was prepared directly from the survived adhesive cells in the wells of the 96-well plates using a simple and inexpensive method, and subjected to digital PCR for measuring relative copy numbers of a target gene NF1 to that of a reference gene RPP30. The NF1 gene is known to be heterozygously deleted in these tumor cells while the RPP30 gene has two copies in both tumor and non-tumor cells. Using the NF1/ RPP30 ratios resulting from the dual digital PCR assay, the proportions of tumor cells were calculated for each drug concentration. RESULTS: Digital PCR confirmed that the tumor cells have only one copy of the NF1 gene while the non-tumor fibroblasts have two copies. By contrast, both types of cells have two copies of the reference gene RPP30. Using the ratio of the two genes, we successfully calculated the proportion of tumor cells which decreased as the dose of the test drug increased up to a certain concentration, indicating that the drug is more effective for the tumor cells than for the non-tumor cells in this dose-range. At the highest dose, we observed a slight increase in the proportion of tumor cells, likely reflecting the toxic effect of the drug on both tumor and non-tumor cells. CONCLUSION: This pilot study demonstrated the feasibility of a genetic- and cell-based tool for testing efficacy and toxicity of anticancer drugs in vitro. The promising results suggest that additional efforts are merited, for further development since such a tool will likely have high application potential (1) in drug discovery where it enables simultaneously assessing therapeutic effect on target cells and toxic effect on non-target cells, and (2) in personalized adjuvant chemotherapy where multiple drugs can be tested in primary cultures derived from surgically removed tumor.
- MeSH
- autoantigeny účinky léků genetika MeSH
- DNA nádorová účinky léků genetika MeSH
- fibroblasty účinky léků MeSH
- geny neurofibromatózy 1 účinky léků MeSH
- inhibitory enzymů farmakologie MeSH
- lidé MeSH
- mutace účinky léků genetika MeSH
- nádorové buněčné linie MeSH
- objevování léků metody MeSH
- pilotní projekty MeSH
- polymerázová řetězová reakce metody MeSH
- protinádorové látky farmakologie toxicita MeSH
- pyrimidiny farmakologie MeSH
- ribonukleasa P účinky léků genetika MeSH
- screeningové testy protinádorových léčiv metody MeSH
- studie proveditelnosti MeSH
- techniky in vitro MeSH
- tyrosinkinasy antagonisté a inhibitory MeSH
- variabilita počtu kopií segmentů DNA účinky léků genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
Poškozující zánět u nemocných s roztroušenou sklerózou mozkomíšní je podmíněn přítomností endogenních signálů nebezpečí, které jsou indukovány nebezpečnými vzory mikroorganismů a jsou důsledkem lokální alterace mozkových struktur. Identifikace signálů nebezpečí prostřednictvím receptorů PRR na buňkách vrozené imunity je následována sestavením inflamasomu, tvorbou prozánětlivých působků a účinnou prezentací antigenů autoreaktivním T-lymfocytům. Ty vyzrávají ve funkčně odlišné subsety. V patogenezi roztroušené sklerózy mozkomíšní se negativně podílejí subsety Th17 a Th1 T-lymfocytů. Imunoregulační subsety Treg, Tr naopak tlumí poškozující zánět a jejich aktivita je zesílena terapií. Poškozující a reparační procesy v RS jsou ovlivněny i abnormálním průběhem buněčné smrti, tj. nekrózy, apoptózy a autofagie.
Inflammatory reaction in patients with multiple sclerosis is induced by the presence of endogenous danger signals caused by both pathogen danger patterns and locally formed altered structures in the brain tissue. Danger signals are identified by the set of PRR receptors expressed on innate immunity cells. Inflammasomes and intracellular signaling pathways are induced in response to danger signals recognition followed by the production of pro-inflammatory mediators and presentation of antigens to auto-reactive T cells, which mature into functionally differentiated subsets. The harmful effects in pathogenesis of multiple sclerosis are associated with activity of Th17 and Th1 subsets. In contrast, Treg and Tr subsets are alleviating inflammatory response and their activity is augmented by therapy. Both harmful and reparative processes associated with multiple sclerosis are influenced by abnormal course of cell death, such as necrosis, apoptosis and autophagy.
- Klíčová slova
- signály nebezpečí, PRR, inflamasom,
- MeSH
- autoantigeny genetika imunologie MeSH
- autofagie MeSH
- biologické markery krev MeSH
- financování organizované MeSH
- inflamasomy genetika MeSH
- interleukiny metabolismus MeSH
- kaspasa 1 metabolismus MeSH
- lidé MeSH
- receptory rozpoznávající vzory genetika imunologie MeSH
- regulační T-lymfocyty imunologie MeSH
- roztroušená skleróza * diagnóza imunologie patofyziologie MeSH
- T-lymfocyty MeSH
- toll-like receptory genetika imunologie MeSH
- zánět genetika imunologie patofyziologie MeSH
- Check Tag
- lidé MeSH
The Apaf-1 interacting protein (APIP) and the uveal autoantigen with coiled coil domains and ankyrin repeats (UACA) belong to endogenous regulators of the apoptosome apparatus, but their role in tumourigenesis and progression of non-small cell lung carcinoma (NSCLC) is not known. Previous studies demonstrated that APIP inhibits the apoptosome-mediated procaspase-9 activation while UACA induces translocation of Apaf-1 from the cytoplasm into the nucleus. Here, we report for the first time that the expression of APIP and UACA genes is down-regulated on the level of both mRNA and protein in NSCLC cells and tumours. In particular, the expression of APIP protein was strikingly decreased and the expression of UACA mRNA and protein was frequently down-regulated in NSCLC tumours of different histopathological types. Moreover, stage IA NSCLC tumours showed significantly lower expression of UACA mRNA compared to higher stage tumours. The weak increase of both APIP and UACA mRNA levels in the 5-aza-2'-deoxycytidine-treated NSCLC cells indicates that mechanisms other than DNA methylation are involved in the regulation of APIP and UACA gene expression in these cancer cells. Taken together, the down-regulation of APIP and UACA expression suggests that the threshold to activate the apoptosome apparatus may be decreased in NSCLC cells due to the lack of APIP-mediated suppression and UACA-assisted Apaf-1 nuclear entry. Moreover, the loss of UACA-assisted Apaf-1 nuclear translocation may underlie the failure of DNA damage checkpoint activation in NSCLC cells leading to their genomic instability.
- MeSH
- apoptóza genetika MeSH
- autoantigeny genetika metabolismus MeSH
- dospělí MeSH
- down regulace MeSH
- genetická transkripce MeSH
- lidé středního věku MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádory plic metabolismus patologie MeSH
- nemalobuněčný karcinom plic metabolismus patologie MeSH
- neparametrická statistika MeSH
- proteiny regulující apoptózu genetika metabolismus MeSH
- regulace genové exprese u nádorů MeSH
- senioři MeSH
- staging nádorů MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
In response to ionizing radiation (IR), cells delay cell cycle progression and activate DNA repair. Both processes are vital for genome integrity, but the mechanisms involved in their coordination are not fully understood. In a mass spectrometry screen, we identified the adenosine triphosphate-dependent chromatin-remodeling protein CHD4 (chromodomain helicase DNA-binding protein 4) as a factor that becomes transiently immobilized on chromatin after IR. Knockdown of CHD4 triggers enhanced Cdc25A degradation and p21(Cip1) accumulation, which lead to more pronounced cyclin-dependent kinase inhibition and extended cell cycle delay. At DNA double-strand breaks, depletion of CHD4 disrupts the chromatin response at the level of the RNF168 ubiquitin ligase, which in turn impairs local ubiquitylation and BRCA1 assembly. These cell cycle and chromatin defects are accompanied by elevated spontaneous and IR-induced DNA breakage, reduced efficiency of DNA repair, and decreased clonogenic survival. Thus, CHD4 emerges as a novel genome caretaker and a factor that facilitates both checkpoint signaling and repair events after DNA damage.
- MeSH
- autoantigeny genetika metabolismus MeSH
- buněčný cyklus genetika MeSH
- CDC geny MeSH
- chromatin * genetika metabolismus MeSH
- chromozomy metabolismus MeSH
- DNA genetika metabolismus MeSH
- dvouřetězcové zlomy DNA MeSH
- fosfatasy cdc25 genetika metabolismus MeSH
- ionizující záření MeSH
- komplex Mi2-NuRD genetika metabolismus MeSH
- lidé MeSH
- malá interferující RNA metabolismus farmakologie MeSH
- nádorové buněčné linie MeSH
- oprava DNA * MeSH
- poškození DNA * fyziologie MeSH
- RNA interference MeSH
- signální transdukce * genetika MeSH
- ubikvitin genetika metabolismus MeSH
- ubikvitinace MeSH
- Check Tag
- lidé MeSH
