Klonální hematopoéza neurčitého potenciálu (clonal hematopoiesis of indeterminate potential; CHIP) nástává během stárnutí lidského organizmu postupnou kumulací somatických a potenciálně preleukemických mutací v hematopoetických kmenových buňkách. Tyto mutace se vyskytují v genech, které mimo jiné hrají důležitou roli v regulaci zánětu. Riziko hematologické malignity u nositelů CHIP mutací je relativně nízké. U těchto pacientů se však vyskytuje 40% riziko celkové mortality způsobené vyšším výskytem infarktu myokardu a CMP. Ačkoli v současnosti přibývají důkazy o vztahu mezi CHIP, zánětem a kardiovaskulárními chorobami, vztah mezi CHIP a CMP dosud nebyl zcela objasněn. Cílem tohoto přehledného referátu je poukázat na aktuální a potenciálně klinicky relevatní problematiku, která je nepochybně výchozí pro další výzkum.
Clonal hematopoiesis of indeterminate potential (CHIP) occurs during human aging through the progressive accumulation of somatic and potentially preleukemic mutations in hematopoietic stem cells. These mutations occur in genes that, among other things, play an important role in the regulation of inflammation. The risk of hematological malignancy in carriers of CHIP mutations is relatively low. However, these patients have a 40% risk of all-cause mortality due to a higher incidence of myocardial infarction and stroke. Although there is recent evidence of a relationship between CHIP, inflammation and cardiovascular diseases, the relationship between CHIP and stroke has not yet been fully elucidated. The aim of this review is to highlight a timely and potentially clinically relevant issue that is undoubtedly a starting point for further research.
- MeSH
- ateroskleróza etiologie genetika komplikace MeSH
- cévní mozková příhoda * etiologie genetika komplikace MeSH
- kardiovaskulární nemoci etiologie genetika komplikace MeSH
- klinická studie jako téma MeSH
- klonální hematopoéza * genetika MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- rizikové faktory kardiovaskulárních chorob MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
BACKGROUND: Surgical treatment of early-stage non-small cell lung cancer (NSCLC) yields highest expectations for recovery. However, the frequency of further disease progression remains high since micro-metastatic disease may be undetected by conventional diagnostic methods. We test the presence and prognostic impact of circulating tumor cells (CTCs) in peripheral blood (PB), tumor-draining pulmonary blood (TDB) and bone marrow (BM) samples from NSCLC patients. METHODS: The presence of circulating/disseminated tumor cells (CTCs/DTCs) was detected by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analysis in PB, TDB and BM samples before surgery in 119 stage IA-IIIA NSCLC patients (Clinical Trial NS10285). RESULTS: NSCLC patients with the presence of carcinoembryonic antigen (CEA) mRNA-positive CTCs/DTCs in TDB and BM had significantly shorter cancer-specific survival (CSS) (P<0.013, resp. P<0.038). Patients with the presence of epithelial cellular adhesion molecule (EpCAM) mRNA-positive CTCs in TDB samples had significantly shorter CSS and disease-free survival (DFS) (P<0.031, resp. P<0.045). A multivariate analysis identified the presence of CEA mRNA-positive CTCs in the PB as an independent negative prognostic factor for DFS (P<0.005). No significant correlation of CTCs/DTCs presence and other prognostic factors was found. CONCLUSIONS: In NSCLC patients undergoing radical surgery, the presence of CEA and EpCAM mRNA-positive CTCs/DTCs is associated with poorer survival.
- Publikační typ
- časopisecké články MeSH
The presence of wild-type RAS alleles, as determined by genotyping codons 12, 13, 59, 61, 117, and 146, is a prerequisite for personalized anti-EGFR treatment of metastatic colorectal cancer (mCRC) patients. Here we describe analytical validation of in-house developed massively parallel sequencing technology (MPS) in comparison to the in vitro diagnostics (IVD) certified qPCR method. DNA extracted from FFPE samples from CRC patients (n=703) and reference standards (n=33) were tested for KRAS and NRAS mutations in 6 codons of exons 2, 3, and 4 using deep amplicon sequencing (DAS) on a MiSeq benchtop sequencer (Illumina). Two different amplicon lengths and two different library preparation methods (long-RAS and short-RAS) were tested in order to evaluate their impact on DAS performance. In parallel, identical tumor DNA was tested by the following IVD assays: therascreen KRAS RGQ PCR Kit (Qiagen), cobas® KRAS Mutation Test (Roche Diagnostics), and SNaPshot assay (Thermo Fisher Scientific). Both DAS assays detected all the mutations present in reference standards and external quality control samples, except for the artificially generated KRAS codon 146 mutation. The DAS assays performed sufficient analytical specificity and sensitivity (≥0.95). The use of shorter amplicons prolonged the preparation steps but significantly improved the sequencing success rate of FFPE-derived DNA. RAS mutation frequencies in the Czech CRC patients were similar to previous reports, although rare mutations were also detected. DAS with short amplicons is a good strategy for routine assessment of somatic mutations in low-quality FFPE-derived DNA.
Cíl: Přesný vztah mezi genovou mutací hematopoetických buněk (clonal hematopoiesis of indeterminate potential; CHIP) a klinickým projevem nebyl doposud jasně popsán. Cílem této akademické multicentrické klinické studie je prozkoumat souvislost mezi přítomností CHIP v periferní krvi a incidencí ischemické CMP (iCMP) a současně tak prokázat, zda CHIP ovlivňuje etiopatogenezi aterosklerózy, což by mohlo vést k určení nového možného rizikového faktoru v diagnostice predispozice k iCMP. Metodika: Jedná se o multicentrickou prospektivní studii, do které je zapojeno 14 iktových a komplexních cerebrovaskulárních center v ČR. Předpokládaný celkový počet je 1 000 zařazených pacientů ve věku 70 a více let a 50 odebraných vzorků aterosklerotických plátů, na jejichž zajištění budou spolupracovat vybraná chirurgická pracoviště. Sledované parametry zahrnují osobní a farmakologickou anamnézu, laboratorní parametry, neurologické parametry a výsledky vybraných zobrazovacích vyšetření. Předpokládaným výstupem bude definování role konkrétních alelických variant a jejich zastoupení jako rizikového faktoru pro rozvoj aterosklerózy a následné manifestace iCMP.
Aim: The exact relationship between the gene mutation of hematopoietic cells (clonal hematopoiesis of indeterminate potential; CHIP) and the clinical manifestation has not yet been clearly described. The aim of this academic multicentric clinical study is to investigate the association between the presence of CHIP in peripheral blood and the incidence of ischemic stroke, and at the same time, to demonstrate whether CHIP contributes to the etiopathogenesis of atherosclerosis. This could lead to the identification of a new potential risk factor in the diagnosis of ischemic stroke predisposition. Methodology: This is a multicenter prospective study involving 14 primary and comprehensive stroke centers in the Czech Republic. The total number of enrolled patients aged ≥ 70 years is expected to be 1,000. The selected surgical centers will collaborate to provide 50 atherosclerotic plaque samples. Monitored parameters include personal and pharmacological history, laboratory parameters, neurological parameters, and results of selected imaging examinations. The expected outcome will be the definition of specific allelic variants and their frequencies as risk factors for the development of atherosclerosis and subsequent manifestation of ischemic stroke.
- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
BACKGROUND: Congenital central hypoventilation syndrome (CCHS) is a rare genetic disorder resulting from mutations in the PHOX2B gene located on chromosome 4p12.3, characterized by hypoventilation secondary to missing responses to both hypercapnia and hypoxia. CASE REPORT: Proband. A girl, hospitalised 5 times for respiratory failure from 6 weeks old, presented at 4 years of age severe cyanosis related to pneumonia. Tracheostomy was done, and she was discharged home using a portable positive pressure ventilator during sleep. Proband's father: The father was retrospectively found out to suffer from severe headache and excessive daytime sleepiness. Molecular genetic evaluation of PHOX2B gene was performed and casual polyalanine repeat expansion mutation c.741_755dup15 in exon 3 was found both in proband and her father in heterozygous form. The proband's grandmother died of respiratory failure after administration of benzodiazepine at the age of fifty years. Considering the grandmother's history, she is highly suspected of having had CCHS as well. CONCLUSION: Repeated respiratory failure of girl was explained by PHOX2B mutation and Ondina curse. Proband´s father has incompletely penetrated PHOX2B heterozygous mutation as well and proband´s grandmother died probably from the consequences of drug interaction with PHOX2B mutated background as well. Both daughter and father currently require overnight mechanical ventilatory support. Although most PHOX2B mutations occur de novo, our case is a rare three generation family affected by autosomal dominant inheritance with incomplete penetrance manifested as the late-form of CCHS and proven PHOX2B mutation in two generations.
- MeSH
- homeodoménové proteiny genetika MeSH
- hypoventilace vrozené genetika MeSH
- lidé MeSH
- mutace genetika MeSH
- peptidy genetika MeSH
- předškolní dítě MeSH
- respirační insuficience genetika MeSH
- spánková apnoe centrální genetika MeSH
- transkripční faktory genetika MeSH
- Check Tag
- lidé MeSH
- předškolní dítě MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
