52 research outputs found
Are plasma concentrations of tamoxifen active metabolites sufficient to ensure therapeutic efficacy for tamoxifen treated women with breast cancer in Poland?
Tamoxifen is the most commonly used drug for treating those patients with breast cancer who are oestrogen receptor positive. The main active metabolite of tamoxifen is (Z)-endoxifen whose therapeutic efficacy depends on its plasma concentration. A therapeutically effective threshold level has indeed been defined for (Z)-endoxifen above which the breast cancer relapse rate is significantly reduced. Such steady-state concentrations are conditional on gene polymorphism, principally cytochrome P450 2D6 (CYP2D6), that modulates the activity of the encoded enzymes that convert tamoxifen to its active metabolites. This drug’s metabolism however may become significantly altered when other medication is concomitantly taken, such as selective serotonin reuptake inhibitors, which inhibit CYP2D6. A recent study have demonstrated that the majority of tamoxifen treated women with breast cancer in Poland, may not in fact attain the therapeutic threshold levels of (Z)-endoxifen. In such cases, personalising optimal treatment should be based on direct monitoring of steady-state plasma concentrations of tamoxifen and its metabolites, which can thereby significantly improve therapeutic efficacy
Czy u leczonych tamoksyfenem kobiet chorych na raka piersi w Polsce stężenie aktywnego metabolitu leku w osoczu osiąga poziom skuteczny terapeutycznie?
Tamoksyfen jest najczęściej stosowanym lekiem u chorych na raka piersi wykazujących ekspresję receptorow steroidowych. Głownym aktywnym metabolitem tamoksyfenu jest (Z)-endoksyfen i to z jego stężeniem wiązana jest ostateczna skuteczność terapii. Opisana została wartość progowa stężenia (Z)-endoksyfenu w osoczu, powyżej ktorej obserwowano istotne obniżenie częstości nawrotu choroby. Zasadniczy wpływ na osiągane w leczeniu stężenie (Z)-endoksyfenu ma polimorfizm genow, głownie cytochromu P450 2D6 (CYP2D6), decydujący o aktywności kodowanych przez nie enzymow metabolizujących tamoksyfen do aktywnych metabolitow. Istotny wpływ modyfikujący na metabolizm leku mogą mieć rownocześnie przyjmowane leki z grupy selektywnych inhibitorow zwrotnego wychwytu serotoniny, będące inhibitorami CYP2D6. Wyniki ostatnio opublikowanych badań wskazują, że u większości kobiet chorych na raka piersi i leczonych tamoksyfenem w Polsce stężenie (Z)-endoksyfenu w osoczu nie osiąga leczniczej wartości progowej. Na podstawie dotychczasowych obserwacji można zakładać, że personalizacja i optymalizacja leczenia w oparciu o bezpośrednie monitorowanie aktualnego stężenia tamoksyfenu i jego metabolitow w osoczu leczonych kobiet może w tych przypadkach w istotny sposob poprawić skuteczność terapii
Genetic diversity of 'Candidatus Phytoplasma mali' strains in Poland
During 2010-2013, samples from 267 apple trees growing in six regions of Poland were tested for phy-
toplasma presence. ‘Candidatus Phytoplasma mali’ was detected in 17 samples from phloem tissue of apple shoots
showing typical apple proliferation symptoms. Molecular characterization of these strains was conducted using
PCR-RFLP and sequence analyses of three regions: ribosomal DNA, ribosomal protein and non-ribosomal DNA
fragment including nitroreductase and rhodanese-like protein genes. Fragments of 16S rDNA plus 16S-23S spacer
region from the ‘Ca. P. mali’ strains showed two restriction profiles: P-I and P-II when digested with HpaII enzyme.
Moreover, based on results of nitroreductase and rhodanese-like protein genes, these phytoplasmas were grouped
into two subtypes: AP-15 and AT-1. The majority of the ‘Ca. P. mali’ strains was classified to the AP-15 subtype.
Two strains were grouped to the AT-1 subtype and another showed both AP-15 and AT-1 profiles. Based on results
of analysis of rpl22 and rps3 ribosomal protein genes, most of the ‘Ca. P. mali’ strains were identified as belonging
to rpX-A. Only one strain was affiliated to rpX-B subgroup
GWAS links variants in neuronal development and actin remodeling related loci with pseudoexfoliation syndrome without glaucoma
Pseudoexfoliation syndrome (PEXS) is an age-related elastosis, strongly associated with the development of secondary glaucoma. It is clearly suggested that PEXS has a genetic component, but this has not been extensively studied. Here, a genome-wide association study (GWAS) using a DNA-pooling approach was conducted to explore the potential association of genetic variants with PEXS in a Polish population, including 103 PEXS patients without glaucoma and 106 perfectly (age- and gender-) matched controls. Individual sample TaqMan genotyping was used to validate GWAS-selected single-nucleotide polymorphism (SNP) associations. Multivariate binary logistic regression analysis was applied to develop a prediction model for PEXS. In total, 15 SNPs representing independent PEXS susceptibility loci were selected for further validation in individual samples. For 14 of these variants, significant differences in the allele and genotype frequencies between cases and controls were identified, of which 12 remained significant after Benjamini-Hochberg adjustment. The minor allele of five SNPs was associated with an increased risk of PEXS development, while for nine SNPs, it showed a protective effect. Beyond the known LOXL1 variant rs2165241, nine other SNPs were located within gene regions, including in OR11L1, CD80, TNIK, CADM2, SORBS2, RNF180, FGF14, FMN1, and RBFOX1 genes. None of these associations with PEXS has previously been reported. Selected SNPs were found to explain nearly 69% of the total risk of PEXS development. The overall risk prediction accuracy for PEXS, expressed by the area under the ROC curve (AUC) value, increased by 0.218, from 0.672 for LOXL1 rs2165241 alone to 0.89 when seven additional SNPs were included in the proposed 8-SNP prediction model. In conclusion, several new susceptibility loci for PEXS without glaucoma suggested that neuronal development and actin remodeling are potentially involved in either PEXS onset or inhibition or delay of its conversion to glaucoma
Genetic diversity of ‘Candidatus Phytoplasma mali’ strains in Poland
During 2010-2013, samples from 267 apple trees growing in six regions of Poland were tested for phytoplasma presence. ‘Candidatus Phytoplasma mali’ was detected in 17 samples from phloem tissue of apple shoots showing typical apple proliferation symptoms. Molecular characterization of these strains was conducted using PCR-RFLP and sequence analyses of three regions: ribosomal DNA, ribosomal protein and non-ribosomal DNA fragment including nitroreductase and rhodanese-like protein genes. Fragments of 16S rDNA plus 16S-23S spacer region from the ‘Ca. P. mali’ strains showed two restriction profiles: P-I and P-II when digested with HpaII enzyme. Moreover, based on results of nitroreductase and rhodanese-like protein genes, these phytoplasmas were grouped into two subtypes: AP-15 and AT-1. The majority of the ‘Ca. P. mali’ strains was classified to the AP-15 subtype. Two strains were grouped to the AT-1 subtype and another showed both AP-15 and AT-1 profiles. Based on results of analysis of rpl22 and rps3 ribosomal protein genes, most of the ‘Ca. P. mali’ strains were identified as belonging to rpX-A. Only one strain was affiliated to rpX-B subgroup
Limited predictive value of achieving beneficial plasma (Z)-endoxifen threshold level by CYP2D6 genotyping in tamoxifen-treated Polish women with breast cancer
Background
Tamoxifen, the most frequently used drug for treating estrogen receptor-positive breast cancer, must be converted into active metabolites to exert its therapeutic efficacy, mainly through CYP2D6 enzymes. The objective of this study was to investigate the impact of CYP2D6 polymorphisms on (Z)-endoxifen-directed tamoxifen metabolism and to assess the usefulness of CYP2D6 genotyping for identifying patients who are likely to have insufficient (Z)-endoxifen concentrations to benefit from standard therapy.
Methods
Blood samples from 279 Polish women with breast cancer receiving tamoxifen 20 mg daily were analyzed for CYP2D6 genotype and drug metabolite concentration. Steady-state plasma levels of tamoxifen and its 14 metabolites were measured by using the ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method.
Results
In nearly 60 % of patients, including over 30 % of patients with fully functional CYP2D6, (Z)-endoxifen concentration was below the predefined threshold of therapeutic efficacy. The most frequently observed CYP2D6 genotype was EM/PM (34.8 %), among which 83.5 % of patients had a combination of wild-type and *4 alleles. Plasma concentration of five metabolites was significantly correlated with CYP2D6 genotype. For the first time, we identified an association between decreased (E/Z)-4-OH-N-desmethyl-tamoxifen-β-D-glucuronide levels (r 2 = 0.23; p < 10 −16 ) and increased CYP2D6 functional impairment. The strongest correlation was observed for (Z)-endoxifen, whose concentration was significantly lower in groups of patients carrying at least one CYP2D6 null allele, compared with EM/EM patients. The CYP2D6 genotype accounted for plasma level variability of (Z)-endoxifen by 27 % (p < 10 −16 ) and for the variability of metabolic ratio indicating (Z)-endoxifen-directed metabolism of tamoxifen by 51 % (p < 10 −43 ).
Conclusions
The majority of breast cancer patients in Poland may not achieve a therapeutic level of (Z)-endoxifen upon receiving a standard dose of tamoxifen. This finding emphasizes the limited value of CYP2D6 genotyping in routine clinical practice for identifying patients who might not benefit from the therapy. In its place, direct monitoring of plasma steady-state (Z)-endoxifen concentration should be performed to personalize and optimize the treatment
Exome sequencing to explore the possibility of predicting genetic susceptibility to the joint occurrence of polycystic ovary syndrome and Hashimoto’s thyroiditis
A large body of evidence indicates that women with polycystic ovary syndrome (PCOS) have a higher risk of developing Hashimoto’s thyroiditis (HT) than healthy individuals. Given the strong genetic impact on both diseases, common predisposing genetic factors are possibly involved but are not fully understood. Here, we performed whole-exome sequencing (WES) for 250 women with sporadic PCOS, HT, combined PCOS and HT (PCOS+HT), and healthy controls to explore the genetic background of the joint occurrence of PCOS and HT. Based on relevant comparative analyses, multivariate logistic regression prediction modeling, and the most informative feature selection using the Monte Carlo feature selection and interdependency discovery algorithm, 77 variants were selected for further validation by TaqMan genotyping in a group of 533 patients. In the allele frequency test, variants in RAB6A, GBP3, and FNDC7 genes were found to significantly (padjusted < 0.05) differentiated the PCOS+HT and PCOS groups, variant in HIF3A differentiated the PCOS+HT and HT groups, whereas variants in CDK20 and CCDC71 differentiated the PCOS+HT and both single disorder groups. TaqMan genotyping data were used to create final prediction models, which differentiated between PCOS+HT and PCOS or HT with a prediction accuracy of AUC = 0.78. Using a 70% cutoff of the prediction score improved the model parameters, increasing the AUC value to 0.87. In summary, we demonstrated the polygenic burden of both PCOS and HT, and many common and intersecting signaling pathways and biological processes whose disorders mutually predispose patients to the development of both diseases
Helicobacter pylori Exploits a Unique Repertoire of Type IV Secretion System Components for Pilus Assembly at the Bacteria-Host Cell Interface
Colonization of the human stomach by Helicobacter pylori is an important risk factor for development of gastric cancer. The H. pylori cag pathogenicity island (cag PAI) encodes components of a type IV secretion system (T4SS) that translocates the bacterial oncoprotein CagA into gastric epithelial cells, and CagL is a specialized component of the cag T4SS that binds the host receptor α5β1 integrin. Here, we utilized a mass spectrometry-based approach to reveal co-purification of CagL, CagI (another integrin-binding protein), and CagH (a protein with weak sequence similarity to CagL). These three proteins are encoded by contiguous genes in the cag PAI, and are detectable on the bacterial surface. All three proteins are required for CagA translocation into host cells and H. pylori-induced IL-8 secretion by gastric epithelial cells; however, these proteins are not homologous to components of T4SSs in other bacterial species. Scanning electron microscopy analysis reveals that these proteins are involved in the formation of pili at the interface between H. pylori and gastric epithelial cells. ΔcagI and ΔcagL mutant strains fail to form pili, whereas a ΔcagH mutant strain exhibits a hyperpiliated phenotype and produces pili that are elongated and thickened compared to those of the wild-type strain. This suggests that pilus dimensions are regulated by CagH. A conserved C-terminal hexapeptide motif is present in CagH, CagI, and CagL. Deletion of these motifs results in abrogation of CagA translocation and IL-8 induction, and the C-terminal motifs of CagI and CagL are required for formation of pili. In summary, these results indicate that CagH, CagI, and CagL are components of a T4SS subassembly involved in pilus biogenesis, and highlight the important role played by unique constituents of the H. pylori cag T4SS
Comparative kinome analysis to identify putative colon tumor biomarkers
Kinase domains are the type of protein domain most commonly found in genes associated with tumorigenesis. Because of this, the human kinome (the protein kinase component of the genome) represents a promising source of cancer biomarkers and potential targets for novel anti-cancer therapies. Alterations in the human colon kinome during the progression from normal colon (NC) through adenoma (AD) to adenocarcinoma (AC) were investigated using integrated transcriptomic and proteomic datasets. Two hundred thirty kinase genes and 42 kinase proteins showed differential expression patterns (fold change ≥ 1.5) in at least one tissue pair-wise comparison (AD vs. NC, AC vs. NC, and/or AC vs. AD). Kinases that exhibited similar trends in expression at both the mRNA and protein levels were further analyzed in individual samples of NC (n = 20), AD (n = 39), and AC (n = 24) by quantitative reverse transcriptase PCR. Individual samples of NC and tumor tissue were distinguishable based on the mRNA levels of a set of 20 kinases. Altered expression of several of these kinases, including chaperone activity of bc1 complex-like (CABC1) kinase, bromodomain adjacent to zinc finger domain protein 1B (BAZ1B) kinase, calcium/calmodulin-dependent protein kinase type II subunit delta (CAMK2D), serine/threonine-protein kinase 24 (STK24), vaccinia-related kinase 3 (VRK3), and TAO kinase 3 (TAOK3), has not been previously reported in tumor tissue. These findings may have diagnostic potential and may lead to the development of novel targeted therapeutic interventions for colorectal cancer
Familial hypercholesterolaemia in children and adolescents from 48 countries: a cross-sectional study
Background: Approximately 450 000 children are born with familial hypercholesterolaemia worldwide every year, yet only 2·1% of adults with familial hypercholesterolaemia were diagnosed before age 18 years via current diagnostic approaches, which are derived from observations in adults. We aimed to characterise children and adolescents with heterozygous familial hypercholesterolaemia (HeFH) and understand current approaches to the identification and management of familial hypercholesterolaemia to inform future public health strategies. Methods: For this cross-sectional study, we assessed children and adolescents younger than 18 years with a clinical or genetic diagnosis of HeFH at the time of entry into the Familial Hypercholesterolaemia Studies Collaboration (FHSC) registry between Oct 1, 2015, and Jan 31, 2021. Data in the registry were collected from 55 regional or national registries in 48 countries. Diagnoses relying on self-reported history of familial hypercholesterolaemia and suspected secondary hypercholesterolaemia were excluded from the registry; people with untreated LDL cholesterol (LDL-C) of at least 13·0 mmol/L were excluded from this study. Data were assessed overall and by WHO region, World Bank country income status, age, diagnostic criteria, and index-case status. The main outcome of this study was to assess current identification and management of children and adolescents with familial hypercholesterolaemia. Findings: Of 63 093 individuals in the FHSC registry, 11 848 (18·8%) were children or adolescents younger than 18 years with HeFH and were included in this study; 5756 (50·2%) of 11 476 included individuals were female and 5720 (49·8%) were male. Sex data were missing for 372 (3·1%) of 11 848 individuals. Median age at registry entry was 9·6 years (IQR 5·8-13·2). 10 099 (89·9%) of 11 235 included individuals had a final genetically confirmed diagnosis of familial hypercholesterolaemia and 1136 (10·1%) had a clinical diagnosis. Genetically confirmed diagnosis data or clinical diagnosis data were missing for 613 (5·2%) of 11 848 individuals. Genetic diagnosis was more common in children and adolescents from high-income countries (9427 [92·4%] of 10 202) than in children and adolescents from non-high-income countries (199 [48·0%] of 415). 3414 (31·6%) of 10 804 children or adolescents were index cases. Familial-hypercholesterolaemia-related physical signs, cardiovascular risk factors, and cardiovascular disease were uncommon, but were more common in non-high-income countries. 7557 (72·4%) of 10 428 included children or adolescents were not taking lipid-lowering medication (LLM) and had a median LDL-C of 5·00 mmol/L (IQR 4·05-6·08). Compared with genetic diagnosis, the use of unadapted clinical criteria intended for use in adults and reliant on more extreme phenotypes could result in 50-75% of children and adolescents with familial hypercholesterolaemia not being identified. Interpretation: Clinical characteristics observed in adults with familial hypercholesterolaemia are uncommon in children and adolescents with familial hypercholesterolaemia, hence detection in this age group relies on measurement of LDL-C and genetic confirmation. Where genetic testing is unavailable, increased availability and use of LDL-C measurements in the first few years of life could help reduce the current gap between prevalence and detection, enabling increased use of combination LLM to reach recommended LDL-C targets early in life
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