Anna Jonkisz

Genetic variants in transforming growth factor-β gene (TGFB1) affect susceptibility to schizophrenia

Immense body of evidence indicates that dysfunction of immune system is implicated in the etiology of schizophrenia. The immune theory of schizophrenia is supported by alterations in cytokine profile in the brain and peripheral blood. Given the strong genetic background of schizophrenia, it might be assumed that aberrant production of cytokines might be the consequence of genetic factors. This study aimed at investigating the association between schizophrenia susceptibility and selected functional polymorphisms in genes encoding cytokines including: interleukin-2 (IL2 −330T>G, rs2069756), interleukin-6 (IL-6 −174G>C, rs1800795), interferon-γ (IFNG +874T>A, rs2430561) as well as for the first time transforming growth factor-β1 (TGFB1 +869T>C, rs1800470 and +916G>C, rs1800471). We recruited 151 subjects with schizophrenia and 279 controls. There was a significant difference in the genotype distribution and allelic frequency of the TGFB1 +869T>C between patients with schizophrenia and healthy controls (pxa0<xa00.05). The risk of schizophrenia was more than two-fold higher in carriers of T allele (CT+TT genotypes) than individuals with CC genotype. Given documented gender differences in incidence of schizophrenia, we conducted separate analyses of male and female participants. We have shown that the association was significant in females, while in males it reached a trend toward statistical significance. To the best of our knowledge, it is the first report showing the association between TGFB1 +869T>C polymorphism and schizophrenia.

Assessment of three epigenotypes in colorectal cancer by combined bisulfite restriction analysis

Recent investigations have demonstrated the clear heterogeneity of sporadic colorectal cancer (CRC) with regard to CpG island methylation. Two unsupervised cluster analyses revealed that CRCs form three distinct DNA methylation subsets, which are referred to as the high‐, intermediate‐, and low‐methylation epigenotypes (HME, IME, and LME, respectively). A recent study by Yagi et al. found a fairly sensitive and specific identification of HME, IME, and LME using two marker panels analyzed by MALDI‐TOF mass spectrometry (MassARRAY). However, the expensive equipment required for this method substantially increases the cost and complexity of the assay. In this article, we demonstrate the assessment of HME, IME, and LME in a group of 233 sporadic CRCs using seven markers proposed by Yagi et al. The DNA methylation of each marker was quantified using combined bisulfite restriction analysis (COBRA) and analyzed along with various genetic factors associated with CRC [the BRAF and KRAS mutations, MLH1 methylation and microsatellite instability (MSI)]. The baseline methylation of each marker was generated from pooled DNA isolated from 50 normal colon tissues. We demonstrate that the correlation of HME, IME, and LME epigenotyped by COBRA using different molecular classifiers is similar to that achieved by MassARRAY. Therefore, epigenotyping CRCs using COBRA is a simple, specific, and cost‐effective method that has the potential to be widely used in CRC research.

CD28/CTLA-4/ICOS haplotypes confers susceptibility to Graves’ disease and modulates clinical phenotype of disease

Graves’ disease, an autoimmune disease with heterogeneous symptoms including Graves’ orbitopathy, has a combined genetic/environmental background, where variations within CD28/CTLA-4/ICOS genes are considered as disease markers. Association of CD28c.17+3T>C(rs3116496), CTLA-4g.319C>T(rs5742909), CTLA-4c.49A>G(rs231775), CTLA-4g.*642AT(8_33), CT60(rs3087243), Jo31(rs11571302), ICOSc.1554+4GT(8_15) polymorphisms with susceptibility to Graves’ disease and clinical outcome was investigated. The study group comprised of 561 Polish Caucasians, including 172 unrelated Graves’ disease patients. CTLA-4c.49A>G, CTLA-4g.319C>T, and CT60 were genotyped by PCR–RFLP; Jo31 and CD28c.17+3C>T by minisequencing; CTLA-4g.*642AT(8_33) and ICOSc.1554+4GT(8_15)—PCR and fluorescence-based technique. CD28c.17+3T>C(rs3116496)T/CTLA-4g.319C>T(rs5742909)C/CTLA-4c.49A>G(rs231775)G/CTLA-4g.*642AT(8_33)(AT16–21)/CT60(rs3087243)G/Jo31(rs11571302)G/ICOSc.1554+4GT(8_15)(m) and TCA(AT<16)GT(m) haplotypes increased risk of Graves’ disease, especially in males, as well as overall Graves’ orbitopathy development with severe outcome. TCG(AT16–21)GG(l) haplotype increased risk of Graves’ disease and reduced the chance of successful medical treatment. Although this haplotype was mainly observed in patients without signs of Graves’ orbitopathy, if Graves’ orbitopathy developed it favored a Graves’ orbitopathy outcome. Haplotype TCA(AT>21)GT(m) increased Graves’ disease risk in women and, in all patients, was linked to Graves’ disease without Graves’ orbitopathy. TCG(AT<16)GG(m) haplotype was predominantly observed in patients without Graves’ orbitopathy, whereas TCA(AT16–21)GG(m) was absent in those patients. TCA(AT16–21)GG(m) occurred in patients with a mild Graves’ orbitopathy outcome. The marker CTLA-4g.*642AT(8_33) was the only independent Graves’ disease risk factor, whereas CT60 was an independent factor for disease progression. Sporadic Graves’ disease was related to presence of CTLA-4c.49A>G[A] and the rare CTLA-4g.319C>T[T] allele variant. Familial background of the disease was exclusively associated with CTLA-4g.*642AT(8_33)[AT>21]/[AT>21] genotype. CD28/CTLA-4/ICOS loci may confer inherited susceptibility to Graves’ disease or may be involved in susceptibility to Graves’ disease and play a pathogenetic role.

Treatment with Ebastine Changes Expression of Histamine H1 and H2 Receptor mRNA on Peripheral Blood Mononuclear Cells

AbstractBackground: Histamine H1 and H2 receptors both belong to a family of G protein-coupled receptors that often show opposite actions on cell activity. Ebastine, a piperidine derivative, and its active metabolite, carebastine, are very potent second-generation histamine H1 receptor antagonists.n Objective: To investigate the influence of treatment with ebastine on the expression of histamine H1 receptor and H2 receptor mRNA on peripheral blood mononuclear cells (PBMC).n Patients: The study was performed in five grass pollen-allergic individuals with seasonal allergic rhinitis. All subjects showed positive skin-prick tests with mixed grass pollen. The study was performed out of the grass pollen season.n Methods: Blood samples were obtained before and after treatment with ebastine (Kestine®, Rhône-Poulenc-Rorer, France) 10 mg/day for 7 days. PBMC were isolated using density-gradient centrifugation. Histamine H1 and H2 receptor mRNA expression were determined using semiquantitative reverse transcription polymerase chain reaction.n Results: Prior to ebastine treatment, mRNA expression measured by the peak area for histamine H1 and H2 receptors was 33, 323 (± 33, 269) relative fluorescence units (RFU)and 59, 511 (± 31, 621) RFU, respectively. After treatment, histamine H1 and H2 receptor mRNA expression increased to 42, 061 (± 28, 263 ) and 89, 913 (+ 13, 053) RFU, respectively, and this difference was statistically significant (p < 0.01) in contrast to the difference prior to the treatment. Ebastine-induced histamine H1 and H2 receptor upregulation, however, moved the balance towards histamine H2 receptor dominance.n Conclusions: In conclusion, ebastine induced H1 and H2 receptor upregulation, but moved the balance towards H2 receptor dominance. This potent H1 receptor antagonist, in addition to reducing allergic symptoms, is able to influence inflammatory responses. Since H2 receptor stimulation shows suppressive effects on cell activation, this can contribute to long-term anti-inflammatory actions of anti-histamines.

KRAS mutation in relation to HER2 overexpression/amplification in colorectal cancer

The development of targeted therapies for KRAS, EGFR and HER2 may increase the range of response in patients with colorectal cancer. Mutation at codons 12 and 13 of the KRAS gene has been shown to be predictive of cetuximab response in colorectal cancer. However, due to the combined effects of multiple oncogenes involved in disease progression of patients with colorectal cancer, it seems to be important to identify the molecular factors that characterize therapy-resistance phenotype of tumors. n nForty six paraffin-embedded colorectal cancer specimens were analyzed for KRAS mutation and HER2 overexpression/amplification. A high–resolution melting (HRM) assay and single-nucleotide polymorphisms (SNPs) were used to detect somatic mutation in exon 2 notably condons 12 and 13 of the KRAS gene. HER2 overexpression was detected using monoclonal antibody and confirmed by fluorescence in situ hybridization (FISH) analysis. n nKRAS mutations for codons 12 and 13 were identified in 16/46 (34.7%) of patients by SNP. The alterations in KRAS gene were observed in similar percentage of both codons. Colorectal cancer showed mainly heterozygous 35G>A and 38G>A KRAS gene mutations. n nHRM analysis showed presence of KRAS exon 2 mutation in 13/46 (28.2%) colorectal cancers. Despite positive SPN results in three cases , HRM technique did not reveal KRAS gene alteration. The concordance rate between the two methodologies was high at 87.5%. KRAS mutation was more frequently observed in poorly differentiated tumors and adenocarcinomas than in other histological types. n nHER2 overexpression was found in 37/46 (80.3%) of all colorectal cancers and in 62.1%, 61.5% of KRAS mutation-positive cases detected by SNP and HRM techniques respectively. HER2 overexpression was accompanied by amplification of HER2 gene. The subgroup of colorectal cancers with KRAS mutation and HER2 overexpression/amplification was poorly differentiated. n nIn summary, the presence of HER2 overexpression/amplification in KRAS mutation–positive colorectal cancers suggests a possible role for the use of specific tyrosine kinase inhibitors in the treatment of disease.

Genetic variability in the system of natriuretic B peptide and principal toxicological parameters in workers exposed to lead

The study was aimed at evaluating the influence of selected polymorphisms of natriuretic peptide B precursor (NPPB) and natriuretic peptide receptor C (NPR3) genes on blood lead concentration (Pb-B) and blood zinc protoporphyrin concentration (ZnPP) in persons occupationally exposed to lead. Investigations were conducted on 360 persons (mean age: 44.49±9.62years), workers exposed to lead compounds. The analysis examined four polymorphisms of BNP gene, i.e.,: rs198388, rs198389, rs632793, and rs6676300; as well as one polymorphism of receptor C for natriuretic peptides, i.e., rs1421811. Heterozygosity in locus rs632793 of NPPB gene may result in higher concentrations of Pb-B, while allele A in locus rs632793 of NPPB gene seems to determine higher concentrations of ZnPP in persons occupationally exposed to lead. Workers exposed to lead and carrying allele C in locus rs198388 of NPPB gene, particularly in the heterozygotic setup, seem to be predisposed to present higher concentrations of ZnPP. Carriership of A allele in locus rs198389 of NPPB gene probably determines higher concentrations of ZnPP in study group. In summary, among persons occupationally exposed to lead, certain relationships were demonstrated between rs632793, rs198388 and rs198389 polymorphisms of NPPB gene and principal toxicological parameters characterizing exposure to lead.