Candidate gene study of HOXB1 in autism spectrum disorder

<p>Abstract</p> <p>Background</p> <p><it>HOXB1 </it>plays a major role in brainstem morphogenesis and could partly determine the cranial circumference in conjunction with <it>HOXA1</it>. In our sample, <it>HOXA1 </it>alleles significantly influence head growth rates both in autistic patients and in population controls. An initial report, suggesting that <it>HOXB1 </it>could confer autism vulnerability in interaction with <it>HOXA1</it>, was not confirmed by five small association studies.</p> <p>Methods</p> <p>Our sample includes 269 autistic individuals, belonging to 219 simplex and 28 multiplex families. A mutational analysis of the two exons and flanking intronic sequences of the <it>HOXB1 </it>gene was carried out in 84 autistic patients by denaturing high performance liquid chromatography, followed by DNA sequencing. Identified rare variants were then searched by a restriction analysis in 236 autistic patients and 325-345 controls. Case-control and family-based association studies were performed on two common variants in 169 Italian patients versus 184 Italian controls and in 247 trios.</p> <p>Results</p> <p>We identified three common polymorphisms, rs72338773 [c.82insACAGCGCCC (INS/nINS)], rs12939811 [c.309A>T (Q103H)], and rs7207109 [c.450G>A (A150A)] and three rare variants, namely IVS1+63G>A, rs35115415 [c.702G>A (V234V)] and c.872_873delinsAA (S291N). SNPs rs72338773 and rs12939811 were not associated with autism, using either a case-control (alleles, exact <it>P </it>= 0.13) or a family-based design [transmission/disequilibrium test (TDT)χ²= 1.774, <it>P </it>= 0.183]. The rare variants, all inherited from one of the parents, were present in two Italian and in two Caucasian-American families. Autistic probands in two families surprisingly inherited a distinct rare variant from each parent. The IVS1+63A allele was present in 3/690 control chromosomes, whereas rare alleles at rs35115415 and c.872_873delinsAA (S291N) were not found in 662 and 650 control chromosomes, respectively. The INS-T309 allele influenced head size, but its effect appears more modest and shows no interaction with <it>HOXA1 </it>alleles. The INS-T309 allele is also associated with more severe stereotypic behaviours, according to ADI-R scores (<it>N </it>= 60 patients, <it>P </it>< 0.01).</p> <p>Conclusions</p> <p><it>HOXB1 </it>mutations do not represent a common cause of autism, nor do <it>HOXB1 </it>common variants play important roles in autism vulnerability. <it>HOXB1 </it>provides minor, albeit detectable contributions to head circumference in autistic patients, with <it>HOXA1 </it>displaying more prominent effects. <it>HOXB1 </it>variants may modulate the clinical phenotype, especially in the area of stereotypic behaviours.</p

Effect of ABCG2/BCRP Expression on Efflux and Uptake of Gefitinib in NSCLC Cell Lines

BCRP/ABCG2 emerged as an important multidrug resistance protein, because it confers resistance to several classes of cancer chemotherapeutic agents and to a number of novel molecularly-targeted therapeutics such as tyrosine kinase inhibitors. Gefitinib is an orally active, selective EGFR tyrosine kinase inhibitor used in the treatment of patients with advanced non small cell lung cancer (NSCLC) carrying activating EGFR mutations. Membrane transporters may affect the distribution and accumulation of gefitinib in tumour cells; in particular a reduced intracellular level of the drug may result from poor uptake, enhanced efflux or increased metabolism

Setup errors in patients with head-neck cancer (HNC), treated using the Intensity Modulated Radiation Therapy (IMRT) technique: how it influences the customised immobilisation systems, patient’s pain and anxiety

Abstract Background In patients with head-neck cancer treated with IMRT, immobility of the upper part of the body during radiation is maintained by means of customised immobilisation devices. The main purpose of this study was to determine how the procedures for preparation of customised immobilisation systems and the patients characteristics influence the extent of setup errors. Methods A longitudinal, prospective study involving 29 patients treated with IMRT. Data were collected before CT simulation and during all the treatment sessions (528 setup errors analysed overall); the correlation with possible risk factors for setup errors was explored using a linear mixed model. Results Setup errors were not influenced by the patient’s anxiety and pain. Temporary removal of the thermoplastic mask before carrying out the CT simulation shows statistically borderline, clinically relevant, increase of setup errors (+24.7%, 95% CI: −0.5% - 55.8%). Moreover, a unit increase of radiation therapists who model the customised thermoplastic mask is associated to a −18% (−29.2% - -4.9%) reduction of the errors. The setup error is influenced by the patient’s physical features; in particular, it increases both in patients in whom the treatment position is obtained with ‘Shoulder down’ (+27.9%, 2.2% - 59.7%) and in patients with ‘Scoliosis/kyphosis’ problems (+65.4%, 2.3% - 164.2%). Using a ‘Small size standard plus customized neck support device’ is associated to a −52.3% (−73.7% - -11.2%) reduction. The increase in number of radiation therapists encountered during the entire treatment cycle does not show associations. Increase in the body mass index is associated with a slight reduction in setup error by (−2.8%, −5% - -0.7%). Conclusion The position of the patient obtained by forcing the shoulders downwards, clinically significant scoliosis or kyphosis and the reduction of the number of radiation therapists who model the thermoplastic mask are found to be statistically significant risk factors that can cause an increase in setup errors, while the use of ‘Small size’ neck support device and patient BMI can diminish them

SYNTHESIS AND CHARACTERIZATION OF ONE-DIMENSIONAL POLYMERIC CHLOROCADMATE(II) SYSTEMS

In this paper we report the synthesis and the structural, thermal, electrical properties of 1-D polymeric chlorocadmates(II), having N-methylpropane-1,3-diammonium (mepnH2) and NN-diethylpropane-1,3-diammonium (et2pnH-2) as countercations. The (mepnH2)2[Cd3CI10] crystallizes in space group P2(1)/c. The unit cell dimensions are a = 10.164 (3) angstrom, b = 13.434 (10) angstrom, c = 10.099 (3) angstrom, beta = 112.04 (3)-degrees, V = 1278 (i) angstrom3, z = 2. The final R value for 2124 reflections (I > 2sigma(I)) is 0.0445 (R(w) = 0.0384). Its structure consists of N-methylpropane-1,3-diamine dications and infinite chains of [(Cd3Cl10)n4n-]n moieties forming 1-D polymer running along the x axis. The (et2pnH2)2[Cd5Cl14].2H2O crystallizes in space group P1BAR. The unit cell dimensions are a = 6.874 (1) angstrom, b = 10.381 (1) angstrom, c = 14.572 (i) angstrom, a = 100.14 (1)-degrees, beta = 99.31 (1)-degrees, gamma = 97.46 (1)-degrees, V = 996.7 (2) angstrom3, Z = 1. The final R value for 3405 reflections (I > 2sigma(I)) is 0.0452 (R(w) = 0.0393). Its structure consists of N,N-diethylpropane-1,3-diamine dications, infinite alternate stacked pseudoplanar trinuclear [Cd3Cl8]2- and dinuclear [Cd2Cl6]2- dianions and uncoordinated water molecules. Both the packings are strengthened by N-H ... Cl hydrogen bonds between anions and cations. A first-order phase transition of (mepnH2)2[Cd3CI10] has been identified by DSC measurements and X-ray powder diffraction, related to the disordering of the hydrocarbon chains at high temperature. This increases the cell volume. The electrical conductivity of the compounds can be envisaged as protonic, and the whole electrical behavior can be correlated to their structures and phase transition

Trastuzumab emtansine is active on HER-2 overexpressing NSCLC cell lines and overcomes gefitinib resistance

BACKGROUND: HER-2 represents a relatively new therapeutic target for non small cell lung cancer (NSCLC) patients. The incidence for reported HER-2 overexpression/amplification/mutations ranges from 2 to 20% in NSCLC. Moreover, HER-2 amplification is a potential mechanism of resistance to tyrosine kinase inhibitors of the epidermal growth factor receptor (EGFR-TKI) (about 10% of cases). T-DM1, trastuzumab emtansine is an antibody-drug conjugate composed by the monoclonal antibody trastuzumab and the microtubule polymerization inhibitor DM1. The activity of T-DM1 has been studied in breast cancer but the role of T-DM1 in lung cancer remains unexplored. METHODS: Antiproliferative and proapoptotic effects of T-DM1 have been investigated in different NSCLC cell lines by MTT, crystal violet staining, morphological study and Western blotting. HER-2 expression and cell cycle were evaluated by flow cytometry and Western blotting. Antibody dependent cell cytotoxicity (ADCC) was measured with a CytoTox assay. Xenografted mice model has been generated using a NSCLC cell line to evaluate the effect of T-DM1 on tumor growth. Moreover, a morphometric and immunohistochemical analysis of tumor xenografts was conducted. RESULTS: In this study we investigated the effect of T-DM1 in a panel of NSCLC cell lines with different HER-2 expression levels, in H1781 cell line carrying HER-2 mutation and in gefitinib resistant HER-2 overexpressing PC9/HER2cl1 cell clone. T-DM1 efficiently inhibited proliferation with arrest in G2-M phase and induced cell death by apoptosis in cells with a significant level of surface expression of HER-2. Antibody-dependent cytotoxicity assay documented that T-DM1 maintained the same activity of trastuzumab. Our data also suggest that targeting HER-2 with T-DM1 potentially overcomes gefitinib resistance. In addition a correlation between cell density/tumor size with both HER-2 expression and T-DM1 activity was established in vitro and in an in vivo xenograft model. CONCLUSIONS: Our results indicate that targeting HER-2 with T-DM1 may offer a new therapeutic approach in HER-2 over-expressing lung cancers including those resistant to EGFR TKIs

Gefitinib Inhibits Invasive Phenotype and Epithelial-Mesenchymal Transition in Drug-Resistant NSCLC Cells with MET Amplification

Despite the initial response, all patients with epidermal growth factor receptor (EGFR)-mutant non-small cell lung cancer (NSCLC) eventually develop acquired resistance to EGFR tyrosine kinase inhibitors (TKIs). The EGFR-T790M secondary mutation is responsible for half of acquired resistance cases, while MET amplification has been associated with acquired resistance in about 5-15% of NSCLCs. Clinical findings indicate the retained addiction of resistant tumors on EGFR signaling. Therefore, we evaluated the molecular mechanisms supporting the therapeutic potential of gefitinib maintenance in the HCC827 GR5 NSCLC cell line harbouring MET amplification as acquired resistance mechanism. We demonstrated that resistant cells can proliferate and survive regardless of the presence of gefitinib, whereas the absence of the drug significantly enhanced cell migration and invasion. Moreover, the continuous exposure to gefitinib prevented the epithelial-mesenchymal transition (EMT) with increased E-cadherin expression and down-regulation of vimentin and N-cadherin. Importantly, the inhibition of cellular migration was correlated with the suppression of EGFR-dependent Src, STAT5 and p38 signaling as assessed by a specific kinase array, western blot analysis and silencing functional studies. On the contrary, the lack of effect of gefitinib on EGFR phosphorylation in the H1975 cells (EGFR-T790M) correlated with the absence of effects on cell migration and invasion. In conclusion, our findings suggest that certain EGFR-mutated patients may still benefit from a second-line therapy including gefitinib based on the specific mechanism underlying tumor cell resistance