Aberrantly methylated genes in human papillary thyroid cancer and their association with BRAF/RAS mutation

Cancer arises through accumulation of epigenetic and genetic alteration. Aberrant promoter methylation is a common epigenetic mechanism of gene silencing in cancer cells. We here performed genome-wide analysis of DNA methylation of promoter regions by Infinium HumanMethylation27 BeadChip, using 14 clinical papillary thyroid cancer samples and 10 normal thyroid samples. Among the 14 papillary cancer cases, 11 showed frequent aberrant methylation, but the other three cases showed no aberrant methylation at all. Distribution of the hypermethylation among cancer samples was non-random, which implied existence of a subset of preferentially methylated papillary thyroid cancer. Among 25 frequently methylated genes, methylation status of six genes (HIST1H3J, POU4F2, SHOX2, PHKG2, TLX3, HOXA7) was validated quantitatively by pyrosequencing. Epigenetic silencing of these genes in methylated papillary thyroid cancer cell lines was confirmed by gene re-expression following treatment with 5-aza-2′-deoxycytidine and trichostatin A, and detected by real-time RT-PCR. Methylation of these six genes was validated by analysis of additional 20 papillary thyroid cancer and 10 normal samples. Among the 34 cancer samples in total, 26 cancer samples with preferential methylation were significantly associated with mutation of BRAF/RAS oncogene (P = 0.04, Fisher's exact test). Thus, we identified new genes with frequent epigenetic hypermethylation in papillary thyroid cancer, two subsets of either preferentially methylated or hardly methylated papillary thyroid cancer, with a concomitant occurrence of oncogene mutation and gene methylation. These hypermethylated genes may constitute potential biomarkers for papillary thyroid cancer

Kinetically Protected Carbon-Bridged Oligo(p-phenylenevinylene) Derivatives for Blue Color Amplified Spontaneous Emission

Carbon-bridged oligo(p-phenylenevinylene)s (COPVn with repeating unit n = 1–6) have demonstrated great success as laser dyes for thin-film organic lasers. The excellent photostability observed in the longer homologues is, however, not present in the blue-emitting shorter compounds COPV1 and COPV2, attributed to the unprotected terminal positions that can degrade by photoreaction in the excited state. Here we report the synthesis of various COPV1 and COPV2 derivatives functionalized at the terminal positions with two types of sterically bulky protecting substituents: Tip (2,4,6-triisopropylphenyl) and tert-butyl (t-Bu) groups. Such molecular designs aim at preventing such photodegradation processes and thus to improve their stability. The efficacy of kinetic isotope effect for stabilization is also examined for COPV2, by the addition at terminal positions of deuterium atoms. Absorption, photoluminescence (PL), including PL quantum yield, and amplified spontaneous emission (ASE) studies have been conducted in polystyrene films doped with each of the derivatives. Significant and slight improvements of the ASE photostability are observed for the compounds with Tip groups and deuterium, respectively. Installation of substituents slightly affects the ASE wavelength within the blue spectral region, that is 385–413 nm and 462–474 nm, for COPV1 and COPV2, respectively.Financial support from Spanish Ministerio de Economía y Competitividad (MINECO) and the European FEDER funds through Grant MAT2015-66586-R is gratefully acknowledged. This work was partially supported by MEXT and JSPS KAKENHI Grant Numbers JP16H04106 and JP19H05716 to HT and JP19H0549 to EN

Blue and Deep‐Blue‐Emitting Organic Lasers with Top‐Layer Distributed Feedback Resonators

All‐solution processed surface‐emitting organic distributed feedback lasers are attractive devices for low‐cost applications. Here, lasers emitting in the spectral region between 375 and 475 nm, in which both active material and resonator (1D relief gratings) are based on solution‐processable polymer films, are reported. Ten different organic compounds dispersed in polystyrene are used as active layers of the prepared devices. They include various carbon‐bridged oligo(p‐phenylenevinylene) (COPVn, with n = 1,2) derivatives and two terfluorene compounds. The synthesis and complete optical and amplified spontaneous emission properties of one of the COPV1 compounds, COPV1(Me)‐t‐Bu, designed for deep‐blue emission, are also included. The feasibility of the resonator fabrication, performed by holographic lithography with a dichromated gelatine photoresist over the active film, is successfully demonstrated for all devices. Remarkably, no resolution limitations are found even for the lowest grating period (235 nm) required for the fabrication of the laser based on COPV1(Me)‐t‐Bu. It is also demonstrated that the rectangular grating profile with duty cycle 0.75:0.25 (hill:valley) is very convenient to optimize the resonator efficacy.The Spanish team acknowledges support from the Spanish Government (MINECO) and the European Community (FEDER) through Grant No. MAT2015-66586-R. H.T. and E.N. thank the financial support from MEXT (JP19H05716 for H.T. and JP19H0549 for E.N.)

Carbon-bridged oligo(p-phenylenevinylene)s for photostable and broadly tunable, solution-processable thin film organic lasers

Thin film organic lasers represent a new generation of inexpensive, mechanically flexible devices for spectroscopy, optical communications and sensing. For this purpose, it is desired to develop highly efficient, stable, wavelength-tunable and solution-processable organic laser materials. Here we report that carbon-bridged oligo(p-phenylenevinylene)s serve as optimal materials combining all these properties simultaneously at the level required for applications by demonstrating amplified spontaneous emission and distributed feedback laser devices. A series of six compounds, with the repeating unit from 1 to 6, doped into polystyrene films undergo amplified spontaneous emission from 385 to 585 nm with remarkably low threshold and high net gain coefficients, as well as high photostability. The fabricated lasers show narrow linewidth (105 pump pulses for oligomers with three to six repeating units) and wavelength tunability across the visible spectrum (408–591 nm).The work in Spain was supported by the Spanish Government (MINECO) and the European Community (FEDER) through grant nos. MAT-2011–28167-C02-01 and CTQ2012-33733 and from the Junta de Andalucía through research project P09-FQM-4708. M.M.-V. has been partly supported by a MINECO FPI fellowship (no. BES-2009-020747). Dr Merino and Dr Retolaza, at Tekniker (Spain), are acknowledged for supplying the NIL fabricated resonators. We also thank I. Garcés for technical assistance. The work in Tokyo was supported by MEXT, Japan (for KAKENHI 15H05754 to E.N. and JST-PRESTO ‘New Materials Science and Element Strategy’ to H.T.)

Three novel NY-ESO-1 epitopes bound to DRB1*0803, DQB1*0401 and DRB1*0901 recognized by CD4 T cells from CHP-NY-ESO-1-vaccinated patients

Three novel NY-ESO-1 CD4 T cell epitopes were identified using PBMC obtained from patients who were vaccinated with a complex of cholesterol-bearing hydrophobized pullulan (CHP) and NY-ESO-1 protein (CHP-NY-ESO-1). The restriction molecules were determined by antibody blocking and using various EBV-B cells with different HLA alleles as APC to present peptides to CD4 T cells. The minimal epitope peptides were determined using various N- and C-termini truncated peptides deduced from 18-mer overlapping peptides originally identified for recognition. Those epitopes were DRB1*0901-restricted NY-ESO-1 87-100. DQB1*0401-restricted NY-ESO-1 95-107 and DRB1*0803-restricted NY-ESO-1 124-134. CD4 T cells used to determine those epitope peptides recognized EBV-B cells or DC that were treated with recombinant NY-ESO-1 protein or NY-ESO-1-expressing tumor cell lysate, suggesting that the epitope peptides are naturally processed. These CD4 T cells showed a cytokine profile with Th1 characteristics. Furthermore, NY-ESO-1 87-100 peptide/HLA-DRB1*0901 tetramer staining was observed. Multiple Th1-type CD4 T cell responses are beneficial for inducing effective anti-tumor responses after NY-ESO-1 protein vaccination

Induction of hepatocyte growth factor production in human dermal fibroblasts and their proliferation by the extract of bitter melon pulp

Hepatocyte growth factor (HGF) is useful as a potential therapeutic agent for hepatic and renal fibrosis and cardiovascular diseases through inducing proliferation of epithelial and endothelial cells. HGF inducers may also be useful as therapeutic agents for these diseases. However, there have been no reports on induction of HGF production by plant extracts or juices. An extract of bitter melon (Momordica charantia L.) pulp markedly induced HGF production. There was a time lag of 72 h before induction of HGF production after the extract addition. Its stimulatory effect was accompanied by upregulation of HGF gene expression. Increases in mitogen-activated protein kinases (MAPKs) were observed from 72 h after the extract addition. Inhibitors of MAPKs suppressed the extract-induced HGF production. The extract also stimulated cell proliferation. Both activities for induction of HGF production and cell proliferation were eluted together in a single peak with 14,000 Da on gel filtration. The results indicate that bitter melon pulp extract induced HGF production and cell proliferation of human dermal fibroblasts and suggest that activation of MAPKs is involved in the HGF induction. Our findings suggest potential usefulness of the extract for tissue regeneration and provide an insight into the molecular mechanism underlying the wound-healing property of bitter melon

CD-DST ホウ オ モチイタ ニュウガン INTRINSIC SUBTYPES ニ オケル コウガンザイ カンジュ セイ シケン ノ ケントウ

乳癌の個別化治療が進む中で,依然としてエストロゲンレセプター (ER) やHER2遺伝子の発現状況と化学療法の薬剤感受性との関係には不明な点が多い.今回我々はCollagen gel Droplet embedded culture Drug Sensitivity Test (CD-DST法) を用いた抗癌剤感受性試験を行い,乳癌のsubtypeと薬剤感受性の関係について検討を行った.対象は乳癌患者60例59検体で,5-FUは34検体,epirubicinは35検体,paclitaxelは29検体で抗癌剤感受性の評価が可能であった.5-FU,epirubicin,paclitaxelいずれの薬剤においてもHER2発現状況と抗癌剤感受性との間に有意な相関は認められなかった.ERの発現を認めるホルモン陽性乳癌ではepirubicinに対する薬剤感受性が有意に低下し,5-FU,paclitaxelに対しても感受性が低い傾向が認められた.術前化学療法施行例では抗癌剤感受性試験の評価可能率が低下する事が判明した.CD-DST法は今後の乳癌化学療法を選択する上で,有用となりうる可能性が示唆された.Breast cancer is classified into four subtypes according to estrogen receptor (ER) and human epidermal growth factor receptor 2( HER2) expression. The biological subtypes have a spectrum of clinical, pathologic, and molecular features. Here, we investigated the correlation between the breast cancer subtype and chemosensitivity using the collagen gel droplet embedded culture drug sensitivity test (CD-DST). The response to 5-fluorouracil( 5-FU) was examined in 34 specimens;the response to epirubicin, in 35; and the response to paclitaxel, in 29. HER2 overexpression was not significantly correlated to the chemotherapeutic response to 5-FU, epirubicin, or paclitaxel. ER-positive breast cancer was significantly less sensitive to epirubicin and tended to be refractory to 5-FU and paclitaxel. The evaluation rate of the anti-drug sensitivity test was significantly lower in the group that received neoadjuvant chemotherapy. CD-DST could be useful in choosing anti-cancer drug agents for breast cancer patients

ラットカテコール-O-メチルトランスフェラーゼアポ、ホロ体の構造生物学的研究及びリガンド複合体の構造解析研究

広島大学(Hiroshima University)博士(薬学)Pharmacologydoctora