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Aurora kinases A and B play a crucial role in the regulation of mitosis, while Aurora C controls meiotic division. These proteins showed controversial behavior upon the development of epithelial tumors. Our aim was to examine if there are any differences in expression of Aurora kinases genes in malignant and non-malignant tumors and non-tumor tissues; to compare their expression with clinical characteristics of patients and expression of other prostate cancer-associated genes. Quantitative RT-PCR was used to determine Aurora A-C genes expression in 33 prostate adenocarcinomas (T), paired conventionally normal tissues (N), and 17 ade­nomas (A). Relative expression values (RE) for genes studied were estimated using 2-ΔCt and 2-ΔΔCt method. The Kruskal-Wallis with correction on multiple comparisons, according to the Benjamini-Hochberg procedure with FDR = 0.2, Dunn-Bonferroni post hoc test and Spearman rank correlation analysis were used for statistical analysis. As turned out, RE values for AURKA were found to be significantly lower in samples of T group in comparison with A group. Moreover, significant up-regulation of AURKC expression levels were detected for T 3-4 stage compared to T 1-2 stage. RE values of AURKC in T group were positively correlated with the tumor stage, while AURKB RE demonstrated a negative correlation with the tumor stage. We also found significant correlations between AURK genes expression levels and prostate cancer-associated genes in T group. We suppose that all these data point to probable involvement of Aurora kinases’ genes in prostate carcinogenesis.

Alcohol misuse is a major public health problem originating from genetic and environmental risk factors. Alterations in the brain epigenome may orchestrate changes in gene expression that lead to alcohol misuse and dependence. Through epigenome-wide association analysis of DNA methylation from human brain tissues, we identified a differentially methylated region, DMR-DLGAP2, associated with alcohol dependence. Methylation within DMR-DLGAP2 was found to be genotype-dependent, allele-specific and associated with reward processing in brain. Methylation at the DMR-DLGAP2 regulated expression of DLGAP2 in vitro, and Dlgap2-deficient mice showed reduced alcohol consumption compared with wild-type controls. These results suggest that DLGAP2 may be an interface for genetic and epigenetic factors controlling alcohol use and dependence.

Aim: To assess relative expression (RE) levels of CAF-, TAM-specific, immune defense-associated genes in prostate tumors and to show correlation of RE with clinical, pathological and molecular characteristics, with the aim to define clinically significant specific alterations in a gene expression pattern. Methods: RE of 23 genes was analyzed by a quantitative polymerase chain reaction in 37 freshly frozen samples of prostate cancer tissues of a different Gleason score (GS) and at various tumor stages, compared with RE in 37 paired conventionally normal prostate tissue (CNT) samples and 20 samples of prostate adenomas. Results: Differences in RE were shown for 11 genes out of 23 studied, when tumor samples were compared with corresponding CNTs. 7 genes, namely ACTA2, CXCL14, CTGF, THY1, FAP, CD163, CCL17 were upregulated in tumors. 4 genes, namely CCR4, NOS2A, MSMB, IL1R1 were downregulated in tumors. 14 genes demonstrated different RE in TNA at different stages: CXCL12, CXCL14, CTGF, FAP, HIF1A, THY1, CCL17, CCL22, CCR4, CD68, CD163, NOS2A, CTLA4, IL1R1. RE changes of 9 genes — CXCL12, CXCL14, HIF1A, CCR4, CCL17, NOS2A, CTLA4, IL1R1, IL2RA — were found in tumors with different GS. Moreover, 9 genes showed differences in RE in TNA, dependently on the presence or absence of the TMPRSS2/ERG fusion and 7 genes showed differences in RE of groups with differential PTEN expression. Significant correlations were calculated between RE of 9 genes in adenocarcinomas and the stage, and GS; also, between RE of 2 genes and the fusion presence; and between RE of 4 genes and PTEN expression. Conclusions: Several gene expression patterns were identified that correlated with the GS, stage and molecular characteristics of tumors, i.e. presence of the TMPRSS2/ERG fusion and alterations in PTEN expression. These expression patterns can be used for molecular profiling of prostate tumors, with the aim to develop personalized medicine approaches. However, the proposed profiling requires a more detailed analysis and a larger cohort of patients with prostate tumor.

Molecular mechanisms that define patterns of neuropeptide expression are essential for the formation and rewiring of neural circuits. The prodynorphin gene (PDYN) gives rise to dynorphin opioid peptides mediating depression and substance dependence. We here demonstrated that PDYN is expressed in neurons in human dorsolateral prefrontal cortex (dlPFC), and identified neuronal differentially methylated region in PDYN locus framed by CCCTC-binding factor binding sites. A short, nucleosome size human-specific promoter CpG island (CGI), a core of this region may serve as a regulatory module, which is hypomethylated in neurons, enriched in 5-hydroxymethylcytosine, and targeted by USF2, a methylation-sensitive E-box transcription factor (TF). USF2 activates PDYN transcription in model systems, and binds to nonmethylated CGI in dlPFC. USF2 and PDYN expression is correlated, and USF2 and PDYN proteins are co-localized in dlPFC. Segregation of activatory TF and repressive CGI methylation may ensure contrasting PDYN expression in neurons and glia in human brain.

Background: Neuropeptide precursors are traditionally viewed as proteins giving rise to small neuropeptide molecules. Prodynorphin (PDYN) is the precursor protein to dynorphins, endogenous ligands for the kappa-opioid receptor. Alternative mRNA splicing of neuropeptide genes may regulate cell- and tissue-specific neuropeptide expression and produce novel protein isoforms. We here searched for novel PDYN mRNA and their protein product in the human brain.Methods: Novel PDYN transcripts were identified using nested PCR amplification of oligo(dT) selected full-length capped mRNA. Gene expression was analyzed by qRT-PCR, PDYN protein by western blotting and confocal imaging, dynorphin peptides by radioimmunoassay. Neuronal nuclei were isolated using fluorescence activated nuclei sorting (FANS) from postmortem human striatal tissue. lmmunofluorescence staining and con focal microscopy was performed for human caudate nucleus.Results: Two novel human PDYN mRNA splicing variants were identified. Expression of one of them was confined to the striatum where its levels constituted up to 30% of total PDYN mRNA. This transcript may be translated into ASP-PDYN protein lacking 13 N-terminal amino acids, a fragment of signal peptide (SP). Delta SP-PDYN was not processed to mature dynorphins and surprisingly, was targeted to the cell nuclei in a model cellular system. The endogenous PDYN protein was identified in the cell nuclei in human striatum by western blotting of isolated neuronal nuclei, and by confocal imaging.Conclusions and general significance: High levels of alternatively spliced Delta SP-PDYN mRNA and nuclear localization of PDYN protein suggests a nuclear function for this isoform of the opioid peptide precursor in human striatum. (C) 2016 Elsevier B.V. All rights reserved.

Aim. To assess relative expression (RE) levels of genes involved in lipid metabolism in prostate tumors. To define clinically significant specific alterations on the basis of the expression pattern. Methods. RE levels were analyzed in 37 samples of prostate cancer tissues by quantitative RT-PCR. The tumors were of a different Gleason score (GS) and various stages; the paired conventionally normal prostate tissue (CNT) samples and 20 samples of prostate adenomas were also analyzed. Results. Increased RE levels of FASN and COX2 were found in an adenocarcinoma group and in adenocarcinomas with GS=7 compared to the adenoma group. Four genes, namely FASN, LDLR, HMGCR and COX2, demonstrated significant RE alterations in the adenocarcinoma groups at different stages compared to the adenoma and CNT groups. Expression of three genes (LDLR, HMGCR, COX2) showed a negative correlation with stage and GS in the adenocarcinoma group. For FASN, LDLR, HMGCR, several positive correlations of RE with levels of the epithelial cell markers were found. CPT1C and COX2 demonstrated positive correlations of RE with expression of mesenchymal, fibroblast and inflammation markers in the adenocarcinoma group. Conclusions. The studied genes controlling lipid metabolism showed differential RE in prostate cancer samples. RE levels of FASN, HMGCR and COX2 might be used as markers of sensitivity and efficacy of inhibitory drugs. Further studies are needed to confirm these data in a larger patient cohort. Мета. Встановити рівні відносної експресії (ВЕ) генів, що задіяні у ліпідному метаболізмі у пухлинах передміхурової залози. На основі цих патернів виявити клінічно-значущі специфічні порушення. Методи. ВЕ була встановлена у 37 зразках раку передміхурової залози (П) методом ПЛР у реальному часі. Пухлини були з різним ступенем Глісону (СГ) та різними стадіями. Крім того ВЕ виявлена у парних умовно-нормальних тканинах (УНТ) та аденомах (А) передміхурової залози. Результати. Підвищені рівні ВЕ FASN та COX2 знайдено у групі аденокарцином та аденокарцином зі СГ=7 порівняно з групою аденом. Чотири гени, а саме FASN, LDLR, HMGCR та COX2 продемонстрували значущі порушення ВЕ у групах аденокарцином з різними стадіями у порівнянні з групою аденом та УНТ. Три гени (LDLR, HMGCR, COX2) показали значущі негативні кореляції зі стадіями та СГ у групі аденокарцином. Для генів FASN, LDLR, HMGCR виявлена низка позитивних кореляцій ВЕ з маркерами епітеліальних клітин, тоді як для генів CPT1С та COX2 знайдено ряд позитивних кореляцій ВЕ з маркерами мезенхімальних клітин, фібробластів та запалення у групі аденокарцином. Висновки. Досліджені гени, що залучені до контролю ліпідного метаболізму показали диференційну експресію у пухлинах передміхурової залози. Рівні ВЕ генів FASN, HMGCR та COX2 можуть бути маркерами чутливості та ефективності препаратів-інгібіторів експресії генів. Для підтвердження отриманих результатів необхідні додаткові дослідження на більшій вибірці пацієнтів. Цель. Установить урони относительной экспрессии (ОЭ) генов, участвующих в липидном метаболизме в опухолях простаты. На основе этих паттернов выявить клинически-значимые специфические нарушения. Методы. ОЭ была установлена в 37 образцах рака простаты (О) методом ПЦР в реальном времени. Были использованы опухоли с разной степенью Глиссона (СГ) и различными стадиями заболевания. Кроме того ОЭ установлена в условно-нормальных тканях (УНТ) и аденомах (А) простаты. Результаты. Повышенные уровни ОЭ FASN и COX2 выявлены в группах аденокарцином и аденокарцином с СГ=7 по сравнению с группой аденом. Четыре гена, а именно FASN, LDLR, HMGCR и COX2 показали значимые изменения ОЭ в группах аденокарцином с разными стадиями по сравнению с группой аденом и УНТ. Три гена (LDLR, HMGCR, COX2) показали значимые негативные корреляции со стадией и СГ в группе аденокарцином. Для генов FASN, LDLR, HMGCR выявлен ряд позитивных корреляций ОЭ с маркерами эпителиальных клеток, тогда как для генов CPT1С и COX2 найден ряд позитивных корреляций с маркерами мезенхимальных клеток, фибробластов и воспаления в группе аденокарцином. Выводы. Исследованные гены, участвующие в контроле липидного метаболизма, показали дифференциальную экспрессию в опухолях простаты. Уровни ОЭ генов FASN, HMGCR и COX2 могут быть маркерами чувствительности и эффективности препаратов-ингибиторов экспрессии. Для подтверждения полученных результатов необходимы дополнительные исследования на большей выборке пациентов.

Molecular changes induced by excessive alcohol consumption may underlie formation of dysphoric state during acute and protracted alcohol withdrawal which leads to craving and relapse. A main molecular addiction hypothesis is that the upregulation of the dynorphin (DYN)/κ-opioid receptor (KOR) system in the nucleus accumbens (NAc) of alcohol-dependent individuals causes the imbalance in activity of D1- and D2 dopamine receptor (DR) expressing neural circuits that results in dysphoria. We here analyzed post-mortem NAc samples of human alcoholics to assess changes in prodynorphin (PDYN) and KOR (OPRK1) gene expression and co-expression (transcriptionally coordinated) patterns. To address alterations in D1- and D2-receptor circuits, we studied the regulatory interactions between these pathways and the DYN/KOR system. No significant differences in PDYN and OPRK1 gene expression levels between alcoholics and controls were evident. However, PDYN and OPRK1 showed transcriptionally coordinated pattern that was significantly different between alcoholics and controls. A downregulation of DRD1 but not DRD2 expression was seen in alcoholics. Expression of DRD1 and DRD2 strongly correlated with that of PDYN and OPRK1 suggesting high levels of transcriptional coordination between these gene clusters. The differences in expression and co-expression patterns were not due to the decline in neuronal proportion in alcoholic brain and thereby represent transcriptional phenomena. Dysregulation of DYN/KOR system and dopamine signaling through both alterations in co-expression patterns of opioid genes and decreased DRD1 gene expression may contribute to imbalance in the activity of D1- and D2-containing pathways which may lead to the negative affective state in human alcoholics. Electronic supplementary material The online version of this article (10.1007/s12035-017-0844-4) contains supplementary material, which is available to authorized users.

Damaged reward areas in human alcoholics : neuronal proportion decline and astrocyte activation

Regulation of the formation and rewiring of neural circuits by neuropeptides may require coordinated production of these signaling molecules and their receptors that may be established at the transcriptional level. Here, we address this hypothesis by comparing absolute expression levels of opioid peptides with their receptors, the largest neuropeptide family, and by characterizing coexpression (transcriptionally coordinated) patterns of these genes. We demonstrated that expression patterns of opioid genes highly correlate within and across functionally and anatomically different areas. Opioid peptide genes, compared with their receptor genes, are transcribed at much greater absolute levels, which suggests formation of a neuropeptide cloud that covers the receptor-expressed circuits. Surprisingly, we found that both expression levels and the proportion of opioid receptors are strongly lateralized in the spinal cord, interregional coexpression patterns are side specific, and intraregional coexpression profiles are affected differently by left- and right-side unilateral body injury. We propose that opioid genes are regulated as interconnected components of the same molecular system distributed between distinct anatomic regions. The striking feature of this system is its asymmetric coexpression patterns, which suggest side-specific regulation of selective neural circuits by opioid neurohormones.-Kononenko, O., Galatenko, V., Andersson, M., Bazov, I., Watanabe, H., Zhou, X. W., Iatsyshyna, A., Mityakina, I., Yakovleva, T., Sarkisyan, D., Ponomarev, I., Krishtal, O., Marklund, N., Tonevitsky, A., Adkins, D. L., Bakalkin, G. Intra- and interregional coregulation of opioid genes: broken symmetry in spinal circuits.

Glutamate is the main excitatory transmitter in the human brain. Drugs that affect the glutamatergic signaling will alter neuronal excitability. Ethanol inhibits glutamate receptors. We examined the expression level of glutamate receptor subunit mRNAs in human post-mortem samples from alcoholics and compared the results to brain samples from control subjects. RNA from hippocampal dentate gyrus (HP-DG), orbitofrontal cortex (OFC), and dorso-lateral prefrontal cortex (DL-PFC) samples from 21 controls and 19 individuals with chronic alcohol dependence were included in the study. Total RNA was assayed using quantitative RT-PCR. Out of the 16 glutamate receptor subunits, mRNAs encoding two AMPA (2-amino-3-(3-hydroxy-5-methyl-isoxazol-4-yl)propanoic acid) receptor subunits GluA2 and GluA3; three kainate receptor subunits GluK2, GluK3 and GluK5 and five NMDA (N-methyl-D-aspartate) receptor subunits GluN1, GluN2A, GluN2C, GluN2D and GluN3A were significantly increased in the HP-DG region in alcoholics. In the OFC, mRNA encoding the NMDA receptor subunit GluN3A was increased, whereas in the DL-PFC, no differences in mRNA levels were observed. Our laboratory has previously shown that the expression of genes encoding inhibitory GABA-A receptors is altered in the HP-DG and OFC of alcoholics (Jin et al., 2011). Whether the changes in one neurotransmitter system drives changes in the other or if they change independently is currently not known. The results demonstrate that excessive long-term alcohol consumption is associated with altered expression of genes encoding glutamate receptors in a brain region-specific manner. It is an intriguing possibility that genetic predisposition to alcoholism may contribute to these gene expression changes.

Acamprosate supports abstinence in some alcohol-dependent subjects, yet predictors of response are unknown. To identify response biomarkers, we investigated associations of abstinence length with polymorphisms in candidate genes in glycine and glutamate neurotransmission pathways and genes previously implicated in acamprosate response. Association analyses were conducted in the discovery sample of 225 alcohol-dependent subjects treated with acamprosate for 3 months in community-based treatment programs in the United States. Data from 110 alcohol-dependent males treated with acamprosate in the study PREDICT were used for replication of the top association findings. Statistical models were adjusted for relevant covariates, including recruitment site and baseline clinical variables associated with response. In the discovery sample, shorter abstinence was associated with increased intensity of alcohol craving and lower number of days between the last drink and initiation of acamprosate treatment. After adjustment for covariates, length of abstinence was associated with the GRIN2B rs2058878 (P = 4.6 x 10(-5)). In the replication sample, shorter abstinence was associated with increased craving, increased depressive mood score and higher alcohol consumption. Association of abstinence length with GRIN2B rs2058878 was marginally significant (P = 0.0675); as in the discovery sample, the minor A allele was associated with longer abstinence. Furthermore, rs2300272, which is in strong linkage disequilibrium with rs2058878, was also associated with abstinence length (P = 0.049). This is the first report of a replicated association of genetic markers with the length of abstinence in acamprosate-treated alcoholics. Investigation of the underlying mechanisms of this association and its usefulness for individualized treatment selection should follow.