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Electroporated Antigen-Encoding mRNA Is Not a Danger Signal to Human Mature Monocyte-Derived Dendritic Cells

JOURNAL OF IMMUNOLOGY RESEARCH 2015年第0期2015卷 952184-952184页

作者： Hoyer, Stefanie Gerer, Kerstin F. Pfeiffer, Isabell A. Prommersberger, Sabrina Hoefflin, Sandra Jaitly, Tanushree Beltrame, Luca Cavalieri, Duccio Schuler, Gerold Vera, Julio Schaft, Niels Doerrie, Jan Univ Klinikum Erlangen Dept Dermatol Erlangen Germany Univ Erlangen Nurnberg Dept Genet D-91054 Erlangen Germany Ist Ric Farmacol Mario Negri Dept Oncol Milan Italy Univ Florence Dept Neurosci Psychol Drug Res & Child Hlth Florence Italy

For therapeutic cancer vaccination, the adoptive transfer of mRNA-electroporated dendritic cells (DCs) is frequently performed, usually with monocyte-derived, cytokine-matured DCs (moDCs). However, DCs are rich in danger-sensing receptors which could recognize the exogenously delivered mRNA and induce DC activation, hence influencing the DCs' immunogenicity. Therefore, we examined whether electroporation of mRNA with a proper cap and a poly-A tail of at least 64 adenosines had any influence on cocktail-matured moDCs. We used 16 different RNAs, encoding tumor antigens (MelanA, NRAS, BRAF, GNAQ, GNA11, and WT1), and variants thereof. None of those RNAs induced changes in the expression of CD25, CD40, CD83, CD86, and CD70 or the secretion of the cytokines IL-8, IL-6, and TNF alpha of more than 1.5-fold compared to the control condition, while an mRNA encoding anNF-kappa B-activation protein as positive control induced massive secretion of the cytokines. To determine whether mRNA electroporation had any effect on the whole transcriptome of the DCs, we performed microarray analyses of DCs of 6 different donors. None of 60,000 probes was significantly different between mock-electroporated DCs and MelanA-transfected DCs. Hence, we conclude that no transcriptional programs were induced within cocktail-matured DCs by electroporation of single tumor-antigen-encoding mRNAs.

关键词： CANCER treatment CANCER vaccines ELECTROPORATION MESSENGER RNA DENDRITIC cells MONOCYTES PHYSIOLOGY CELL culture CELL differentiation CYTOKINES CYTOLOGICAL techniques IMMUNIZATION RNA TUMOR antigens microarray technology GENE expression profiling TRANSPLANTATION

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MicroRNA Profiling Response to Acupuncture Therapy in Spontaneously Hypertensive Rats

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EVIDENCE-BASED COMPLEMENTARY AND ALTERNATIVE MEDICINE 2015年第1期2015卷 1-9页

作者： Wang, Jia-You Li, Hui Ma, Chun-Mei Wang, Jia-Lu Lai, Xin-Sheng Zhou, Shu-Feng Guangzhou Univ Chinese Med Dept Human Anat Coll Fundamental Med Sci Guangzhou 510006 Guangdong Peoples R China Univ S Florida Dept Pharmaceut Sci Coll Pharm Tampa FL 33612 USA Guangzhou Univ Chinese Med Dept Acupuncture & Moxibust Coll Acupuncture & Moxibust Guangzhou 510006 Guangdong Peoples R China Guiyang Med Univ Guizhou Prov Key Lab Regenerat Med Stem Cell & Tissue Engn Res Ctr Guiyang 550004 Guizhou Peoples R China Guiyang Med Univ Sino US Joint Lab Med Sci Guiyang 550004 Guizhou Peoples R China

MicroRNAs (miRNAs) are a group of endogenous noncoding RNAs that play important roles in many biological processes. This study aimed to check if miRNAs were involved in the response to acupuncture in rats. microarray analysis was performed to compare the miRNA expression profiles of medulla in spontaneously hypertensive rats (SHRs) treated with or without acupuncture. Our microarray analysis identified 222 differentially expressed miRNAs in the medulla of SHRs treated with acupuncture at taichong acupoint. Among these miRNAs, 23 miRNAs with a significant difference were found in acupuncture-treated SHRs compared to untreated rats. These 23 miRNAs could regulate 2963 target genes which were enriched in at least 14 pathways based on our bioinformatic analysis. miRNA-339, miR-223, and miR-145 were downregulated in the medulla of SHRs compared to normotensive rats. Notably, these miRNAs were upregulated to basal levels in the medulla of SHRs treated with acupuncture at taichong in comparison with SHRs receiving acupuncture at nonacupoint group or SHRs without any treatment. Our findings have revealed significant changes of a panel of selective miRNAs in hypertensive rats treated at taichong acupoint. These data provide insights into how acupuncture elicits beneficial effects on hypertension.

关键词： HYPERTENSION -- Treatment ACUPUNCTURE ACUPUNCTURE points ANIMAL experimentation POLYMERASE chain reaction RATS RESEARCH -- Finance RNA BIOINFORMATICS REVERSE transcriptase polymerase chain reaction DATA analysis -- Software microarray technology GENE expression profiling ONE-way analysis of variance

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Prediction of Metabolic Gene Biomarkers for Neurodegenerative Disease by an Integrated Network-Based Approach

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BIOMED RESEARCH INTERNATIONAL 2015年第0期2015卷 432012-432012页

作者： Ni, Qi Su, Xianming Chen, Jingqi Tian, Weidong Fudan Univ Sch Life Sci Dept Biostat & Computat Biol State Key Lab Genet Engn Shanghai 200438 Peoples R China Xi An Jiao Tong Univ Affiliated Hosp 1 Dept Geriatr Cardiol Xian 710000 Shaanxi Peoples R China

Neurodegenerative diseases (NDs), such as Parkinson's disease (PD) and Huntington's disease (HD), have become more and more common among aged people worldwide. One hallmark of NDs is the presence of intracellular accumulation of specific pathogenic proteins that may result from abnormal function of metabolic processes. Previously, we have developed a computational method named Met-express that predicted key enzyme-coding genes in cancer development by integrating cancer gene coexpression network with the metabolic network. Here, we applied Met-express to predict key enzyme-coding genes in both PD and HD. Functional enrichment analysis and literature review of predicted genes suggested that there might be some common pathogenic metabolic pathways for PD and HD. We further found that the predicted genes had significant functional association with known disease genes, with some of them already documented as biomarkers or therapeutic targets for NDs. As such, the predicted metabolic genes may be of use as novel biomarkers not only for ND diagnosis but also for potential therapeutic treatments.

关键词： NEURODEGENERATION PROTEIN metabolism CORRELATION (Statistics) FISHER exact test GENE expression GENES HUNTINGTON'S chorea PARKINSON'S disease FINANCING of research TUMORS microarray technology GENETIC aspects

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Carbohydrate microarrays Identify Blood Group Precursor Cryptic Epitopes as Potential Immunological Targets of Breast Cancer

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JOURNAL OF IMMUNOLOGY RESEARCH 2015年第0期2015卷 510810-510810页

作者： Wang, Denong Tang, Jin Liu, Shaoyi Huang, Jiaoti SRI Int Tumor Glyc Lab Biosci Div Menlo Pk CA 94025 USA Cornell Univ Dept Pharmacol Weill Med Coll New York NY 10065 USA Univ Calif Los Angeles David Geffen Sch Med Dept Pathol Los Angeles CA 90095 USA

Using carbohydrate microarrays, we explored potential natural ligands of antitumor monoclonal antibody HAE3. This antibody was raised against a murine mammary tumor antigen but was found to cross-react with a number of human epithelial tumors in tissues. Our carbohydrate microarray analysis reveals that HAE3 is specific for an O-glycan cryptic epitope that is normally hidden in the cores of blood group substances. Using HAE3 to screen tumor cell surface markers by flow cytometry, we found that the HAE3 glycoepitope, gp(HAE3), was highly expressed by a number of human breast cancer cell lines, including some triple-negative cancers that lack the estrogen, progesterone, and Her2/neu receptors. Taken together, we demonstrate that HAE3 recognizes a conserved cryptic glycoepitope of blood group precursors, which is nevertheless selectively expressed and surface-exposed in certain breast tumor cells. The potential of this class of O-glycan cryptic antigens in breast cancer subtyping and targeted immunotherapy warrants further investigation.

关键词： BREAST cancer immunology IMMUNOTHERAPY CARBOHYDRATES microarray technology CHEMICAL precursors ANTIGENIC determinants BLOOD groups THERAPEUTIC use

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MicroRNA and mRNA Features of Malignant Pleural Mesothelioma and Benign Asbestos-Related Pleural Effusion

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BIOMED RESEARCH INTERNATIONAL 2015年第0期2015卷 635748-635748页

作者： Ak, Guntulu Tomaszek, Sandra C. Kosari, Farhad Metintas, Muzaffer Jett, James R. Metintas, Selma Yildirim, Huseyin Dundar, Emine Dong, Jie Aubry, Marie Christine Wigle, Dennis A. Thomas, Charles F., Jr. Eskisehir Osmangazi Univ Fac Med Dept Chest Dis TR-26480 Eskisehir Turkey Mayo Clin Dept Surg Div Thorac Surg Rochester MN 55905 USA Mayo Clin Dept Mol Med Rochester MN 55905 USA Natl Jewish Hlth Div Oncol Denver CO 80206 USA Eskisehir Osmangazi Univ Fac Med Dept Publ Hlth TR-26480 Eskisehir Turkey Eskisehir Osmangazi Univ Fac Med Dept Pathol TR-26480 Eskisehir Turkey Mayo Clin Dept Lab Med & Pathol Rochester MN 55905 USA Mayo Clin Div Pulm & Crit Care Med Rochester MN 55905 USA

Introduction. We investigated the expression of microRNAs and mRNAs in pleural tissues from patients with either malignant pleural mesothelioma or benign asbestos-related pleural effusion. Methods. Fresh frozen tissues from a total of 18 malignant pleural mesothelioma and 6 benign asbestos-related pleural effusion patients were studied. Expression profiling of mRNA and microRNA was performed using standard protocols. Results. We discovered significant upregulation of multiple microRNAs in malignant pleural mesothelioma compared to benign asbestos-related pleural effusion. Hsa-miR-484, hsa-miR-320, hsa-let-7a, and hsa-miR-125a-5p were able to discriminate malignant from benign disease. Dynamically regulated mRNAs were also identified. MET was the most highly overexpressed gene in malignant pleural mesothelioma compared to benign asbestos-related pleural effusion. Integrated analyses examining microRNA-mRNA interactions suggested multiple altered targets within the Notch signaling pathway. Conclusions. Specific microRNAs and mRNAs may have diagnostic utility in differentiating patients with malignant pleural mesothelioma from benign asbestos-related pleural effusion. These studies may be particularly helpful in patients who reside in a region with a high incidence of mesothelioma.

关键词： MESOTHELIOMA PLEURA TUMORS PLEURAL effusions ASBESTOS GENE expression FINANCING of research RNA T-test (Statistics) RECEIVER operating characteristic curves DATA analysis software microarray technology GENETIC aspects

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Profiling of Selected MicroRNAs in Proliferative Eutopic Endometrium of Women with Ovarian Endometriosis

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BIOMED RESEARCH INTERNATIONAL 2015年第0期2015卷 760698-760698页

作者： Laudanski, P. Charkiewicz, R. Tolwinska, A. Szamatowicz, J. Charkiewicz, A. Niklinski, J. Med Univ Bialystok Dept Perinatol PL-15276 Bialystok Poland Med Univ Bialystok Dept Clin Mol Biol PL-15269 Bialystok Poland Clin EDMED Bialystok PL-15044 Bialystok Poland Med Univ Bialystok Dept Gynecol & Gynecol Oncol PL-15276 Bialystok Poland Med Univ Bialystok Dept Med Pathomorphol PL-15269 Bialystok Poland

It has been well documented that aberrant expression of selected microRNAs (miRNAs) might contribute to the pathogenesis of disease. The aim of the present study is to compare miRNA expression by the most comprehensive locked-nucleic acid (LNA) miRNA microarray in eutopic endometrium of patients with endometriosis and control. In the study we recruited 21 patients with endometriosis and 25 were disease-free women. The miRNA expression profiles were determined using the LNA miRNA microarray and validated for selected molecules by real-time PCR. We identified 1198 human miRNAs significantly differentially altered in endometriosis versus control samples using false discovery rate of <5%. However only 136 miRNAs showed differential regulation by fold change of at least 1.3. By the use of selected statistical analysis we obtained 45 potential pathways that might play a role in the pathogenesis of endometriosis. We also found that natural killer cell mediated cytotoxicity pathway was found to be inhibited which is consistent with previous studies. There are several pathways that may be potentially dysregulated, due to abnormal miRNA expression, in eutopic endometrium of patients with endometriosis and in this way contribute to its pathogenesis.

关键词： MICRORNA ENDOMETRIUM microarray technology ENDOMETRIOSIS CONTROL groups (Research) GENE expression KILLER cells CELL-mediated cytotoxicity

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Correcting positional correlations in Affymetrix® Genome Chips

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SCIENTIFIC REPORTS 2015年第1期5卷 9078-9078页

作者： Homouz, Dirar Chen, Gang Kudlicki, Andrzej S. Khalifa Univ Sci Technol & Res Abu Dhabi U Arab Emirates BGI Tech Shenzhen Peoples R China Univ Texas Med Branch Dept Biochem & Mol Biol Galveston TX 77555 USA Univ Texas Med Branch Inst Translat Sci Galveston TX 77555 USA Univ Texas Med Branch Sealy Ctr Mol Med Galveston TX 77555 USA

We report and model a previously undescribed systematic error causing spurious excess correlations that depend on the distance between probes on Affymetrix (R) microarrays. The phenomenon affects pairs of features with large chip separations, up to over 100 probes apart. The effect may have a significant impact on analysis of correlations in large collections of expression data, where the systematic experimental errors are repeated in many data sets. Examples of such studies include analysis of functions and interactions in groups of genes, as well as global properties of genomes. We find that the average correlations between probes on Affymetrix microarrays are larger for smaller chip distances, which points out to a previously undescribed positional artifact. The magnitude of the artifact depends on the design of the chip, and we find it to be especially high for the yeast S98 microarray, where spurious excess correlations reach 0.1 at a distance of 50 probes. We have designed an algorithm to correct this bias and provide new data sets with the corrected expression values. This algorithm was successfully implemented to remove the positional artifact from the S98 chip data while preserving the integrity of the data.

关键词： GENOMES DATA analysis microarray technology ALGORITHMS INTEGRATED circuits

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Herb-Partitioned Moxibustion and the miRNAs Related to Crohn's Disease: A Study Based on Rat Models

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EVIDENCE-BASED COMPLEMENTARY AND ALTERNATIVE MEDICINE 2015年第1期2015卷 1-13页

作者： Wei, Kai Zhang, Dan Hong, Jue Zhang, Cuihong Feng, Xiaoming Huang, Yan Liu, Jie Wu, Lingxiang Wu, Huangan Ma, Xiaopeng Fudan Univ Huashan Hosp Dept Integrated Tradit Chinese & Western Med Shanghai 200030 Peoples R China Shanghai Univ Tradit Chinese Med Shanghai 201203 Peoples R China Shanghai Res Inst Acupuncture Moxibust & Meridian Shanghai 200030 Peoples R China

Crohn's disease (CD) is a major subtype of inflammatory bowel disease (IBD). Herb-partitioned moxibustion (HPM) has been proven to be effective in treating CD by a large amount of clinical and experimental researches. MiRNAs (microRNAs) are increasingly recognized as important posttranscriptional regulators of inflammatory genes. In this study, we established experimental CD rat models and investigated the miRNAs associated with the onset of experimental CD;then, we further identified CD-related miRNAs that were regulated by HPM and explored the relationship between CD and the potential target genes of involved miRNAs. We found that miR-147 and miR-205 were significantly downregulated in colons of experimental CD rats and may be closely associated with the onset of experimental CD. HPM may extenuate inflammatory responses in colons and ameliorate colonic damages in CD via upregulating the expression of miR-147 and miR-205 and then further downregulating the expression of inflammation-related mRNAs, negatively regulating inflammatory signal pathways, and reducing the production of downstream inflammatory cytokines.

关键词： CROHN'S disease -- Treatment ANIMAL experimentation BIOLOGICAL models COLON (Anatomy) CYTOKINES IN situ hybridization INTESTINAL mucosa MOXIBUSTION NONPARAMETRIC statistics POLYMERASE chain reaction RATS RESEARCH -- Finance RNA TREATMENT effectiveness REVERSE transcriptase polymerase chain reaction DATA analysis -- Software microarray technology ONE-way analysis of variance

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Shikonin Inhibits Inflammatory Response in Rheumatoid Arthritis Synovial Fibroblasts via lncRNA-NR024118

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EVIDENCE-BASED COMPLEMENTARY AND ALTERNATIVE MEDICINE 2015年第1期2015卷 n/a页

作者： Yang, Ke-ya Chen, Dong-liang Henan Univ Huaihe Hosp Cadres Ward Kaifeng 475000 Peoples R China Henan Univ Huaihe Hosp Tradit Med Ward Kaifeng 475000 Peoples R China

Background. Shikonin is a major chemical component of zicao that possesses anti-inflammatory properties and the ability to mediate cellular and humoral immunity, especially in rheumatoid arthritis (RA). We investigated the impact of shikonin on inflammatory response in RA synovial fibroblasts using the CAIA model. Methods. Severe polyarticular arthritis was induced in Balb/c female mice. Expressions of lncRNA-NR024118, SOCS3, proinflammatory cytokines, and MMPs were evaluated using RT-RCR. Histone acetylation and SOCS3 protein expression were assessed by ChIP assay and western blot, respectively. Results. Mice treated with shikonin showed an abrogation of soft tissue and bone lesions. Shikonin remarkably enhanced the expression of NR024118 and SOCS3 and suppressed the secretion and expression of IL-6, IL-8, and MMPs. Proliferation of cultured RA synovial fibroblasts in the presence of IL-1 beta was also significantly inhibited by shikonin. Moreover, shikonin dose-dependently increased acetylation of histone H3 at the promoter of NR024118. Finally, NR024118 overexpression and interference significantly changed SOCS3 expression and NR024118 interference could reverse regulation of shikonin on SOCS3, proinflammatory cytokines, and MMPs expression level in MH7A cells. Conclusion. Our results reveal that, in the CAIA mouse model of RA, shikonin has disease modifying activity that is attributable to the inhibition of inflammatory response via lncRNA-NR024118.

关键词： ANIMAL experimentation BIOLOGICAL models CELL culture CYTOKINES DRUGS -- Dose-response relationship FIBROBLASTS HERBAL medicine IMMUNITY INTERLEUKINS MEDICINAL plants CHINESE medicine MICE POLYMERASE chain reaction RHEUMATOID arthritis RNA T-test (Statistics) WESTERN immunoblotting REVERSE transcriptase polymerase chain reaction DATA analysis -- Software microarray technology ADMINISTRATION of drugs DOSAGE of drugs

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Clinical Significance of SASH1 Expression in Glioma

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DISEASE MARKERS 2015年第1期2015卷 1-7页

作者： Yang, Liu Zhang, Haitao Yao, Qi Yan, Yingying Wu, Ronghua Liu, Mei Nantong Univ Affiliated Hosp Dept Neurosurg Nantong 226001 Peoples R China Nantong Univ Key Lab Neuroregenerat Nantong 226001 Peoples R China

Objective. SAM and SH3 domain containing 1 (SASH1) is a recently discovered tumor suppressor gene. The role of SASH1 in glioma has not yet been described. We investigated SASH1 expression in glioma cases to determine its clinical significance on glioma pathogenesis and prognosis. Methods. We produced tissue microarrays using 121 patient-derived glioma samples and 30 patient-derived nontumor cerebral samples. Immunohistochemistry and Western blotting were used to evaluate SASH1 expression. We used Fisher's exact tests to determine relationships between SASH1 expression and clinicopathological characteristics;Cox regression analysis to evaluate the independency of different SASH1 expression;Kaplan-Meier analysis to determine any correlation of SASH1 expression with survival rate. Results. SASH1 expression was closely correlated with the WHO glioma grade. Of the 121 cases, 66.9% with low SASH1 expression were mostly grade III-IV cases, whereas 33.1% with high SASH1 expression were mostly grades I-II. Kaplan-Meier analysis revealed a significant positive correlation between SASH1 expression and postoperative survival. Conclusions. SASH1 was widely expressed in normal and low-grade glioma tissues. SASH1 expression strongly correlated with glioma grades, showing higher expression at a lower grade, which decreased significantly as grade increased. Furthermore, SASH1 expression was positively correlated with better postoperative survival in patients with glioma.

关键词： GLIOMAS PATIENTS ANTIONCOGENES GENE expression PROGNOSIS microarray technology IMMUNOHISTOCHEMISTRY

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