TY - JOUR A1 - Radermacher, Kim A. A1 - Wingler, Kirstin A1 - Kleikers, Pamela A1 - Altenhöfer, Sebastian A1 - Hermans, Johannes J. R. A1 - Kleinschnitz, Christoph A1 - Schmidt, Harald H. H. W. T1 - The 1027th target candidate in stroke: Will NADPH oxidase hold up? JF - Experimental and Translational Stroke Medicine N2 - As recently reviewed, 1026 neuroprotective drug candidates in stroke research have all failed on their road towards validation and clinical translation, reasons being quality issues in preclinical research and publication bias. Quality control guidelines for preclinical stroke studies have now been established. However, sufficient understanding of the underlying mechanisms of neuronal death after stroke that could be possibly translated into new therapies is lacking. One exception is the hypothesis that cellular death is mediated by oxidative stress. Oxidative stress is defined as an excess of reactive oxygen species (ROS) derived from different possible enzymatic sources. Among these, NADPH oxidases (NOX1-5) stand out as they represent the only known enzyme family that has no other function than to produce ROS. Based on data from different NOX knockout mouse models in ischemic stroke, the most relevant isoform appears to be NOX4. Here we discuss the state-of-the-art of this target with respect to stroke and open questions that need to be addressed on the path towards clinical translation. KW - NADPH oxidases (NOX) KW - stroke therapy KW - oxidative stress Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-124197 VL - 4 IS - 11 ER - TY - JOUR A1 - Klotz, Barbara A1 - Mentrup, Birgit A1 - Regensburger, Martina A1 - Zeck, Sabine A1 - Schneidereit, Jutta A1 - Schupp, Nicole A1 - Linden, Christian A1 - Merz, Cornelia A1 - Ebert, Regina A1 - Jakob, Franz T1 - 1,25-Dihydroxyvitamin D3 Treatment Delays Cellular Aging in Human Mesenchymal Stem Cells while Maintaining Their Multipotent Capacity JF - PLoS ONE N2 - 1,25-dihydroxyvitamin D3 (1,25D3) was reported to induce premature organismal aging in fibroblast growth factor-23 (Fgf23) and klotho deficient mice, which is of main interest as 1,25D3 supplementation of its precursor cholecalciferol is used in basic osteoporosis treatment. We wanted to know if 1,25D3 is able to modulate aging processes on a cellular level in human mesenchymal stem cells (hMSC). Effects of 100 nM 1,25D3 on hMSC were analyzed by cell proliferation and apoptosis assay, beta-galactosidase staining, VDR and surface marker immunocytochemistry, RT-PCR of 1,25D3-responsive, quiescence-and replicative senescence-associated genes. 1,25D3 treatment significantly inhibited hMSC proliferation and apoptosis after 72 h and delayed the development of replicative senescence in long-term cultures according to beta-galactosidase staining and P16 expression. Cell morphology changed from a fibroblast like appearance to broad and rounded shapes. Long term treatment did not induce lineage commitment in terms of osteogenic pathways but maintained their clonogenic capacity, their surface marker characteristics (expression of CD73, CD90, CD105) and their multipotency to develop towards the chondrogenic, adipogenic and osteogenic pathways. In conclusion, 1,25D3 delays replicative senescence in primary hMSC while the pro-aging effects seen in mouse models might mainly be due to elevated systemic phosphate levels, which propagate organismal aging. KW - perspectives KW - bone marrow KW - mutant mice KW - oxidative stress KW - transcription factors KW - vitamin-D-receptor KW - differentiation KW - tissue KW - 2',7'-dichlorofluorescin KW - homeostasis Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-133392 VL - 7 IS - 1 ER - TY - JOUR A1 - Cox-Limpens, Kimberly E. M. A1 - Vles, Johan S. H. A1 - van den Hove, Daniel L. A. A1 - Zimmermann, Luc Ji A1 - Gavilanes, Antonio W. D. T1 - Fetal asphyctic preconditioning alters the transcriptional response to perinatal asphyxia JF - BMC Neuroscience N2 - Background: Genomic reprogramming is thought to be, at least in part, responsible for the protective effect of brain preconditioning. Unraveling mechanisms of this endogenous neuroprotection, activated by preconditioning, is an important step towards new clinical strategies for treating asphyctic neonates. Therefore, we investigated whole-genome transcriptional changes in the brain of rats which underwent perinatal asphyxia (PA), and rats where PA was preceded by fetal asphyctic preconditioning (FAPA). Offspring were sacrificed 6 h and 96 h after birth, and whole-genome transcription was investigated using the Affymetrix Gene1.0ST chip. Microarray data were analyzed with the Bioconductor Limma package. In addition to univariate analysis, we performed Gene Set Enrichment Analysis (GSEA) in order to derive results with maximum biological relevance. Results: We observed minimal, 25% or less, overlap of differentially regulated transcripts across different experimental groups which leads us to conclude that the transcriptional phenotype of these groups is largely unique. In both the PA and FAPA group we observe an upregulation of transcripts involved in cellular stress. Contrastingly, transcripts with a function in the cell nucleus were mostly downregulated in PA animals, while we see considerable upregulation in the FAPA group. Furthermore, we observed that histone deacetylases (HDACs) are exclusively regulated in FAPA animals. Conclusions: This study is the first to investigate whole-genome transcription in the neonatal brain after PA alone, and after perinatal asphyxia preceded by preconditioning (FAPA). We describe several genes/pathways, such as ubiquitination and proteolysis, which were not previously linked to preconditioning-induced neuroprotection. Furthermore, we observed that the majority of upregulated genes in preconditioned animals have a function in the cell nucleus, including several epigenetic players such as HDACs, which suggests that epigenetic mechanisms are likely to play a role in preconditioning-induced neuroprotection. KW - Perinatal Asphyxia KW - oxidative stress KW - microarray KW - cerebral artery occlusion KW - ischemic brain injury KW - genomic response KW - protein aggregation KW - immediate early genes KW - neuroprotection KW - tolerance KW - rat KW - expression KW - transient global ischemia KW - ubiquitination KW - epigenetics KW - fetal preconditioning KW - neonatal brain Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-116185 VL - 15 ER - TY - JOUR A1 - Brand, Susanne A1 - Amann, Kerstin A1 - Mandel, Philipp A1 - Zimnol, Anna A1 - Schupp, Nicole T1 - Oxidative DNA Damage in Kidneys and Heart of Hypertensive Mice Is Prevented by Blocking Angiotensin II and Aldosterone Receptors JF - PLOS ONE N2 - INTRODUCTION: Recently, we could show that angiotensin II, the reactive peptide of the blood pressure-regulating renin-angiotensin-aldosterone-system, causes the formation of reactive oxygen species and DNA damage in kidneys and hearts of hypertensive mice. To further investigate on the one hand the mechanism of DNA damage caused by angiotensin II, and on the other hand possible intervention strategies against end-organ damage, the effects of substances interfering with the renin-angiotensin-aldosterone-system on angiotensin II-induced genomic damage were studied. METHODS: In C57BL/6-mice, hypertension was induced by infusion of 600 ng/kg • min angiotensin II. The animals were additionally treated with the angiotensin II type 1 receptor blocker candesartan, the mineralocorticoid receptor blocker eplerenone and the antioxidant tempol. DNA damage and the activation of transcription factors were studied by immunohistochemistry and protein expression analysis. RESULTS: Administration of angiotensin II led to a significant increase of blood pressure, decreased only by candesartan. In kidneys and hearts of angiotensin II-treated animals, significant oxidative stress could be detected (1.5-fold over control). The redox-sensitive transcription factors Nrf2 and NF-κB were activated in the kidney by angiotensin II-treatment (4- and 3-fold over control, respectively) and reduced by all interventions. In kidneys and hearts an increase of DNA damage (3- and 2-fold over control, respectively) and of DNA repair (3-fold over control) was found. These effects were ameliorated by all interventions in both organs. Consistently, candesartan and tempol were more effective than eplerenone. CONCLUSION: Angiotensin II-induced DNA damage is caused by angiotensin II type 1 receptor-mediated formation of oxidative stress in vivo. The angiotensin II-mediated physiological increase of aldosterone adds to the DNA-damaging effects. Blocking angiotensin II and mineralocorticoid receptors therefore has beneficial effects on end-organ damage independent of blood pressure normalization. KW - aldosterone KW - oxidative stress KW - transcription factors KW - kidneys KW - heart KW - hypertension KW - DNA damage KW - blood pressure Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-118011 SN - 1932-6203 VL - 9 IS - 12 ER - TY - JOUR A1 - Schoen, Christoph A1 - Kischkies, Laura A1 - Elias, Johannes A1 - Ampattu, Biju Joseph T1 - Metabolism and virulence in Neisseria meningitidis N2 - A longstanding question in infection biology addresses the genetic basis for invasive behavior in commensal pathogens. A prime example for such a pathogen is Neisseria meningitidis. On the one hand it is a harmless commensal bacterium exquisitely adapted to humans, and on the other hand it sometimes behaves like a ferocious pathogen causing potentially lethal disease such as sepsis and acute bacterial meningitis. Despite the lack of a classical repertoire of virulence genes in N. meningitidis separating commensal from invasive strains, molecular epidemiology suggests that carriage and invasive strains belong to genetically distinct populations. In recent years, it has become increasingly clear that metabolic adaptation enables meningococci to exploit host resources, supporting the concept of nutritional virulence as a crucial determinant of invasive capability. Here, we discuss the contribution of core metabolic pathways in the context of colonization and invasion with special emphasis on results from genome-wide surveys. The metabolism of lactate, the oxidative stress response, and, in particular, glutathione metabolism as well as the denitrification pathway provide examples of how meningococcal metabolism is intimately linked to pathogenesis. We further discuss evidence from genome-wide approaches regarding potential metabolic differences between strains from hyperinvasive and carriage lineages and present new data assessing in vitro growth differences of strains from these two populations. We hypothesize that strains from carriage and hyperinvasive lineages differ in the expression of regulatory genes involved particularly in stress responses and amino acid metabolism under infection conditions. KW - Neisseria meningitidis KW - virulence KW - pathometabolism KW - oxidative stress KW - glutathione KW - γ-glutamyl cycle KW - glutamate dehydrogenase KW - nitrite respiration Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-113118 ER - TY - JOUR A1 - Remes, Bernhard A1 - Berghoff, Bork A. A1 - Förstner, Konrad U. A1 - Klug, Gabriele T1 - Role of oxygen and the OxyR protein in the response to iron limitation in Rhodobacter sphaeroides JF - BMC Genomics N2 - Background: High intracellular levels of unbound iron can contribute to the production of reactive oxygen species (ROS) via the Fenton reaction, while depletion of iron limits the availability of iron-containing proteins, some of which have important functions in defence against oxidative stress. Vice versa increased ROS levels lead to the damage of proteins with iron sulphur centres. Thus, organisms have to coordinate and balance their responses to oxidative stress and iron availability. Our knowledge of the molecular mechanisms underlying the co-regulation of these responses remains limited. To discriminate between a direct cellular response to iron limitation and indirect responses, which are the consequence of increased levels of ROS, we compared the response of the alpha-proteobacterium Rhodobacter sphaeroides to iron limitation in the presence or absence of oxygen. Results: One third of all genes with altered expression under iron limitation showed a response that was independent of oxygen availability. The other iron-regulated genes showed different responses in oxic or anoxic conditions and were grouped into six clusters based on the different expression profiles. For two of these clusters, induction in response to iron limitation under oxic conditions was dependent on the OxyR regulatory protein. An OxyR mutant showed increased ROS production and impaired growth under iron limitation. Conclusion: Some R. sphaeroides genes respond to iron limitation irrespective of oxygen availability. These genes therefore reflect a "core iron response" that is independent of potential ROS production under oxic, iron-limiting conditions. However, the regulation of most of the iron-responsive genes was biased by oxygen availability. Most strikingly, the OxyR-dependent activation of a subset of genes upon iron limitation under oxic conditions, including many genes with a role in iron metabolism, revealed that elevated ROS levels were an important trigger for this response. OxyR thus provides a regulatory link between the responses to oxidative stress and to iron limitation in R. sphaeroides. KW - oxidative stress KW - Rhodobacter sphaeroides KW - RNAseq KW - OxyR KW - iron limitation KW - transcriptomics KW - dependent gene-expression KW - hydrogen-peroxide KW - escherichia coli Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-115357 SN - 1471-2164 VL - 15 IS - 794 ER - TY - JOUR A1 - Leonhardt, Ines A1 - Spielberg, Steffi A1 - Weber, Michael A1 - Albrecht-Eckardt, Daniela A1 - Bläss, Markus A1 - Claus, Ralf A1 - Barz, Dagmar A1 - Scherlach, Kirstin A1 - Hertweck, Christian A1 - Löffler, Jürgen A1 - Hünniger, Kerstin A1 - Kurzai, Oliver T1 - The fungal quorum-sensing molecule farnesol activates innate immune cells but suppresses cellular adaptive immunity JF - mBio N2 - Farnesol, produced by the polymorphic fungus Candida albicans, is the first quorum-sensing molecule discovered in eukaryotes. Its main function is control of C. albicans filamentation, a process closely linked to pathogenesis. In this study, we analyzed the effects of farnesol on innate immune cells known to be important for fungal clearance and protective immunity. Farnesol enhanced the expression of activation markers on monocytes (CD86 and HLA-DR) and neutrophils (CD66b and CD11b) and promoted oxidative burst and the release of proinflammatory cytokines (tumor necrosis factor alpha [TNF-\(\alpha\)] and macrophage inflammatory protein 1 alpha [MIP-1 \(\alpha\)]). However, this activation did not result in enhanced fungal uptake or killing. Furthermore, the differentiation of monocytes to immature dendritic cells (iDC) was significantly affected by farnesol. Several markers important for maturation and antigen presentation like CD1a, CD83, CD86, and CD80 were significantly reduced in the presence of farnesol. Furthermore, farnesol modulated migrational behavior and cytokine release and impaired the ability of DC to induce T cell proliferation. Of major importance was the absence of interleukin 12 (IL-12) induction in iDC generated in the presence of farnesol. Transcriptome analyses revealed a farnesol-induced shift in effector molecule expression and a down-regulation of the granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor during monocytes to iDC differentiation. Taken together, our data unveil the ability of farnesol to act as a virulence factor of C. albicans by influencing innate immune cells to promote inflammation and mitigating the Th1 response, which is essential for fungal clearance. KW - human dendritic cells KW - Pseudomonas aeruginosa KW - induced apoptosis KW - cytokine production KW - biofilm formation KW - Candida albicans KW - mouse model KW - systemic candidiasis KW - oxidative stress KW - carcinoma cells Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-143756 VL - 6 IS - 2 ER - TY - JOUR A1 - Blein, Sophie A1 - Bardel, Claire A1 - Danjean, Vincent A1 - McGuffog, Lesley A1 - Healay, Sue A1 - Barrowdale, Daniel A1 - Lee, Andrew A1 - Dennis, Joe A1 - Kuchenbaecker, Karoline B. A1 - Soucy, Penny A1 - Terry, Mary Beth A1 - Chung, Wendy K. A1 - Goldgar, David E. A1 - Buys, Saundra S. A1 - Janavicius, Ramunas A1 - Tihomirova, Laima A1 - Tung, Nadine A1 - Dorfling, Cecilia M. A1 - van Rensburg, Elizabeth J. A1 - Neuhausen, Susan L. A1 - Ding, Yuan Chun A1 - Gerdes, Anne-Marie A1 - Ejlertsen, Bent A1 - Nielsen, Finn C. A1 - Hansen, Thomas V. O. A1 - Osorio, Ana A1 - Benitez, Javier A1 - Andreas Conejero, Raquel A1 - Segota, Ena A1 - Weitzel, Jeffrey N. A1 - Thelander, Margo A1 - Peterlongo, Paolo A1 - Radice, Paolo A1 - Pensotti, Valeria A1 - Dolcetti, Riccardo A1 - Bonanni, Bernardo A1 - Peissel, Bernard A1 - Zaffaroni, Daniela A1 - Scuvera, Giulietta A1 - Manoukian, Siranoush A1 - Varesco, Liliana A1 - Capone, Gabriele L. A1 - Papi, Laura A1 - Ottini, Laura A1 - Yannoukakos, Drakoulis A1 - Konstantopoulou, Irene A1 - Garber, Judy A1 - Hamann, Ute A1 - Donaldson, Alan A1 - Brady, Angela A1 - Brewer, Carole A1 - Foo, Claire A1 - Evans, D. Gareth A1 - Frost, Debra A1 - Eccles, Diana A1 - Douglas, Fiona A1 - Cook, Jackie A1 - Adlard, Julian A1 - Barwell, Julian A1 - Walker, Lisa A1 - Izatt, Louise A1 - Side, Lucy E. A1 - Kennedy, M. John A1 - Tischkowitz, Marc A1 - Rogers, Mark T. A1 - Porteous, Mary E. A1 - Morrison, Patrick J. A1 - Platte, Radka A1 - Eeles, Ros A1 - Davidson, Rosemarie A1 - Hodgson, Shirley A1 - Cole, Trevor A1 - Godwin, Andrew K A1 - Isaacs, Claudine A1 - Claes, Kathleen A1 - De Leeneer, Kim A1 - Meindl, Alfons A1 - Gehrig, Andrea A1 - Wappenschmidt, Barbara A1 - Sutter, Christian A1 - Engel, Christoph A1 - Niederacher, Dieter A1 - Steinemann, Doris A1 - Plendl, Hansjoerg A1 - Kast, Karin A1 - Rhiem, Kerstin A1 - Ditsch, Nina A1 - Arnold, Norbert A1 - Varon-Mateeva, Raymonda A1 - Schmutzler, Rita K. A1 - Preisler-Adams, Sabine A1 - Markov, Nadja Bogdanova A1 - Wang-Gohrke, Shan A1 - de Pauw, Antoine A1 - Lefol, Cedrick A1 - Lasset, Christine A1 - Leroux, Dominique A1 - Rouleau, Etienne A1 - Damiola, Francesca A1 - Dreyfus, Helene A1 - Barjhoux, Laure A1 - Golmard, Lisa A1 - Uhrhammer, Nancy A1 - Bonadona, Valerie A1 - Sornin, Valerie A1 - Bignon, Yves-Jean A1 - Carter, Jonathan A1 - Van Le, Linda A1 - Piedmonte, Marion A1 - DiSilvestro, Paul A. A1 - de la Hoya, Miguel A1 - Caldes, Trinidad A1 - Nevanlinna, Heli A1 - Aittomäki, Kristiina A1 - Jager, Agnes A1 - van den Ouweland, Ans M. W. A1 - Kets, Carolien M. A1 - Aalfs, Cora M. A1 - van Leeuwen, Flora E. A1 - Hogervorst, Frans B. L. A1 - Meijers-Heijboer, Hanne E. J. A1 - Oosterwijk, Jan C. A1 - van Roozendaal, Kees E. P. A1 - Rookus, Matti A. A1 - Devilee, Peter A1 - van der Luijt, Rob B. A1 - Olah, Edith A1 - Diez, Orland A1 - Teule, Alex A1 - Lazaro, Conxi A1 - Blanco, Ignacio A1 - Del Valle, Jesus A1 - Jakubowska, Anna A1 - Sukiennicki, Grzegorz A1 - Gronwald, Jacek A1 - Spurdle, Amanda B. A1 - Foulkes, William A1 - Olswold, Curtis A1 - Lindor, Noralene M. A1 - Pankratz, Vernon S. A1 - Szabo, Csilla I. A1 - Lincoln, Anne A1 - Jacobs, Lauren A1 - Corines, Marina A1 - Robson, Mark A1 - Vijai, Joseph A1 - Berger, Andreas A1 - Fink-Retter, Anneliese A1 - Singer, Christian F. A1 - Rappaport, Christine A1 - Geschwantler Kaulich, Daphne A1 - Pfeiler, Georg A1 - Tea, Muy-Kheng A1 - Greene, Mark H. A1 - Mai, Phuong L. A1 - Rennert, Gad A1 - Imyanitov, Evgeny N. A1 - Mulligan, Anna Marie A1 - Glendon, Gord A1 - Andrulis, Irene L. A1 - Tchatchou, Andrine A1 - Toland, Amanda Ewart A1 - Pedersen, Inge Sokilde A1 - Thomassen, Mads A1 - Kruse, Torben A. A1 - Jensen, Uffe Birk A1 - Caligo, Maria A. A1 - Friedman, Eitan A1 - Zidan, Jamal A1 - Laitman, Yael A1 - Lindblom, Annika A1 - Melin, Beatrice A1 - Arver, Brita A1 - Loman, Niklas A1 - Rosenquist, Richard A1 - Olopade, Olufunmilayo I. A1 - Nussbaum, Robert L. A1 - Ramus, Susan J. A1 - Nathanson, Katherine L. A1 - Domchek, Susan M. A1 - Rebbeck, Timothy R. A1 - Arun, Banu K. A1 - Mitchell, Gillian A1 - Karlan, Bethy Y. A1 - Lester, Jenny A1 - Orsulic, Sandra A1 - Stoppa-Lyonnet, Dominique A1 - Thomas, Gilles A1 - Simard, Jacques A1 - Couch, Fergus J. A1 - Offit, Kenenth A1 - Easton, Douglas F. A1 - Chenevix-Trench, Georgia A1 - Antoniou, Antonis C. A1 - Mazoyer, Sylvie A1 - Phelan, Catherine M. A1 - Sinilnikova, Olga M. A1 - Cox, David G. T1 - An original phylogenetic approach identified mitochondrial haplogroup T1a1 as inversely associated with breast cancer risk in BRCA2 mutation carriers JF - Breast Cancer Research N2 - Introduction: Individuals carrying pathogenic mutations in the BRCA1 and BRCA2 genes have a high lifetime risk of breast cancer. BRCA1 and BRCA2 are involved in DNA double-strand break repair, DNA alterations that can be caused by exposure to reactive oxygen species, a main source of which are mitochondria. Mitochondrial genome variations affect electron transport chain efficiency and reactive oxygen species production. Individuals with different mitochondrial haplogroups differ in their metabolism and sensitivity to oxidative stress. Variability in mitochondrial genetic background can alter reactive oxygen species production, leading to cancer risk. In the present study, we tested the hypothesis that mitochondrial haplogroups modify breast cancer risk in BRCA1/2 mutation carriers. Methods: We genotyped 22,214 (11,421 affected, 10,793 unaffected) mutation carriers belonging to the Consortium of Investigators of Modifiers of BRCA1/2 for 129 mitochondrial polymorphisms using the iCOGS array. Haplogroup inference and association detection were performed using a phylogenetic approach. ALTree was applied to explore the reference mitochondrial evolutionary tree and detect subclades enriched in affected or unaffected individuals. Results: We discovered that subclade T1a1 was depleted in affected BRCA2 mutation carriers compared with the rest of clade T (hazard ratio (HR) = 0.55; 95% confidence interval (CI), 0.34 to 0.88; P = 0.01). Compared with the most frequent haplogroup in the general population (that is, H and T clades), the T1a1 haplogroup has a HR of 0.62 (95% CI, 0.40 to 0.95; P = 0.03). We also identified three potential susceptibility loci, including G13708A/rs28359178, which has demonstrated an inverse association with familial breast cancer risk. Conclusions: This study illustrates how original approaches such as the phylogeny-based method we used can empower classical molecular epidemiological studies aimed at identifying association or risk modification effects. KW - single-nucleotide polymorphisms KW - genetic modifiers KW - oxidative stress KW - consortium KW - multiple diseases KW - DNA KW - haplogroups KW - susceptibility KW - Ovarian KW - variants Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-145458 VL - 17 IS - 61 ER - TY - JOUR A1 - Kim, Jae Ho A1 - Franck, Julien A1 - Kang, Taewook A1 - Heinsen, Helmut A1 - Ravid, Rivka A1 - Ferrer, Isidro A1 - Cheon, Mi Hee A1 - Lee, Joo-Yong A1 - Yoo, Jong Shin A1 - Steinbusch, Harry W. A1 - Salzet, Michel A1 - Fournier, Isabelle A1 - Park, Young Mok T1 - Proteome-wide characterization of signalling interactions in the hippocampal CA4/DG subfield of patients with Alzheimer's disease JF - Scientific Reports N2 - Alzheimer's disease (AD) is the most common form of dementia; however, mechanisms and biomarkers remain unclear. Here, we examined hippocampal CA4 and dentate gyrus subfields, which are less studied in the context of AD pathology, in post-mortem AD and control tissue to identify possible biomarkers. We performed mass spectrometry-based proteomic analysis combined with label-free quantification for identification of differentially expressed proteins. We identified 4,328 proteins, of which 113 showed more than 2-fold higher or lower expression in AD hippocampi than in control tissues. Five proteins were identified as putative AD biomarkers (MDH2, PCLO, TRRAP, YWHAZ, and MUC19 isoform 5) and were cross-validated by immunoblotting, selected reaction monitoring, and MALDI imaging. We also used a bioinformatics approach to examine upstream signalling interactions of the 113 regulated proteins. Five upstream signalling (IGF1, BDNF, ZAP70, MYC, and cyclosporin A) factors showed novel interactions in AD hippocampi. Taken together, these results demonstrate a novel platform that may provide new strategies for the early detection of AD and thus its diagnosis. KW - imaging mass spectrometry KW - neuron navigator 3 KW - dentate gyrus KW - growth factor KW - mouse model KW - neurotrophic factor KW - entorhinal cortex KW - factor expression KW - oxidative stress KW - memory deficits Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-151727 VL - 5 IS - 11138 ER - TY - JOUR A1 - Kleikers, Pamela W. M. A1 - Hooijmans, Carlijn A1 - Göb, Eva A1 - Langhauser, Friederike A1 - Rewell, Sarah S. J. A1 - Radermacher, Kim A1 - Ritskes-Hoitinga, Merel A1 - Howells, David W. A1 - Kleinschnitz, Christoph A1 - Schmidt, Harald H. H. W. T1 - A combined pre-clinical meta-analysis and randomized confirmatory trial approach to improve data validity for therapeutic target validation JF - Scientific Reports N2 - Biomedical research suffers from a dramatically poor translational success. For example, in ischemic stroke, a condition with a high medical need, over a thousand experimental drug targets were unsuccessful. Here, we adopt methods from clinical research for a late-stage pre-clinical meta-analysis (MA) and randomized confirmatory trial (pRCT) approach. A profound body of literature suggests NOX\(_{2}\) to be a major therapeutic target in stroke. Systematic review and MA of all available NOX\(_{2}\)\(^{-/y}\) studies revealed a positive publication bias and lack of statistical power to detect a relevant reduction in infarct size. A fully powered multi-center pRCT rejects NOX\(_{2}\) as a target to improve neurofunctional outcomes or achieve a translationally relevant infarct size reduction. Thus stringent statistical thresholds, reporting negative data and a MA-pRCT approach can ensure biomedical data validity and overcome risks of bias. KW - focal cerebral ischemia KW - darbepoetin alpha KW - mice KW - translational stroke research KW - colony-stimulating factor KW - NADPH oxidase inhibitors KW - chronic kidney disease KW - diabetes mellitus KW - oxidative stress KW - search filter Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-151401 VL - 5 IS - 13428 ER -