Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 45
Filter
1.
Biomolecules ; 13(7)2023 07 04.
Article in English | MEDLINE | ID: mdl-37509110

ABSTRACT

Popliteal artery aneurysm (PAA) is the most frequent peripheral aneurysm, primarily seen in male smokers with a prevalence below 1%. This exploratory study aims to shed light on cellular mechanisms involved in PAA progression. Sixteen human PAA and eight non-aneurysmatic popliteal artery samples, partially from the same patients, were analyzed by immunohistochemistry, fluorescence imaging, Affymetrix mRNA expression profiling, qPCR and OLink proteomics, and compared to atherosclerotic (n = 6) and abdominal aortic aneurysm (AAA) tissue (n = 19). Additionally, primary cell culture of PAA-derived vascular smooth muscle cells (VSMC) was established for modulation and growth analysis. Compared to non-aneurysmatic popliteal arteries, VSMCs lose the contractile phenotype and the cell proliferation rate increases significantly in PAA. Array analysis identified APOE higher expressed in PAA samples, co-localizing with VSMCs. APOE stimulation of primary human PAA VSMCs significantly reduced cell proliferation. Accordingly, contractile VSMC markers were significantly upregulated. A single case of osseous mechanically induced PAA with a non-diseased VSMC profile emphasizes these findings. Carefully concluded, PAA pathogenesis shows similar features to AAA, yet the mechanisms involved might differ. APOE is specifically higher expressed in PAA tissue and could be involved in VSMC phenotype rescue.


Subject(s)
Aortic Aneurysm, Abdominal , Popliteal Artery Aneurysm , Humans , Male , Aortic Aneurysm, Abdominal/metabolism , Phenotype , Myocytes, Smooth Muscle/metabolism , Apolipoproteins E/genetics , Apolipoproteins E/metabolism , Apolipoproteins/metabolism
2.
Euro Surveill ; 27(43)2022 10.
Article in English | MEDLINE | ID: mdl-36305336

ABSTRACT

BackgroundTracking person-to-person SARS-CoV-2 transmission in the population is important to understand the epidemiology of community transmission and may contribute to the containment of SARS-CoV-2. Neither contact tracing nor genomic surveillance alone, however, are typically sufficient to achieve this objective.AimWe demonstrate the successful application of the integrated genomic surveillance (IGS) system of the German city of Düsseldorf for tracing SARS-CoV-2 transmission chains in the population as well as detecting and investigating travel-associated SARS-CoV-2 infection clusters.MethodsGenomic surveillance, phylogenetic analysis, and structured case interviews were integrated to elucidate two genetically defined clusters of SARS-CoV-2 isolates detected by IGS in Düsseldorf in July 2021.ResultsCluster 1 (n = 67 Düsseldorf cases) and Cluster 2 (n = 36) were detected in a surveillance dataset of 518 high-quality SARS-CoV-2 genomes from Düsseldorf (53% of total cases, sampled mid-June to July 2021). Cluster 1 could be traced back to a complex pattern of transmission in nightlife venues following a putative importation by a SARS-CoV-2-infected return traveller (IP) in late June; 28 SARS-CoV-2 cases could be epidemiologically directly linked to IP. Supported by viral genome data from Spain, Cluster 2 was shown to represent multiple independent introduction events of a viral strain circulating in Catalonia and other European countries, followed by diffuse community transmission in Düsseldorf.ConclusionIGS enabled high-resolution tracing of SARS-CoV-2 transmission in an internationally connected city during community transmission and provided infection chain-level evidence of the downstream propagation of travel-imported SARS-CoV-2 cases.


Subject(s)
COVID-19 , Communicable Diseases, Imported , Humans , SARS-CoV-2/genetics , Travel , Communicable Diseases, Imported/epidemiology , COVID-19/epidemiology , Phylogeny , Contact Tracing , Germany/epidemiology , Genomics
3.
Nutr Res ; 107: 12-25, 2022 11.
Article in English | MEDLINE | ID: mdl-36162275

ABSTRACT

In this cross-sectional study, we hypothesized that a high dietary ratio of omega-6 (n-6) to omega-3 (n-3) fatty acids could be associated with an altered gut bacterial composition and with the disease severity in patients with nonalcoholic fatty liver disease (NAFLD). A total of 101 NAFLD patients were included in the study, of which 63 underwent a liver biopsy. All 101 patients completed a 14-day food and activity record. Ebispro 2016 professional software was used to calculate individual macronutrients and micronutrients consumed. Patients were grouped into 3 quantiles (Q) according to a low (Q1: <6.1, n = 34), moderate (Q2: 6.1-7.8, n = 33), or high (Q3: >7.8, n = 34) dietary n-6/n-3 ratio. Stool samples were analyzed using 16S rRNA gene sequencing. Spearman correlation coefficients and principal coordinate analysis were used to detect differences in the bacterial composition of the gut microbiota. The median dietary n-6/n-3 ratio of all patients was 6.7 (range, 3.1-14.9). No significant associations between the dietary n-6/n-3 ratio and the gut microbiota composition or disease severity were observed. However, the abundance of specific bacteria such as Catenibacterium or Lactobacillus ruminis were found to be positively correlated and the abundance of Clostridium were negatively correlated with dietary n-6 fatty acid intake. The results indicate that a high dietary n-6/n-3 ratio is probably not a highly relevant factor in the pathogenesis of human NAFLD. Further studies are needed to clarify the importance of interactions between gut bacterial taxa and n-6 fatty acids in the pathophysiology of NAFLD.


Subject(s)
Fatty Acids, Omega-3 , Gastrointestinal Microbiome , Non-alcoholic Fatty Liver Disease , Humans , Non-alcoholic Fatty Liver Disease/etiology , Gastrointestinal Microbiome/physiology , RNA, Ribosomal, 16S/genetics , Cross-Sectional Studies , Bacteria/genetics , Severity of Illness Index
4.
J Atten Disord ; 25(4): 572-583, 2021 02.
Article in English | MEDLINE | ID: mdl-30623719

ABSTRACT

Objective: The DIRAS2 gene is associated with ADHD, but its function is largely unknown. Thus, we aimed to explore the genes and molecular pathways affected by DIRAS2. Method: Using short hairpin RNAs, we downregulated Diras2 in murine hippocampal primary cells. Gene expression was analyzed by microarray and affected pathways were identified. We used quantitative real-time polymerase chain reaction (qPCR) to confirm expression changes and analyzed enrichment of differentially expressed genes in an ADHD GWAS (genome-wide association studies) sample. Results:Diras2 knockdown altered expression of 1,612 genes, which were enriched for biological processes involved in neurodevelopment. Expression changes were confirmed for 33 out of 88 selected genes. These 33 genes showed significant enrichment in ADHD patients in a gene-set-based analysis. Conclusion: Our findings show that Diras2 affects numerous genes and thus molecular pathways that are relevant for neurodevelopmental processes. These findings may further support the hypothesis that DIRAS2 is linked to etiological processes underlying ADHD.


Subject(s)
Attention Deficit Disorder with Hyperactivity , Genome-Wide Association Study , Animals , Attention Deficit Disorder with Hyperactivity/genetics , GTP Phosphohydrolases , Gene Knockdown Techniques , Hippocampus , Humans , Mice
5.
Haematologica ; 105(9): 2316-2326, 2020 09 01.
Article in English | MEDLINE | ID: mdl-33054056

ABSTRACT

Oncogenic RAS provides crucial survival signaling for up to half of multiple myeloma cases, but has so far remained a clinically undruggable target. RAL is a member of the RAS superfamily of small GTPases and is considered to be a potential mediator of oncogenic RAS signaling. In primary multiple myeloma, we found RAL to be overexpressed in the vast majority of samples when compared with pre-malignant monoclonal gammopathy of undetermined significance or normal plasma cells. We analyzed the functional effects of RAL abrogation in myeloma cell lines and found that RAL is a critical mediator of survival. RNAi-mediated knockdown of RAL resulted in rapid induction of tumor cell death, an effect which was independent from signaling via mitogen-activated protein kinase, but appears to be partially dependent on Akt activity. Notably, RAL activation was not correlated with the presence of activating RAS mutations and remained unaffected by knockdown of oncogenic RAS. Furthermore, transcriptome analysis yielded distinct RNA expression signatures after knockdown of either RAS or RAL. Combining RAL depletion with clinically relevant anti-myeloma agents led to enhanced rates of cell death. Our data demonstrate that RAL promotes multiple myeloma cell survival independently of oncogenic RAS and, thus, this pathway represents a potential therapeutic target in its own right.


Subject(s)
GTP Phosphohydrolases , Multiple Myeloma , Cell Survival/genetics , Genes, ras , Humans , Multiple Myeloma/genetics , ral GTP-Binding Proteins/genetics , ral GTP-Binding Proteins/metabolism
6.
J Gastroenterol Hepatol ; 35(11): 1969-1977, 2020 Nov.
Article in English | MEDLINE | ID: mdl-32267559

ABSTRACT

BACKGROUND AND AIM: Several studies observed alterations in the gut microbiota in patients with non-alcoholic fatty liver disease (NAFLD). However, analyzed patient populations and methods strongly differ among these studies. The aim of this study was to prove the reproducibility of published results and to provide a detailed overview of all findings in our NAFLD cohort using next generation sequencing methods. METHODS: The individual taxonomic microbiota composition of fecal samples from 90 NAFLD patients and 21 healthy controls was analyzed using 16S rRNA gene sequencing. Study participants were grouped according to their disease stage and compared regarding their gut microbiota composition. Studies were identified from PubMed listed publications, and the results were compared with the findings in our cohort. RESULTS: Results from 13 identified studies were compared with our data. A decreased abundance of the Bacteroidetes and Ruminococcaceae as well as an increased abundance of Lactobacillaceae and Veillonellaceae and Dorea were the most frequently reported changes among NAFLD patients in 4/13, 5/13, 4/13, 2/13, and 3/13 studies, respectively. Even though these alterations in the gut microbiota composition were also observed in our patient cohort, the majority of published differences could not be reproduced, neither in our own nor in other NAFLD cohort studies. CONCLUSION: Despite repeatedly reproduced abundance patterns of specific bacteria, the heterogeneous study results did not reveal a consistent disease specific gut microbiota signature. Further prospective studies with homogenous patient cohorts and standardized methods are necessary to phenotype NAFLD by the gut microbiota.


Subject(s)
Gastrointestinal Microbiome , Non-alcoholic Fatty Liver Disease/genetics , Non-alcoholic Fatty Liver Disease/microbiology , Phenotype , Adult , Bacteroidetes , Cross-Sectional Studies , Female , Gastrointestinal Microbiome/genetics , High-Throughput Nucleotide Sequencing/methods , Humans , Lactobacillaceae , Male , Prospective Studies , RNA, Ribosomal, 16S , Ruminococcus , Veillonella , Young Adult
7.
World J Biol Psychiatry ; 20(6): 462-475, 2019 07.
Article in English | MEDLINE | ID: mdl-29067888

ABSTRACT

Objectives: The aim of our study was to investigate molecular mechanisms of lithium action by studying the gene expression profile of peripheral cell models generated from bipolar patients (BD) and healthy controls (HC). Methods: EBV-immortalised lymphoblastoid cells (LCLs) and fibroblast cells from BD and HC were incubated with either lithium chloride or plain medium for 3 weeks. We first conducted a microarray gene expression study. The most promising differentially regulated genes in terms of lithium-associated or disorder-associated pathways were then replicated by quantitative real-time PCR (qRT-PCR). Results: The pooled microarray analysis showed 459 genes to be differentially regulated in BD compared to HC and 58 due to lithium treatment in LCLs, and 295 genes to be differentially regulated in BD compared to HC and five due to lithium treatment in fibroblasts. After correction for multiple comparison, EPHB1 disorder × treatment interactions remained significant in LCLs validated by qRT-PCR. In the control group, lithium influenced the expression of ANP32E, PLEKHA2, KCNK1, PRKCH, ST3GAL6 and AIF1. In bipolar and control fibroblast cells lithium treatment decreased FGF9 expression. Conclusions: The differentially regulated genes in our study add evidence for the relevance of inflammation, neuronal/glial development, phosphatidylinositol second-messenger pathway and ion channels in the mode of action of lithium.


Subject(s)
Antimanic Agents/pharmacology , Bipolar Disorder/drug therapy , Bipolar Disorder/pathology , Gene Expression/drug effects , Lithium Compounds/pharmacology , Cell Line , Cells, Cultured , Fibroblasts/drug effects , Humans , Lymphocytes/drug effects , Microarray Analysis , RNA, Messenger/metabolism
8.
Hypertension ; 73(1): 102-111, 2019 01.
Article in English | MEDLINE | ID: mdl-30543467

ABSTRACT

Myocardial infarction (MI) is a major cause of death worldwide. Here, we identify the macrophage MR (mineralocorticoid receptor) as a crucial pathogenic player in cardiac wound repair after MI. Seven days after left coronary artery ligation, mice with myeloid cell-restricted MR deficiency compared with WT (wild type) controls displayed improved cardiac function and remodeling associated with enhanced infarct neovascularization and scar maturation. Gene expression profiling of heart-resident and infarct macrophages revealed that MR deletion drives macrophage differentiation in the ischemic microenvironment toward a phenotype outside the M1/M2 paradigm, with regulation of multiple interrelated factors controlling wound healing and tissue repair. Mechanistic and functional data suggest that inactivation of the macrophage MR promotes myocardial infarct healing through enhanced efferocytosis of neutrophils, the suppression of free radical formation, and the modulation of fibroblast activation state. Crucially, targeted delivery of MR antagonists to macrophages, with a single administration of RU28318 or eplerenone-containing liposomes at the onset of MI, improved the healing response and protected against cardiac remodeling and functional deterioration, offering an effective and unique therapeutic strategy for cardiac repair.


Subject(s)
Eplerenone/pharmacology , Heart/physiopathology , Myocardial Infarction , Myocardium , Receptors, Mineralocorticoid , Wound Healing , Animals , Cell Differentiation/drug effects , Cellular Microenvironment/drug effects , Disease Models, Animal , Gene Expression Profiling/methods , Liposomes , Macrophages/drug effects , Macrophages/metabolism , Mice , Mineralocorticoid Receptor Antagonists/pharmacology , Myocardial Infarction/drug therapy , Myocardial Infarction/metabolism , Myocardial Infarction/pathology , Myocardial Infarction/physiopathology , Myocardium/metabolism , Myocardium/pathology , Receptors, Mineralocorticoid/deficiency , Receptors, Mineralocorticoid/metabolism , Ventricular Remodeling/drug effects , Ventricular Remodeling/physiology , Wound Healing/drug effects , Wound Healing/physiology
9.
J Virol ; 92(17)2018 09 01.
Article in English | MEDLINE | ID: mdl-29925665

ABSTRACT

We found earlier that ectopic expression of the cytidine deaminase APOBEC3G (A3G) in Vero cells inhibits measles virus (MV), respiratory syncytial virus, and mumps virus, while the mechanism of inhibition remained unclear. A microarray analysis revealed that in A3G-transduced Vero cells, several cellular transcripts were differentially expressed, suggesting that A3G regulates the expression of host factors. One of the most upregulated host cell factors, REDD1 (regulated in development and DNA damage response-1, also called DDIT4), reduced MV replication ∼10-fold upon overexpression in Vero cells. REDD1 is an endogenous inhibitor of mTORC1 (mammalian target of rapamycin complex-1), the central regulator of cellular metabolism. Interestingly, rapamycin reduced the MV replication similarly to REDD1 overexpression, while the combination of both did not lead to further inhibition, suggesting that the same pathway is affected. REDD1 silencing in A3G-expressing Vero cells abolished the inhibitory effect of A3G. In addition, silencing of A3G led to reduced REDD1 expression, confirming that its expression is regulated by A3G. In primary human peripheral blood lymphocytes (PBL), expression of A3G and REDD1 was found to be stimulated by phytohemagglutinin (PHA) and interleukin-2. Small interfering RNA (siRNA)-mediated depletion of A3G in PHA-stimulated PBL reduced REDD1 expression and increased viral titers, which corroborates our findings in Vero cells. Silencing of REDD1 also increased viral titers, confirming the antiviral role of REDD1. Finally, pharmacological inhibition of mTORC1 by rapamycin in PHA-stimulated PBL reduced viral replication to the level found in unstimulated lymphocytes, indicating that mTORC1 activity supports MV replication as a proviral host factor.IMPORTANCE Knowledge about host factors supporting or restricting virus replication is required for a deeper understanding of virus-cell interactions and may eventually provide the basis for therapeutic intervention. This work was undertaken predominantly to explain the mechanism of A3G-mediated inhibition of MV, a negative-strand RNA virus that is not affected by the deaminase activity of A3G acting on single-stranded DNA. We found that A3G regulates the expression of several cellular proteins, which influences the capacity of the host cell to replicate MV. One of these, REDD1, which modulates the cellular metabolism in a central position by regulating the kinase complex mTORC1, was identified as the major cellular factor impairing MV replication. These findings show interesting aspects of the function of A3G and the dependence of the MV replication on the metabolic state of the cell. Interestingly, pharmacological inhibition of mTORC1 can be utilized to inhibit MV replication in Vero cells and primary human peripheral blood lymphocytes.


Subject(s)
APOBEC-3G Deaminase/genetics , Measles virus/physiology , Mechanistic Target of Rapamycin Complex 1/antagonists & inhibitors , Transcription Factors/antagonists & inhibitors , Virus Replication/genetics , APOBEC-3G Deaminase/metabolism , Animals , Antiviral Agents/pharmacology , Cell Line , Chlorocebus aethiops , DNA Replication , Host-Pathogen Interactions/genetics , Humans , Interleukin-2/pharmacology , Leukocytes, Mononuclear/drug effects , Leukocytes, Mononuclear/virology , Lymphocytes/drug effects , Lymphocytes/virology , Measles virus/drug effects , Mechanistic Target of Rapamycin Complex 1/drug effects , Phytohemagglutinins/pharmacology , RNA, Small Interfering , Sirolimus/pharmacology , Transcription Factors/deficiency , Transcription Factors/drug effects , Vero Cells , Virus Replication/drug effects
10.
J Neural Transm (Vienna) ; 125(1): 77-87, 2018 01.
Article in English | MEDLINE | ID: mdl-29027019

ABSTRACT

Alzheimer's disease (AD) is a neurodegenerative disorder that preferentially affects individuals of advanced age. Heritability estimates for AD range between 60 and 80%, but only few genetic risk factors have been identified so far. In the present explorative study, we aimed at characterizing the genetic contribution to late-onset AD in participants of the Vienna Transdanube Aging (VITA) longitudinal birth cohort study in a two-step approach. First, we performed a genome-wide screen of pooled DNA samples (n = 588) to identify allele frequency differences between AD patients and non-AD individuals using life-time diagnoses made at the age of 80 (t = 60 months). This analysis suggested a high proportion of brain-expressed genes required for cell adhesion, cell signaling and cell morphogenesis, and also scored in known AD risk genes. In a second step, we confirmed associations using individual genotypes of top-ranked markers examining AD diagnoses as well as the dimensional scores: FULD and MMSE determined up to the age of 82.5 (t = 90 months). Taken together, our study proposes genes ANKS1B, ENST00000414107, LOC100505811, SLC22A14, QRFPR, ZDHHC8P1, ADAMTS3 and PPFIA1 as possible new candidates involved in the etiology of late-onset AD, with further research being needed to clarify their exact roles.


Subject(s)
Aging/genetics , Alzheimer Disease/diagnosis , Alzheimer Disease/genetics , Genetic Loci/genetics , Genetic Predisposition to Disease/genetics , Genetic Testing/methods , Aged , Aged, 80 and over , Alzheimer Disease/epidemiology , Austria/epidemiology , Cohort Studies , Female , Genetic Predisposition to Disease/epidemiology , Humans , Longitudinal Studies , Male
11.
Am J Med Genet B Neuropsychiatr Genet ; 174(7): 740-750, 2017 Oct.
Article in English | MEDLINE | ID: mdl-28777493

ABSTRACT

In a previous study, we identified the single nucleotide polymorphism (SNP) rs4500567, located in the upstream region of tetraspanin 8 (TSPAN8), to be associated with bipolar disorder (BD). Due to its proximal position, the SNP might have an impact on promoter activity, thus on TSPAN8 gene expression. We investigated the impact of rs4500567 on TSPAN8 expression in vitro with luciferase-based promoter assays in human embryonic kidney (HEK293) and neuroblastoma cells (SH-SY5Y), and its effect on expression of downstream associated genes by microarray-based transcriptome analyses. Immunohistochemical localization studies on murine brain slices served to identify possible target regions of altered TSPAN8 expression in the brain. Promoter assays revealed decreased TSPAN8 expression in presence of the minor allele. Transcriptome analyses of TSPAN8-knockdown cells, mirroring the effects of putatively reduced TSPAN8 expression in minor allele carriers, resulted in 231 differentially expressed genes with enrichments of relevant signaling pathways for psychiatric disorders and neuronal development. Finally, we demonstrate Tspan8 abundance in mouse cerebellum and hippocampus. These findings point to a role of TSPAN8 in neuronal function or development. Considering a rather protective effect of the minor allele of rs4500567, our findings reveal a possible novel mechanism that contributes to the development of BD.


Subject(s)
Bipolar Disorder/pathology , Brain/pathology , Gene Expression Regulation , Neuroblastoma/pathology , Polymorphism, Single Nucleotide , Tetraspanins/metabolism , Animals , Basic-Leucine Zipper Transcription Factors/genetics , Basic-Leucine Zipper Transcription Factors/metabolism , Bipolar Disorder/genetics , Bipolar Disorder/metabolism , Brain/metabolism , Female , Gene Expression Profiling , HEK293 Cells , Humans , Luciferases/metabolism , Mice , Mice, Inbred C57BL , Neuroblastoma/genetics , Neuroblastoma/metabolism , Promoter Regions, Genetic , Signal Transduction , Tetraspanins/genetics , Tumor Cells, Cultured
12.
FASEB J ; 31(11): 5122-5132, 2017 11.
Article in English | MEDLINE | ID: mdl-28768721

ABSTRACT

Cell- and tissue-specific actions of glucocorticoids are mediated by the glucocorticoid receptor. Here, we demonstrate that the glucocorticoid receptor (GR) in macrophages is essential for cardiac healing after myocardial infarction. Compared with GRflox (wild-type controls), GRLysMCre mice that lacked GR in myeloid cells showed increased acute mortality as a result of cardiac rupture. Seven days after left coronary artery ligation, GRLysMCre mice exhibited worse cardiac function and adverse remodeling associated with impaired scar formation and angiogenic response to ischemic injury. Inactivation of GR altered the functional differentiation/maturation of monocyte-derived macrophages in the infarcted myocardium. Mechanistically, CD45+/CD11b+/Ly6G-/F4/80+ macrophages isolated from GRLysMCre infarcts showed deregulation of factors that control inflammation, neovascularization, collagen degradation, and scar tissue formation. Moreover, we demonstrate that cardiac fibroblasts sorted from the ischemic myocardium of GRLysMCre mice compared with cells isolated from injured GRflox hearts displayed higher matrix metalloproteinase 2 expression, and we provide evidence that the macrophage GR regulates myofibroblast differentiation in the infarct microenvironment during the early phase of wound healing. In summary, GR signaling in macrophages, playing a crucial role in tissue-repairing mechanisms, could be a potential therapeutic target during wound healing after ischemic myocardial injury.-Galuppo, P., Vettorazzi, S., Hövelmann, J., Scholz, C.-J., Tuckermann, J. P., Bauersachs, J., Fraccarollo, D. The glucocorticoid receptor in monocyte-derived macrophages is critical for cardiac infarct repair and remodeling.


Subject(s)
Macrophages/metabolism , Monocytes/metabolism , Myocardial Infarction/metabolism , Myocardium/metabolism , Receptors, Glucocorticoid/metabolism , Signal Transduction , Animals , Antigens, Differentiation/genetics , Antigens, Differentiation/metabolism , Macrophages/pathology , Mice , Mice, Transgenic , Monocytes/pathology , Myocardial Infarction/genetics , Myocardial Infarction/pathology , Myocardium/pathology , Myofibroblasts/metabolism , Myofibroblasts/pathology , Receptors, Glucocorticoid/genetics
13.
Acta Neuropathol ; 134(2): 281-295, 2017 08.
Article in English | MEDLINE | ID: mdl-28620692

ABSTRACT

Multiple sclerosis (MS) is a chronic autoimmune disease of the central nervous system (CNS) in young adults that has serious negative socioeconomic effects. In addition to symptoms caused by CNS pathology, the majority of MS patients frequently exhibit gastrointestinal dysfunction, which was previously either explained by the presence of spinal cord lesions or not directly linked to the autoimmune etiology of the disease. Here, we studied the enteric nervous system (ENS) in a B cell- and antibody-dependent mouse model of MS by immunohistochemistry and electron microscopy at different stages of the disease. ENS degeneration was evident prior to the development of CNS lesions and the onset of neurological deficits in mice. The pathology was antibody mediated and caused a significant decrease in gastrointestinal motility, which was associated with ENS gliosis and neuronal loss. We identified autoantibodies against four potential target antigens derived from enteric glia and/or neurons by immunoprecipitation and mass spectrometry. Antibodies against three of the target antigens were also present in the plasma of MS patients as confirmed by ELISA. The analysis of human colon resectates provided evidence of gliosis and ENS degeneration in MS patients compared to non-MS controls. For the first time, this study establishes a pathomechanistic link between the well-established autoimmune attack on the CNS and ENS pathology in MS, which might provide a paradigm shift in our current understanding of the immunopathogenesis of the disease with broad diagnostic and therapeutic implications.


Subject(s)
Autoantibodies/blood , Gastrointestinal Diseases/etiology , Multiple Sclerosis , Animals , Central Nervous System/metabolism , Central Nervous System/pathology , Cytokines/metabolism , Disease Models, Animal , Enteric Nervous System/metabolism , Enteric Nervous System/pathology , Enteric Nervous System/ultrastructure , Female , Freund's Adjuvant/toxicity , Humans , Male , Mice , Mice, Inbred C57BL , Multiple Sclerosis/complications , Multiple Sclerosis/immunology , Multiple Sclerosis/pathology , Muscle, Smooth/pathology , Muscle, Smooth/ultrastructure , Myelin Basic Protein/immunology , Myelin Basic Protein/metabolism , Myelin Basic Protein/toxicity , Myelin-Oligodendrocyte Glycoprotein/immunology , Myelin-Oligodendrocyte Glycoprotein/toxicity , Myenteric Plexus/pathology , Myenteric Plexus/ultrastructure , Recombinant Fusion Proteins/immunology , Recombinant Fusion Proteins/toxicity , Tubulin/metabolism
14.
Proc Natl Acad Sci U S A ; 114(16): E3295-E3304, 2017 04 18.
Article in English | MEDLINE | ID: mdl-28381594

ABSTRACT

Despite continuous interest in multiple sclerosis (MS) research, there is still a lack of neuroprotective strategies, because the main focus has remained on modulating the immune response. Here we performed in-depth analysis of neurodegeneration in experimental autoimmune encephalomyelitis (EAE) and in in vitro studies regarding the effect of the well-established L-type calcium channel antagonist nimodipine. Nimodipine treatment attenuated clinical EAE and spinal cord degeneration and promoted remyelination. Surprisingly, we observed calcium channel-independent effects on microglia, resulting in apoptosis. These effects were cell-type specific and irrespective of microglia polarization. Apoptosis was accompanied by decreased levels of nitric oxide (NO) and inducible NO synthase (iNOS) in cell culture as well as decreased iNOS and reactive oxygen species levels in EAE. In addition, increased numbers of Olig2+APC+ oligodendrocytes were detected. Overall, nimodipine application seems to generate a favorable environment for regenerative processes and therefore could be a treatment option for MS, because it combines features of immunomodulation with beneficial effects on neuroregeneration.


Subject(s)
Apoptosis/drug effects , Disease Models, Animal , Encephalomyelitis, Autoimmune, Experimental/pathology , Microglia/pathology , Multiple Sclerosis/pathology , Nimodipine/pharmacology , Remyelination/physiology , Animals , Calcium Channels, L-Type/chemistry , Cells, Cultured , Encephalomyelitis, Autoimmune, Experimental/drug therapy , Encephalomyelitis, Autoimmune, Experimental/metabolism , Mice , Microglia/drug effects , Microglia/metabolism , Multiple Sclerosis/drug therapy , Multiple Sclerosis/metabolism , Nitric Oxide Synthase Type II/metabolism , Oligodendroglia/drug effects , Oligodendroglia/metabolism , Oligodendroglia/pathology , Reactive Oxygen Species/metabolism , Remyelination/drug effects , Spinal Cord/drug effects , Spinal Cord/metabolism , Spinal Cord/pathology
15.
Biol Lett ; 12(12)2016 12.
Article in English | MEDLINE | ID: mdl-28003518

ABSTRACT

A painful event establishes two opponent memories: cues that are associated with pain onset are remembered negatively, whereas cues that coincide with the relief at pain offset acquire positive valence. Such punishment- versus relief-memories are conserved across species, including humans, and the balance between them is critical for adaptive behaviour with respect to pain and trauma. In the fruit fly, Drosophila melanogaster as a study case, we found that both punishment- and relief-memories display natural variation across wild-derived inbred strains, but they do not covary, suggesting a considerable level of dissociation in their genetic effectors. This provokes the question whether there may be heritable inter-individual differences in the balance between these opponent memories in man, with potential psycho-clinical implications.


Subject(s)
Drosophila melanogaster/genetics , Animals , Association Learning , Conditioning, Psychological/physiology , Drosophila melanogaster/physiology , Electroshock , Genetic Variation , Memory , Odorants , Punishment , Reward , Smell
16.
Oncotarget ; 7(22): 32956-68, 2016 May 31.
Article in English | MEDLINE | ID: mdl-27121059

ABSTRACT

The pocket protein (PP) family consists of the three members RB1, p107 and p130 all possessing tumor suppressive properties. Indeed, the PPs jointly control the G1/S transition mainly by inhibiting E2F transcription factors. Notably, several viral oncoproteins are capable of binding and inhibiting PPs. Merkel cell polyomavirus (MCPyV) is considered as etiological factor for Merkel cell carcinoma (MCC) with expression of the viral Large T antigen (LT) harboring an intact PP binding domain being required for proliferation of most MCC cells. Therefore, we analyzed the interaction of MCPyV-LT with the PPs. Co-IP experiments indicate that MCPyV-LT binds potently only to RB1. Moreover, MCPyV-LT knockdown-induced growth arrest in MCC cells can be rescued by knockdown of RB1, but not by p107 or p130 knockdown. Accordingly, cell cycle arrest and E2F target gene repression mediated by the single PPs can only in the case of RB1 be significantly reverted by MCPyV-LT expression. Moreover, data from an MCC patient indicate that loss of RB1 rendered the MCPyV-positive MCC cells LT independent. Thus, our results suggest that RB1 is the dominant tumor suppressor PP in MCC, and that inactivation of RB1 by MCPyV-LT is largely sufficient for its growth supporting function in established MCPyV-positive MCC cells.


Subject(s)
Antigens, Viral, Tumor/metabolism , Carcinoma, Merkel Cell/metabolism , Retinoblastoma Binding Proteins/metabolism , Skin Neoplasms/metabolism , Ubiquitin-Protein Ligases/metabolism , Antigens, Viral, Tumor/genetics , Carcinoma, Merkel Cell/genetics , Carcinoma, Merkel Cell/pathology , Carcinoma, Merkel Cell/virology , Humans , Merkel cell polyomavirus/metabolism , Polyomavirus Infections/genetics , Polyomavirus Infections/metabolism , Retinoblastoma Binding Proteins/genetics , Skin Neoplasms/genetics , Skin Neoplasms/pathology , Skin Neoplasms/virology , Transfection , Ubiquitin-Protein Ligases/genetics
17.
PLoS One ; 11(3): e0151335, 2016.
Article in English | MEDLINE | ID: mdl-26964064

ABSTRACT

BACKGROUND: Enteric glial cells (EGCs) are the main constituent of the enteric nervous system and share similarities with astrocytes from the central nervous system including their reactivity to an inflammatory microenvironment. Previous studies on EGC pathophysiology have specifically focused on mucosal glia activation and its contribution to mucosal inflammatory processes observed in the gut of inflammatory bowel disease (IBD) patients. In contrast knowledge is scarce on intestinal inflammation not locally restricted to the mucosa but systemically affecting the intestine and its effect on the overall EGC network. METHODS AND RESULTS: In this study, we analyzed the biological effects of a systemic LPS-induced hyperinflammatory insult on overall EGCs in a rat model in vivo, mimicking the clinical situation of systemic inflammation response syndrome (SIRS). Tissues from small and large intestine were removed 4 hours after systemic LPS-injection and analyzed on transcript and protein level. Laser capture microdissection was performed to study plexus-specific gene expression alterations. Upon systemic LPS-injection in vivo we observed a rapid and dramatic activation of Glial Fibrillary Acidic Protein (GFAP)-expressing glia on mRNA level, locally restricted to the myenteric plexus. To study the specific role of the GFAP subpopulation, we established flow cytometry-purified primary glial cell cultures from GFAP promotor-driven EGFP reporter mice. After LPS stimulation, we analyzed cytokine secretion and global gene expression profiles, which were finally implemented in a bioinformatic comparative transcriptome analysis. Enriched GFAP+ glial cells cultured as gliospheres secreted increased levels of prominent inflammatory cytokines upon LPS stimulation. Additionally, a shift in myenteric glial gene expression profile was induced that predominantly affected genes associated with immune response. CONCLUSION AND SIGNIFICANCE: Our findings identify the myenteric GFAP-expressing glial subpopulation as particularly susceptible and responsive to acute systemic inflammation of the gut wall and complement knowledge on glial involvement in mucosal inflammation of the intestine.


Subject(s)
Myenteric Plexus/pathology , Neuroglia/pathology , Acute Disease , Animals , Gene Expression Profiling , Glial Fibrillary Acidic Protein/metabolism , Humans , Inflammation/pathology , Lipopolysaccharides/pharmacology , Male , Mice , Neuroglia/drug effects , Neuroglia/metabolism , Rats , Sepsis/pathology
18.
Int J Mol Sci ; 17(1)2016 Jan 12.
Article in English | MEDLINE | ID: mdl-26771601

ABSTRACT

Limited comprehension of aneurysm pathology has led to inconclusive results from clinical trials. miRNAs are key regulators of post-translational gene modification and are useful tools in elucidating key features of aneurysm pathogenesis in distinct entities of abdominal and popliteal aneurysms. Here, surgically harvested specimens from 19 abdominal aortic aneurysm (AAA) and 8 popliteal artery aneurysm (PAA) patients were analyzed for miRNA expression and histologically classified regarding extracellular matrix (ECM) remodeling and inflammation. DIANA-based computational target prediction and pathway enrichment analysis verified our results, as well as previous ones. miRNA-362, -19b-1, -194, -769, -21 and -550 were significantly down-regulated in AAA samples depending on degree of inflammation. Similar or inverse regulation was found for miR-769, 19b-1 and miR-550, -21, whereas miR-194 and -362 were unaltered in PAA. In situ hybridization verified higher expression of miR-550 and -21 in PAA compared to AAA and computational analysis for target genes and pathway enrichment affirmed signal transduction, cell-cell-interaction and cell degradation pathways, in line with previous results. Despite the vague role of miRNAs for potential diagnostic and treatment purposes, the number of candidates from tissue signature studies is increasing. Tissue morphology influences subsequent research, yet comparison of distinct entities of aneurysm disease can unravel core pathways.


Subject(s)
Aorta, Abdominal/metabolism , Aortic Aneurysm, Abdominal/genetics , MicroRNAs/genetics , Popliteal Artery/metabolism , Aorta, Abdominal/pathology , Aorta, Abdominal/surgery , Aortic Aneurysm, Abdominal/metabolism , Aortic Aneurysm, Abdominal/pathology , Aortic Aneurysm, Abdominal/surgery , Extracellular Matrix/genetics , Extracellular Matrix/metabolism , Gene Expression Regulation , Humans , In Situ Hybridization , Inflammation , MicroRNAs/metabolism , Organ Specificity , Popliteal Artery/pathology , Popliteal Artery/surgery , Signal Transduction , Transcriptome
19.
PLoS One ; 10(5): e0126986, 2015.
Article in English | MEDLINE | ID: mdl-25992709

ABSTRACT

Electric shock is a common stimulus for nociception-research and the most widely used reinforcement in aversive associative learning experiments. Yet, nothing is known about the mechanisms it recruits at the periphery. To help fill this gap, we undertook a genome-wide association analysis using 38 inbred Drosophila melanogaster strains, which avoided shock to varying extents. We identified 514 genes whose expression levels and/ or sequences co-varied with shock avoidance scores. We independently scrutinized 14 of these genes using mutants, validating the effect of 7 of them on shock avoidance. This emphasizes the value of our candidate gene list as a guide for follow-up research. In addition, by integrating our association results with external protein-protein interaction data we obtained a shock avoidance-associated network of 38 genes. Both this network and the original candidate list contained a substantial number of genes that affect mechanosensory bristles, which are hair-like organs distributed across the fly's body. These results may point to a potential role for mechanosensory bristles in shock sensation. Thus, we not only provide a first list of candidate genes for shock avoidance, but also point to an interesting new hypothesis on nociceptive mechanisms.


Subject(s)
Avoidance Learning , Drosophila melanogaster/physiology , Electroshock , Genetic Association Studies , Genome-Wide Association Study , Animals , DNA Transposable Elements , Gene Deletion , Gene Expression Profiling , Gene Regulatory Networks , Locomotion , Mutagenesis, Insertional , Reproducibility of Results
20.
Cell Signal ; 27(9): 1781-8, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26004136

ABSTRACT

Colon cancer is one of the most common tumors in the human population. Recent studies have shown a reduced risk for colon cancer in patients given the antidepressant fluoxetine (FLX). The exact mechanism by which FLX might protect from colon cancer remains however controversial. Here, FLX reduced the development of different colon tumor xenografts, as well as proliferation in hypoxic tumor areas within them. FLX treatment also decreased microvessel numbers in tumors. Although FLX did not increase serum and tumor glucose levels as much as the colon chemotherapy gold standard Fluorouracil did, lactate levels were significantly augmented within tumors by FLX treatment. The gene expression of the MCT4 lactate transporter was significantly downregulated. Total protein amounts from the third and fifth mitochondrial complexes were significantly decreased by FLX in tumors. Cell culture experiments revealed that FLX reduced the mitochondrial membrane potential significantly and disabled the reactive oxygen species production of the third mitochondrial complex. Furthermore, FLX arrested hypoxic colon tumor cells in the G0/G1 phase of the cell-cycle. The expression of key cell-cycle-related checkpoint proteins was enhanced in cell culture and in vivo experiments. Therefore, we suggest FLX impairs energy generation, cell cycle progression and proliferation in tumor cells, especially under condition of hypoxia. This then leads to reduced microvessel formation and tumor shrinkage in xenograft models.


Subject(s)
Colonic Neoplasms/drug therapy , Fluoxetine/pharmacology , Neoplasms, Experimental/drug therapy , Animals , Caco-2 Cells , Cell Hypoxia/drug effects , Colonic Neoplasms/metabolism , Colonic Neoplasms/pathology , G1 Phase/drug effects , Humans , Mice , Mice, Inbred NOD , Mice, SCID , Neoplasms, Experimental/metabolism , Neoplasms, Experimental/pathology , Resting Phase, Cell Cycle/drug effects , Xenograft Model Antitumor Assays
SELECTION OF CITATIONS
SEARCH DETAIL
...