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NOD2-and disease-specific gene expression profiles of peripheral blood mononuclear cells from Crohn's disease patients

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World Journal of Gastroenterology 2018年第11期24卷 1196-1205页

作者： Holger Schufler maria rohde Sarah rohde Astrid Huth Nicole Gittel Hannes Hollborn Dirk Koczan Ane Glass Georg Lamprecht Robert Jaster Department of Medicine II Division of GastroenterologyRostock University Medical CenterRostock 18057Germany Institute of Immunology Rostock University Medical CenterRostock 18057Germany Institute for Biostatistics and Informatics in Medicine and Ageing Research Rostock 18057Germany

AIM To investigate disease-specific gene expression profiles of peripheral blood mononuclear cells(PBMCs) from Crohn's disease(CD) patients in clinical *** Patients with CD in clinical remission or with very low disease activity according to the Crohn's disease activity index were genotyped regarding nucleotidebinding oligomerization domain 2(NOD2),and PBMCs from wild-type(WT)-NOD2 patients,patients with homozygous or heterozygous NOD2 mutations and healthy donors were isolated for further *** cells were cultured with vitamin D,peptidoglycan(PGN) and lipopolysaccharide(LPS) for defined periods of time before RNA was isolated and subjected to microarray analysis using Clariom S assays and quantitative realtime ***2-and disease-specific gene expression profiles were evaluated with repeated measure ANOVA by a general linear *** Employing microarray assays,a total of 267 genes were identified that were significantly up-or downregulated in PBMCs of WT-NOD2 patients,compared to healthy donors after challenge with vitamin D and/or a combination of LPS and PGN(P < 0.05;threshold:≥ 2-fold change).For further analysis by real-time PCR,genes with known impact on inflammation and immunity were selected that fulfilled predefined expression *** a larger cohort of patients and controls,a disease-associated expression pattern,with higher transcript levels in vitamin D-treated PBMCs from patients,was observed for three of these genes,CLEC5 A(P < 0.030),lysozyme(LYZ;P < 0.047) and TREM1(P < 0.023).Six genes were found to be expressed in a NOD2-dependent manner(CD101,P < 0.002;CLEC5 A,P < 0.020;CXCL5,P < 0.009;IL-24,P < 0.044;ITGB2,P < 0.041;LYZ,P < 0.042).Interestingly,the highest transcript levels were observed in patients with heterozygous NOD2 *** Our data identify CLEC5 A and LYZ as CD-and NOD2-associated genes of PBMCs and encourage further studies on their pathomechanistic roles.

关键词： Peripheral blood mononuclear cells Gene expression NOD2 Lysozyme Crohn's disease CLEC5A

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Absent MicroRNAs in Different Tissues of Patients with Acquired Cardiomyopathy

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Genomics, Proteomics & Bioinformatics 2016年第4期14卷 224-234页

作者： Christine S. Siegismund maria rohde Uwe Kiihl Felicitas Escher Heinz Peter Schultheiss Dirk Lassner Institute for Cardiac Diagnostics and Therapy(IKDT) 12203 BerlinGermany Department of Cardiology Campus VirchowCharite-University Hospital Berlin13353 BerlinGermany

MicroRNAs （miRNAs） can be found in a wide range of tissues and body fluids, and their specific signatures can be used to determine diseases or predict clinical courses. The miRNA profiles in biological samples （tissue, serum, peripheral blood mononuelear cells or other body fluids） differ significantly even in the same patient and therefore have their own specificity for the presented condition. Complex profiles of deregulated miRNAs are of high interest, whereas the importance of non-expressed miRNAs was ignored. Since miRNAs regulate gene expression rather negatively, absent miRNAs could indicate genes with unaltered expression that therefore are normally expressed in specific compartments or under specific disease situations. For the first time, non-detectable miRNAs in different tissues and body fluids from patients with different diseases （cardiomyopathies, Alzheimer＇s disease, bladder cancer, and ocular cancer） were analyzed and compared in this study, miRNA expression data were generated by microarray or TaqMan PCR-based platforms. Lists of absent miRNAs of primarily cardiac patients （myocardium, blood cells, and serum） were clustered and analyzed for potentially involved pathways using two prediction platforms, i.e., miRNA enrichment analysis and annotation tool （miEAA） and DIANA miRPath. Extensive search in biomedical publication databases for the relevance of non-expressed miRNAs in predicted pathways revealed no evidence for their involvement in heart-related pathways as indicated by software tools, confirming proposed approach.

关键词： Cardiomyopathy Heart muscle biopsy Absent mi RNAs Peripheral blood mononuclear cell Serum

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Myocarditis—Personalized Medicine by Expanded Endomyocardial Biopsy Diagnostics

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World Journal of Cardiovascular Diseases 2014年第6期4卷 325-340页

作者： Dirk Lassner maria rohde Christine Sabine Siegismund Uwe Kuhl Ulrich MichaelGross Felicitas Escher Carsten Tschope Heinz-Peter Schultheiss Institute of Cardiac Diagnostics and Therapy BerlinGermany Department of Cardiology and Pneumonology Charite—University Medicine BerlinCampus Benjamin FranklinBerlinGermany

Myocarditis and dilated cardiomyopathy (DCM) are acute or chronic disorders of myocardium. The gold standard for final confirmation of causative reasons of these heart muscle diseases is the endomyocardial biopsy (EMB) analysis. Due to focal pathology, diagnostics are failing if the EMB does not contain the area of interest. Personalized medicine comprises the genetic information together with the phenotypic and environmental factors to yield a tailored healthcare for each individual and removes the limitations of the “one-size-fits-all” therapy approach. This provides the opportunity to translate therapies from bench to bedside, to diagnose and predict disease, and to improve patient-tailored treatments based on the unique signatures of a patient’s disease. Furthermore, novel treatment schedules can be identified which have eventually the chance to enhance long-term survivals. Global biomarkers such as specific gene expression signatures or miRNA profiles not only have the potential to reduce this problem but also add valuable information for individualized therapy decisions. In future, multiplex approaches allowing rapid and absolutely reliable identification of inflammatory or virally-induced myocardial diseases will replace singleplex methods such as direct detection of viral genomes in one single biopsy. Gene or miRNA profiles are upcoming diagnostic biomarkers for cardiomyopathies which are not only detectable in tissue samples but in body fluids as well. Consequently, a systemic diagnostic approach by determination of distinct expression pattern in e.g., peripheral blood samples will support the characterization of distinct cardiomyopathies by means of non-invasive methods.

关键词： Cardiomyopathy Endomyocardial Biopsy Personalized Medicine Biomarker MicroRNAs

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Recent Advances in Molecular Diagnostics and Treatment of Heart Muscle Diseases--From Biopsy-Focused to Systemic Diagnostics

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Journal of Analytical Sciences, Methods and Instrumentation 2013年第2期3卷 98-109页

作者： Dirk Lassner Christine Sabine Siegismund Jenny Stehr maria rohde Felicitas Escher Carsten Tschope Ulrich Michael Gross Uwe Kühl Heinz-Peter Schultheiss Institute of Cardiac Diagnostics and Therapy BerlinGermany Department of Cardiology and Pneumonology Charité-Universityhospital BerlinCampus Benjamin FranklinBerlinGermany

Cardiomyopathies are acute or chronic disorders of myocardium. Diagnostic characterization of disease entities demands endomyocardial biopsy analyses with histological, immunohistochemical and molecular biological tissue evaluations to establish a final diagnosis. Only such biopsy-based information allows so far a specific treatment of distinct cardiomyopathy subgroups. In order to reduce sampling error, tissue specimens have to be obtained and analyzed to get clinically relevant data for specific treatment options. Specific gene expression and microRNA (miRNAs) profiles as well as genetic markers add additional valuable information which not only reduce the sampling error but also improve patient management. Advantages of such biomarkers result from their general expression within the entire altered myocardium. Thus, obtained information does not depend on small tissue areas reached by biopsy. This very fact allows prediction of a myocardial infection even in virus-negative areas adjoining positive biopsy specimen. The combination of multiple deregulated miRNAs or genes into one disease specific diagnostic profile demands the integration of new profiling technologies in the routine workflows of cardiological laboratories. In future, multiplex approaches allowing rapid and absolutely reliable identification of inflammatory or virally-induced myocardial diseases will replace singleplex methods such as direct detection of viral genomes in one single biopsy. miRNAs are stable biomarkers which are not only detectable in tissue samples but also in body fluids. Consequently, the determination of distinct miRNA patterns in e.g. peripheral blood samples will provide a systemic diagnostic approach for the characterization of distinct cardiomyopathies by means of non-invasive methods. This will reduce the number of undiagnosed patients who have to undergo endomyocardial biopsy for final confirmation of their myocardial complaints. The resulting molecular diagnostics will pave

关键词： Biomarker Cardiomyopathy Endomyocardial Biopsy Microrna Systemic Diagnostics

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