Voelklingen Heart Center

Saar, Germany

Voelklingen Heart Center

Saar, Germany

Time filter

Source Type

Keller A.,Biomarker Discovery Center | Keller A.,Saarland University | Leidinger P.,Saarland University | Bauer A.,German Cancer Research Center | And 44 more authors.
Nature Methods | Year: 2011

In a multicenter study, we determined the expression profiles of 863 microRNAs by array analysis of 454 blood samples from human individuals with different cancers or noncancer diseases, and validated this 'miRNome' by quantitative real-time PCR. We detected consistently deregulated profiles for all tested diseases; pathway analysis confirmed disease association of the respective microRNAs. We observed significant correlations (P = 0.004) between the genomic location of disease-associated genetic variants and deregulated microRNAs. © 2011 Nature America, Inc. All rights reserved.


Leidinger P.,Saarland University | Keller A.,Biomarker Discovery Center | Keller A.,Febit Biomedical GmbH | Keller A.,Saarland University | And 6 more authors.
Lung Cancer | Year: 2011

Recently we reported differential miRNA signatures in blood cells of lung cancer patients and healthy controls. With the present study we wanted to investigate if miRNA blood signatures are also suited to differentiate lung cancer patients from COPD patients. We compared the expression of 863 human miRNAs in blood cells of lung cancer patients, COPD patients, and healthy controls. The miRNA pattern from patients with lung cancer and COPD were more similar to each other than to the healthy controls. However, we were able to discriminate lung cancer patients and COPD patients with 90.4% accuracy, 89.2% specificity, and 91.7% sensitivity. In total, 140 miRNAs were significant for the comparison COPD and controls, 61 miRNAs were significant for the comparison lung cancer and controls, and 14 miRNAs were significant for the comparison lung cancer and COPD. Screening target databases yielded over 400 putative targets for those 14 miRNAs. The predicted mRNA targets of three of the 14 miRNAs were significantly up-regulated in PBMCs of lung cancer patients compared to patients with non-malignant lung diseases. In conclusion, we showed that blood miRNA signatures are suitable to distinguish lung cancer from COPD. © 2011 Elsevier Ireland Ltd.


PubMed | University of Oslo, Saarland University, InSCREENeX GmbH, Helmholtz Institute for Pharmaceutical Research Saarland HIPS and 2 more.
Type: Journal Article | Journal: ALTEX | Year: 2016

This paper describes a new human alveolar epithelial cell line (hAELVi - human Alveolar Epithelial Lentivirus immortalized) with type I-like characteristics and functional tight junctions, suitable to model the air-blood barrier of the peripheral lung. Primary human alveolar epithelial cells were immortalized by a novel regimen, grown as monolayers on permeable filter supports and characterized morphologically, biochemically and biophysically. hAELVi cells maintain the capacity to form tight intercellular junctions, with high trans-epithelial electrical resistance (> 1000 *cm). The cells could be kept in culture over several days, up to passage 75, under liquid-liquid as well as air-liquid conditions. Ultrastructural analysis and real time PCR revealed type I-like cell properties, such as the presence of caveolae, expression of caveolin-1, and absence of surfactant protein C. Accounting for the barrier properties, inter-digitations sealed with tight junctions and desmosomes were also observed. Low permeability of the hydrophilic marker sodium fluorescein confirmed the suitability of hAELVi cells for in vitro transport studies across the alveolar epithelium. These results suggest that hAELVi cells reflect the essential features of the air-blood barrier, as needed for an alternative to animal testing to study absorption and toxicity of inhaled drugs, chemicals and nanomaterials.


Keller A.,Biomarker Discovery Center Heidelberg | Keller A.,Saarland University | Keller A.,Siemens AG | Backes C.,Biomarker Discovery Center Heidelberg | And 13 more authors.
Molecular BioSystems | Year: 2011

MicroRNAs (miRNAs) are increasingly envisaged as biomarkers for various tumor and non-tumor diseases. MiRNA biomarker identification is, as of now, mostly performed in a candidate approach, limiting discovery to annotated miRNAs and ignoring unknown ones with potential diagnostic value. Here, we applied high-throughput SOLiD transcriptome sequencing of miRNAs expressed in human peripheral blood of patients with lung cancer. We developed a bioinformatics pipeline to generate profiles of miRNA markers and to detect novel miRNAs with diagnostic information. Applying our approach, we detected 76 previously unknown miRNAs and 41 novel mature forms of known precursors. In addition, we identified 32 annotated and seven unknown miRNAs that were significantly altered in cancer patients. These results demonstrate that deep sequencing of small RNAs bears high potential to quantify miRNAs in peripheral blood and to identify previously unknown miRNAs serving as biomarker for lung cancer. © 2011 The Royal Society of Chemistry.


Leidinger P.,Saarland University | Backes C.,Saarland University | Dahmke I.N.,Saarland University | Galata V.,Saarland University | And 5 more authors.
Oncotarget | Year: 2014

There is evidence of blood-borne miRNA signatures for various human diseases. To dissect the origin of disease-specific miRNA expression in human blood, we separately analyzed the miRNome of different immune cell subtypes, each in lung cancer patients and healthy individuals. Each immune cell type revealed a specific miRNA expression pattern also dependinging on the cell origin, line of defense, and function. The overall expression pattern of each leukocyte subtype showed great similarities between patients and controls. However, for each cell subtype we identified miRNAs that were deregulated in lung cancer patients including hsa-miR-21, a well-known oncomiR associated with poor lung cancer prognosis that was up-regulated in all leukocyte subtype comparisons of cancer versus controls. While the miRNome of cells of the adaptive immune system allowed only a weak separation between patients and controls, cells of the innate immune system allowed perfect or nearly perfect classification. Leukocytes of lung cancer patients show a cancer-specific miRNA expression profile. Our data also show that cancer specific miRNA expression pattern of whole blood samples are not determined by a single cell type. The data indicate that additional blood components, like erythrocytes, platelets, or exosomes might contribute to the disease specificity of a miRNA signature.


Leidinger P.,Saarland University | Backes C.,Saarland University | Blatt M.,Saarland University | Keller A.,Saarland University | And 4 more authors.
Molecular Cancer | Year: 2015

Objectives: In our previous studies we reported a panel of 24 miRNAs that allowed discrimination between blood of lung tumor patients independent of the histological subtype and blood of healthy controls with an accuracy of 95.4% [94.9%-95.9%]. Here, we now separately analyzed the miRNA expression in blood of non-small cell lung cancer (NSCLC), including squamous cell lung cancer and adenocarcinoma, and small cell lung cancer (SCLC) patients. Patients and methods: In total, we examined the expression levels of 1,205 miRNAs in blood samples from 20 patients from each of the three histological groups and determined differentially expressed miRNAs between histological subtypes and metastatic and non-metastatic lung cancer. We further determined the overlap of miRNAs expressed in each subgroup with the 24-miRNA signature of lung tumor patients. Results: Based on a raw p-value < 0.05, only 18 blood-borne miRNAs were differentially expressed between patients with adenocarcinoma and with squamous cell lung carcinoma, 11 miRNAs between adenocarcinoma and SCLC, and 2 between squamous cell lung carcinoma and SCLC. Likewise, the comparison based on a fold change of 1.5 did not reveal major differences of the blood-borne miRNA expression pattern between NSCLC and SCLC. In addition, we found a large overlap between the blood-borne miRNAs detected in the three histological subgroups and the previously described 24-miRNA signature that separates lung cancer patients form controls. We identified several miRNAs that allowed differentiating between metastatic and non-metastatic tumors both in blood of patients with adenocarcinoma and in blood of patients with SCLC. Conclusion: There is a common miRNA expression pattern in blood of lung cancer patients that does not allow a reliable further subtyping into NSCLC or SCLC, or into adenocarcinoma and squamous cell lung cancer. The previously described 24-miRNA signature for lung cancer appears not primarily dependent on histological subtypes. However, metastatic adenocarcinoma and SCLC can be predicted with 75% accuracy. © 2014 Leidinger et al.; licensee BioMed Central Ltd.


Leidinger P.,Saarland University | Keller A.,Saarland University | Keller A.,Siemens AG | Backes C.,Saarland University | And 2 more authors.
RNA Biology | Year: 2012

MiRNAs are powerful biomarkers for detecting various diseases from tissue and body fluids. The potential of these molecules to monitor patients over time has, however, been less explored. We followed the fate of the plasma miRNome of lung cancer patients starting prior to surgery and ending 18 mo after surgery, with blood taken at three-month intervals. Principal component and clustering analysis showed that the differences of the overall miRNA patterns between the different time points were significantly smaller than between patients. For each patient we found a rather specific fluctuating miRNA pattern. We identified miRNAs that showed a significant correlation between expression level and time distance from surgery. A network analysis revealed 12 correlated miRNAs regulating 48 genes that were deregulated in lung cancer tissue. Our data underline the importance of studies that follow the fate of miRNAs over time, both to further our understanding of the biology of miRNA signatures and to establish these signatures as biomarkers. © 2012 Landes Bioscience.


Hoppstadter J.,Saarland University | Seif M.,Korea Advanced Institute of Science and Technology | Dembek A.,Saarland University | Cavelius C.,Leibniz Institute for New Materials | And 3 more authors.
Frontiers in Pharmacology | Year: 2015

While silica nanoparticles have enabled numerous industrial and medical applications, their toxicological safety requires further evaluation. Macrophages are the major cell population responsible for nanoparticle clearance in vivo. The prevailing macrophage phenotype largely depends on the local immune status of the host. Whereas M1-polarized macrophages are considered as pro-inflammatory macrophages involved in host defense, M2 macrophages exhibit anti-inflammatory and wound-healing properties, but also promote tumor growth. We employed different models of M1 and M2 polarization: granulocyte-macrophage colony-stimulating factor/lipopolysaccharide (LPS)/interferon (IFN)-γ was used to generate primary human M1 cells and macrophage colony-stimulating factor (M-CSF)/interleukin (IL)-10 to differentiate M2 monocyte-derived macrophages (MDM). PMA-differentiated THP-1 cells were polarized towards an M1 type by LPS/IFN-γ and towards M2 by IL-10. Uptake of fluorescent silica nanoparticles (Ø26 and 41 nm) and microparticles (Ø1.75 μm) was quantified. At the concentration used (50 μg/ml), silica nanoparticles did not influence cell viability as assessed by MTT assay. Nanoparticle uptake was enhanced in M2-polarized primary human MDM compared with M1 cells, as shown by flow cytometric and microscopic approaches. In contrast, the uptake of microparticles did not differ between M1 and M2 phenotypes. M2 polarization was also associated with increased nanoparticle uptake in the macrophage-like THP-1 cell line. In accordance, in vivo polarized M2-like primary human tumor-associated macrophages obtained from lung tumors took up more nanoparticles than M1-like alveolar macrophages isolated from the surrounding lung tissue. In summary, our data indicate that the M2 polarization of macrophages promotes nanoparticle internalization. Therefore, the phenotypical differences between macrophage subsets should be taken into consideration in future investigations on nanosafety, but might also open up therapeutic perspectives allowing to specifically target M2 polarized macrophages. © 2015 Hoppstädter, Seif, Dembek, Cavelius, Huwer, Kraegeloh and Kiemer.


PubMed | Saarland University and Voelklingen Heart Center
Type: Journal Article | Journal: Oncotarget | Year: 2014

There is evidence of blood-borne miRNA signatures for various human diseases. To dissect the origin of disease-specific miRNA expression in human blood, we separately analyzed the miRNome of different immune cell subtypes, each in lung cancer patients and healthy individuals. Each immune cell type revealed a specific miRNA expression pattern also dependinging on the cell origin, line of defense, and function. The overall expression pattern of each leukocyte subtype showed great similarities between patients and controls. However, for each cell subtype we identified miRNAs that were deregulated in lung cancer patients including hsa-miR-21, a well-known oncomiR associated with poor lung cancer prognosis that was up-regulated in all leukocyte subtype comparisons of cancer versus controls. While the miRNome of cells of the adaptive immune system allowed only a weak separation between patients and controls, cells of the innate immune system allowed perfect or nearly perfect classification. Leukocytes of lung cancer patients show a cancer-specific miRNA expression profile. Our data also show that cancer specific miRNA expression pattern of whole blood samples are not determined by a single cell type. The data indicate that additional blood components, like erythrocytes, platelets, or exosomes might contribute to the disease specificity of a miRNA signature.


PubMed | Voelklingen Heart Center, Leibniz Institute for New Materials, Saarland University and Korea Advanced Institute of Science and Technology
Type: | Journal: Frontiers in pharmacology | Year: 2015

While silica nanoparticles have enabled numerous industrial and medical applications, their toxicological safety requires further evaluation. Macrophages are the major cell population responsible for nanoparticle clearance in vivo. The prevailing macrophage phenotype largely depends on the local immune status of the host. Whereas M1-polarized macrophages are considered as pro-inflammatory macrophages involved in host defense, M2 macrophages exhibit anti-inflammatory and wound-healing properties, but also promote tumor growth. We employed different models of M1 and M2 polarization: granulocyte-macrophage colony-stimulating factor/lipopolysaccharide (LPS)/interferon (IFN)- was used to generate primary human M1 cells and macrophage colony-stimulating factor (M-CSF)/interleukin (IL)-10 to differentiate M2 monocyte-derived macrophages (MDM). PMA-differentiated THP-1 cells were polarized towards an M1 type by LPS/IFN- and towards M2 by IL-10. Uptake of fluorescent silica nanoparticles (26 and 41 nm) and microparticles (1.75 m) was quantified. At the concentration used (50 g/ml), silica nanoparticles did not influence cell viability as assessed by MTT assay. Nanoparticle uptake was enhanced in M2-polarized primary human MDM compared with M1 cells, as shown by flow cytometric and microscopic approaches. In contrast, the uptake of microparticles did not differ between M1 and M2 phenotypes. M2 polarization was also associated with increased nanoparticle uptake in the macrophage-like THP-1 cell line. In accordance, in vivo polarized M2-like primary human tumor-associated macrophages obtained from lung tumors took up more nanoparticles than M1-like alveolar macrophages isolated from the surrounding lung tissue. In summary, our data indicate that the M2 polarization of macrophages promotes nanoparticle internalization. Therefore, the phenotypical differences between macrophage subsets should be taken into consideration in future investigations on nanosafety, but might also open up therapeutic perspectives allowing to specifically target M2 polarized macrophages.

Loading Voelklingen Heart Center collaborators
Loading Voelklingen Heart Center collaborators