111 results on '"Wruck W"'
Search Results
2. Use of Stem Cells in Toxicology
- Author
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Matz, P., primary, Spitzhorn, L.-S., additional, Otte, J., additional, Kawala, M.-A., additional, Woestmann, J., additional, Yigit, H., additional, Wruck, W., additional, and Adjaye, J., additional
- Published
- 2017
- Full Text
- View/download PDF
3. Entschlüsselung molekularer und (epi-)genetischer Mechanismen während der Differenzierung von Embryonalkarzinomen zu Dottersacktumoren
- Author
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Kotthoff, M, Wruck, W, Adjaye, JA, Nettersheim, D, Skowron, M, Bremmer, F, Fichtner, A, Schönberger, S, Calaminus, G, Vokuhl, C, Pfister, D, Heidenreich, A, Albers, P, Kotthoff, M, Wruck, W, Adjaye, JA, Nettersheim, D, Skowron, M, Bremmer, F, Fichtner, A, Schönberger, S, Calaminus, G, Vokuhl, C, Pfister, D, Heidenreich, A, and Albers, P
- Published
- 2022
4. New Technology Closes Micro-Annular Flow Paths in the Wellbore, Stopping Downhole Gas from Escaping to the Surface
- Author
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Green, C., primary, Evans, R., additional, Fry, B., additional, and Wruck, W. S., additional
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- 2021
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5. Gene expression profiling in aggressive digital papillary adenocarcinoma sheds light on the architecture of a rare sweat gland carcinoma
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Surowy, H. M., Giesen, A. K., Otte, J., Buettner, R., Falkenstein, D., Friedl, H., Meier, F., Petzsch, P., Wachtmeister, T., Westphal, D., Wieczorek, D., Wruck, W., Adjaye, J., Ruetten, A., Redler, S., Surowy, H. M., Giesen, A. K., Otte, J., Buettner, R., Falkenstein, D., Friedl, H., Meier, F., Petzsch, P., Wachtmeister, T., Westphal, D., Wieczorek, D., Wruck, W., Adjaye, J., Ruetten, A., and Redler, S.
- Abstract
Background Sweat gland carcinomas are rare cutaneous adnexal malignancies. Aggressive digital papillary adenocarcinoma (ADPA) represents a very rare subentity, thought to arise almost exclusively from the sweat glands of the fingers and toes. The aetiology of sweat gland carcinomas and ADPA is largely unknown. ADPAs are most likely driven by somatic mutations. However, somatic mutation patterns are largely unexplored, creating barriers to the development of effective therapeutic approaches to the treatment of ADPA. Objectives To investigate the transcriptome profile of ADPA using a sample of eight formalin-fixed, paraffin-embedded tissue samples of ADPA and healthy control tissue. Methods Transcriptome profiling was performed using the Affymetrix PrimeView Human Gene Expression Microarray and findings were validated via reverse transcription of RNA and real-time quantitative polymerase chain reaction. Results Transcriptome analyses showed increased tumour expression of 2266 genes, with significant involvement of cell cycle, ribosomal and crucial cancer pathways. Our results point to tumour overexpression of FGFR2 (P = 0 center dot 001). Conclusions The results indicate the involvement of crucial oncogenic driver pathways, highlighting cell cycle and ribosomal pathways in the aetiology of ADPA. Suggested tumour overexpression of FGFR2 raises the hope that targeting the fibroblast growth factor (FGF)/FGF receptor axis might be a promising treatment for ADPA and probably for the overall group of sweat gland carcinomas.
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- 2019
6. Gene expression profiling in aggressive digital papillary adenocarcinoma sheds light on the architecture of a rare sweat gland carcinoma
- Author
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Surowy, H.M., primary, Giesen, A.K., additional, Otte, J., additional, Büttner, R., additional, Falkenstein, D., additional, Friedl, H., additional, Meier, F., additional, Petzsch, P., additional, Wachtmeister, T., additional, Westphal, D., additional, Wieczorek, D., additional, Wruck, W., additional, Adjaye, J., additional, Rütten, A., additional, and Redler, S., additional
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- 2019
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7. Combined ultra-low input mRNA and whole-genome sequencing of human embryonic stem cells
- Author
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Mertes, F., Lichtner, B., Kuhl, H., Blattner, M., Otte, J., Wruck, W., Timmermann, B., Lehrach, H., and Adjaye, J.
- Subjects
Embryonic Stem Cells ,Next Generation Sequencing ,Pluripotency ,Rna-seq ,Single Cell ,Ultra-low Input Sequencing ,Whole-genome Sequencing ,Genetics ,Biotechnology - Abstract
Background: Next Generation Sequencing has proven to be an exceptionally powerful tool in the field of genomics and transcriptomics. With recent development it is nowadays possible to analyze ultra-low input sample material down to single cells. Nevertheless, investigating such sample material often limits the analysis to either the genome or transcriptome. We describe here a combined analysis of both types of nucleic acids from the same sample material. Methods: The method described enables the combined preparation of amplified cDNA as well as amplified whole-genome DNA from an ultra-low input sample material derived from a sub-colony of in-vitro cultivated human embryonic stem cells. cDNA is prepared by the application of oligo-dT coupled magnetic beads for mRNA capture, first strand synthesis and 3'-tailing followed by PCR. Whole-genome amplified DNA is prepared by Phi29 mediated amplification. Illumina sequencing is applied to short fragment libraries prepared from the amplified samples. Results: We developed a protocol which enables the combined analysis of the genome as well as the transcriptome by Next Generation Sequencing from ultra-low input samples. The protocol was evaluated by sequencing sub-colony structures from human embryonic stem cells containing 150 to 200 cells. The method can be adapted to any available sequencing system. Conclusions: To our knowledge, this is the first report where sub-colonies of human embryonic stem cells have been analyzed both at the genomic as well as transcriptome level. The method of this proof of concept study may find useful practical applications for cases where only a limited number of cells are available, e.g. for tissues samples from biopsies, tumor spheres, circulating tumor cells and cells from early embryonic development. The results we present demonstrate that a combined analysis of genomic DNA and messenger RNA from ultra-low input samples is feasible and can readily be applied to other cellular systems with limited material available.
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- 2015
8. Comparative induction of pluripotency in human umbilical vein endothelial cells and dermal fibroblasts and further differentiation into Hepatocytes
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Matz, P, primary, Wruck, W, additional, and Adjaye, J, additional
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- 2015
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9. Modelling of human nonalcoholic fatty liver disease with hepatocyte like cells derived from pluripotent stem cells
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Graffmann, N, primary, Kawala, MA, additional, Ring, S, additional, Wruck, W, additional, and Adjaye, J, additional
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- 2015
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10. Regenerative potential of human pluripotent stem cell-derived MSCs in a Gunn rat liver injury model
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Spitzhorn, LS, primary, Megges, M, additional, Kordes, C, additional, Sawitza, I, additional, Götze, S, additional, Kawala, MA, additional, Wruck, W, additional, Oreffo, R, additional, Häussinger, D, additional, and Adjaye, J, additional
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- 2015
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11. Construction of a 'unigene' cDNA clone set by oligonucleotide fingerprinting allows access to 25 000 potential sugar beet genes
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Herwig, R., Schulz, B., Weisshaar, Bernd, Hennig, S., Steinfath, M., Drungowski, M., Stahl, D., Wruck, W., Menze, A., O'Brien, J., Lehrach, H., and Radelof, U.
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Expressed Sequence Tags ,DNA, Complementary ,Transcription, Genetic ,Arabidopsis ,Gene Dosage ,Oligonucleotides ,Genes, Plant ,DNA Fingerprinting ,Plant Roots ,Sensitivity and Specificity ,Automation ,Multigene Family ,Cluster Analysis ,Beta vulgaris ,Cloning, Molecular ,Genome, Plant ,Gene Library - Abstract
Access to the complete gene inventory of an organism is crucial to understanding physiological processes like development, differentiation, pathogenesis, or adaptation to the environment. Transcripts from many active genes are present at low copy numbers. Therefore, procedures that rely on random EST sequencing or on normalisation and subtraction methods have to produce massively redundant data to get access to low-abundance genes. Here, we present an improved oligonucleotide fingerprinting (ofp) approach to the genome of sugar beet (Beta vulgaris), a plant for which practically no molecular information has been available. To identify distinct genes and to provide a representative 'unigene' cDNA set for sugar beet, 159 936 cDNA clones were processed utilizing large-scale, high-throughput data generation and analysis methods. Data analysis yielded 30 444 ofp clusters reflecting the number of different genes in the original cDNA sample. A sample of 10 961 cDNA clones, each representing a different cluster, were selected for sequencing. Standard sequence analysis confirmed that 89% of these EST sequences did represent different genes. These results indicate that the full set of 30 444 ofp clusters represent up to 25 000 genes. We conclude that the ofp analysis pipeline is an accurate and effective way to construct large representative 'unigene' sets for any plant of interest with no requirement for prior molecular sequence data.
- Published
- 2002
12. A systems biology approach to deciphering the etiology of steatosis employing patient-derived dermal fibroblasts and iPS cells.
- Author
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Jozefczuk, J., Kashofer, K., Ummani, R., Henjes, F., Rehman, S., Geenen, S., Wruck, W., Regenbrecht, C., Daskalaki, A., Wierling, C., Turano, P., Bertini, I., Korf, U., Zatloukal, K., Westerhoff, H.V., Lehrach, H., Adjaye, J., Jozefczuk, J., Kashofer, K., Ummani, R., Henjes, F., Rehman, S., Geenen, S., Wruck, W., Regenbrecht, C., Daskalaki, A., Wierling, C., Turano, P., Bertini, I., Korf, U., Zatloukal, K., Westerhoff, H.V., Lehrach, H., and Adjaye, J.
- Abstract
Non-alcoholic fatty liver disease comprises a broad spectrum of disease states ranging from simple steatosis to non-alcoholic steatohepatitis. As a result of increases in the prevalences of obesity, insulin resistance, and hyperlipidemia, the number of people with hepatic steatosis continues to increase. Differences in susceptibility to steatohepatitis and its progression to cirrhosis have been attributed to a complex interplay of genetic and external factors all addressing the intracellular network. Increase in sugar or refined carbohydrate consumption results in an increase of insulin and insulin resistance that can lead to the accumulation of fat in the liver. Here we demonstrate how a multidisciplinary approach encompassing cellular reprogramming, transcriptomics, proteomics, metabolomics, modeling, network reconstruction, and data management can be employed to unveil the mechanisms underlying the progression of steatosis. Proteomics revealed reduced AKT/mTOR signaling in fibroblasts derived from steatosis patients and further establishes that the insulin-resistant phenotype is present not only in insulin-metabolizing central organs, e.g., the liver, but is also manifested in skin fibroblasts. Transcriptome data enabled the generation of a regulatory network based on the transcription factor SREBF1, linked to a metabolic network of glycerolipid, and fatty acid biosynthesis including the downstream transcriptional targets of SREBF1 which include LIPIN1 (LPIN) and low density lipoprotein receptor. Glutathione metabolism was among the pathways enriched in steatosis patients in comparison to healthy controls. By using a model of the glutathione pathway we predict a significant increase in the flux through glutathione synthesis as both gamma-glutamylcysteine synthetase and glutathione synthetase have an increased flux. We anticipate that a larger cohort of patients and matched controls will confirm our preliminary findings presented here. © 2012 Jozefczuk, Kashofer
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- 2012
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13. Data management strategies for multinational large-scale systems biology projects
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Wruck, W., primary, Peuker, M., additional, and Regenbrecht, C. R. A., additional
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- 2012
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14. ChemInform Abstract: Use of Conductive Materials to Enhance Lead-Acid Battery Formation.
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BULLOCK, K. R., primary, MAHATO, B. K., additional, and WRUCK, W. J., additional
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- 2010
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15. New Tools for Oligonucleotide Fingerprinting
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Guerasimova, A., primary, Nyarsik, L., additional, Girnus, I., additional, Steinfath, M., additional, Wruck, W., additional, Griffiths, H., additional, Herwig, R., additional, Wierling, C., additional, O’Brien, J., additional, Eickhoff, H., additional, Lehrach, H., additional, and Radelof, U., additional
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- 2001
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16. Rechargeable Zn ‐ MnO2 Alkaline Batteries
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Wruck, W. J., primary, Reichman, B., additional, Bullock, K. R., additional, and Kao, W. ‐H., additional
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- 1991
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17. Use of Conductive Materials to Enhance Lead‐Acid Battery Formation
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Bullock, K. R., primary, Mahato, B. K., additional, and Wruck, W. J., additional
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- 1991
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18. Automated image analysis for array hybridization experiments.
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Steinfath, M, Wruck, W, Seidel, H, Lehrach, H, Radelof, U, and O'Brien, J
- Abstract
Image analysis is a major part of data evaluation for array hybridization experiments in molecular biology. The program presented here is designed to analyze automatically images from hybridization experiments with various arrangements: different kinds of probes (oligonucleotides or complex probes), different supports (nylon filters or glass slides), different labeling of probes (radioactively or fluorescently). The program is currently applied to oligonucleotide fingerprinting projects and complex hybridizations. The only precondition for the use of the program is that the targets are arrayed in a grid, which can be approximately transformed to an orthogonal equidistant grid by a projective mapping.
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- 2001
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19. Cross-species hybridisation of human and bovine orthologous genes on high density cDNA microarrays
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Hultschig Claus, Carnwath Joseph W, Nowak Monika, Brink Thore C, BenKahla Alia, Wruck Wasco, Herrmann Doris, Herwig Ralf, Adjaye James, Niemann Heiner, and Lehrach Hans
- Subjects
Biotechnology ,TP248.13-248.65 ,Genetics ,QH426-470 - Abstract
Abstract Background Cross-species gene-expression comparison is a powerful tool for the discovery of evolutionarily conserved mechanisms and pathways of expression control. The usefulness of cDNA microarrays in this context is that broad areas of homology are compared and hybridization probes are sufficiently large that small inter-species differences in nucleotide sequence would not affect the analytical results. This comparative genomics approach would allow a common set of genes within a specific developmental, metabolic, or disease-related gene pathway to be evaluated in experimental models of human diseases. The objective of this study was to investigate the feasibility and reproducibility of cross-species analysis employing a human cDNA microarray as probe. Results As a proof of principle, total RNA derived from human and bovine fetal brains was used as a source of labelled targets for hybridisation onto a human cDNA microarray composed of 349 characterised genes. Each gene was spotted 20 times representing 6,980 data points thus enabling highly reproducible spot quantification. Employing high stringency hybridisation and washing conditions, followed by data analysis, revealed slight differences in the expression levels and reproducibility of the signals between the two species. We also assigned each of the genes into three expression level categories- i.e. high, medium and low. The correlation co-efficient of cross hybridisation between the orthologous genes was 0.94. Verification of the array data by semi-quantitative RT-PCR using common primer sequences enabled co-amplification of both human and bovine transcripts. Finally, we were able to assign gene names to previously uncharacterised bovine ESTs. Conclusions Results of our study demonstrate the harnessing and utilisation power of comparative genomics and prove the feasibility of using human microarrays to facilitate the identification of co-expressed orthologous genes in common tissues derived from different species.
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- 2004
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20. ChemInform Abstract: Use of Conductive Materials to Enhance Lead-Acid Battery Formation.
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BULLOCK, K. R., MAHATO, B. K., and WRUCK, W. J.
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- 1992
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21. Dual battery system
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Wruck, W
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- 1993
22. Molecular and epigenetic ex vivo profiling of testis cancer-associated fibroblasts and their interaction with germ cell tumor cells and macrophages.
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Stephan A, Suhrmann JH, Skowron MA, Che Y, Poschmann G, Petzsch P, Kresbach C, Wruck W, Pongratanakul P, Adjaye J, Stühler K, Köhrer K, Schüller U, and Nettersheim D
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- Male, Humans, Gene Expression Regulation, Neoplastic, DNA Methylation, Cell Line, Tumor, Cell Communication, Cisplatin pharmacology, THP-1 Cells, Cell Proliferation, Testicular Neoplasms metabolism, Testicular Neoplasms genetics, Testicular Neoplasms pathology, Tumor Microenvironment, Cancer-Associated Fibroblasts metabolism, Cancer-Associated Fibroblasts pathology, Neoplasms, Germ Cell and Embryonal genetics, Neoplasms, Germ Cell and Embryonal metabolism, Neoplasms, Germ Cell and Embryonal pathology, Macrophages metabolism, Macrophages pathology, Epigenesis, Genetic
- Abstract
Germ cell tumors (GCT) are the most common solid tumors in young men of age 15 - 40. In previous studies, we profiled the interaction of GCT cells with cells of the tumor microenvironment (TM), which showed that especially the 3D interaction of fibroblasts (FB) or macrophages with GCT cells influenced the growth behavior and cisplatin response as well as the transcriptome and secretome of the tumor cells, suggesting that the crosstalk of these cells with GCT cells is crucial for tumor progression and therapy outcome. In this study, we shed light on the mechanisms of activation of cancer-associated fibroblasts (CAF) in the GCT setting and their effects on GCT cells lines and the monocyte cell line THP-1. Ex vivo cultures of GCT-derived CAF were established and characterized molecularly and epigenetically by performing DNA methylation arrays, RNA sequencing, and mass spectrometry-based secretome analysis. We demonstrated that the activation state of CAF is influenced by their former prevailing tumor environment in which they have resided. Hereby, we postulate that seminoma (SE) and embryonal carcinoma (EC) activate CAF, while teratoma (TER) play only a minor role in CAF formation. In turn, CAF influence proliferation and the expression of cisplatin sensitivity-related factors in GCT cells lines as well as polarization of in vitro-induced macrophages by the identified effector molecules IGFBP1, LGALS3BP, LYVE1, and PTX3. Our data suggests that the vital interaction of CAF with GCT cells and with macrophages has a huge influence on shaping the extracellular matrix as well as on recruitment of immune cells to the TM. In conclusion, therapeutically interfering with CAF and / or macrophages in addition to the standard therapy might slow-down progression of GCT and re-shaping of the TM to a tumor-promoting environment. Significance: The interaction of CAF with GCT and macrophages considerably influences the microenvironment. Thus, therapeutically interfering with CAF might slow-down progression of GCT and re-shaping of the microenvironment to a tumor-promoting environment., Competing Interests: Conflict of interest statement All authors declare to have no conflict of interest, (Copyright © 2024. Published by Elsevier B.V.)
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- 2024
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23. Gene expression profiling in porocarcinoma indicates heterogeneous tumor development and substantiates poromas as precursor lesions.
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Holst S, Weber AK, Meier F, Otte J, Petzsch P, Reifenberger J, Wachtmeister T, Westphal D, Ziemer M, Wruck W, Adjaye J, Betz RC, Rütten A, Surowy HM, and Redler S
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- Humans, Precancerous Conditions genetics, Precancerous Conditions pathology, Female, Male, Cell Transformation, Neoplastic genetics, Gene Expression Regulation, Neoplastic, Aged, Middle Aged, Gene Expression Profiling, Eccrine Porocarcinoma genetics, Eccrine Porocarcinoma pathology, Sweat Gland Neoplasms genetics, Sweat Gland Neoplasms pathology, Sweat Gland Neoplasms metabolism
- Abstract
Background and Objectives: Malignant sweat gland tumors are rare, with the most common being eccrine porocarcinoma (EP). Approximately 18% of benign eccrine poroma (EPO) transit to EP. Previous research has provided first insights into the mutational landscape of EP. However, only few studies have performed gene expression analyses. This leaves a gap in the understanding of EP biology and potential drivers of malignant transformation from EPO to EP., Methods: Transcriptome profiling of 23 samples of primary EP and normal skin (NS). Findings from the EP samples were then tested in 17 samples of EPO., Results: Transcriptome profiling revealed diversity in gene expression and indicated biologically heterogeneous sub-entities as well as widespread gene downregulation in EP. Downregulated genes included CD74, NDGR1, SRRM2, CDC42, ANXA2, KFL9 and NOP53. Expression levels of CD74, NDGR1, SRRM2, ANXA2, and NOP53 showed a stepwise-reduction in expression from NS via EPO to EP, thus supporting the hypothesis that EPO represents a transitional state in EP development., Conclusions: We demonstrated that EP is molecularly complex and that evolutionary trajectories correspond to tumor initiation and progression. Our results provide further evidence implicating the p53 axis and the EGFR pathway. Larger samples are warranted to confirm our findings., (© 2024 The Authors. Journal der Deutschen Dermatologischen Gesellschaft published by Wiley‐VCH GmbH on behalf of Deutsche Dermatologische Gesellschaft.)
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- 2024
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24. Cockayne Syndrome Patient iPSC-Derived Brain Organoids and Neurospheres Show Early Transcriptional Dysregulation of Biological Processes Associated with Brain Development and Metabolism.
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Szepanowski LP, Wruck W, Kapr J, Rossi A, Fritsche E, Krutmann J, and Adjaye J
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- Humans, DNA Helicases genetics, DNA Repair Enzymes metabolism, Poly-ADP-Ribose Binding Proteins genetics, Poly-ADP-Ribose Binding Proteins metabolism, Brain metabolism, Organoids metabolism, Induced Pluripotent Stem Cells metabolism, Cockayne Syndrome genetics, Cockayne Syndrome metabolism
- Abstract
Cockayne syndrome (CS) is a rare hereditary autosomal recessive disorder primarily caused by mutations in Cockayne syndrome protein A (CSA) or B (CSB). While many of the functions of CSB have been at least partially elucidated, little is known about the actual developmental dysregulation in this devasting disorder. Of particular interest is the regulation of cerebral development as the most debilitating symptoms are of neurological nature. We generated neurospheres and cerebral organoids utilizing Cockayne syndrome B protein (CSB)-deficient induced pluripotent stem cells derived from two patients with distinct severity levels of CS and healthy controls. The transcriptome of both developmental timepoints was explored using RNA-Seq and bioinformatic analysis to identify dysregulated biological processes common to both patients with CS in comparison to the control. CSB-deficient neurospheres displayed upregulation of the VEGFA-VEGFR2 signalling pathway, vesicle-mediated transport and head development. CSB-deficient cerebral organoids exhibited downregulation of brain development, neuron projection development and synaptic signalling. We further identified the upregulation of steroid biosynthesis as common to both timepoints, in particular the upregulation of the cholesterol biosynthesis branch. Our results provide insights into the neurodevelopmental dysregulation in patients with CS and strengthen the theory that CS is not only a neurodegenerative but also a neurodevelopmental disorder.
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- 2024
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25. Free Bilirubin Induces Neuro-Inflammation in an Induced Pluripotent Stem Cell-Derived Cortical Organoid Model of Crigler-Najjar Syndrome.
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Pranty AI, Wruck W, and Adjaye J
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- Humans, Brain, Cytokines, Bilirubin, Crigler-Najjar Syndrome, Induced Pluripotent Stem Cells, Kernicterus
- Abstract
Bilirubin-induced neurological damage (BIND), which might progress to kernicterus, occurs as a consequence of defects in the bilirubin conjugation machinery, thus enabling albumin-unbound free bilirubin (BF) to cross the blood-brain barrier and accumulate within. A defect in the UGT1A1 enzyme-encoding gene, which is directly responsible for bilirubin conjugation, can cause Crigler-Najjar syndrome (CNS) and Gilbert's syndrome. We used human-induced pluripotent stem cell (hiPSC)-derived 3D brain organoids to model BIND in vitro and unveil the molecular basis of the detrimental effects of BF in the developing human brain. Healthy and patient-derived iPSCs were differentiated into day-20 brain organoids, and then stimulated with 200 nM BF. Analyses at 24 and 72 h post-treatment point to BF-induced neuro-inflammation in both cell lines. Transcriptome, associated KEGG, and Gene Ontology analyses unveiled the activation of distinct inflammatory pathways, such as cytokine-cytokine receptor interaction, MAPK signaling, and NFκB activation. Furthermore, the mRNA expression and secretome analysis confirmed an upregulation of pro-inflammatory cytokines such as IL-6 and IL-8 upon BF stimulation. This novel study has provided insights into how a human iPSC-derived 3D brain organoid model can serve as a prospective platform for studying the etiology of BIND kernicterus.
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- 2023
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26. JNK Signalling Regulates Self-Renewal of Proliferative Urine-Derived Renal Progenitor Cells via Inhibition of Ferroptosis.
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Nguyen L, Thewes L, Westerhoff M, Wruck W, Reichert AS, Berndt C, and Adjaye J
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- Humans, Kidney, Renal Dialysis, Stem Cells, Ferroptosis, Renal Insufficiency, Chronic, MAP Kinase Signaling System
- Abstract
With a global increase in chronic kidney disease patients, alternatives to dialysis and organ transplantation are needed. Stem cell-based therapies could be one possibility to treat chronic kidney disease. Here, we used multipotent urine-derived renal progenitor cells (UdRPCs) to study nephrogenesis. UdRPCs treated with the JNK inhibitor-AEG3482 displayed decreased proliferation and downregulated transcription of cell cycle-associated genes as well as the kidney progenitor markers-SIX2, SALL1 and VCAM1. In addition, levels of activated SMAD2/3, which is associated with the maintenance of self-renewal in UdRPCs, were decreased. JNK inhibition resulted in less efficient oxidative phosphorylation and more lipid peroxidation via ferroptosis, an iron-dependent non-apoptotic cell death pathway linked to various forms of kidney disease. Our study is the first to describe the importance of JNK signalling as a link between maintenance of self-renewal and protection against ferroptosis in SIX2-positive renal progenitor cells.
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- 2023
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27. Generation of two Alpha-I antitrypsin deficiency patient-derived induced pluripotent stem cell lines ISRM-AATD-iPSC-1 (HHUUKDi011-A) and ISRM-AATD-iPSC-2 (HHUUKDi012-A).
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Ncube A, Bewersdorf L, Spitzhorn LS, Loerch C, Bohndorf M, Graffmann N, May L, Amzou S, Fromme M, Wruck W, Strnad P, and Adjaye J
- Subjects
- Humans, Male, Female, Kruppel-Like Factor 4, Cell Differentiation, Cell Line, Embryonic Stem Cells, Induced Pluripotent Stem Cells metabolism
- Abstract
SIX2-positive urine derived renal progenitor cells were isolated from a male and female alpha1-antitrypsin deficiency (AATD) patients both harboring the homozygous PiZZ genotype. The cells were reprogrammed to generate two integration-free induced pluripotent stem cell (iPSC) lines by transfecting episomal-based plasmids expressing OCT4, SOX2, NANOG, c-MYC, KLF4 and LIN28. Pluripotency was confirmed by immunocytochemistry for associated markers and embryoid body-based differentiation into the three germ layers. The iPSC lines carried the parental PiZZ genotype. Comparative transcriptome analyses with human embryonic stem cell line H9 revealed a Pearson correlation of 0.945 for ISRM-AATD-iPSC-1 and 0.939 for ISRM-AATD-iPSC-2 respectively., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)
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- 2023
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28. Natural Products in Renal-Associated Drug Discovery.
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Wruck W, Genfi AKA, and Adjaye J
- Abstract
The global increase in the incidence of kidney failure constitutes a major public health problem. Kidney disease is classified into acute and chronic: acute kidney injury (AKI) is associated with an abrupt decline in kidney function and chronic kidney disease (CKD) with chronic renal failure for more than three months. Although both kidney syndromes are multifactorial, inflammation and oxidative stress play major roles in the diversity of processes leading to these kidney malfunctions. Here, we reviewed various publications on medicinal plants with antioxidant and anti-inflammatory properties with the potential to treat and manage kidney-associated diseases in rodent models. Additionally, we conducted a meta-analysis to identify gene signatures and associated biological processes perturbed in human and mouse cells treated with antioxidants such as epigallocatechin gallate (EGCG), the active ingredient in green tea, and the mushroom Ganoderma lucidum (GL) and in kidney disease rodent models. We identified EGCG- and GL-regulated gene signatures linked to metabolism; inflammation (NRG1, E2F1, NFKB1 and JUN); ion signalling; transport; renal processes (SLC12A1 and LOX) and VEGF, ERBB and BDNF signalling. Medicinal plant extracts are proving to be effective for the prevention, management and treatment of kidney-associated diseases; however, more detailed characterisations of their targets are needed to enable more trust in their application in the management of kidney-associated diseases.
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- 2023
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29. Unveiling Angiotensin II and Losartan-Induced Gene Regulatory Networks Using Human Urine-Derived Podocytes.
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Thimm C, Erichsen L, Wruck W, and Adjaye J
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- Humans, Losartan pharmacology, Losartan metabolism, Angiotensin II metabolism, Gene Regulatory Networks, Receptor, Angiotensin, Type 1 genetics, Receptor, Angiotensin, Type 1 metabolism, Podocytes metabolism, Hypertension metabolism
- Abstract
Podocytes are highly specialized cells that play a pivotal role in the blood filtration process in the glomeruli of the kidney, and their dysfunction leads to renal diseases. For this reason, the study and application of this cell type is of great importance in the field of regenerative medicine. Hypertension is mainly regulated by the renin-angiotensin-aldosterone system (RAAS), with its main mediator being angiotensin II (ANG II). Elevated ANG II levels lead to a pro-fibrotic, inflammatory, and hypertrophic milieu that induces apoptosis in podocytes. The activation of RAAS is critical for the pathogenesis of podocyte injury; as such, to prevent podocyte damage, patients with hypertension are administered drugs that modulate RAAS signaling. A prime example is the orally active, non-peptide, selective angiotensin-II-type I receptor (AGTR1) blocker losartan. Here, we demonstrate that SIX2-positive urine-derived renal progenitor cells (UdRPCs) and their immortalized counterpart (UM51-hTERT) can be directly differentiated into mature podocytes. These podocytes show activation of RAAS after stimulation with ANG II, resulting in ANG II-dependent upregulation of the expression of the angiotensin-II-type I receptor, AGTR1, and the downregulated expression of the angiotensin-II-type II receptor 2 (AGTR2). The stimulation of podocytes with losartan counteracts ANG II-dependent changes, resulting in a dependent favoring of the specific receptor from AGTR1 to AGTR2. Transcriptome analysis revealed 94 losartan-induced genes associated with diverse biological processes and pathways such as vascular smooth muscle contraction, the oxytocin signaling pathway, renin secretion, and ECM-receptor interaction. Co-stimulation with losartan and ANG II induced the exclusive expression of 106 genes associated with DNA methylation or demethylation, cell differentiation, the developmental process, response to muscle stretch, and calcium ion transmembrane transport. These findings highlight the usefulness of UdRPC-derived podocytes in studying the RAAS pathway and nephrotoxicity in various kidney diseases.
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- 2023
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30. Secreted Cytokines within the Urine of AKI Patients Modulate TP53 and SIRT1 Levels in a Human Podocyte Cell Model.
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Erichsen L, Thimm C, Wruck W, Kaierle D, Schless M, Huthmann L, Dimski T, Kindgen-Milles D, Brandenburger T, and Adjaye J
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- Humans, Male, Female, Cytokines, Sirtuin 1, Kidney, Creatinine, Biomarkers urine, Tumor Suppressor Protein p53, Podocytes, Acute Kidney Injury etiology
- Abstract
Acute kidney injury (AKI) is a major kidney disease with a poor clinical outcome. It is a common complication, with an incidence of 10-15% of patients admitted to hospital. This rate even increases for patients who are admitted to the intensive care unit, with an incidence of >50%. AKI is characterized by a rapid increase in serum creatinine, decrease in urine output, or both. The associated symptoms include feeling sick or being sick, diarrhoea, dehydration, decreased urine output (although occasionally the urine output remains normal), fluid retention causing swelling in the legs or ankles, shortness of breath, fatigue and nausea. However, sometimes acute kidney injury causes no signs or symptoms and is detected by lab tests. Therefore, the identification of cytokines for the early detection and diagnosis of AKI is highly desirable, as their application might enable the prevention of the progression from AKI to chronic kidney disease (CKD). In this study, we analysed the secretome of the urine of an AKI patient cohort by employing a kidney-biomarker cytokine assay. Based on these results, we suggest ADIPOQ, EGF and SERPIN3A as potential cytokines that might be able to detect AKI as early as 24 h post-surgery. For the later stages, as common cytokines for the detection of AKI in both male and female patients, we suggest VEGF, SERPIN3A, TNFSF12, ANPEP, CXCL1, REN, CLU and PLAU. These cytokines in combination might present a robust strategy for identifying the development of AKI as early as 24 h or 72 h post-surgery. Furthermore, we evaluated the effect of patient and healthy urine on human podocyte cells. We conclude that cytokines abundant in the urine of AKI patients trigger processes that are needed to repair the damaged nephron and activate TP53 and SIRT1 to maintain the balance between proliferation, angiogenesis, and cell cycle arrest.
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- 2023
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31. Gene expression studies of WT1 mutant Wilms tumor cell lines in the frame work of published kidney development data reveals their early kidney stem cell origin.
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Royer-Pokora B, Wruck W, Adjaye J, and Beier M
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- Humans, WT1 Proteins genetics, Kidney pathology, Stem Cells metabolism, Cell Line, Tumor, Epithelial Cells metabolism, Gene Expression, Genes, Wilms Tumor, Wilms Tumor pathology, Kidney Neoplasms pathology
- Abstract
In order to get a better insight into the timing of WT1 mutant Wilms tumor development, we compared the gene expression profiles of nine established WT1 mutant Wilms tumor cell lines with published data from different kidney cell types during development. Publications describing genes expressed in nephrogenic precursor cells, ureteric bud cells, more mature nephrogenic epithelial cells and interstitial cell types were used. These studies uncovered that the WT1 mutant Wilms tumor cells lines express genes from the earliest nephrogenic progenitor cells, as well as from more differentiated nephron cells with the highest expression from the stromal/interstitial compartment. The expression of genes from all cell compartments points to an early developmental origin of the tumor in a common stem cell. Although variability of the expression of specific genes was evident between the cell lines the overall expression pattern was very similar. This is likely dependent on their different genetic backgrounds with distinct WT1 mutations and the absence/presence of mutant CTNNB1., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Royer-Pokora et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
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- 2023
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32. Derivation of the Immortalized Cell Line UM51-PrePodo-hTERT and Its Responsiveness to Angiotensin II and Activation of the RAAS Pathway.
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Erichsen L, Kloss LDF, Thimm C, Bohndorf M, Schichel K, Wruck W, and Adjaye J
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- Male, Humans, Aged, Middle Aged, Cell Line, Cells, Cultured, Cellular Senescence genetics, Renin-Angiotensin System, Angiotensin II pharmacology
- Abstract
Recent demographic studies predict there will be a considerable increase in the number of elderly people within the next few decades. Aging has been recognized as one of the main risk factors for the world's most prevalent diseases such as neurodegenerative disorders, cancer, cardiovascular disease, and metabolic diseases. During the process of aging, a gradual loss of tissue volume and organ function is observed, which is partially caused by replicative senescence. The capacity of cellular proliferation and replicative senescence is tightly regulated by their telomere length. When telomere length is critically shortened with progressive cell division, cells become proliferatively arrested, and DNA damage response and cellular senescence are triggered, whereupon the "Hayflick limit" is attained at this stage. Podocytes are a cell type found in the kidney glomerulus where they have major roles in blood filtration. Mature podocytes are terminal differentiated cells that are unable to undergo cell division in vivo. For this reason, the establishment of primary podocyte cell cultures has been very challenging. In our present study, we present the successful immortalization of a human podocyte progenitor cell line, of which the primary cells were isolated directly from the urine of a 51-year-old male. The immortalized cell line was cultured over the course of one year (~100 passages) with high proliferation capacity, endowed with contact inhibition and P53 expression. Furthermore, by immunofluorescence-based expression and quantitative real-time PCR for the podocyte markers CD2AP, LMX1B, NPHS1, SYNPO and WT1, we confirmed the differentiation capacity of the immortalized cells. Finally, we evaluated and confirmed the responsiveness of the immortalized cells on the main mediator angiotensin II (ANGII) of the renin-angiotensin system (RAAS). In conclusion, we have shown that it is possible to bypass cellular replicative senescence (Hayflick limit) by TERT-driven immortalization of human urine-derived pre-podocyte cells from a 51-year-old African male.
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- 2023
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33. Urine-Based Detection of Biomarkers Indicative of Chronic Kidney Disease in a Patient Cohort from Ghana.
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Wruck W, Boima V, Erichsen L, Thimm C, Koranteng T, Kwakyi E, Antwi S, Adu D, and Adjaye J
- Abstract
Chronic kidney disease (CKD) is a global health burden with a continuously increasing prevalence associated with an increasing incidence of diabetes and hypertension in aging populations. CKD is characterized by low glomerular filtration rate (GFR) and other renal impairments including proteinuria, thus implying that multiple factors may contribute to the etiology this disease. While there are indications of ethnic differences, it is hard to disentangle these from confounding social factors. Usually, CKD is detected in later stages of the disease when irreversible renal damage has already occurred, thus suggesting a need for early non-invasive diagnostic markers. In this study, we explored the urine secretome of a CKD patient cohort from Ghana with 40 gender-matched patients and 40 gender-matched healthy controls employing a kidney injury and a more general cytokine assay. We identified panels of kidney-specific cytokine markers, which were also gender-specific, and a panel of gender-independent cytokine markers. The gender-specific markers are IL10 and MME for male and CLU, RETN, AGER, EGFR and VEGFA for female. The gender-independent cytokine markers were APOA1, ANGPT2, C5, CFD, GH1, ICAM1, IGFBP2, IL8, KLK4, MMP9 and SPP1 (up-regulated) and FLT3LG, CSF1, PDGFA, RETN and VEGFA (down-regulated). APOA1-the major component of HDL particles-was up-regulated in Ghanaian CKD patients and its co-occurrence with APOL1 in a subpopulation of HDL particles may point to specific CKD-predisposing APOL1 haplotypes in patients of African descent-this, however, needs further investigation. The identified panels, though preliminary, lay down the foundation for the development of robust CKD-diagnostic assays.
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- 2022
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34. Correction: Nguyen et al. The Nephrotoxin Puromycin Aminonucleoside Induces Injury in Kidney Organoids Differentiated from Induced Pluripotent Stem Cells. Cells 2022, 11 , 635.
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Nguyen L, Wruck W, Erichsen L, Graffmann N, and Adjaye J
- Abstract
In the original publication [...].
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- 2022
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35. Activation of the Renin-Angiotensin System Disrupts the Cytoskeletal Architecture of Human Urine-Derived Podocytes.
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Erichsen L, Thimm C, Bohndorf M, Rahman MS, Wruck W, and Adjaye J
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- Angiotensin II metabolism, Angiotensin II pharmacology, Humans, Kidney metabolism, Losartan pharmacology, Renin-Angiotensin System, Kidney Diseases metabolism, Podocytes metabolism
- Abstract
High blood pressure is one of the major public health problems that causes severe disorders in several tissues including the human kidney. One of the most important signaling pathways associated with the regulation of blood pressure is the renin-angiotensin system (RAS), with its main mediator angiotensin II (ANGII). Elevated levels of circulating and intracellular ANGII and aldosterone lead to pro-fibrotic, -inflammatory, and -hypertrophic milieu that causes remodeling and dysfunction in cardiovascular and renal tissues. Furthermore, ANGII has been recognized as a major risk factor for the induction of apoptosis in podocytes, ultimately leading to chronic kidney disease (CKD). In the past, disease modeling of kidney-associated diseases was extremely difficult, as the derivation of kidney originated cells is very challenging. Here we describe a differentiation protocol for reproducible differentiation of sine oculis homeobox homolog 2 (SIX2)-positive urine-derived renal progenitor cells (UdRPCs) into podocytes bearing typical cellular processes. The UdRPCs-derived podocytes show the activation of the renin-angiotensin system by being responsive to ANGII stimulation. Our data reveal the ANGII-dependent downregulation of nephrin ( NPHS1) and synaptopodin ( SYNPO) , resulting in the disruption of the podocyte cytoskeletal architecture, as shown by immunofluorescence-based detection of α-Actinin. Furthermore, we show that the cytoskeletal disruption is mainly mediated through angiotensin II receptor type 1 (AGTR1) signaling and can be rescued by AGTR1 inhibition with the selective, competitive angiotensin II receptor type 1 antagonist, losartan. In the present manuscript we confirm and propose UdRPCs differentiated to podocytes as a unique cell type useful for studying nephrogenesis and associated diseases. Furthermore, the responsiveness of UdRPCs-derived podocytes to ANGII implies potential applications in nephrotoxicity studies and drug screening.
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- 2022
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36. Impaired p53-Mediated DNA Damage Response Contributes to Microcephaly in Nijmegen Breakage Syndrome Patient-Derived Cerebral Organoids.
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Martins S, Erichsen L, Datsi A, Wruck W, Goering W, Chatzantonaki E, de Amorim VCM, Rossi A, Chrzanowska KH, and Adjaye J
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- DNA Damage, Humans, Organoids metabolism, Tumor Suppressor Protein p53 genetics, Tumor Suppressor Protein p53 metabolism, Microcephaly genetics, Nijmegen Breakage Syndrome genetics, Nijmegen Breakage Syndrome metabolism
- Abstract
Nijmegen Breakage Syndrome (NBS) is a rare autosomal recessive genetic disorder caused by mutations within nibrin ( NBN ), a DNA damage repair protein. Hallmarks of NBS include chromosomal instability and clinical manifestations such as growth retardation, immunodeficiency, and progressive microcephaly. We employed induced pluripotent stem cell-derived cerebral organoids from two NBS patients to study the etiology of microcephaly. We show that NBS organoids carrying the homozygous 657del5 NBN mutation are significantly smaller with disrupted cyto-architecture. The organoids exhibit premature differentiation, and Neuronatin (NNAT) over-expression. Furthermore, pathways related to DNA damage response and cell cycle are differentially regulated compared to controls. After exposure to bleomycin, NBS organoids undergo delayed p53-mediated DNA damage response and aberrant trans-synaptic signaling, which ultimately leads to neuronal apoptosis. Our data provide insights into how mutations within NBN alters neurogenesis in NBS patients, thus providing a proof of concept that cerebral organoids are a valuable tool for studying DNA damage-related disorders.
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- 2022
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37. The Nephrotoxin Puromycin Aminonucleoside Induces Injury in Kidney Organoids Differentiated from Induced Pluripotent Stem Cells.
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Nguyen L, Wruck W, Erichsen L, Graffmann N, and Adjaye J
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- Humans, Kidney, Organoids metabolism, Puromycin Aminonucleoside metabolism, Puromycin Aminonucleoside pharmacology, Acute Kidney Injury chemically induced, Acute Kidney Injury metabolism, Induced Pluripotent Stem Cells
- Abstract
Kidney diseases, including acute kidney injury (AKI) and chronic kidney disease (CKD), which can progress to end stage renal disease (ESRD), are a worldwide health burden. Organ transplantation or kidney dialysis are the only effective available therapeutic tools. Therefore, in vitro models of kidney diseases and the development of prospective therapeutic options are urgently needed. Within the kidney, the glomeruli are involved in blood filtration and waste excretion and are easily affected by changing cellular conditions. Puromycin aminonucleoside (PAN) is a nephrotoxin, which can be employed to induce acute glomerular damage and to model glomerular disease. For this reason, we generated kidney organoids from three iPSC lines and treated these with PAN in order to induce kidney injury. Morphological observations revealed the disruption of glomerular and tubular structures within the kidney organoids upon PAN treatment, which were confirmed by transcriptome analyses. Subsequent analyses revealed an upregulation of immune response as well as inflammatory and cell-death-related processes. We conclude that the treatment of iPSC-derived kidney organoids with PAN induces kidney injury mediated by an intertwined network of inflammation, cytoskeletal re-arrangement, DNA damage, apoptosis and cell death. Furthermore, urine-stem-cell-derived kidney organoids can be used to model kidney-associated diseases and drug discovery.
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- 2022
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38. Detailed phylogenetic analysis tracks transmission of distinct SARS-COV-2 variants from China and Europe to West Africa.
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Wruck W and Adjaye J
- Subjects
- Africa, Western, Biodiversity, China, Europe, Gambia, Genetic Variation, Genome, Viral, Geography, Ghana, Humans, Nigeria, Phylogeny, Senegal, Time Factors, COVID-19 transmission, COVID-19 virology, Computational Biology methods, Mutation, SARS-CoV-2
- Abstract
SARS-CoV-2, the virus causing the COVID-19 pandemic emerged in December 2019 in China and raised fears it could overwhelm healthcare systems worldwide. Mutations of the virus are monitored by the GISAID database from which we downloaded sequences from four West African countries Ghana, Gambia, Senegal and Nigeria from February 2020 to April 2020. We subjected the sequences to phylogenetic analysis employing the nextstrain pipeline. We found country-specific patterns of viral variants and supplemented that with data on novel variants from June 2021. Until April 2020, variants carrying the crucial Europe-associated D614G amino acid change were predominantly found in Senegal and Gambia, and combinations of late variants with and early variants without D614G in Ghana and Nigeria. In June 2021 all variants carried the D614G amino acid substitution. Senegal and Gambia exhibited again variants transmitted from Europe (alpha or delta), Ghana a combination of several variants and in Nigeria the original Eta variant. Detailed analysis of distinct samples revealed that some might have circulated latently and some reflect migration routes. The distinct patterns of variants within the West African countries point at their global transmission via air traffic predominantly from Europe and only limited transmission between the West African countries., (© 2021. The Author(s).)
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- 2021
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39. Transcriptome-based analysis of blood samples reveals elevation of DNA damage response, neutrophil degranulation, cancer and neurodegenerative pathways in Plasmodium falciparum patients.
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Karikari AA, Wruck W, and Adjaye J
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- Dried Blood Spot Testing, Malaria, Falciparum complications, Malaria, Falciparum parasitology, Neoplasms complications, Neurodegenerative Diseases complications, Cell Degranulation, DNA Damage, Malaria, Falciparum physiopathology, Neutrophils physiology, Plasmodium falciparum physiology, Transcriptome
- Abstract
Background: Malaria caused by Plasmodium falciparum results in severe complications including cerebral malaria (CM) especially in children. While the majority of falciparum malaria survivors make a full recovery, there are reports of some patients ending up with neurological sequelae or cognitive deficit., Methods: An analysis of pooled transcriptome data of whole blood samples derived from two studies involving various P. falciparum infections, comprising mild malaria (MM), non-cerebral severe malaria (NCM) and CM was performed. Pathways and gene ontologies (GOs) elevated in the distinct P. falciparum infections were determined., Results: In all, 2876 genes were expressed in common between the 3 forms of falciparum malaria, with CM having the least number of expressed genes. In contrast to other research findings, the analysis from this study showed MM share similar biological processes with cancer and neurodegenerative diseases, NCM is associated with drug resistance and glutathione metabolism and CM is correlated with endocannabinoid signalling and non-alcoholic fatty liver disease (NAFLD). GO revealed the terms biogenesis, DNA damage response and IL-10 production in MM, down-regulation of cytoskeletal organization and amyloid-beta clearance in NCM and aberrant signalling, neutrophil degranulation and gene repression in CM. Differential gene expression analysis between CM and NCM showed the up-regulation of neutrophil activation and response to herbicides, while regulation of axon diameter was down-regulated in CM., Conclusions: Results from this study reveal that P. falciparum-mediated inflammatory and cellular stress mechanisms may impair brain function in MM, NCM and CM. However, the neurological deficits predominantly reported in CM cases could be attributed to the down-regulation of various genes involved in cellular function through transcriptional repression, axonal dysfunction, dysregulation of signalling pathways and neurodegeneration. It is anticipated that the data from this study, might form the basis for future hypothesis-driven malaria research., (© 2021. The Author(s).)
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- 2021
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40. Human Induced Pluripotent Stem Cell-Derived Mesenchymal Stem Cells Acquire Rejuvenation and Reduced Heterogeneity.
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Wruck W, Graffmann N, Spitzhorn LS, and Adjaye J
- Abstract
Despite the uniform selection criteria for the isolation of human mesenchymal stem cells (MSCs), considerable heterogeneity exists which reflects the distinct tissue origins and differences between individuals with respect to their genetic background and age. This heterogeneity is manifested by the variabilities seen in the transcriptomes, proteomes, secretomes, and epigenomes of tissue-specific MSCs. Here, we review literature on different aspects of MSC heterogeneity including the role of epigenetics and the impact of MSC heterogeneity on therapies. We then combine this with a meta-analysis of transcriptome data from distinct MSC subpopulations derived from bone marrow, adipose tissue, cruciate, tonsil, kidney, umbilical cord, fetus, and induced pluripotent stem cells derived MSCs (iMSCs). Beyond that, we investigate transcriptome differences between tissue-specific MSCs and pluripotent stem cells. Our meta-analysis of numerous MSC-related data sets revealed markers and associated biological processes characterizing the heterogeneity and the common features of MSCs from various tissues. We found that this heterogeneity is mainly related to the origin of the MSCs and infer that microenvironment and epigenetics are key drivers. The epigenomes of MSCs alter with age and this has a profound impact on their differentiation capabilities. Epigenetic modifications of MSCs are propagated during cell divisions and manifest in differentiated cells, thus contributing to diseased or healthy phenotypes of the respective tissue. An approach used to reduce heterogeneity caused by age- and tissue-related epigenetic and microenvironmental patterns is the iMSC concept: iMSCs are MSCs generated from induced pluripotent stem cells (iPSCs). During iMSC generation epigenetic and chromatin remodeling result in a gene expression pattern associated with rejuvenation thus allowing to overcome age-related shortcomings (e.g., limited differentiation and proliferation capacity). The importance of the iMSC concept is underlined by multiple clinical trials. In conclusion, we propose the use of rejuvenated iMSCs to bypass tissue- and age-related heterogeneity which are associated with native MSCs., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Wruck, Graffmann, Spitzhorn and Adjaye.)
- Published
- 2021
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41. Generation of a Crigler-Najjar Syndrome Type I patient-derived induced pluripotent stem cell line CNS705 (HHUUKDi005-A).
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Graffmann N, Martins S, Ljubikj T, Matte JC, Bohndorf M, Wruck W, and Adjaye J
- Subjects
- Cell Differentiation, Cell Line, Cellular Reprogramming, Embryoid Bodies, Fibroblasts, Humans, Kruppel-Like Factor 4, Crigler-Najjar Syndrome, Induced Pluripotent Stem Cells
- Abstract
Human fibroblasts cells from a Crigler-Najjar Syndrome (CNS) patient were used to generate integration-free induced pluripotent stem cells (iPSCs) by over-expressing episomal-based plasmids expressing OCT4, SOX2, NANOG, KLF4, c-MYC and LIN28. The derived CNS705-iPSC line is homozygous for the UGT1A1 c.877_890delTACATTAATGCTTCinsA mutation. Pluripotency was confirmed by the expression of associated markers and embryoid body-based differentiation into cell types from all three germ layers. Comparative transcriptome analysis of the iPSC and the human embryonic stem cell line H9 revealed a Pearson's correlation of 0.9468., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)
- Published
- 2021
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42. The pioneer and differentiation factor FOXA2 is a key driver of yolk-sac tumour formation and a new biomarker for paediatric and adult yolk-sac tumours.
- Author
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Wruck W, Bremmer F, Kotthoff M, Fichtner A, Skowron MA, Schönberger S, Calaminus G, Vokuhl C, Pfister D, Heidenreich A, Albers P, Adjaye J, and Nettersheim D
- Subjects
- Age Factors, Cell Line, Tumor, DNA Methylation, Disease Susceptibility, Endodermal Sinus Tumor pathology, Humans, Immunohistochemistry, Models, Biological, Biomarkers, Tumor, Endodermal Sinus Tumor genetics, Endodermal Sinus Tumor metabolism, Hepatocyte Nuclear Factor 3-beta genetics, Hepatocyte Nuclear Factor 3-beta metabolism, Oncogene Proteins genetics, Oncogene Proteins metabolism
- Abstract
Yolk-sac tumours (YSTs), a germ cell tumour subtype, occur in newborns and infants as well as in young adults of age 14-44 years. In clinics, adult patients with YSTs face a poor prognosis, as these tumours are often therapy-resistant and count for many germ cell tumour related deaths. So far, the molecular and (epi)genetic mechanisms that control development of YST are far from being understood. We deciphered the molecular and (epi)genetic mechanisms regulating YST formation by meta-analysing high-throughput data of gene and microRNA expression, DNA methylation and mutational burden. We validated our findings by qRT-PCR and immunohistochemical analyses of paediatric and adult YSTs. On a molecular level, paediatric and adult YSTs were nearly indistinguishable, but were considerably different from embryonal carcinomas, the stem cell precursor of YSTs. We identified FOXA2 as a putative key driver of YST formation, subsequently inducing AFP, GPC3, APOA1/APOB, ALB and GATA3/4/6 expression. In YSTs, WNT-, BMP- and MAPK signalling-related genes were up-regulated, while pluripotency- and (primordial) germ cell-associated genes were down-regulated. Expression of FOXA2 and related key factors seems to be regulated by DNA methylation, histone methylation / acetylation and microRNAs. Additionally, our results highlight FOXA2 as a promising new biomarker for paediatric and adult YSTs., (© 2021 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.)
- Published
- 2021
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43. A stem cell based in vitro model of NAFLD enables the analysis of patient specific individual metabolic adaptations in response to a high fat diet and AdipoRon interference.
- Author
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Graffmann N, Ncube A, Martins S, Fiszl AR, Reuther P, Bohndorf M, Wruck W, Beller M, Czekelius C, and Adjaye J
- Subjects
- Adiponectin metabolism, Biomarkers, Cell Differentiation, Cells, Cultured, Gene Expression Regulation, Gluconeogenesis, Hepatocytes metabolism, Humans, Induced Pluripotent Stem Cells cytology, Induced Pluripotent Stem Cells metabolism, Lipid Metabolism, Non-alcoholic Fatty Liver Disease pathology, Piperidines therapeutic use, Signal Transduction, Adaptation, Biological, Diet, High-Fat, Energy Metabolism, Non-alcoholic Fatty Liver Disease etiology, Non-alcoholic Fatty Liver Disease metabolism, Piperidines pharmacology, Stem Cells metabolism
- Abstract
Non-alcoholic fatty liver disease (NAFLD) is a multifactorial disease. Its development and progression depend on genetically predisposed susceptibility of the patient towards several 'hits' that induce fat storage first and later inflammation and fibrosis. Here, we differentiated induced pluripotent stem cells (iPSCs) derived from four distinct donors with varying disease stages into hepatocyte like cells (HLCs) and determined fat storage as well as metabolic adaptations after stimulations with oleic acid. We could recapitulate the complex networks that control lipid and glucose metabolism and we identified distinct gene expression profiles related to the steatosis phenotype of the donor. In an attempt to reverse the steatotic phenotype, cells were treated with the small molecule AdipoRon, a synthetic analogue of adiponectin. Although the responses varied between cells lines, they suggest a general influence of AdipoRon on metabolism, transport, immune system, cell stress and signalling., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2021. Published by The Company of Biologists Ltd.)
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- 2021
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44. SARS-CoV-2 receptor ACE2 is co-expressed with genes related to transmembrane serine proteases, viral entry, immunity and cellular stress.
- Author
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Wruck W and Adjaye J
- Subjects
- Angiotensin-Converting Enzyme 2 genetics, COVID-19 pathology, Computational Biology, Humans, Interleukin-1beta immunology, Interleukin-33 immunology, Interleukin-6 immunology, Interleukins immunology, Lung cytology, Membrane Proteins genetics, Nuclear Proteins immunology, Phosphoproteins immunology, Reactive Oxygen Species metabolism, Receptors, Virus genetics, Receptors, Virus metabolism, Respiratory Mucosa cytology, Serine Endopeptidases genetics, Transcriptome genetics, Angiotensin-Converting Enzyme 2 metabolism, Epithelial Cells metabolism, Membrane Proteins metabolism, Respiratory Mucosa metabolism, SARS-CoV-2 metabolism, Serine Endopeptidases metabolism
- Abstract
The COVID-19 pandemic resulting from the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) which emerged in December 2019 in Wuhan in China has placed immense burden on national economies and global health. At present neither vaccination nor therapies are available. Here, we performed a meta-analysis of RNA-sequencing data from three studies employing human lung epithelial cells. Of these one focused on lung epithelial cells infected with SARS-CoV-2. We aimed at identifying genes co-expressed with angiotensin I converting enzyme 2 (ACE2) the human cell entry receptor of SARS-CoV-2, and unveiled several genes correlated or inversely correlated with high significance, among the most significant of these was the transmembrane serine protease 4 (TMPRSS4). Serine proteases are known to be involved in the infection process by priming the virus spike protein. Pathway analysis revealed virus infection amongst the most significantly correlated pathways. Gene Ontologies revealed regulation of viral life cycle, immune responses, pro-inflammatory responses- several interleukins such as IL6, IL1, IL20 and IL33, IFI16 regulating the interferon response to a virus, chemo-attraction of macrophages, and cellular stress resulting from activated Reactive Oxygen Species. We believe that this dataset will aid in a better understanding of the molecular mechanism(s) underlying COVID-19.
- Published
- 2020
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45. IPSC-Derived Neuronal Cultures Carrying the Alzheimer's Disease Associated TREM2 R47H Variant Enables the Construction of an Aβ-Induced Gene Regulatory Network.
- Author
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Martins S, Müller-Schiffmann A, Erichsen L, Bohndorf M, Wruck W, Sleegers K, Van Broeckhoven C, Korth C, and Adjaye J
- Subjects
- Aged, Alzheimer Disease metabolism, Amyloid beta-Peptides genetics, Amyloid beta-Peptides metabolism, Brain metabolism, Cell Differentiation genetics, Endoplasmic Reticulum genetics, Endoplasmic Reticulum metabolism, Female, Gene Regulatory Networks genetics, Genetic Predisposition to Disease genetics, Genetic Variation genetics, Humans, Male, Membrane Glycoproteins metabolism, Models, Biological, Mutation genetics, Myeloid Cells metabolism, Neurons metabolism, Pluripotent Stem Cells metabolism, Pluripotent Stem Cells physiology, Receptors, Immunologic metabolism, Alzheimer Disease genetics, Membrane Glycoproteins genetics, Receptors, Immunologic genetics
- Abstract
Genes associated with immune response and inflammation have been identified as genetic risk factors for late-onset Alzheimer´s disease (LOAD). The rare R47H variant within triggering receptor expressed on myeloid cells 2 (TREM2) has been shown to increase the risk for developing Alzheimer's disease (AD) 2-3-fold. Here, we report the generation and characterization of a model of late-onset Alzheimer's disease (LOAD) using lymphoblast-derived induced pluripotent stem cells (iPSCs) from patients carrying the TREM2 R47H mutation, as well as from control individuals without dementia. All iPSCs efficiently differentiated into mature neuronal cultures, however AD neuronal cultures showed a distinct gene expression profile. Furthermore, manipulation of the iPSC-derived neuronal cultures with an Aβ-S8C dimer highlighted metabolic pathways, phagosome and immune response as the most perturbed pathways in AD neuronal cultures. Through the construction of an Aβ-induced gene regulatory network, we were able to identify an Aβ signature linked to protein processing in the endoplasmic reticulum (ER), which emphasized ER-stress, as a potential causal role in LOAD. Overall, this study has shown that our AD-iPSC based model can be used for in-depth studies to better understand the molecular mechanisms underlying the etiology of LOAD and provides new opportunities for screening of potential therapeutic targets.
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- 2020
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46. Transcriptomic analysis of marine endophytic fungi extract identifies highly enriched anti-fungal fractions targeting cancer pathways in HepG2 cell lines.
- Author
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Blessie EJ, Wruck W, Abbey BA, Ncube A, Graffmann N, Amarh V, Arthur PK, and Adjaye J
- Subjects
- Antifungal Agents chemistry, Antineoplastic Agents chemistry, Apoptosis drug effects, Candida albicans drug effects, Cell Proliferation drug effects, Hep G2 Cells, Humans, Microbial Sensitivity Tests, Protein Interaction Maps, Saccharomyces cerevisiae drug effects, Seaweed chemistry, Antifungal Agents pharmacology, Antineoplastic Agents pharmacology, Endophytes chemistry, Transcriptome genetics
- Abstract
Background: Marine endophytic fungi (MEF) are good sources of structurally unique and biologically active secondary metabolites. Due to the increase in antimicrobial resistance, the secondary metabolites from MEF ought to be fully explored to identify candidates which could serve as lead compounds for novel drug development. These secondary metabolites might also be useful for development of new cancer drugs. In this study, ethyl acetate extracts from marine endophytic fungal cultures were tested for their antifungal activity and anticancer properties against C. albicans and the human liver cancer cell line HepG2, respectively. The highly enriched fractions were also analyzed by high performance liquid chromatography coupled with high resolution mass spectrometry (HPLC-HRMS) and their effect on the HepG2 cells was assessed via transcriptomics and with a proliferation assay., Results: We demonstrated that the fractions could reduce proliferation in HepG2 cells. The detailed transcriptome analysis revealed regulation of several cancer- and metabolism-related pathways and gene ontologies. The down-regulated pathways included, cell cycle, p53 signaling, DNA replication, sphingolipid metabolism and drug metabolism by cytochrome P450. The upregulated pathways included HIF-1 signaling, focal adhesion, necroptosis and transcriptional mis-regulation of cancer. Furthermore, a protein interaction network was constructed based on the 26 proteins distinguishing the three treatment conditions from the untreated cells. This network was composed of central functional components associated with metabolism and cancer such as TNF, MAPK, TRIM21 and one component contained APP., Conclusions: The purified fractions from MEF investigated in this study showed antifungal activity against C. albicans and S. cerevisiae alone or both and reduced proliferation of the human liver cancer cell line HepG2 implicating regulation of several cancer- and metabolism-related pathways. The data from this study could be instrumental in identifying new pathways associated with liver cancer anti-proliferative processes which can be used for the development of novel antifungal and anti-cancer drugs.
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- 2020
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47. Meta-analysis of human prefrontal cortex reveals activation of GFAP and decline of synaptic transmission in the aging brain.
- Author
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Wruck W and Adjaye J
- Subjects
- Adult, Age Factors, Aged, Aging metabolism, Cadmium, Calcium-Calmodulin-Dependent Protein Kinase Type 4 genetics, Catecholamines metabolism, Cluster Analysis, Cyclic AMP Response Element-Binding Protein genetics, Dendritic Spines metabolism, Extracellular Signal-Regulated MAP Kinases genetics, Female, Gene Expression Profiling, Gene Ontology, Glial Fibrillary Acidic Protein genetics, Humans, Inflammation genetics, Male, Middle Aged, Morphogenesis genetics, Neurogenesis genetics, Principal Component Analysis, Sex Factors, Signal Transduction genetics, Synaptic Transmission genetics, Transcriptome, Zinc, Aging genetics, Prefrontal Cortex metabolism
- Abstract
Despite ongoing research efforts, mechanisms of brain aging are still enigmatic and need to be elucidated for a better understanding of age-associated cognitive decline. The aim of this study is to investigate aging in the prefrontal cortex region of human brain in a meta-analysis of transcriptome datasets. We analyzed 591 gene expression datasets pertaining to female and male human prefrontal cortex biopsies of distinct ages. We used hierarchical clustering and principal component analysis (PCA) to determine the influence of sex and age on global transcriptome levels. In sex-specific analysis we identified genes correlating with age and differentially expressed between groups of young, middle-aged and aged. Pathways and gene ontologies (GOs) over-represented in the resulting gene sets were calculated. Potential causal relationships between genes and between GOs were explored employing the Granger test of gene expression time series over the range of ages. The most outstanding results were the age-related decline of synaptic transmission and activated expression of glial fibrillary acidic protein (GFAP) in both sexes. We found an antagonistic relationship between calcium/calmodulin dependent protein kinase IV (CAMK4) and GFAP which may include regulatory mechanisms involving cAMP responsive element binding protein (CREB) and mitogen-activated protein kinase (MAPK, alias ERK). Common to both sexes was a decline in synaptic transmission, neurogenesis and an increased base-level of inflammatory and immune-related processes. Furthermore, we detected differences in dendritic spine morphogenesis, catecholamine signaling and cellular responses to external stimuli, particularly to metal (Zinc and cadmium) ions which were higher in female brains.
- Published
- 2020
- Full Text
- View/download PDF
48. Functional omics analyses reveal only minor effects of microRNAs on human somatic stem cell differentiation.
- Author
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Schira-Heinen J, Czapla A, Hendricks M, Kloetgen A, Wruck W, Adjaye J, Kögler G, Werner Müller H, Stühler K, and Trompeter HI
- Subjects
- Cell Differentiation, Cell Proliferation, Chromatography, Liquid, Computational Biology, Gene Expression Profiling, Gene Expression Regulation, High-Throughput Nucleotide Sequencing, Humans, Peptides chemistry, Phenotype, Proteome, Proteomics, Tandem Mass Spectrometry, Transcriptome, Fetal Blood cytology, MicroRNAs metabolism, Stem Cells cytology, Tretinoin pharmacology
- Abstract
The contribution of microRNA-mediated posttranscriptional regulation on the final proteome in differentiating cells remains elusive. Here, we evaluated the impact of microRNAs (miRNAs) on the proteome of human umbilical cord blood-derived unrestricted somatic stem cells (USSC) during retinoic acid (RA) differentiation by a systemic approach using next generation sequencing analysing mRNA and miRNA expression and quantitative mass spectrometry-based proteome analyses. Interestingly, regulation of mRNAs and their dedicated proteins highly correlated during RA-incubation. Additionally, RA-induced USSC demonstrated a clear separation from native USSC thereby shifting from a proliferating to a metabolic phenotype. Bioinformatic integration of up- and downregulated miRNAs and proteins initially implied a strong impact of the miRNome on the XXL-USSC proteome. However, quantitative proteome analysis of the miRNA contribution on the final proteome after ectopic overexpression of downregulated miR-27a-5p and miR-221-5p or inhibition of upregulated miR-34a-5p, respectively, followed by RA-induction revealed only minor proportions of differentially abundant proteins. In addition, only small overlaps of these regulated proteins with inversely abundant proteins in non-transfected RA-treated USSC were observed. Hence, mRNA transcription rather than miRNA-mediated regulation is the driving force for protein regulation upon RA-incubation, strongly suggesting that miRNAs are fine-tuning regulators rather than active primary switches during RA-induction of USSC.
- Published
- 2020
- Full Text
- View/download PDF
49. The FGF, TGFβ and WNT axis Modulate Self-renewal of Human SIX2 + Urine Derived Renal Progenitor Cells.
- Author
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Rahman MS, Wruck W, Spitzhorn LS, Nguyen L, Bohndorf M, Martins S, Asar F, Ncube A, Erichsen L, Graffmann N, and Adjaye J
- Subjects
- Cell Differentiation genetics, Female, Fibroblast Growth Factor 2 metabolism, Gene Expression, Humans, Male, Pluripotent Stem Cells metabolism, Transcriptome, Transforming Growth Factor beta metabolism, Cell Self Renewal genetics, Cell Self Renewal physiology, Fibroblast Growth Factor 2 physiology, Kidney cytology, Pluripotent Stem Cells physiology, Signal Transduction genetics, Signal Transduction physiology, Transforming Growth Factor beta physiology, Urine cytology, Wnt Signaling Pathway genetics, Wnt Signaling Pathway physiology
- Abstract
Human urine is a non-invasive source of renal stem cells with regeneration potential. Urine-derived renal progenitor cells were isolated from 10 individuals of both genders and distinct ages. These renal progenitors express pluripotency-associated proteins- TRA-1-60, TRA-1-81, SSEA4, C-KIT and CD133, as well as the renal stem cell markers -SIX2, CITED1, WT1, CD24 and CD106. The transcriptomes of all SIX2
+ renal progenitors clustered together, and distinct from the human kidney biopsy-derived epithelial proximal cells (hREPCs). Stimulation of the urine-derived renal progenitor cells (UdRPCs) with the GSK3β-inhibitor (CHIR99021) induced differentiation. Transcriptome and KEGG pathway analysis revealed upregulation of WNT-associated genes- AXIN2, JUN and NKD1. Protein interaction network identified JUN- a downstream target of the WNT pathway in association with STAT3, ATF2 and MAPK1 as a putative negative regulator of self-renewal. Furthermore, like pluripotent stem cells, self-renewal is maintained by FGF2-driven TGFβ-SMAD2/3 pathway. The urine-derived renal progenitor cells and the data presented should lay the foundation for studying nephrogenesis in human.- Published
- 2020
- Full Text
- View/download PDF
50. FGF Signalling in the Self-Renewal of Colon Cancer Organoids.
- Author
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Otte J, Dizdar L, Behrens B, Goering W, Knoefel WT, Wruck W, Stoecklein NH, and Adjaye J
- Subjects
- Aged, Aged, 80 and over, Cell Differentiation drug effects, Cell Differentiation genetics, Cell Proliferation drug effects, Colonic Neoplasms genetics, Colonic Neoplasms metabolism, Female, Gene Expression Regulation, Neoplastic drug effects, Humans, Male, Middle Aged, Neoplasm Staging, Neoplastic Stem Cells drug effects, Neoplastic Stem Cells pathology, Neoplastic Stem Cells physiology, Organoids metabolism, Organoids pathology, Primary Cell Culture, Signal Transduction drug effects, Spheroids, Cellular drug effects, Spheroids, Cellular metabolism, Spheroids, Cellular pathology, Tumor Cells, Cultured, Cell Self Renewal drug effects, Colonic Neoplasms pathology, Fibroblast Growth Factor 2 pharmacology, Organoids drug effects
- Abstract
The progression of colorectal cancer (CRC) is supposedly driven by cancer stem cells (CSC) which are able to self-renew and simultaneously fuel bulk tumour mass with highly proliferative and differentiated tumour cells. However, the CSC-phenotype in CRC is unstable and dependent on environmental cues. Fibroblast growth factor 2 (FGF2) is essential and necessary for the maintenance of self-renewal in adult and embryonic stem cells. Investigating its role in self-renewal in advanced CRC patient-derived organoids, we unveiled that FGF-receptor (FGFR) inhibition prevents organoid formation in very early expanding cells but induces cyst formation when applied to pre-established organoids. Comprehensive transcriptome analyses revealed that the induction of the transcription factor activator-protein-1 (AP-1) together with MAPK activation was most prominent after FGFR-inhibition. These effects resemble mechanisms of an acquired resistance against other described tyrosine kinase inhibitors such as EGF-receptor targeted therapies. Furthermore, we detected elevated expression levels of several self-renewal and stemness-associated genes in organoid cultures with active FGF2 signalling. The combined data assume that CSCs are a heterogeneous population while self-renewal is a common feature regulated by distinct but converging pathways. Finally, we highlight FGF2 signalling as one of numerous components of the complex regulation of stemness in cancer.
- Published
- 2019
- Full Text
- View/download PDF
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