Spelling suggestions: "subject:"epigenetic"" "subject:"epigenetics""
51 |
Analysis of DNA methylation changes and behavioural outcomes in adulthood induced by prenatal exposure to a mixture of endocrine disrupting chemicalsKamil, Shane January 2022 (has links)
Endocrine disrupting chemicals, or EDCs, are some of the most prevalent toxic chemicals found in the environment because of human activity and they have a variety of adverse effects on both humans and wildlife. A proposed mechanism through which EDCs can negatively affect an organism is via an epigenetic mechanism known as DNA methylation, which can affect the development of the organism with negative outcomes later in life. The aim of this project was to investigate the effect of a prenatal exposure to mixture of EDCs, called Mixture N1, and its adverse effects. Mixture N1 has in a previous study been detected in the first semester of mothers, and linked to language delay in the offspring in the SELMA cohort study. We investigated relationships with DNA methylation pattern changes in key genes with exposure, gene expression and behaviour in the adult brains of the exposed mice. Our results showed correlative as well as linear relationships between methylation and different behavioural outcomes for target genes. One gene in particular - Nr3c1 - stood out among the results, having links to both stress and sociability. Specifically, DNA methylation of this gene correlates to active stress coping behaviours as well as sociability, but with no mediation component in these relationships. These results are promising for the use of methylation analysis as a biomarker of EDC mixture exposure, but more so as a predictor of negative behavioural outcomes later in life. More research could strengthen this use, and uncover the mechanism through which methylation alone might affect changes in behaviour.
|
52 |
Evaluation of Epigenetic Biomarkers in Primary and Iatrogenic Immune DeficienciesSchulze, Janika 03 December 2021 (has links)
Ein neuartiger Ansatz für die Immunphänotypisierung wird vorgestellt. Die Durchflusszytometrie (FACS) ist die übliche Methode für die Charaketrisierung des Immunsystems. Jedoch ist die Verfügbarkeit von frischem Vollblut, sowie ein schnelle Probenlogistik Vorraussetzung für die Analyse. Als potentialle Alternative werden epigentische qPCR Assays vorgestellt.
Für die Quantifizierung von B- und NK-Zellen wurden epigenetische qPCR Systeme etabliert. Anhand eines erweiterten epigenetischen Markerpanels wurde die klinische Anwendung in drei Kohorten getestet: a) 41 Patienten mit primären Immundefizienzen (PID); b) 19 Neugeborene mit und ohne PID und c) 28 Patienten nach einer Stammzelltransplantation (SZT).
In Kohorte a) und c) konnte die Äquivalenz der Ergebnisse mit FACS bestätigt werden. Diskrepanzen bei der regulatorischen T-Zell Quantifizierung in einzelnen PID Patienten wurde festgestellt, welche durch Mutationen verursacht wurden, die die Integrität der analysierten Proteine beeinflussen. Zudem konnte die Anwendung der epigentischen Quantifizierung in Trockenblutkarten von Neugeborenen gezeigt werden. Dies würde die Anwendung auch im Neugeborenen-Screening für die Erkennung von PIDs ermöglichen.
Für die Anwendung in der SZT konnte gezeigt werden, dass das epigenetische System eine frühe Analyse der Immunrekonstitution ermöglicht, welche eine prädiktive Aussage über das Überleben der Patienten erlaubt. Patienten, welche eine Immunantwort der Lymphozyten gegen eine Virusinfektion bereits am Tag 26 nach Transplantation aufwiesen, hatten eine signifikant höhere Überlebenschance als Patienten ohne Immunantwort.
Zusammengefassend zeigen die Daten, dass die epigenetische Systeme für klinische Anwendungen eine zuverlässige Methode darstellt. Die Aussagekraft der Daten ist aufgrund der Studiengröße noch limitiert, und komplizierte klinische Szenarien erschweren die Evaluierung. Deshalb sind weitere Studien erforderlich, um das gezeigte Potenzial zu validieren. / A novel approach for immunophenotyping for clinical applications is presented here. Flow cytometry is currently a common method to characterize the immune system but requiring fresh whole blood and good sample logistics which is not always available. To overcome this limitations, epigenetic qPCR assays are introduced as potential alternative.
New epigenetic qPCR systems to quantiy B and NK cells have been established. Using an extended epigenetic marker panel, clinical applications were tested in three patient cohorts: a) 41 patients with different primary immunodeficiencies (PID); b) 19 newborns with and without PID and c) 28 patients after stem cell transplantation (SCT).
In cohort a) and c) the equivalence of the epigenetic quantification with flow cytometry was confirmed. However, discrepancies between both methods for regulatory T-cell quantification were found in individual PID patients caused by disease-associated mutations affecting the integrity of the respective protein. Furthermore, the epigenetic quantification using dried blood spots from newborns was demonstrated. This would allow the implementation of epigenetic immunophenotyping in neonatal screening for the detection of congenital immunodeficiencies.
For the application in SCT, it was shown that the epigenetic system allows an early analysis of immune reconstitution, which may allow a prediction of the patients' overall survival. Patients who showed an immune response of lymphocytes against viral infections at day 26 after transplantation had a significantly higher survival rate.
In summary, the available data show that epigenetic immunophenotyping is a reliable analytical method for various clinical applications. The significance of the data is still limited due to the size of the study and complicated clinical scenarios make the evaluation of individual measurements difficult. Therefore, further extensive investigations are needed to clinically validate the demonstrated potential.
|
53 |
Algorithms for non-coding transcriptome analysis and their application to study the germ-layers developmentHita Ardiaca, Andrea 09 July 2024 (has links)
Next-generation sequencing (NGS) ermöglicht das molekulare Profiling von Zellen mit beispiellos hohem Durchsatz. Allerdings ist der Fokus oftmals auf proteinkodierende Proteine beschränkt, wodurch die vollständige Diversität des Transkriptoms übersehen wird. Nicht-kodierende RNA-Moleküle variieren stark in ihrer Biogenese, Struktur und Funktion, wodurch ihre unverzerrte Inklusion in die Analyse erschwert wird. Diese Promotion fokussiert sich auf das Verständnis nicht-kodierender RNA und navigiert durch drei aufeinander aufbauende Säulen in der Analyse, um Beobachtungen in Wissen zu verwandeln: Generierung von Daten, Quantifizierung und Interpretation. Diese drei Säulen werden in den drei Kapiteln der Dissertation aus der bioinformatischen Perspektive adressiert, indem Schlüsselherausforderungen beschrieben und neue Lösungen vorgestellt werden, um die Analyse des gesamten Transkriptoms mit NGS-Techniken zu verbessern. Zunächst wird ein vollautomatischer Algorithmus vorgestellt, welcher die verschiedenen Quellen von aus der Vorberei- tung von Bibliotheken resultierenden Artefakten mittels unüberwachtes Lernen erkennt, was anschließend zur Optimierung der Protokolle zur Vorbereitung von total-RNA-seq-Bibliotheken genutzt werden kann. Zudem werden die primären Herausforderungen der Quantifizierung von total-RNA-seq behandelt: die Prozessierung von Reads, die mehreren, möglicherweise überlappenden Loci zugeordnet werden können, wie auch die Tatsache, dass manche Loci mehrfach im Genom vorkommen und ein Read zu all diesen Loci passen kann. Diese beiden Fälle können auch gleichzeitig vorkommen, was die Analyse von nicht-kodierender RNA mit üblichen Methoden erschwert. Um diese Problematik anzugehen, wird eine neue Software namens Multi-Graph count (MGcount) vorgestellt. Diese ordnet hierarchisch Reads Transkripten zu, um unter anderem eine Diskrepanz zwischen der Loci-Länge von small und long RNA zu berücksichtigen. Wenn Reads konsistent mehrfach alignieren, fasst MGcount Loci in Communitys zusammen. Es wird gezeigt, dass die Beurteilung der Expression auf der Community-Ebene eine genauere Quantifizierung von biologisch bedeutsamen RNA-Einheiten (Einfachtranskript oder Locusfamilien) ermöglicht. Schließlich wird MGcount angewandt, um nicht-kodierende RNA während der Differenzierung von induzierten pluripotenten Stammzellen in die Keimblätter Mesoderm, Endoderm und Ektoderm zu analysieren. In dieser Dissertation wird eine Multi-Omics-Analyse erfolgreich angewandt, um sowohl die Expressionsverläufe von verschiedenen RNA-Biotypen während der Determination zu charakterisieren als auch einen Zusammenhang bezüglich Chromatin-Remodellierung (“chromatin remodeling“) und DNA-Methylierung an den jeweiligen Loci herzustellen. Schlussendlich dient diese Dissertation als Ratgeber für alle Forschenden, die neue Einsichten in das nicht-kodierende Transkriptom gewinnen wollen. / Next-generation sequencing (NGS) techniques enable the molecular profiling of cells with unprecedented high throughput. Yet, in transcriptome analysis, the focus is often restricted to protein-coding RNA, overlooking the transcriptome in its entire diversity. Non-coding RNA molecules largely vary in biogenesis, structure and function and this challenges their unbiased inclusion into the analyses. This doctoral research places non-coding RNA understanding at the focus spot and navigates through the three workflow pillars that must align effectively to turn observations into knowledge: data generation, quantification, and interpretation. Throughout three chapters, this Thesis addresses these pillars from a Bioinformatics perspective, by outlining key challenges and introducing novel solutions to improve whole-transcriptome analysis through NGS techniques. First, we introduce a fully automatic algorithm that identifies sources of library preparation artifacts in an unsupervised manner and we demonstrate its utility within the development and optimization of total-RNA-seq library preparation protocols. Secondly, we address a major challenge in total-RNA-seq quantification; processing reads that align to multiple loci that overlap within the same genomic region or/and multiple loci that are present in high copy numbers. Such ambiguous alignments commonly arise due to the inherent characteristics of non-coding RNA. To tackle this, we introduce a novel software, named Multi-Graph count (MGcount), that hierarchically assigns reads to transcripts to account for loci length disparity between small-RNA and long-RNA and subsequently collapses loci where reads consistently multi-map into communities defined in a data-driven fashion. We show that these cohesive communities allow the quantification of biologically meaningful RNA entities (single-transcripts or locus-families) and estimate their abundance more accurately. Finally, we apply the developed method to investigate non-coding RNA in early development, specifically during the differentiation of Induced Pluripotent Stem Cells into the three germ-layer lineages, namely, mesoderm, endoderm, and ectoderm. In this study, we leverage a multi-omics analysis to characterize the expression trajectories of diverse RNA biotypes along cell-commitment and the interplay with chromatin remodeling and DNA methylation patterns at the locus surroundings. Ultimately, this work is intended to serve as a guide for all those who want to gain new insights from the non-coding transcriptome.
|
54 |
Die Einzelnen und ihre Energie: Der Blick auf den Menschen in der Sicht der Wissenschaft Das Familienstellen, die Verschränkung und die EpigenetikFischer, Ernst Peter 29 January 2019 (has links)
Das Familienstellen, das Stellen von Familienkonstellationen, das auch als System-
Aufstellung bezeichnet wird, stellt ein therapeutisches Verfahren dar, das seit den 1970er Jahren immer mehr Zuspruch und Anwendung in der Psychiatrie findet und inzwischen auch in Unternehmen eingesetzt wird, um Entscheidungen in komplexen Situationen und in sich permanent wandelnden Kontexten zu treffen oder den Sand im Getriebe ausfindig zu machen, der die Betriebsabläufe stört. Der vielfach angemerkte Erfolg des Familienstellens bringt die Herausforderung von wissenschaftlichen Erklärungen mit sich, wobei in diesem Beitrag Vorschläge gemacht werden, die sich vor allem in der Quantenphysik umschauen und bei der Epigenetik bedienen. Es gehört zu den spannenden Fragen der Gegenwart, wie man „Von der Quantenphysik zum Bewusstsein“ und damit zu den Einflüssen der Familienkonstellation auf
den Einzelnen in der Gruppe kommt. Eine wichtige Rolle spielt dabei das Konzept der Energie, deren Eigenschaft, unzerstörbar zu sein, mehr Aufmerksamkeit im humanen Bereich verdient, als ihr bisher zugestanden wird.
|
55 |
Analysis of an epigenetic regulator in mouse embryonic stem cell self-renewal and differentiation / Analyse eines epigenetischen Regulators bei der Selbsterneuerung und Differenzierung muriner embryonaler StammzellenLubitz, Sandra 10 January 2006 (has links) (PDF)
Mammals have two orthologs, Mll and Trx2, for the Drososphila protein Trithorax (TRX), which is the founding member of the trithorax group (TrxG) of epigenetic regulators. TrxG proteins are characterized by an evolutionary conserved SET domain. A major function of all SET domain- containing proteins is to modulate gene activity, but the underlying mechanisms are poorly understood. Apparently TRX, Mll and Trx2 are histone H3 lysine 4 specific methyltransferases. So far all evidence points to roles in expression of specific target genes. However, target genes and function of the epigenetic regulator Trx2 were still unknown. Homozygous trx2 mutant embryos arrest in development because of severe and widespread defects {Glaser, 2005 #296}. Thus mouse embryonic stem (ES) cells carrying a null mutation of trx2 were used as an alternative model system to address the implication of Trx2 in differentiation. This study showed that Trx2 is redundant for ES cell self-renewal. Homozygous trx2 knockout ES cells did not exhibit cell cycle defects. However, loss of Trx2 resulted in reduced proliferation and increased apoptosis rates in trx2-/- ES cells. Due to the fact that differentiation requires an appropriate rate of population growth, trx2-/- cells were affected adversely upon in vitro differentiation. Neurogeneic differentiation of trx2 mutant cells generated fewer mature neurons than wild type cells. Moreover a temporal delay in the developmental progression to differentiation became apparent. Cardiac differentiation of trx2-/- cells confirmed the developmental defect and temporal delay. Notably differentiation of trx2-/- cells was merely delayed or impaired but it was not absent, implying that Trx2 is not required for gene expression programs specific for neurons or cardiac myocytes. We propose that differentiation of trx2-/- ES cells is impaired because apoptosis is disturbing differentiation. Apart from analyzing the phenotype of trx2 mutant cells, this work was focused on the identification of Trx2 target genes. Oligonucleotide expression arrays were used to identify genes whose expression levels were affected by the absence of Trx2. In general, loss of Trx2 function resulted in more genes with decreased than increased expression levels. This is consistent with the hypothesis that Trx2 functions as a transcriptional activator. Comparison of gene expression profiles for constitutive and conditional trx2 mutant cells enabled a distinction between direct and indirect target genes for Trx2. As a result Magoh2 was identified as the key candidate target gene for Trx2. Interaction between Trx2 and Magoh2 suggested a potential regulatory role for Trx2 in alternative splicing. Furthermore this work provided evidence that Trx2 could be involved in the maintenance of CpG island promoter gene expression, thus providing a potent regulatory mechanism for ubiquitously expressed genes.
|
56 |
Investigations into the regulation of histone H2B monoubiquitination / Investigations into the regulation of histone H2B monoubiquitinationShchebet, Andrei 18 April 2011 (has links)
No description available.
|
57 |
Die Funktionsanalyse und Pharmakomodulation des Amyloid-Vorläufer-Proteins (APP) in vitro und in vivo - Eine neue Zielstruktur zur Behandlung maligner Tumore / The functional analysis and pharmacomodulation of the β-amyloid precursor protein (APP) in vitro and in vivo - a novel molecular target for cancer therapyVenkataramani, Vivek 05 March 2012 (has links)
No description available.
|
58 |
Pluripotency of multipotent adult germ-line stem cells: analysis of apoptotic and epigenetic features / Pluripotenz der multipotenten adulten Keimstammzellen: Analyse der apoptotischer und epigenetischer MerkmaleKhromov, Tatjana 29 November 2011 (has links)
No description available.
|
59 |
Epigenetische Suppression von RASAL1 und ATP2A2 als Biomarker und Therapieansatz bei kardialer Fibrogenese / Epigenetic suppression of RASAL1 and ATP2A2 as biomarker and therapeutic approach in cardiac fibrosisGosch, Jonas 12 July 2021 (has links)
No description available.
|
60 |
Analysis of an epigenetic regulator in mouse embryonic stem cell self-renewal and differentiationLubitz, Sandra 06 December 2005 (has links)
Mammals have two orthologs, Mll and Trx2, for the Drososphila protein Trithorax (TRX), which is the founding member of the trithorax group (TrxG) of epigenetic regulators. TrxG proteins are characterized by an evolutionary conserved SET domain. A major function of all SET domain- containing proteins is to modulate gene activity, but the underlying mechanisms are poorly understood. Apparently TRX, Mll and Trx2 are histone H3 lysine 4 specific methyltransferases. So far all evidence points to roles in expression of specific target genes. However, target genes and function of the epigenetic regulator Trx2 were still unknown. Homozygous trx2 mutant embryos arrest in development because of severe and widespread defects {Glaser, 2005 #296}. Thus mouse embryonic stem (ES) cells carrying a null mutation of trx2 were used as an alternative model system to address the implication of Trx2 in differentiation. This study showed that Trx2 is redundant for ES cell self-renewal. Homozygous trx2 knockout ES cells did not exhibit cell cycle defects. However, loss of Trx2 resulted in reduced proliferation and increased apoptosis rates in trx2-/- ES cells. Due to the fact that differentiation requires an appropriate rate of population growth, trx2-/- cells were affected adversely upon in vitro differentiation. Neurogeneic differentiation of trx2 mutant cells generated fewer mature neurons than wild type cells. Moreover a temporal delay in the developmental progression to differentiation became apparent. Cardiac differentiation of trx2-/- cells confirmed the developmental defect and temporal delay. Notably differentiation of trx2-/- cells was merely delayed or impaired but it was not absent, implying that Trx2 is not required for gene expression programs specific for neurons or cardiac myocytes. We propose that differentiation of trx2-/- ES cells is impaired because apoptosis is disturbing differentiation. Apart from analyzing the phenotype of trx2 mutant cells, this work was focused on the identification of Trx2 target genes. Oligonucleotide expression arrays were used to identify genes whose expression levels were affected by the absence of Trx2. In general, loss of Trx2 function resulted in more genes with decreased than increased expression levels. This is consistent with the hypothesis that Trx2 functions as a transcriptional activator. Comparison of gene expression profiles for constitutive and conditional trx2 mutant cells enabled a distinction between direct and indirect target genes for Trx2. As a result Magoh2 was identified as the key candidate target gene for Trx2. Interaction between Trx2 and Magoh2 suggested a potential regulatory role for Trx2 in alternative splicing. Furthermore this work provided evidence that Trx2 could be involved in the maintenance of CpG island promoter gene expression, thus providing a potent regulatory mechanism for ubiquitously expressed genes.
|
Page generated in 0.0566 seconds