Dokument: Label-free detection of brain tumour cell immunogenicity upon mesenchymal transformation using Raman technology

Titel:	Label-free detection of brain tumour cell immunogenicity upon mesenchymal transformation using Raman technology
URL für Lesezeichen:	https://docserv.uni-duesseldorf.de/servlets/DocumentServlet?id=55262
URN (NBN):	urn:nbn:de:hbz:061-20210127-105057-3
Kollektion:	Dissertationen
Sprache:	Englisch
Dokumententyp:	Wissenschaftliche Abschlussarbeiten » Dissertation
Medientyp:	Text
Autor:	Tsiampali, Julia [Autor]
Dateien:	[Dateien anzeigen] Adobe PDF [Details] 11,73 MB in einer Datei [ZIP-Datei erzeugen] Dateien vom 21.01.2021 / geändert 21.01.2021
Beitragende:	Prof. Dr. med. Maciaczyk Jaroslaw [Gutachter] Jun.-Prof. Dr. Span, Ingrid [Gutachter]
Stichwörter:	CD73, epithelial–mesenchymal transition, glioblastoma, monocytes, Raman spectroscopy, T-cells
Dewey Dezimal-Klassifikation:	500 Naturwissenschaften und Mathematik » 570 Biowissenschaften; Biologie
Beschreibung:	Glioblastomas (GBMs) are the most aggressive primary brain tumours in adults and despite strong efforts current therapeutic options have limited impact on glioblastoma stem-like cells (GSCs). GSCs contribute to GBM’s progression and chemoresistance and are enriched by epithelial mesenchymal transition (EMT). EMT has been recently correlated with CD73 that has emerged as an interesting target in the treatment of GBMs. However, the role of CD73 in GSC progression remains elusive. This work aims to investigate whether GSC enrichment is dependent on the enzymatic and/or the non-enzymatic activity of CD73 and further to investigate the impact of EMT activator ZEB1 and CD73 inhibition on GSC immunogenicity using Raman spectroscopy. Chapter 2.1 studies the effect of CD73 inhibition on GSC growth. More specifically, it is shown that the effects on GSC proliferation and clonogenicity were independent from its enzymatic activity but dependent on the CD73 protein level. Selective inhibition of CD73 enzymatic activity by APCP inhibitor affects significantly only on the invasiveness of GSCs, indicating the involvement of adenosine signalling as one of the ways of the invasive regulation. Our results suggested that CD73 and the adenosine receptor A3 are downstreams of EMT which would be helpful to monitor the invasive properties of GBMs and enhance the anti-EMT therapy. Chapter 2.2 studies the ability of Raman spectroscopy to discriminate T-cells and monocytes with different phenotypes after incubation with media conditioned by GSCs with ZEB1 and CD73 inhibition. It is shown that discriminatory analysis using linear discriminant analysis (LDA) and support vector machine classification (SVM) yielded sensitivities and specificities of over 70 and 67% respectively upon validation against an independent test set. Principal component analysis (PCA) of the Raman spectra showed that T-cells and monocytes incubated with tumour-conditioned media of GSCs with inhibited ZEB1 and CD73 formed distinct clusters comparing to controls. To further evaluate the effect of ZEB1 and CD73 inhibition of GSCs on T-cells and monocytes flow cytometry analysis was performed and our data suggested that GSCs with ZEB1 and CD73 inhibition can actively influence the phenotype of T-cells and monocytes, explaining the clear separation of clusters in PCA scores of Raman spectra. These findings highlight the use of Raman spectroscopy for the detection of molecular differences in immune and cancer cells.
Lizenz:	Urheberrechtsschutz
Fachbereich / Einrichtung:	Mathematisch- Naturwissenschaftliche Fakultät » WE Biologie
Dokument erstellt am:	27.01.2021
Dateien geändert am:	27.01.2021
Promotionsantrag am:	05.11.2020
Datum der Promotion:	11.12.2020