FRET-Bildgebung zum Nachweis der Phopshorylierung der anaplastic lymhoma kinase (ALK) im nicht-kleinzelligen Bronchialkarzinom

Wagner, Tabea

dc.contributor.advisor	Kroemer, Heyo K. Prof. Dr.
dc.contributor.author	Wagner, Tabea
dc.date.accessioned	2018-05-22T06:33:03Z
dc.date.available	2018-06-12T22:50:07Z
dc.date.issued	2018-05-22
dc.identifier.uri	http://hdl.handle.net/11858/00-1735-0000-002E-E3F0-F
dc.identifier.uri	http://dx.doi.org/10.53846/goediss-6879
dc.language.iso	deu	de
dc.rights.uri	http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject.ddc	610	de
dc.title	FRET-Bildgebung zum Nachweis der Phopshorylierung der anaplastic lymhoma kinase (ALK) im nicht-kleinzelligen Bronchialkarzinom	de
dc.type	doctoralThesis	de
dc.title.translated	FRET-Imaging to proof the phosphorylation of the anaplastic lymphoma kinase in NSCLC	de
dc.contributor.referee	Ströbel, Philipp Prof. Dr.
dc.date.examination	2018-06-05
dc.description.abstracteng	Background: Anaplastic lymphoma kinase (ALK) is an oncogenic driver mutation in T-cell lymphoma and NSCLC. Small molecule inhibitors are available. Treatment decisions are mostly made on ALK expression but their efficacy widely varies from complete response to progression under therapy. I regard ALK activity, judged by its phosphorylation, a highly relevant factor in the evaluation of therapeutic sensitivity. Phosphorylation can be quantified by Förster Resonance Energy Transfer (FRET), here shown by confocal acceptor photobleaching micorscopy, and could be used to select patients for targeted therapies. Methods: Confocal laser microscopy, cell culture, FFPE-tissue, Immunocytofluorescence, Immunohistofluorescence, Westernblots, Immunoprecipitation, overall-survival-measurements Results: I show ALK phosphorylation in human cancer cell lines. FRET strongly correlates with the pharmacological ALK inhibition sensitivity of the cell lines, implying a promising role as companion diagnostic test for ALK activity drug. The same cell lines were subjected to cytospin and block preparation. I could closely reproduce the results, demonstrating the utility of the method in routine cytological work. Also I could implement Immunohistofluorescence to apply the method in formalin-fixed, paraffin-embedded (FFPE) human tissue. Conclusions: I present a quantitative cell ALK phosphorylation assay, based on confocal acceptor photobleaching FRET microscopy that holds promise for future diagnostic applications on FFPE samples of human cancer. This method provides both information on ALK expression, comparable to immunohistochemistry and FISH methods, and on ALK phosphorylation, relfecting its activity and likely predicting sensitivity to targeted therapies.	de
dc.contributor.coReferee	Wouters-Bunt, Freddy Prof. Dr.
dc.subject.ger	EML4-ALK	de
dc.subject.ger	Bronchialkarzinom	de
dc.subject.ger	Konfokale Mikroskopie	de
dc.subject.eng	EML4-ALK	de
dc.subject.eng	FRET	de
dc.subject.eng	Acceptor Photobleaching	de
dc.subject.eng	Lung cancer	de
dc.subject.eng	Confocal microscopy	de
dc.identifier.urn	urn:nbn:de:gbv:7-11858/00-1735-0000-002E-E3F0-F-3
dc.affiliation.institute	Medizinische Fakultät	de
dc.subject.gokfull	Medizin (PPN619874732)	de
dc.subject.gokfull	Pathologie / Pathologische Anatomie / Histopathologie / Zytopathologie - Allgemein- und Gesamtdarstellungen (PPN619875674)	de
dc.description.embargoed	2018-06-12
dc.identifier.ppn	1023072777

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