The Effects of Conventional Chemotherapeutics on the Activation of Type I Interferons (IFN-α/β) in Hepatic Cancer Cells in vitro: The Significance of Endogenous Type III Interferons (IFN-λ)

Grothe, Luca Maria

dc.contributor.advisor	Mihm, Sabine Prof. Dr.
dc.contributor.author	Grothe, Luca Maria
dc.date.accessioned	2023-04-24T15:46:47Z
dc.date.available	2023-05-10T00:50:10Z
dc.date.issued	2023-04-24
dc.identifier.uri	http://resolver.sub.uni-goettingen.de/purl?ediss-11858/14639
dc.identifier.uri	http://dx.doi.org/10.53846/goediss-9849
dc.format.extent	XXX Seiten	de
dc.language.iso	eng	de
dc.rights.uri	http://creativecommons.org/licenses/by/4.0/
dc.subject.ddc	610	de
dc.title	The Effects of Conventional Chemotherapeutics on the Activation of Type I Interferons (IFN-α/β) in Hepatic Cancer Cells in vitro: The Significance of Endogenous Type III Interferons (IFN-λ)	de
dc.type	doctoralThesis	de
dc.contributor.referee	Mihm, Sabine Prof. Dr.
dc.date.examination	2023-05-02	de
dc.description.abstracteng	HCC is characterized by limited therapy options, poor response rates to systemic chemotherapy, and high lethality. Preclinical cancer models have revealed the efficacy of genotoxic drugs to rely on the release of type I IFNs by neoplastic cells boosting an antitumor response. This thesis elucidates the capacity of the transplantable hepatoma cancer cell line Hepa 1-6 to activate type I IFNs (IFN-α/β), type III IFNs (IFN-λ) and IFN effectors, and compares the parental Hepa 1-6 cell line to CRISPR/Cas9-engineered Ifnl2/3-deficient isogenic clones in view of preclinical models. Methods: Isogenic Ifnl2/3-deficient clones were generated from murine Hepa 1-6 hepatoma cells (DMSZ Braunschweig) using CRISPR/Cas9 genome engineering. Knockout was validated by conventional PCR assay on gDNA and mRNA level as well as by sequencing. Parental and deficient lines were stimulated with the RNA analogue poly(I:C) and treated with gemcitabine, doxorubicin or oxaliplatin. The expression of selected IFNs and IFN effectors was quantified via qRT-PCR. Cell viability was assessed by MTS reduction assay. Results: Hepa 1-6 cells were shown to respond to various stimulatory regimens based on the pattern recognition receptor agonist poly(I:C) with both type I IFN and type III IFN (Ifnl2/3) gene expression. Beyond that, type III IFN gene activation could also be achieved by a short exposure to gemcitabine if combined with poly(I:C) priming in a synergistic manner. This induction was diminished by timed culture medium exchange or addition of RNA targeting nucleases, suggesting release of a mediating compound. Moreover, gemcitabine, rather than doxorubicin or oxaliplatin, was found to be a potent inducer of the IFN effector Cxcl10. However, chemokine inducibility by and chemosensitivity towards gemcitabine appeared to be independent of Ifnl2/3 gene expression as these biological responses were comparable in extent in CRISPR/Cas9 engineered Ifnl2/3-deficient isogenic clones. Conclusions: Chemotherapy was demonstrated to be able to modulate poly(I:C)-primed Ifnl2/3 activation and subsequently IFN effector induction. The nucleoside analogue gemcitabine was demonstrated to independently promote Ifnl2/3 and Cxcl10 activation in transplantable hepatoma cells. Ifnl2/3-deficiency was shown to not influence in vitro chemosensitivity of murine Hepa 1-6 hepatoma cells. Nevertheless, both by-effect properties are expected to impact interactive immunoediting in hepatocellular carcinoma in vivo and thus the establishment of an antitumoral immune response.	de
dc.contributor.coReferee	Kube, Dieter Prof. Dr.
dc.subject.eng	hepatocellular carcinoma	de
dc.subject.eng	interferons	de
dc.subject.eng	immunogenic cell death	de
dc.subject.eng	CRISPR/Cas9	de
dc.subject.eng	IFNL	de
dc.subject.eng	Hepa 1-6	de
dc.subject.eng	type III interferons	de
dc.subject.eng	gemcitabine	de
dc.subject.eng	doxorubicin	de
dc.subject.eng	oxaliplatin	de
dc.subject.eng	poly(I:C)	de
dc.subject.eng	antitumoral immune response	de
dc.subject.eng	DAMPs	de
dc.subject.eng	CXCL10	de
dc.subject.eng	immunoediting	de
dc.identifier.urn	urn:nbn:de:gbv:7-ediss-14639-9
dc.affiliation.institute	Medizinische Fakultät	de
dc.subject.gokfull	Medizin (PPN619874732)	de
dc.subject.gokfull	Gentherapie (PPN619875194)	de
dc.subject.gokfull	Innere Medizin - Allgemein- und Gesamtdarstellungen (PPN619875747)	de
dc.subject.gokfull	Immunologie / Allergologie / Umweltmedizin / Medizinische Ökologie - Allgemein- und Gesamtdarstellungen (PPN619875445)	de
dc.subject.gokfull	Methoden und Techniken in der Medizin (PPN619875143)	de
dc.description.embargoed	2023-05-10	de
dc.identifier.ppn	1843556936
dc.notes.confirmationsent	Confirmation sent 2023-04-24T19:45:02	de

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