Publikationsdatenbank

Functional and (ultra)morphological characterization of patient-derived organoids from head and neck cancer (2024)

Art

Poster

Autoren

Fisch, Anne-Sophie (WE 1)

Plendl, Johanna (WE 1)

Tinhofer-Keilholz, Ingeborg

Kongress

12th Meeting of the Young Generation of Veterinary Anatomists (YGVA)

Zagreb, Croatia, 17. – 19.07.2024

Quelle

Anatomia, histologia, embryologia

Bandzählung: 53

Heftzählung: S1

Seiten: 17

ISSN: 0340-2096

Sprache

Englisch

Verweise

URL (Volltext): https://onlinelibrary.wiley.com/doi/epdf/10.1111/ahe.13066

DOI: 10.1111/ahe.13066

Kontakt

Institut für Veterinär-Anatomie

Koserstr. 20
14195 Berlin
+49 30 838 75784
anatomie@vetmed.fu-berlin.de

Abstract / Zusammenfassung

Intrinsic and environmental tumour characteristics increase therapy resistance, resulting in treatment failures and high recurrence rates in human head and neck squamous cell carcinomas (HNSCC). This study aims at the (ultra)morphological and functional characterization of patient-derived
organoids (PDOs). It explores their applicability to investigate individual resistance mechanisms to radiation. Ultrastructural features of two PDOs and their changes upon irradiation were explored with transmission electron microscopy.
Hypoxia was assessed using Pimonidazole in one PDO. ATP-dependent viability, clonogenic survival and organoid volumes were used as functional read outs for radiosensitivity in three models. Immunofluorescent gH2AX foci were stained for visualization of DNA double strand breaks in one PDO. Cell death and proliferation was assessed using live dead fixable green reagent and Ki67 with confocal fluorescence microscopy.
High nuclear to cytoplasmatic ratios, prominent cell organelles and wide intercellular spaces, correspond to the ultrastructure of poorly to moderately differentiated squamous cell carcinomas. Upon irradiation, cells in the outer rim of the organoid formed protective barriers and ejected cell debris in an organized manner. Interestingly, extent of damage between organoids was heterogeneous within the same model. Similar to this observation was the abundance of proliferative and dead cells. Compared to initial gH2AX foci, assessed 1 h after irradiation, fewer foci were detected after 24 h, due to functioning DNA repair mechanisms. The inner core of several organoids had a positive fluorescence signal for hypoxia. PDOs showed a dose-dependent decrease in metabolic activity, proliferation and clonogenic survival, and a considerable interpatient variability in survival. PDOs might be valuable models to investigate individual responses to radiation therapy and the molecular mechanisms underlying radioresistance. In future experiments, we plan to establish a protocol for the automated quantification of gH2AX in organoids. Furthermore, we will investigate the impact of hypoxia on radiosensitivity of the models in more detail.