Publikationsdatenbank

Patient-derived organoids from head and neck cancer facilitate the investigation of individual responses to irradiation (2023)

Art

Vortrag

Autor

Fisch, Anne-Sophie (WE 1)

Kongress

International Biotech Innovation Days 2023 (IBID)

Senftenberg, 17. – 19.10.2023

Quelle

Sprache

Englisch

Kontakt

Institut für Veterinär-Anatomie

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

Abstract / Zusammenfassung

Introduction & aim
Worldwide, head and neck squamous cell carcinomas (HNSCC) account for more than 500 000 deaths per year. Intrinsic and environmental tumor characteristics increase therapy resistance, resulting in treatment failures and high recurrence rates (1). This study aims for the establishment of representative patient-derived organoid (PDO) models to study individual responses to irradiation.
Materials & methods
The ex vivo assessment of radiosensitivity was established using 3D cultures of radioresistant / -sensitive subclones from the FaDu cell line cultured in Matrigel. This irradiation protocol was then applied to PDO models. Formalin-fixed and paraffin-embedded organoid sections were stained with haematoxylin-eosin and immunohistochemical markers p40, CK5/6 and Ki67. Fluorescent Pimonidazole was used to visualize hypoxia in PDO cryosections. Ultrastructural changes upon irradiation were explored with transmission electron microscopy.
Results
With our irradiation protocol, we were able to distinguish radiosensitive from resistant FaDu models. Preliminary results from PDOs (n=3) showed a dose-dependent decrease in proliferation, cell-viability (CellTiterGlo®) and clonogenic potential, and a considerable interpatient variability in radiosensitivity. Histopathological analysis confirmed their squamous cell carcinoma phenotype. The inner core of several organoids had a positive fluorescence signal for hypoxia. Signs of cell death characterized ultrastructural changes in irradiated organoids: cell fragmentation, vacuolization, membrane blebbing and loss of cell-cell contacts. A double dose was necessary to induce comparable effects seen in the radiosensitive subclone in the more resistant FaDu clone. Additionally cells in the irradiated PDOs formed protective barriers as well as ejecting cell debris in an organized manner.
Conclusion & outlook
PDOs might be valuable models to investigate individual responses to radiation therapy and the molecular mechanisms underlying radioresistance. In future experiments, we will investigate the impact of hypoxia on radiosensitivity of the models in more detail.