Live Imaging to Quantify Cellular Radiosensitivity in Patient-Derived Tumor Organoids

Maud Charpentier, Norma Bloy, Silvia C. Formenti, Lorenzo Galluzzi, Sandra Demaria

Research output: Contribution to journalArticlepeer-review

Abstract

Radiation therapy (RT) is one of the mainstays of modern clinical cancer management. However, not all cancer types are equally sensitive to irradiation, often (but not always) because of differences in the ability of malignant cells to repair oxidative DNA damage as elicited by ionizing rays. Clonogenic assays have been employed for decades to assess the sensitivity of cultured cancer cells to ionizing irradiation, largely because irradiated cancer cells often die in a delayed manner that is difficult to quantify with short-term flow cytometry-or microscopy-assisted techniques. Unfortunately, clonogenic assays cannot be employed as such for more complex tumor models, such as patient-derived tumor organoids (PDTOs). Indeed, irradiating established PDTOs may not necessarily abrogate their growth as multicellular units, unless their stem-like compartment is completely eradicated. Moreover, irradiating PDTO-derived single-cell suspensions may not properly recapitulate the sensitivity of malignant cells to RT in the context of established PDTOs. Here, we detail an adaptation of conventional clonogenic assays that involves exposure of established PDTOs to ionizing radiation, followed by single-cell dissociation, replating in suitable culture conditions and live imaging. Non-irradiated (control) PDTO-derived stem-like cells reform growing PDTOs with a PDTO-specific efficiency, which is negatively influenced by irradiation in a dose-dependent manner. In these conditions, PDTO-forming efficiency and growth rate can be quantified as a measure of radiosensitivity on time-lapse images collected until control PDTOs achieve a predefined space occupancy.

Original languageEnglish
Article numbere66680
JournalJournal of Visualized Experiments
Volume2024
Issue number206
DOIs
StatePublished - Apr 2024
Externally publishedYes

Keywords

  • Humans
  • Neoplasms/radiotherapy
  • Organoids/radiation effects
  • Radiation Tolerance

Fingerprint

Dive into the research topics of 'Live Imaging to Quantify Cellular Radiosensitivity in Patient-Derived Tumor Organoids'. Together they form a unique fingerprint.

Cite this