Using non-small cell lung cancer organoids to investigate responses to radiation

Velasco Martinez, Carmen 2023. Using non-small cell lung cancer organoids to investigate responses to radiation. PhD Thesis, Cardiff University. Item availability restricted.

Preview	PDF (PhD thesis) - Accepted Post-Print Version Download (74MB) | Preview
	PDF (Cardiff University Electronic Publication Form) - Supplemental Material Restricted to Repository staff only Download (1MB)

Abstract

Background Cancer organoids preserve the histological, genetic, cellular, spatial, and functional diversity of the tumours from which they are derived and have been shown to accurately reproduce clinical responses to drug treatments. In addition, organoids represent relatively simple cultures that can be adapted for high-throughput research purposes. The first Non-Small Cell Lung Cancer (NSCLC) long-term organoids were described in 2019 and at the time this work was started, only this one protocol had been published. A range of cancers including NSCLC are treated in the clinic by radiotherapy and at the start of this project, it was unknown whether organoids would be able to model tumour responses to radiation. In this work, I aimed to examine the response of NSCLC organoids to radiation while investigating the potential of organoid systems to emulate combination therapies, specifically combining radiotherapy with DNA damage response inhibitors (DDRis) to enhance the therapeutic effects of radiation-induced DNA damage. Methods Adapting a range of novel assay formats (i.e. ATP-based, image-based), I established radiation-responses assays in 3D formats (including organoids and spheroids) and analysed responses to radiation and DDRi combination therapies. Results The direct evaluation of post-irradiation reproductive cell death through 3D re-plating assays emerged as a pivotal assay for radiation organoid-response assessment. This could be combined with less sensitive but more rapid methodologies, such as ATP assays, when results were regularly cross-verified. Furthermore, a preliminary investigation corroborated the existence of intra-organoid ‘cell type’ variability within NSCLC organoids. This intraorganoid heterogeneity may have profound implications for comprehending intra-tumoral NSCLC variability in vivo, a phenomenon often implicated in cancer recurrence and patient mortality. Conclusions My findings highlight the need for employing a combination of techniques to accurately assess organoid responses to radiation. Specifically, the re-plating assay stands out as a critically sensitive method for evaluating organoid radiation-response.

Item Type:	Thesis (PhD)
Date Type:	Completion
Status:	Unpublished
Schools:	Biosciences
Subjects:	Q Science > Q Science (General)
Date of First Compliant Deposit:	29 February 2024
Last Modified:	01 Mar 2024 09:37
URI:	https://orca.cardiff.ac.uk/id/eprint/166728

Actions (repository staff only)

Edit Item