Scriptie

The Photodynamic Therapy of Mesothelioma Cell Lines

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Vak
Cancer Biology

Instelling
Sheffield Hallam University (SHU)

A variety of cell lines were utilised in this research including MSTO-211H, MRC-5 and NCI-H28. WST-8 assay was performed to determine cell viability of multiple cancer cell lines under different treatment conditions with SHU-C5 photosensitiser and vehicle control. Relative WST-8 activity was determ...

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Type Scriptie
Begeleider(s) Dr sarah haywood-small
Jaar Onbekend

mesothelioma
cancer
photodynamic therapy
pdt
phototherapy
mpm

Instelling
Sheffield Hallam University (SHU)
Studie Onbekend
Vak
Cancer Biology

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Effectiveness of Novel SHU Photosensitisers for treatment

of Mesothelioma Cell Lines and Headspace VOC Analysis for

Disease and Treatment Monitoring.

By Elliott Fox

This report is presented in partial fulfilment of Cancer Biology MSc

requirements

Date of Submission: 08/09/2022

1

,Abstract

Mesothelioma is a rare neoplasm of the pleural and peritoneal mesothelial cells. 85% of
cases are associated with asbestos and a latency period of 30-40 years before diagnosis,
contributing to the poor overall survival of 12 months with chemotherapy. PDT is a
promising treatment option, however, there are currently no photosensitisers that have
been approved for application on mesothelioma. There is also a lack of treatment
monitoring of mesothelioma; volatile organic compounds have shown promise as a
diagnostics tool in mesothelioma and as a monitoring tool in lung cancer.

A variety of cell lines were utilised in this research including MSTO-211H, MRC-5 and
NCI-H28. WST-8 assay was performed to determine cell viability of multiple cancer cell lines
under different treatment conditions with SHU-C5 photosensitiser and vehicle control.
Relative WST-8 activity was determined in excel and significant reductions were identified
by a two-tailed equal variance t-test, p<0.05. Volatile organic compounds were collected
from the headspace of NCI-H28 cells in various treatment conditions sing an SPME fiber and
VOC profiles were analysed using GC-MS, SIMCA and Metaboanalyst.

There was a significant decrease in relative WST-8 activity in the MSTO-211H 48-hour
light SHU-C5 group at concentrations of 5 and 15 g/mL by 18% (sd, 0.110; p<0.05) and 26%
(sd, 0.110; p<0.05). There were no significant differences in relative WST-8 activity between
any group for the MRC-5 cell line (p<0.05). There was a significant decrease in relative WST-
8 activity in the NCI-H28 24-hour light SHU-C5 group at concentrations of 10 and 15 g/mL
by 13% (sd, 0.175; p<0.05) and 10% (sd, 0.098; p<0.05) and a significant increase at 20
g/mL by 43% (sd, 0.396, p<0.05). There was a significant decrease in relative WST-8 activity
in the NCI-H28 48-hour dark SHU-C5 group at a concentration of 15 g/mL by 17% (sd,
0.034; p<0.001).

The SHU-C5 photosensitiser demonstrated a light-dependent cytotoxic response to
MSTO-211H and NCI-H28 cell lines, therefore, showing potential as a photosensitising agent
for the PDT application of mesothelioma. Upon light treatment with SHU-C5, the VOC profile
was significantly altered, indicating that breathomics could be used as a monitoring tool for

2

,mesothelioma. The research analyses the efficiency of the SHU-C5 photosensitiser and
provides a platform for future research into the monitoring of treatment of mesothelioma.
Index

1.0 Introduction
1.1 Mesothelioma
1.2 Current Therapies
1.3 Photodynamic Therapy
1.4 The Use of Breathomics to Monitor Treatment Methods
1.5 Aims and Objectives
2.0 Materials and Methods
2.1 Cell Lines
2.2 Cculture Cconditions
2.3 Ppassaging of Ccells
2.4 Ccell Ccounting
2.5 P porphyrin prepPreparation
2.6 wWSTet-8 Ccell Vviability
2.7 wetWST-8 Ddata Aanalysis
2.8 vocVOC Eextraction
2.9 gcmsGas Chromatography-Mass Spectrometry
2.10 gemsGCMS Data Analysisd/anal
3.0 Rresults
3.1 Ccellular Vviability of SHU-C5 to Determine Toxicity
3.2 Headspace Analysis of Volatile Organic Compounds
4.0 Discussion
4.1 Efficacy of SHU-C5 Photosensitiser
4.2 Headspace Analysis of NCI-H28 Cell Line
5.0 Conclusion
6.0 References

3

, 1.0 Introduction

1.1 Mesothelioma

Mesothelioma is an aggressive rare cancer of the pleural and peritoneal mesothelial cells;

85% of malignant pleural mesothelioma cases are associated with asbestos exposure,

therefore, contributing to the ban of asbestos in various countries, including the UK (Park et

al., 2012; Chen et al., 2019; Murphy et al., 2021). Due to latency period of 30-40 years

between first exposure to asbestos and mesothelioma development, there is a continuing

rise in cases in the UK (Murphy et al., 2021; HSE, 2022). Malignant mesothelioma has a poor

prognosis when left untreated, resulting in a median overall survival (OS) of ~6 months (Bei

et al., 2022)

Malignant mesothelioma can be histologically identified by three different subtypes:

epithelioid (~60-70%), sarcomatoid (10-20%) or biphasic (15-25%), biphasic represents a

mixture of both epithelioid and sarcomatoid cells (Meyerhoff et al., 2015; Wadowski et al.,

2019; Janssens et al., 2022). The epithelioid subtype tends to have a more favourable

prognosis and overall life expectancy prognosis, whereas the sarcomatoid subtype is

associated with the least favourable prognosis (Nuvoli et al., 2018; Little et al., 2021).

Other factors contributing to the poor OS of malignant mesothelioma include the

limited treatment options, absence of malignant mesothelioma blood and tissue markers

4

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