Repurposing Itraconazole as an Adjuvant for the Treatment of Glioblastoma Multiforme

Tez Türü: Doktora

Tezin Yürütüldüğü Kurum: University of Leicester, Cancer Research, Cancer Research, İngiltere

Tez Danışmanı: Prof. Dr. Don Jones

Tezin Onay Tarihi: 2022

Tezin Dili: İngilizce

Desteklendiği Program: Diğer

Özet:

Glioblastoma multiforme (GBM) is one of the most aggressive human cancers, with a poor

prognosis. Despite recent advances in the understanding of GBM's molecular biology,

patient outcome continues to be poor due to the disease's incurable, aggressive, and

recurrent nature. As a result, novel therapeutic strategies for the management of GBM

patients are urgently needed. Itraconazole (ITC), a systemic antifungal, is a key regulator of

hedgehog (HH) signalling, which is frequently deregulated in GBM. Because of its inhibitory

effects on cell proliferation and tumour angiogenesis, ITC has been identified as a novel

potential anticancer agent and is herein investigated in the context of GBM.

In the present study, cell viability and reproductive integrity of GBM cells were assessed by

AlamarBlue cell viability and clonogenic assays. Spheroid growth and relative colony cell

growth were assessed by spheroid and associated absorbance reading assays. DNA damage

levels and cell cycle distribution of the GBM cells were assessed by single cell gel

electrophoresis (COMET) and flow cytometry, respectively, and O6-methylguanine DNA

methyltransferase (MGMT) and HH protein levels were assessed by western blot analysis.

Using these techniques we show that ITC, both alone and in combination treatments,

inhibits GBM cell and spheroid growth in vitro, increased IR-induced DNA damage and

caused G1 cell cycle arrest. Furthermore, ITC treatment decreased the levels of HH

signalling pathway proteins sonic hedgehog (SHH) and smoothened (SMO). Together, these

findings indicate that ITC inhibition of HH signalling pathway may increase TMZ, radiation

and radiochemotherapy (RCT) responses and so serve as a potential adjunct to RCT in the

management of GBM.