MICA: GSK-3 as a multifunctional target for glioblastoma treatment; Hitting multiple tumour hallmarks with a single drug.

High grade gliomas are an aggressive type of brain tumour affecting both adults and children. These tumours are
characterised by extensive infiltration of normal brain by tumour cells. These migrating cells render it impossible to
completely remove the tumour surgically and as a consequence the survival for victims of this kind of tumour is particularly
poor, with the vast majority of patients not surviving beyond two years after diagnosis. It is thought that an effective therapy
that targets the tumour cells that invade the brain would be of great use in treating this disease. Dr Lawler has identified a
strategy that blocks glioblastoma cell migration by acting on a specific cellular protein called GSK-3. Because GSK-3 is
implicated in common illnesses including diabetes and Alzheimer's disease, a large number of drugs have been
developed that will block its action.
Dr Lawler has shown that these drugs also potently block adult brain tumour cell migration. Astra-Zeneca have developed
drugs that target GSK-3 for potential use in the treatment of Alzheimer's disease. One of these drugs, AZD1080
has been tested in humans, and here we will carry out studies that will determine whether it is suitable for the
treatment of glioblastoma - a new application for this drug, which may represent the fastest way of getting
this approach to the clinic.
AZD1080 will be tested in a variety of laboratory models of glioblastoma alongside existing treatments.
Evidence of activity in these models will be used to support future clinical trials.It is hoped that this study will
ultimately contribute to the development of an effective clinical approach for the treatment of glioblastoma.

New treatments are badly needed for the aggressive brain tumour glioblastoma multiforme. We have identified a potential strategy based on inhibition of GSK-3 that may be of benefit to patients through effects on multiple tumour hallmarks, including invasion and angiogenesis. There is also evidence that GSK-3 inhibition may enhance the effectiveness and increase the tumour selectivity of the current standard of care, temozolomide plus radiation. The potential of the novel GSK-3 inhibitor AZD1080 for glioblastoma will be tested using a range of in vitro assays, molecular readouts, and studies on animal models.

In Aim 1, a panel of migration assays (transwell, 3D spheroid, and a novel 3D nanofiber-based assay) will be used in glioblastoma cell lines to determine the effects of AZD1080 on tumour cell migration. We will also examine cell viability parameters in combination with radiation and temozolomide, as well as assays on endothelial cell migration/tube formation and glioma initiating cell differentiation. In Aim 2, analysis of signaling pathways will be performed to determine the downstream effectors of AZD1080 treatment in glioblastoma cells. This will be done by combining global gene expression profiling data and phosphoproteomic data with readouts from luciferase reporter constructs measuring the activities of specific transcription factors regulated by GSK-3 in cancer cells. Finally in Aim 3, we will perform studies on intracranial mouse tumour models of glioblastoma. Dosing will be established followed by survival studies, using AZD1080 alone, and also in combination with radiation and temozolomide. Detailed analysis of the phenotypes of treated tumours will be carried out using immunostaining for tumour proliferation, apoptosis and tumour associated blood vessels.

These experiments will determine whether AZD1080 is a suitable drug to bring forward for the treatment of glioblastoma in human patients.

The research described in this proposal is ultimately designed to provide a benefit to paediatric patients with high grade
gliomas by identifying and developing a novel anti-invasive therapeutic approach; a recognised area of need. Our proposal
is a pre-clinical study which will provide data that will be used in future to support a clinical trial in human glioblastoma patients if appropriate.
We anticipate that four main groups (academic scientists, pharmaceutical companies, clinicians, and patients) will benefit from the ongoing development of
methods/processes as well as final outcomes of this research.
Academic Impact
No anti-invasive therapies are available for the treatment of glioblastoma. Yet this is a vital process in the growth of brain tumours, and also in the
ability of tumours to evade treatments. Moreover, by attacking multiple tumour hallmarks in addition to invasion inhibition of GSK-3 may provide
a level of biological synergy that may be highly efficacious clinically. This study will provide new mechanistic information on the use of GSK-3 inhibitors
in glioblastoma biology. In addition, the use of novel migration assays will help to promote some recently developed methods, that may have broader
research applications. Brain tumours are known as an underfunded/under-researched area (Burnet et al., Br J. Cancer
2005).
Economic and Societal Impact
BUSINESS/INDUSTRY - The new knowledge and scientific advancement gained during this study may benefit
Pharmaceutical companies as it will highlight a novel application of GSK-3 inhibitors. A number of companies in additiona to Astra-Zeneca
Astra-Zeneca, have developed GSK-3 inhibitors for other applications, which may find a use in the treatment of glioblastoma following on from this study.
This grant will enable closer links between our Insitution and Astra-Zeneca, so that further collaborations may also arise involving our unique combination
of basic research in neurooncology, a large patient base, clinical trials expertise and pharmaceutical drug development.
Mechanistic understanding of the role of GSK-3 in glioma cell migration developed in this project will also allow the identification of potentially novel targets,
which may be of commercial interest.
MEDICAL PROFESSIONALS - Clinicians and nurses who work with paediatric brain tumour patients on a daily basis may
become an important beneficiary of this work - more effective therapeutic approaches with fewer side-effects will improve
their work activities.
PATIENTS AND FAMILIES - The current mainstay of treatment for glioblastoma is maximal surgical resection,
radiotherapy and chemotherapy. However, despite this intensive approach mortality is inevitable and median surivival is little more than one year.
Quality of life for patients is compromised by the immediate and late effects of this aggressive treatment, and in particular for young children, the
effects of current management on neurocognitive, social and emotional development can in themselves be devastating.
Novel treatment strategies are therefore very much needed and this project aims to provide pre-clinical data to bring a new treatment forward,
and by further understanding mechinsms involved provide biomarkers that could be used in the clinical arena for patient benefit, and
improve patients overall health and quality of life