The development of genomics has led to a comprehensive delineation of Diffuse Intrinsic Pontine Glioma (DIPG) heterogenous mutational landscape. This includes several interconnected cellular processes that can be pursued by drugs targeting cellular copper called chelating agents. These drugs bind copper and prevent its use in the cell. Copper chelating agents are already clinically approved and used to treat certain genetic disorders. Moreover, they have been proved to accumulate in the brain and have been successfully tested in various cancers, including high-grade gliomas. Copper is an essential transition metal frequently increased in cancer known to influence critical cellular processes. Targeted therapy utilizing both novel and repurposed drugs initially demonstrate strong efficacy, before failing in advanced cancers as drug resistance develops and relapse occurs. Overcoming this limitation involves the development of strategies and protocols aimed at a wider targeting of the underlying molecular changes. Intracellular signalling pathways, epigenetic mechanisms and cell metabolism are among the most common therapeutic targets, with molecular investigations increasingly demonstrating the strong influence each mechanism exerts on the others. Interestingly, all these mechanisms can be influenced by intracellular copper. We propose that copper chelating agents may simultaneously target these mechanisms in DIPG, serving as an excellent candidate for combination therapy.
DIPG is the most dismal paediatric brain tumour, characterised by universal mortality and an average survival of less than one year. With an average age of diagnosis of 5-7 years, this paediatric brain tumour is characterised by its severe symptoms, limited treatment options and universal mortality. Over 250 clinical trials have failed to improve survival, leaving palliative radiotherapy as the only currently available treatment regimen. This therapy only provides a temporary relief from symptoms and does not represent a cure. A novel therapeutic approach to this disease is therefore essential. Several factors contribute to the unique challenges in treating DIPG, including localisation, infiltrative pattern of growth and chemoresistance.
In DIPG, a wide range of dysregulated signalling mechanisms, epigenetic mutations and metabolic alterations drive cell growth, proliferation and survival. Hence, we hypothesise that targeting copper-dependent cellular processes using copper chelating agents will result in DIPG cell death, reduced invasiveness and improved survival of DIPG mouse models. Overall, our final goal is to establish a promising pre-clinical protocol that could rapidly be translated into a clinical trial to improve outcomes for DIPG patients. This will be done by utilising already approved and non-toxic drugs targeting copper, an essential co-factor of cellular pathway that are typically altered in DIPG.
Our preliminary results demonstrate changes in the expression and activation of specific targets in response to copper supplementation and copper chelation, as well as activity of copper chelating agents against DIPG cells. In this novel research proposal, the actual mechanisms and effects of this phenomenon will be established. In detail, the first two aims will be focused on studying expression at both RNA and protein levels of key enzymes in response to copper supplementation as well as copper deprivation in DIPG cells. This will establish the dependence of specific cellular pathways driving DIPG to copper levels. In the third part of the study, we will use a screening methodology to determine specific candidate genes that are influenced by copper levels and copper chelator treatment and are essential to support the DIPG copper-dependent cancer-driving phenotype. This information will be used to better guide targeted therapy for future drug combination tests both in vitro and in vivo. Lastly, we will test the efficacy of copper chelators in vivo in xenograft patient derived DIPG mouse models. Given the already existing clinical data in both human and mice, we anticipate that copper chelation therapy will be well-tolerated in animal models. We will compare control mice against those receiving the copper chelating agents in reducing neuropathic symptoms as well as improving survival. At endpoint, comprehensive histological analysis will be conducted to accurately establish the effect of copper chelation on global tumour growth, induction of tumour cell death and invasion in surrounding healthy brain tissue.
We anticipate that a successful pre-clinical study will pave the way to clinical trials testing copper chelating agents in DIPG patients as both single agents and in combination with radiotherapy. In a devastating disease that have not seen any significant improvement to patient treatment for over 30 years, this would be of outmost importance.
Scientific Merit: With this project, we will take advantage of our deep understanding of DIPG biology, by targeting not a specific molecular driver of tumour progression, but a common co-factor central to a multitude of cellular processes, copper. This will be done utilising an already approved, non-toxic, copper chelating agent that is known to reach the brain in the presence of an intact blood-brain barrier. Importantly, by impairing multiple cellular pathways simultaneously, we anticipate that DIPG cells will not be able to switch the aberrant signalling to an alternative one, avoiding the acquisition of drug resistance.
Disease Impact: Despite some encouraging pre-clinical studies, most clinical trials have failed to significantly improve DIPG patients’ survival, compared to standard palliative radiotherapy. This is usually due to drug insufficient brain bioavailability, high toxicity or acquired resistance. Today DIPG is still universally lethal and patients are in desperate need of alternative therapies. We believe that copper chelation could represent one such therapy.
Copper chelators are a consolidated treatment for copper accumulation syndromes, like Wilson’s Disease. As such, copper lowering agents are well known to cross the blood-brain barrier and alleviating neurological symptoms due to copper accumulation. Being characterised by expected low toxicity, this approach will allow us to test copper chelation in combination with other chemotherapeutic agents targeting more specific pathways of interest.
Innovation: This project represents a novel way to target several DIPG driving pathways, a significant chance to better characterise copper chelation mechanisms in cancer treatment and a unique opportunity to design a clinically significant study with a unique translational potential.
Feasibility: The high feasibility of this proposal derives from the location where it will take place and the unique expertise of the team members. Our research group is part of the Children Cancer Institute (CCI), the largest Australian program dedicated to developing new treatments for malignancies of childhood. The institute is clearly already equipped all the techniques and infrastructures to carry out the research proposal. We have assembled a motivated team of talented molecular biologists who are not only capable to conduct the proposed project, but deeply driven by the common vision to alleviate patients suffering and succeed in the fight against DIPG. Moreover, substantial biostatistics and bioinformatics support will be made available to us through the in-house CCI data science team. Importantly, we work in close relationship with paediatric oncologists and scientists at the Sydney Children’s Hospitals Network and Kids Cancer Alliance and therefore are well positioned to undertake high-impact translational research.
Expertise: Being part of the CCI, we work in close relationship with the Zero Childhood Cancer (ZCC) paediatric precision medicine program that has enrolled hundreds of children nationally, 40% with malignant brain tumours, providing in-depth, personalised tumour analysis. As such, we already contributed globally to the deep understanding of DIPG genetics and biology. We are now using this wealth of knowledge to map DIPG driving pathways and targeting them using a novel thinking-out-of-the-box approach.
Dr Orazio Vittorio is the team-leader of the Metal-Targeted Therapy (MTTI) and Immunology group. He has a growing reputation in understanding the role of metals, such as copper, in paediatric malignancies progression as well as its targeting using chelating agents.
Dr Federica Saletta is a senior research officer in the MTTI group. She comes not only with a plethora of molecular biology skills, but also with an outstanding track records in histopathology technique development for molecular diagnostics.
Associate Professor David Ziegler is a senior paediatric oncologist and a world-renowned brain cancer expert. He is leading the Brain Tumour Group at CCI, conducting in vitro drug screening and in vivo personalised xenograft (PDX) brain tumour models.
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Lorem ipsum dolor sit amet, consectetur adipiscing elit. Integer gravida non felis non euismod. Fusce finibus aliquet consequat. Nam ac metus bibendum, iaculis purus sed, suscipit ligula. Proin et nisi libero. Mauris non urna urna. Nullam augue eros, fringilla sed mauris vitae, porta tincidunt risus. Aliquam sed tincidunt sem. Quisque lacinia quam tortor, imperdiet efficitur odio iaculis in. Sed ultricies condimentum volutpat. Vivamus dignissim faucibus porta.
Curabitur ut ipsum non odio malesuada vulputate. Morbi maximus, est eu lobortis molestie, tortor sapien hendrerit nisi, in cursus odio diam ut odio. Fusce pulvinar volutpat velit. Aliquam erat volutpat. Integer rhoncus mollis suscipit. Praesent non ipsum mollis, finibus nunc a, scelerisque nibh. In feugiat iaculis velit, eu semper lacus dignissim nec. Praesent vitae nisi leo. Cras venenatis dictum magna ut semper. Sed eget eros nibh. Sed vitae quam sed dolor faucibus elementum. Curabitur interdum porttitor finibus. Nullam tincidunt odio lectus, sit amet rhoncus libero dapibus sed. Sed mollis egestas enim, vel porta tortor volutpat eget.
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Lorem ipsum dolor sit amet, consectetur adipiscing elit. Integer gravida non felis non euismod. Fusce finibus aliquet consequat. Nam ac metus bibendum, iaculis purus sed, suscipit ligula. Proin et nisi libero. Mauris non urna urna. Nullam augue eros, fringilla sed mauris vitae, porta tincidunt risus. Aliquam sed tincidunt sem. Quisque lacinia quam tortor, imperdiet efficitur odio iaculis in. Sed ultricies condimentum volutpat. Vivamus dignissim faucibus porta.
Curabitur ut ipsum non odio malesuada vulputate. Morbi maximus, est eu lobortis molestie, tortor sapien hendrerit nisi, in cursus odio diam ut odio. Fusce pulvinar volutpat velit. Aliquam erat volutpat. Integer rhoncus mollis suscipit. Praesent non ipsum mollis, finibus nunc a, scelerisque nibh. In feugiat iaculis velit, eu semper lacus dignissim nec. Praesent vitae nisi leo. Cras venenatis dictum magna ut semper. Sed eget eros nibh. Sed vitae quam sed dolor faucibus elementum. Curabitur interdum porttitor finibus. Nullam tincidunt odio lectus, sit amet rhoncus libero dapibus sed. Sed mollis egestas enim, vel porta tortor volutpat eget.