Cancer biology: identification of new treatment options for EML4-ALK driven lung cancer through understanding mechanisms of cell division and migration.

Lung cancer is the largest cause of cancer-related deaths in the UK and worldwide. While little progress has been made in long-term survival rates, we now have a good understanding of the genetic drivers raising the prospect of earlier diagnosis and more effective treatments. Around 5% of lung cancers are driven by expression of the EML4-ALK oncogenic fusion and this genetic change is commonly seen in younger patients without a history of smoking. While many of these patients initially respond well to targeted ALK inhibitors, some patients show little or no response and all patients inevitably relapse. Hence, there is an urgent need to better understand resistance mechanisms to ALK inhibitors in lung cancer patients and develop new therapeutic and diagnostic approaches.

We have recently discovered a novel pathway that promotes changes in cell morphology and accelerated cell migration and potentially drives the metastatic dissemination of EML4-ALK positive cancers. This pathway involves formation of a complex between EML4-ALK and the cell cycle-regulated NEK9 and NEK7 kinases, normally involved in the regulation of microtubule organization and cell division. In this project, the PhD student will be trained in cutting edge discovery and translational cancer research. They will explore the cellular mechanisms involved in changes driven by EML4-ALK that could lead to identification of new targetable pathways, test whether activation of these pathways may lead to increased sensitivity to particular drugs, and develop a novel diagnostic approaches to the identification of EML4-ALK oncogenic fusions. These will then be tested using patient samples obtained from those who have EML4-ALK lung cancers. Together, these approaches will support an evidence-based rationale for stratification of EML4-ALK positive lung cancer patients to particular treatment protocols.

Background

Advances in molecular profiling together with deeper understanding of the pathways that drive tumour formation have led to a more personalised – or ‘precision medicine’ - approach to the treatment of cancer patients. For many cancers this is reflected in substantial improvements in overall survival rates. However, patients often still relapse and more research is required to understand and overcome resistance mechanisms. Furthermore, survival rates for some major cancers, such as lung cancer, remain stubbornly low.

While significant progress in long-term survival of lung cancer is yet to be made, we now have good understanding of the genetic drivers and this is leading to clinical application of targeted treatments. 5% of lung cancers are driven by EML4-ALK oncogenic fusions and many of these initially respond well to targeted ALK inhibitors. However, patients with some EML4-ALK variants respond poorly and all patients eventually relapse.

We have discovered a novel pathway involving the cell cycle-regulated NEK protein kinases that contribute to microtubule organization and cell division. Moreover, we have shown that this pathway promotes accelerated cell migration and potentially drives the metastatic dissemination of EML4-ALK tumours. Our goal is to better understand this pathway, examine whether it sensitises these tumours to particular drug combinations, and develop novel diagnostic approaches that will enable patient stratification to the most effective treatments.

Aims & Objectives

The aims of this project are to examine a novel signaling pathway that drives metastasis of lung cancer cells expressing EML4-ALK oncogenic fusion variants, determine whether activation of this pathway sensitizes cells to particular combinations of chemotherapeutic agents and establish novel approaches to the diagnosis of EML4-ALK lung cancers.

The specific objectives are to:

1. Examine how EML4-ALK variants, together with NEK9 and NEK7, modify the cytoskeleton to promote migration and invasion of cells, with a focus on proteins involved in microtubule and actin organization.
2. Test whether drugs that interfere with microtubule dynamics (vinorelbine, paclitaxel) or actin organization (FAK, SRC, Rho signaling inhibitors) inhibit the mitotic division or survival of EML4-ALK expressing lung cancer cells, alone or in combination with clinically approved targeted ALK inhibitors (e.g. crizotinib, alectinib, ceritinib, lorlatinib).
3. Develop novel diagnostic approaches, including the use of mass spectrometry and liquid biopsies, to determination of specific EML4-ALK variant expression in patient samples.

Experimental techniques

Dependent on the direction in which the project develops, key experimental techniques in which the student is likely to be trained include:

• 2D and 3D cell culture, including of lung cancer patient-derived cell lines, specifically looking at cell division and cell migration
• high resolution imaging of fixed and live cells, including confocal fluorescence and tomographic electron microscopy, for protein localization and cytoskeleton analysis
• protein expression analysis by Western blot following transfection and siRNA-mediated depletion
• flow cytometry and clonogenic assays for cell death and survival in response to drug treatments
• immunohistochemistry of patient samples
• mass spectrometry for protein and post-translational modification identification
• CRISPR-mediated gene editing

What the student will gain from the project

The student will gain a detailed understanding of the fundamental and translational research that underlies the development of novel anti-cancer precision medicine drugs and diagnostic approaches. They will learn cutting-edge molecular cell biology techniques and the importance of obtaining quantified data that meets statistical significance. They will also gain an appreciation of the challenges associated with applying laboratory-based experimental data to patient treatment and the legal and ethical frameworks in place when using patient samples e.g. HTA requirements.

Translational Implications

The translational goals of this project are to (i) identify clinically relevant drug combinations that might be effective in treatment of EML4-ALK driven lung cancers, and (ii) develop novel approaches to EML4-ALK variant diagnosis that could be applied to other oncogenic fusion proteins.

Enquiries email name and address:

Professor Andrew M. Fry

Department of Molecular and Cell Biology

University of Leicester

Leicester LE1 7RH

U.K.

[Email Address Removed]