Project management and coordination
Work Package 1

This Work Package is tasked with the effective administration, management and governance of the GLINT Project across all GLINT project partners. This incorporates duties covering all aspects of project monitoring, reporting, financial and contractual administration according to the Commission’s rules, ensuring proper communication within the consortium and implementing the project’s governance and decision-making structures as laid out in Description of Action. The specific objectives of this Work Package are:

• To ensure the efficient and effective management of GLINT through the implementation of the project’s governance and decision-making structures;
  
• To ensure effective communications both internal (within the consortium) and external (liaison with the European Commission and third parties);
  
• To monitor and report on the project’s progress and identify any aspects requiring attention to the appropriate bodies;
  
• To ensure that the work and tasks are completed on time, within budget and according to high quality standards;
  
• To monitor progress between planned and actual activities, identify any aspects requiring attention to ensure goals are achieved within the stated time frame;
  
• To ensure that the contractual, project management and financial aspects of GLINT are carried out in an efficient, transparent and correct manner and in accordance with European Commission rules;
  
• To provide effective linkage and communication between the European Commission, partners and other interested parties;
  
• To ensure that gender equality matters are adequately addressed.

Development of advances and novel imaging sequences across European Centres
Work Package 2

This work package is dedicated to the development of advanced MR imaging techniques across the European centres involved in the project, allowing robust detection of the small exchange-related signal on clinical scanners (3T Siemens PRISMA and MMR scanners and experimental 9.4T human scanner) and for animal scanners in WP4. The methods developed within the WP will take advantage of the rapid exchange protons of the hydroxyl groups of glucose molecules with the water and its effects on all physical relaxation rates present in an MRI experiment, including T1, T2, and T1ρ. 

Objectives of the work package will thus include:

• Implementation of n-pool Bloch-McConnell simulation environment for field strengths ranging from 3T to 14T to calculate CEST, T2 and T1ρ effects;
• Increase and optimize labelling and readout efficiency under in vivo conditions at various field strengths (7T/9.4T human brain and 3T human body);
  
• Evaluation and optimization of alternative approaches combining multiple contrasts such as T1ρ contrast and quantitative T1 and T2 measurements will be investigated under in vivo conditions at various field strengths (7T/9.4T human brain and 3T human body);
• Evaluation and integration of main field homogeneity effects (B0) and variation of the RF transmit field (B1). The final goal is to apply this package of techniques and sequences together with quantitative data analysis techniques to be developed in WP3 to human studies as described in WP7, as well as for preclinical animal experiments described in WP4. The sequences developed here will play an important role not only for the GLINT proposal, but also for the development of endogenous CEST techniques beyond the end of GLINT, which interest the major MRI manufacturers, including Philips and Siemens, both located within Europe. Siemens in particular, on which all of the work present in this application will be implemented, will have access through existing collaboration agreements both at UCL and MPG.

Pharmacokinetic analyses
Work Package 3

The main objectives of this Work Package are to establish the entire post-processing pipeline for all MRI experiments undertaken within the GLINT consortium. This will be lead by OM in collaboration with 5 other partners of the GLINT consortium, to ensure that all algorithms will be tested on real data prior to the distribution of the Software through the Olea-SphereTM package to every group in Europe and the world who wants to use it. The development of new applications for the post-processing of emerging MR sequences is a strategic issue for OM, since the company intends to keep up competitively with the development of MR physics. Therefore the primary outcome of this work package will help one of the most successful SME within Europe, OM, to increase its offer and therefore bring them some definitive advantage over the competition. For this to happen, the following three objectives are set:

• Detection of the signal stemming from GlucoCEST and other glucose-sensing experiments, and separation from the background signal;
• Development a set of non-linear differential equations to appropriately account for the Michaelis-Menten processes involved;
• Development of a physician-friendly method requires a software tool directly available on any radiological PACS systems to provide rapid image-processing analysis for WP2, WP5 and WP7, making use of the results from WP3, tasks 1 & 2.

Origin of GlucoCEST contrast in glucose and glucose analogues
Work Package 4

The main objective of WP 4 will be to assess the potential biochemical pathways and the sources of the GlucoCEST signal for native and methylated glucose analogues. The results from this WP will allow to us to properly define the pK used in WP 3. The following sub-aims are also defined:

• Uptake and metabolism of 3OMG will be studied by following the CEST effects but also by 13C NMR spectroscopy of extracts of tumours after administration of [13C]3OMG as well as in vivo localized 13C NMR spectroscopy, both in brain and solid tumour models.
• Other glucose analogues GlucoCEST signal will be tested to avoid risking the whole programme on a single molecule pair (Glc and 3OMG). These glucose analogues will need to pass the following three criteria: high take by tumours, exhibit high CEST or exchange-related effect and have low or no toxic effects.
• Genetically-encoded FRET sensors specific for energy metabolites measured by fluorescence optical imaging, e.g. glucose, glutamate, ATP, NADH, lactate and pyruvate will be used in tumour brain cells and in tumour-bearing brain slices to establish a clear biochemical origin of the measured GlucoCEST signal.
• As chemical exchange rates of hydroxyl mobile protons belonging to glucose and glucose derivatives are strongly affected by the solution pH where the molecules are dissolved, the influence of the tumour extracellular pH on the measured MRI GlucoCEST contrast will be evaluated.

Detection thresholds and response to therapy through coordinated EU-wide collaborative work
Work Package 5

The main objective of WP5 is to establish the detection limits of the candidate molecules selected in WP3 for both in vitro and in vivo conditions. In in vitro conditions, the following effects will be evaluated for discerning and optimizing the achievable CEST contrast: probe concentration; pH; temperature; magnetic field strength. For in vivo evaluation, representative tumour models will be selected and the influence of the following parameters will be evaluated: injected dose; administration route; magnetic field strength.
A second objective is devoted to exploit these molecules as early reporter of a therapeutic effect. This includes the administration of conventional therapies and new anticancer therapies (inhibitors of tumour metabolism) on selected murine tumour models for assessing the capability of the GlucoCEST contrast to report on treatment response. The GlucoCEST derived response will be compared with the gold-standard [18F]FDG-PET imaging assessment to understand the role that this new proposed approach may have as an alternative imaging method. Moreover, other MRIbased modalities, such as changes in tissue cellularity, vascularisation and acidosis will be evaluated and compared to the GlucoCEST readout for its ability to assess the therapeutic response.

Toxicology studies and preparing for regulatory approval across Europe
Work Package 6

In Work Package 6 the following objectives are addressed:

• To perform bio-distribution, excretion and toxicity studies in healthy and tumour rodents models on 3OMG, and possibly other derivatives selected from WP4.
• From these results of, one or more of the glucose derivatives will be submitted to the Regulatory agencies in order to take the project further into clinical trials.
• In vivo pK-pD assessment of glucose analogues will be performed using radio-labelled isotopes.
• Glucose receptor binding affinities will be assessed to estimate the risks of patient hypoglycaemia, and detailed analysis of systemic effects due to the injection of glucose derivatives will be undertaken, such as e.g. insulin response, metabolic rate, and glycaemic levels.

First-in-man studies of GlucoCEST in patients from two European centres
Work Package 7

Data from first-in-man studies in cancer patients will be acquired, based on the optimised imaging techniques developed jointly with MPG in WP 2 and the data analysed quantitatively using the models developed in collaboration with OM within WP 3. Main objective: for GlucoCEST to bridge the 1st translational gap as outlined in the world's largest independent cancer research charity (Cancer Research UK - CRUK) imaging biomarker roadmap by providing a detailed validation of GlucoCEST in patients in the EU, and quantify its utility in detecting and staging cancer, as well as predicting response to therapy. This will be done through:

• technical validation of defining a glucose administration regimen and assessing reproducibility,
• biological validation against [18F]FDG-PET, multi-parametric MRI, optical / digital histopathology and genomic markers,
• biomarker qualification in proof of concept patient cohorts establishing potential: detection of tumour, staging of the disease, early prediction of response to therapy and surveillance post-therapy.

To address the above, three different cohorts of cancer patients will be recruited from established referral pathways at the UCLH MacMillan Cancer Centre (£110m purpose built centre integrating care of cancer patients with cutting-edge clinical and radiological research) and the National Hospital of Neurology and Neurosurgery (NHNN) within UCL. In addition, patients suffering from primary brain tumour (glioma) will also be recruited from the Tübingen University Hospital through MPG. Each cohort will provide the substrate for validation; biomarker qualification will be targeted to address unmet clinical need to provide maximum clinical impact:

• Head and neck squamous cell carcinoma,
• Paediatric lymphoma,
• Primary gliomas

Dissemination, knowledge management and exploitation
Work Package 8

WP8 will develop and pursue the overall external project communication, dissemination and exploitation activities as well as knowledge management to raise scientific and public awareness and ensure uptake of the project work and outcomes across Europe. WP8 will demonstrate the benefits for all end-users. Input to this horizontal task will be required from all project partners.
In detail the objectives of this work package are to

• Develop a communication and dissemination strategy and material to reach the European arena;
• Disseminate project results to relevant stakeholders: scientists, clinicians, healthcare providers, industry, funding agencies, decision makers, patient organisations, the wider public and future venture capitalists;
• Set up a knowledge management system to monitor potential IP;
• Establish an exploitation strategy to ensure that GLINT project results impact the European diagnostics sector and detail post-project financing for further development and future commercialisation;
• Coordinate and support the activities of the External International Advisory Board (EIAB) and the Exploitation Advisory Board (ExB).