Dynamic Activatable Fluorophores

Informations projet

DYNAFLUORS

N° de convention de subvention: 771443

DOI

10.3030/771443

Projet clôturé

Date de signature de la CE 17 Avril 2018

Date de début 1 Juin 2018

Date de fin 31 Decembre 2024

Financé au titre de

• EXCELLENT SCIENCE - European Research Council (ERC)

Coût total

€ 1 986 650,00

Contribution de l’UE

€ 1 986 650,00

Coordonné par

THE UNIVERSITY OF EDINBURGH
United Kingdom

Periodic Reporting for period 5 - DYNAFLUORS (Dynamic Activatable Fluorophores)

Période du rapport: 2024-06-01 au 2024-12-31

Résumé du contexte et des objectifs généraux du projet

The vision for DYNAFLUORS was to develop the first chemical imaging probes able to monitor in real time the activity of immune cells. The research and technological achievements for each of the 3 Aims are detailed below.

1. Imaging immune cells in tumours. We produced new chemical methodologies and invented a new rapid methodology for the production of highly photostable NIR tricarbocyanine N-triazole dyes. The latter are protected in a patent and have been tested in translational model of ocular inflammation. We have also invented SCOTfluors as some of the smallest NIR fluorophores to label biomolecules without impairing their biological activity. SCOTfluors can also be used for light-guided therapy such as photodynamic therapy, and be converted into amino acids for the preparation of fluorogenic peptides. Another key objective of Aim 1 was the synthesis and characterisation of chemokine-profluorophores to target metastasis-associated macrophages. We have used chemokines (e.g. CCL2) to target recruited malignant macrophages. Our novel AND-construct mCCL2-MAF specifically labels in vivo subpopulations of malignant macrophages in a preclinical model of breast cancer. Fluorescent chemokines can be also applied to detect subpopulations of B cells and offer opportunities to visualise subsets of immune cells that were not ‘visible’ to date as well as to modulate their function in tumours and other diseases.

2. Functional probes for adaptive immune cells. The lack of direct biomarkers to monitor T cell activity in tumours hampers the rational design of effective immunotherapies. We have designed new granzyme B-reactive peptide structures and reported the first chemiluminescent granzyme B probe for imaging NK and T cell activity in vivo in tumours. We have also prepared fluorescent analogues for granzyme A and employed it to image natural killer cells in tumours. This preliminary work led to two ERC PoC grant (IBDIMAGE and FLUOROTRAP) and the EIC Transition IBDSENSE to translate granzyme B-reactive constructs as disruptive technologies for the diagnosis of immune-related diseases.

3. Translation. We have applied our probes to clinical samples (e.g. tumour biopsies, stool supernatants, urine) through national and international collaborations to translate the lead probes from Aims 1 and 2 for biomedical use.
We have established collaborations with multiple clinical groups to translate our technologies to biomedical applications. These include teams in Edinburgh (Dockrell, Akram, Ho), in Glasgow (Knight), in Barcelona (Guirado) and in Groningen (Nagengast).
Examples of translation include 10 technologies licensed and commercialised worldwide:
• Merck Millipore: commercialisation of Trp-BODIPY.
• Cambridge Research Biochemicals: license for use of Trp-BODIPY in Discovery® Peptides.
• BioLegend: commercialisation of ApoTrackerTM reagents.
• TOCRIS: commercialisation of SCOTfluors: Se-NADA, SCOTfluor lactic acid 510, SCOTfluor glucose 510, SCOTfluor 510 fluoro, ADC Tracker Green.
• IRIS Biotech: commercialisation of PotM fluorogenic amino acids: Fmoc-L-Dap(NBSD)-OH and Fmoc-L-Dap(NBTD)-OH.

Travail effectué depuis le début du projet jusqu’à la fin de la période considérée dans le rapport et principaux résultats atteints jusqu’à présent

The research performed in DYNAFLUORS has resulted in innovations and new methodologies with high impact in optical probe development and bioimaging and translational studies. Since the beginning of DYNAFLUORS, the team has delivered numerous high-impact outputs for knowledge transfer, with excellent innovation, creativity and productivity, including 50+ scientific publications, 4 patents on fluorescent technologies for biomedical imaging, fluorescent reagents commercialised under exclusive or non-exclusive licenses. The PI has been invited to give many national and international conferences/seminars, including 20+ keynote lectures.

Selected 5 key publications:
1. N. Barth, …, M. Vendrell*. A fluorogenic cyclic peptide for imaging and quantification of drug-induced apoptosis. Nat. Commun. 2020, 11, 4027. [Development of Apotrackers as fluorescent peptides for universal detection of apoptotic cells in vivo. Commercialised worldwide by BioLegend (exclusive license)].
2. J. Scott, ... M. Vendrell*. A fluorogenic probe for granzyme B enables in-biopsy evaluation and screening of response to anticancer immunotherapies. Nat. Commun. 2022, 13, 2366. [An enzyme-activatable peptide technology to monitor T cell function in biosamples. Led to formation of the spin-out company IDxSense].
3. F. de Moliner, …, M. Vendrell*. Small fluorogenic for peptide-guided background-free imaging. Angew. Chem. Int. Ed. 2023, e202216231. [3 fluorescent amino acids commercialised by Tocris and Iris Biotech].
4. E. Kuru*, …, M. Vendrell*, G. M. Church*. Rapid discovery and evolution of nanosensors containing fluorogenic amino acids. Nat. Commun. 2024, 15, 7531. [An evolution-guided strategy to generate fluorescent protein biosensors as low-cost and real-time diagnostics].
5. M. Bertolini, …, M. Vendrell*. Chemo-click: receptor-controlled and bioorthogonal chemokine ligation for real-time imaging of drug-resistant leukemic B cells. J. Am. Chem. Soc. 2024, 146, 30565. [First-in-class platform combining chemokine proteins and bioorthogonal reactions to label disease-relevant subpopulations of immune cells, with potential as diagnostics for inflammatory diseases and cancer patient stratification].

Progrès au-delà de l’état des connaissances et impact potentiel prévu (y compris l’impact socio-économique et les conséquences sociétales plus larges du projet jusqu’à présent)

The team has developed a very strong network of collaborations with industrial partners for technology transfer. As detailed above, the team has filed 4 patents, some of which have led to licenses for technologies that are currently commercialised by global manufacturers in the field of fluorescence reagents and imaging (BioLegend, Merck/MilliporeSigma, TOCRIS, IRIS Biotech). Furthermore, the team has drawn widely to collaborate with pharmaceutical companies and biotechs to establish collaborations that add value to the project and have led to MTAs for testing new technologies and student placements. The team is also sharing technologies and collaborating widely with academic groups within the scientific community. Many of these collaborations have led to additional research funding (2 ERC PoC, EIC Transition) to explore other collaborative projects, unilateral or bilateral student exchanges and participation in European research consortia (NOVAMRI, PIANO).

In summary, DYNAFLUORS has delivered outstanding progress at multiple levels:
1. Several of the technologies developed in DYNAFLUORS are currently evaluated in clinical studies to advance in the diagnostic of inflammatory diseases. The translation of these technologies would have a positive impact in personalised medicine.
2. Some early-career researchers have gone on to independent positions in academia or industry. They have all found their own niche and are fully independent (details above). The PI has consolidated as Chair of Translational Chemistry and Biomedical Imaging and head of IRR Chemistry Hub at the University of Edinburgh.
3. The PI has secured funds for ERC PoC projects and EIC Transition grants to translate the best technologies developed in DYNAFLUORS. The PI has also submitted applications to other ERC schemes and intends to secure additional ERC funding building on the preliminary data generated in DYNAFLUORS.

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