Introducing deuterium for next generation chemical biology probes and direct imaging

Informazioni relative al progetto

deuterON

ID dell’accordo di sovvenzione: 101042046

DOI

10.3030/101042046

Data della firma CE 17 Maggio 2022

Data di avvio 1 Settembre 2022

Data di completamento 31 Agosto 2028

Finanziato da

• European Research Council (ERC)

Costo totale

€ 1 499 469,00

Contributo UE

€ 1 499 469,00

Coordinato da

FORSCHUNGSVERBUND BERLIN EV
Germany

Periodic Reporting for period 1 - deuterON (Introducing deuterium for next generation chemical biology probes and direct imaging)

Periodo di rendicontazione: 2022-09-01 al 2025-02-28

Sintesi del contesto e degli obiettivi generali del progetto

The project's main objective is to explore isotope effects, in particular heavy hydrogen (deuterium) in the area of Chemical Biology to break interdisciplinary new ground with implications to combat diseases. This is subdivided into 3 parts: 1) The organic synthesis of a new generation of dyes for fluorescent microscopy. Fluorescence microscopy is of vital importance to understand cellular function, however, small organic dyes often limit sophisticated applications. As such, we aim to introduce deuterium on a new class of fluorophores to overcome such roadblocks, making these chemicals more stable, that is to last longer. 2) Organic synthesis and exploration of small molecule photoswitches by means of deuteration. Such compounds may be used to control biological function with light, with the main advantage that it is remote-controllable with respect to traditional pharmacology. To progress the field, faster and more stable switches with enhanced pharmacological profiles are needed, which will be achieved by using deuterium. 3) In imaging, a main desire is to not perturb the studied cellular system at hand. By using deuterium, we aim to image drug distribution and catabolism directly using Raman microscopy. Combined, these aims steer toward a generalizable big impact in the life sciences.

Lavoro eseguito dall’inizio del progetto fino alla fine del periodo coperto dalla relazione e principali risultati finora ottenuti

Main activities have been rooted in organic synthesis and photophysical characterizations. The need to synthesize deuterium containing molecules is challenging due to their change in reactivity with respect to non-deuterated species. We have tackled this successfully. High purity is achieved by reverse phase high performance liquid chromatography. Photophysical measurements include UV/Vis spectroscopy, fluorescence quantum yield. Labelling of proteins has been achieved in vitro, with biomolecules from recombinant sources. Cell culture has been performed, including molecular biology (e.g. cloning, transfection), in HEK293, CHO-K1 and MIN6 cells, and is combined with sophisticated imaging by widefield, confocal, lifetime and super-resolution stimulated emission by depletion microscopy. Main achievements inlcude:
1) Roßmann; […], Levitz*; Broichhagen*, Deuteration as a general strategy to enhance azobenzene-based photopharmacology, Angew. Chem. Int. Ed. 2024. Highlighting that deuteration of photoswitches enhances efficiency and speed for the optical control of ion channels and GPCRs.
2) Ast; […], Broichhagen*; Hodson*, Revealing the tissue-level complexity of endogenous glucagon-like peptide 1-receptor expression and signaling, Nat. Commun. 2023. Created a CRISPR/Cas9 SNAP-tag knock-in mouse to study the glucagon-like peptide 1 receptor using custom-tailored dyes for sensitive microscopy.
3) Invited speaker to the "Chemiedozententagung", the 128th International Titisee Conference, the 9th European Synapse Meeting, and the 5th Meeting of the Incretin Study Group, as well as the 28th and 29th International Workshop of PicoQuant.

Progressi oltre lo stato dell’arte e potenziale impatto previsto (incluso l’impatto socioeconomico e le implicazioni sociali più ampie del progetto fino ad ora)

We have applied for IP protecting our peptide labelling technology and are in the process of filing a patent.