Introducing deuterium for next generation chemical biology probes and direct imaging

Projektinformationen

deuterON

ID Finanzhilfevereinbarung: 101042046

DOI

10.3030/101042046

EK-Unterschriftsdatum 17 Mai 2022

Startdatum 1 September 2022

Enddatum 31 August 2028

Finanziert unter

• European Research Council (ERC)

Gesamtkosten

€ 1 499 469,00

EU-Beitrag

€ 1 499 469,00

Koordiniert durch

FORSCHUNGSVERBUND BERLIN EV
Germany

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

Berichtszeitraum: 2022-09-01 bis 2025-02-28

Zusammenfassung vom Kontext und den Gesamtzielen des Projekts

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.

Arbeit, die ab Beginn des Projekts bis zum Ende des durch den Bericht erfassten Berichtszeitraums geleistet wurde, und die wichtigsten bis dahin erzielten Ergebnisse

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.

Fortschritte, die über den aktuellen Stand der Technik hinausgehen und voraussichtliche potenzielle Auswirkungen (einschließlich der bis dato erzielten sozioökonomischen Auswirkungen und weiter gefassten gesellschaftlichen Auswirkungen des Projekts)

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