Introducing deuterium for next generation chemical biology probes and direct imaging

Información del proyecto

deuterON

Identificador del acuerdo de subvención: 101042046

DOI

10.3030/101042046

Fecha de la firma de la CE 17 Mayo 2022

Fecha de inicio 1 Septiembre 2022

Fecha de finalización 31 Agosto 2028

Financiado con arreglo a

European Research Council (ERC)

Coste total

€ 1 499 469,00

Aportación de la UE

€ 1 499 469,00

1 499 469,00

Coordinado por

FORSCHUNGSVERBUND BERLIN EV
Germany

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

Período documentado: 2022-09-01 hasta 2025-02-28

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.

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.

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

Descargar Descargar el contenido de la página