Molecular probes for lysosomal accumulation, release and environmental sensing.

The project will operate in the EPSRC research theme of Healthcare Technologies, in the priority research area of Technology Touching Life, and the areas of Chemical Biology, Analytical Science, and Synthetic Organic Chemistry.

Lysosomes are membrane bound organelles characterised by a low pH, which contain hydrolytic enzymes that function as part of the cellular recycling system. The low pH associated with these organelles provides a unique opportunity for accumulation, release, and sensing using small molecules, permitting an understanding of this chemical environment and its relationship to disease.

We will design, synthesise, and monitor small molecules that are able to examine each of these areas using bioinformatics to process and interpret the imaging data generated. The Project will be conducted in three distinct areas that will feedback to enable probe development.

Area 1: Synthetic chemistry. The design and synthesis of small molecules that will accumulate in the lysosomal region of a cell. The probes will contain three distinct regions (i) an environmentally sensitive group that is responsive to either the pH or redox environment of a cell. (ii) a Raman reporter consisting of a conjugated bisarylbutadiyne moiety. (iii) an lysosome targeting group that will initially comprise of a secondary amine, or protected secondary amine that will be unmasked in the lysosome.

Area 2: Analytical Chemistry. The ability of the compounds to report back on either pH or redox potential as ratiometric probes will be determined. If successful, the ability of probes to accumulate with lysosomes will be established.

Area 3. On identification of an appropriate probe, we will establish chemical biology workflows to monitor the effect of exogenous molecules on lysosome environment in a spatiotemporal manner, identifying screens to monitor phenotypic readouts in models of disease.