A Multifunctional Genetic Probe for Networking Lysosomal Calcium Biology (360G-Wellcome-222541_Z_21_Z)

The lysosome, once thought to be a cellular ‘trash-can’, is now found to be the key to understanding the signalling controlling human ageing, neurodegeneration, and a range of diseases from diabetes to cancer. Developing tools to understand how the lysosome works will be key to future biological studies. In this project, we will develop and analyse a new imaging probe which will allow us to examine calcium signalling in functional lysosomes in live-imaging, proteomic and lipidomic modalities to develop a fine-grained understanding of how lysosomes function in health and disease. The genetic probe we are developing makes it easy to isolate, image, and manipulate lysosomes in living cells and in cellular extracts using one transfectable tool, offering many avenues of application to basic and industry-focused research. One key regulator of lysosomal calcium storage is the messenger NAADP, which we will examine in detail. We will produce coordinated maps of the behaviour of calcium levels inside the lysosome under conditions known to alter NAADP signalling, coupled with the metabolic and proteomic profiles of lysosomes in the same conditions and in mutants of the muscular dystrophy gene INPP5K: giving a comprehensive map of the regulation of this key signalling axis.