Project description DEENESFRITPL Zooming in on misdirected protein traffic and mapping neuropathological detours Large-scale studies of all the proteins produced by a cell or organism (proteomics) have significantly advanced our understanding of diseases. However, we need increased resolution at the subcellular level to identify protein trafficking deficits, which cause many diseases. Neurological diseases account for a large proportion of rare diseases and can have devastating effects. Neurons are unusual compared to other cells in that their structure is compartmentalized into dendrites, cell body and axon. This results in relatively ‘long’ cells, so efficient protein trafficking is particularly important in neurons. The EU-funded RARE MAPS project is exploiting a pioneering technique called Dynamic Organellar Maps. It will provide proteomic data with subcellular (organelle-level) localization in human induced pluripotent stem cell (hiPSC)-derived neurons and brains from a mouse model of a rare neurodegenerative disorder, called AP-4 deficiency syndrome. The study will boost the utility of the technique, while shedding light on the role of axonal protein trafficking in a rare neurological disease. The approach will provide a discovery pipeline that can be widely applied to rare neurological disorders. Show the project objective Hide the project objective Objective Rare diseases are a major unmet medical need, as is the definition of the relevant disease mechanisms. Many rare diseases affect the nervous system. These are challenging to treat, and mechanistic studies are difficult due to the inaccessibility of patient tissue. Global proteomic studies have provided insight into whole tissue or cell changes in protein abundance but lose information on protein subcellular localisation, which is important because defects in protein trafficking are implicated in many neurological disorders. In ‘RARE MAPS’ I propose an unbiased mechanistic discovery pipeline combining human induced pluripotent stem cells (hiPSCs) with advanced spatial proteomics. I will use a method developed by Dr. Borner called ‘dynamic organellar maps’, which provides quantitative protein subcellular localisation information at the whole proteome level. Used comparatively, it can detect changes in protein localisation due to a perturbation, allowing unbiased screening for phenotypic changes. To develop this workflow, I will apply it to the rare neurodegenerative disorder AP-4 deficiency syndrome. AP-4 knockout hiPSCs will be differentiated into cortical neurons and maps will be made of intermediate cortical stem cells and mature cortical neurons. Comparison to control cells will enable the detection of changes to protein localisation and abundance. I will also apply the maps to brain tissue from an AP-4 deficient mouse model to detect protein mislocalisation in vivo. I will then use CRISPR/Cas9 technology to investigate the role of novel and known AP-4-associated proteins in neuronal autophagy and axonal health. This project will demonstrate the utility of dynamic organellar maps to reveal molecular mechanisms of rare neurological disorders as well as provide new insights into the pathogenesis of AP-4 deficiency and the role of protein trafficking and autophagy in the axon. Fields of science natural sciencesbiological sciencesneurobiologynatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsproteomicsmedical and health sciencesmedical biotechnologycells technologiesstem cellsmedical and health sciencesbasic medicineneurology Keywords mass spectrometry induced pluripotent stem cell neuron axon gene editing CRISPR adaptor protein complex autophagy neurodegeneration hereditary spastic paraplegia vesicles Programme(s) H2020-EU.1.3. - EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions Main Programme H2020-EU.1.3.2. - Nurturing excellence by means of cross-border and cross-sector mobility Topic(s) MSCA-IF-2019 - Individual Fellowships Call for proposal H2020-MSCA-IF-2019 See other projects for this call Funding Scheme MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF) Coordinator MAX-PLANCK-GESELLSCHAFT ZUR FORDERUNG DER WISSENSCHAFTEN EV Net EU contribution € 162 806,40 Address HOFGARTENSTRASSE 8 80539 Munchen Germany See on map Region Bayern Oberbayern München, Kreisfreie Stadt Activity type Research Organisations Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Total cost € 162 806,40
