Determining the signatures of pan-ubiquitylation in the immune proteome as highly sensitive and effective markers for neurodegenerative disease

SUMMARYAlzheimerandapos s diseaseADis the most devastating age related disease due to its damage to cognitive and motor capacityheavily care dependent natureand lack of early diagnostics and effective therapeuticsThe first step toward its cure is to discover early stage molecular markers based on its fundamental pathogenesisaccumulation of protein aggregates in plaques and chronic proinflammationThe main biochemical regulatory mechanism governing these two cellular events is post translational modification of proteins by ubiquitinUband ubiquitin like proteinsUblsCirculating immune cells in blood and cerebrospinal fluidCSFprovide a means of liquid biopsy to monitor protein degeneration in the brainWe hypothesize that the signature of the Ub and Ubl immune proteome serves as a marker for ADHoweverthe precise information encoded in these Ub Ubl modification sites is poorly understoodThis is mainly due to a lack of reagents to selectively isolate Ubor Ubl conjugated peptides from protease treated cell lysatesWe will develop methods to allow the specifically isolation and identification of Ub or Ubl conjugation sites at the proteome levelThis will be accomplished by developing a novel affinity matrix to specifically enrich the ubiquitin remnants generated from protease digestionresulting in optimal quantification by mass spectrometry analysisUsing this technologywe will identify the bona fide pan ubiquitylation patterns of the immune proteome as a pathological fingerprint for neurodegenerationThe success of this project will provide the first comprehensive pan ubiquitin immune proteome to monitor and diagnose AD Profiling the spectrum of diseases through the immune proteome is an attractive approach to sense brain degeneration through immune cells as a means of liquid biopsyTo datethe tools for identifying the ubiquitin specific proteome have not been availableLifeSensors will develop reagents for ubiquitin and ubiquitin like proteins to establish such a proteomics analysiswhich can be applied to identify the suitability of druggable candidates to treat age associated neurodegenerative diseases