The precise tracking of protein molecules is expected to reveal a depth and richness about the B-cell differentiation pathway and deviations from it. It will provide a window into the developmental transformations from human stem cell through to B-cells and will define the major steps in between. We will employ a 3-pronged approach to survey proteins that can all contribute to the formation of cell-specific barcodes. First, targeting the chromatin template, we will track each of 75 histone marks with 2-5% precision, and follow that up with targeted ChIP-seq on those marks most changed to reveal the best gene-based classifiers of cellular differentiation. Secondly, top-down proteomics will be used to study the intracellular proteins on an isoform-specific basis. Using unsupervised clustering and advanced biostatistical methods, we will convert quantitative proteomics data into exquisitely sensitive and proteoform-resolved molecular species for inclusion in refined barcodes that can be established efficiently and used subsequently within a cell-based framework extensible to an entire system. Thirdly, the cell surface proteome of B-cells at different differentiation steps will be interrogated for eventual biomarker targeting. This study will provide us with unmatched knowledge about proteins as exquisitely sensitive reporters of cell differentiation and relatedness.
|Effective start/end date||3/1/14 → 10/1/18|
- Paul G. Allen Family Foundation (11715)