Single cell analysis of the kinome

Project Summary
Profiling kinases and their activities in single cells will enable exploration of cell type specific
functional and biochemical pathways. While we have learned a great deal about transcriptional
states of different cell types in the past decade through single cell genomics, we still understand
little about proteins and in particular kinases networks in diverse cell populations. This is largely
because single-cell kinome analysis faces major technical challenges. Unlike nucleic acids,
proteins cannot be amplified, making detection of minute quantities from single cells difficult. We
recently demonstrated a proof-of-principle experiment to accurately detect protein PTM isoforms
in single cells. Here we propose to scale this method up to profile kinases at the global level in
single cells in both cell culture and in tissues. We will integrate the single cell kinome analysis
with RNA measurements to identify cell type specific kinase profiles. Ultimately, we will map
the kinome with spatial context in the mouse and human brain to examine the kinase pathways
involved in aging and neurodegeneration which could identify druggable targets. This project is a
major departure from our current research. Its goals are ambitious but achievable. The project
will leverage our expertise in single cell technology development and in application of new
technology to diverse biological systems.

Grant Number: 4DP1NS131408-04
NIH Institute/Center: NIH
Principal Investigator: Long Cai