New ionization source technology for single-cell multi-omics

About This Grant

This project will develop, optimize and evaluate the analytical performance of a new tool, termed Thermal Ink Jet Spray (TIJ Spray). TIJ Spray will enable the direct injection of low input samples, including individual cells, into mass spectrometers for multi-omics analysis to enable discovery of cell phenotypic heterogeneity of drug response. TIJ Spray in conjunction with ion mobility spectrometry and integrated with high-resolution tandem mass spectrometers (IMMS) promises of providing the necessary throughput, sensitivity and specificity for detecting and characterizing cell type heterogeneity, subpopulation heterogeneity and drug response based on molecular ion profiles. Comprehensive characterization of the proteome and metabolome of individual cells allows inferring the functional state of individual cells and reveals functional phenotype heterogeneity. Knowledge of existence of functionally heterogeneous phenotypes is of relevance for the biology of tumors and tumor microenvironments because of the presence of highly heterogeneous cell populations and possible presence of cells of the same type but displaying different functional and metabolic phenotypes. Distinct phenotypes are masked in bulk cell populations, which hinders the discovery and validation of phenotype-specific biomarkers as well as confounds drug discovery. In this high risk/high reward project, we will develop this new technology and demonstrate its analytical performance for multi-omic analysis of single cells by accomplishing the following specific aims: Aim 1: Develop and optimize TIJ spray and its technical integration with IMMS. We will develop TIJ spray and demonstrate its the analytical capabilities in conjunction with IMMS for detection of different classes of analytes including those found in individual cells. Aim 2: Demonstrate TIJ Spray IMMS for multi-omics of single cells for cellular phenotyping and detection of phenotype-specific drug response. We will demonstrate and evaluate performance of TIJ Spray IMMS for obtaining multi-omics data in a pathobiologically relevant context and evaluate its analytical performance for detecting multi-omics changes, and differential drug uptake and response. When successful, we aim for achieving throughput of several hundreds of cells per day resulting in high quality multi-omic profiles for single-cell functional phenotyping. When we can show “fitness for purpose”, TIJ Spray IMMS has the potential to democratize single-cell mass spectrometry making it available for a wider range of laboratories and the biomedical community.