How does the molecular composition of cells and cell-cell contacts change over time or after experimental manipulations? This fundamental question is asked across all fields of biomedical science and is at the heart of most studies focused on determining cellular and molecular mechanisms of biological phenomenon. Yet, our ability to answer this question is severely constrained by the limitations of current methods to detect the protein composition of particular cellular structures. Imagine the rapid progress biological science could make if we could detect more than a handful of proteins at specified locations on cells in each experiment. The objective of our project is to develop a new approach to achieve this goal?called ?context-rich mass cytometry? ?a technology that enables multi-parametric analysis of proteins in the context of cellular interactions. Mass spectrometry has recently emerged as a powerful tool for cell analysis. One version of this technology called mass cytometry or CyToF is used for multi-parametric analysis of protein expression in single cells. It is similar to flow cytometry in that cells are labeled with antibodies and analyzed on cell-by-cell basis in high- throughput manner. However, unlike flow cytometry which employs fluorescently-labeled antibodies and is limited to ~12 parameters, mass cytometry employs rare earth metals as antibody labels and may be used to analyze up to 60 intracellular or cell surface markers. While an exciting technology, mass cytometry requires that cells be extracted from their native microenvironment and homogenized into single cell suspension prior to analysis. This makes connecting the protein signature to the microenvironment context very challenging. The objective of our project is to develop ?context-rich mass cytometry? ?a technology that enables multi- parametric analysis of proteins in the context of cellular interactions. Once developed this mass spectrometry approach will be used to determine the composition and organization of proteins within immune synapses. This new knowledge enabled by our technology may, in the future, be parlayed into strategies and therapies for preventing infections.

Public Health Relevance

In a variety of biological systems information is exchanged via synapse ? a junction formed between adjacent cells. For example, such synapses are formed by immune cells to convey information about pathogens invading the host. What happens at these synapses and what kind of molecules are involved in forming and shaping synapses is a critically important but poorly understood question. Our goal is to remedy this by developing a novel technology for profiling and quantifying proteins involve in neural and immune synapses.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM123757-03S1
Application #
9956224
Study Section
Enabling Bioanalytical and Imaging Technologies Study Section (EBIT)
Program Officer
Krepkiy, Dmitriy
Project Start
2017-09-06
Project End
2021-07-31
Budget Start
2019-08-01
Budget End
2020-07-31
Support Year
3
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Mayo Clinic, Rochester
Department
Type
DUNS #
006471700
City
Rochester
State
MN
Country
United States
Zip Code
55905