We propose to acquire a new CyTOF mass cytometer from DVS Sciences to complement the available instrumentation at the University of Rochester's Flow Cytometry Core. Further, this instrument is being acquired to serve as a regional resource for the upstate New York region. The heart of the new CyTOF mass cytometer is an induced coupled plasma Mass spectrometer (ICP--MS), that can very precisely measure and quantify the atomic mass of the elemental tags. These tags, labeled to antibodies, are similar to fluorochromes in traditional fluorescence based cytometry, with several important advantages including more markers can be probed simultaneously (33 at present, with the potential of up to 100), no 'mass- overlap', thus no Need for compensation, which limits sensitivity in traditional fluorescent cytometry and no cellular background/autofluorescence, as the chosen elements do not exist normally in cells. Thus, the CyTOF will allow researchers the unparalleled ability to probe the complex cellular pathways of normal and diseased cells in great detail at both the surface antigen level, allowing for traditional cellular sub-setting, as well as offering a complex understanding of signaling pathways in these subsets. The interconnection between these signaling pathways will be able to be understood at the single cell level allowing researchers to design and target new therapies. The proposed studies will be conducted initially by 11 major PIs who oversee over 20 NIH grants. Additional collaborations and outreach will target, and recruit those researchers of the URMC Flow Core (over 100), as well as those in Syracuse and Buffalo who could benefit from this technology. By obtaining the technology proposed in this grant, the Western NY regional researchers will be uniquely positioned to advance the knowledge of treatment for several forms of human disease. Conditions pertaining to our studies include autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and lupus;viral infection, parasites, lymphoma and leukemia.
Identification of changes in signaling pathways of cells is critical for our understanding of how diseases, such as cancer and lupus, perturb the normal cellular responses. Understanding these perturbations can lead to a better way to selectively and specifically target the diseased state cells, thus resulting in a better understanding of how t tackle and cure a patient. Such targeted therapies remain elusive because of limitations of current tools that make such complex, cellular network studies difficult and cumbersome. The CyTOF mass cytometer will allow researcher in the upstate New York regions (including Buffalo, Syracuse and Rochester) to being to rapidly identify these signaling pathways in well defined populations, at the single cell level, thus offering the opportunity to understand and treat diseases more efficiently and more accurately.