The ability to visualize the presence, abundance, location and functional state of many proteins at once in cells and tissues with preserved morphology has long been a goal. This proposal seeks to optimize recently demonstrated mass-spec-based imaging methods that can, when fully developed, detect as many as 40 to 100+ molecular targets in cells and tissues using simultaneously applied mass-tag-labeled antibodies, with outstanding morphology. The technique is analogous to laser-scanning confocal microscopy, except that the scanning is performed using a tightly focused ion beam, rather than a similarly sized point of laser light. And instead of fluorophores, the labels detected are clustes of metal atoms attached to any given probe. Preliminary data using a highly focused, scanning oxygen ion beam and 10 antibodies applied simultaneously to clinical breast cancer tissues demonstrate histology-like images that reveal bound antibody location with subcellular resolution. Going forward, we would like to optimize sensitivity and reliability, with the goal of making this a standard, high-performance, imaging approach for basic and translational science discovery, drug development, and possible clinical deployment. The present project will address: reliable pre-analytical sample preparation methods and labeling techniques for protein analytes; ion-beam imaging method improvements based on current and projected instrumentation; powerful, easy-to-use software for equipment control, display and analysis; and validation in several important biological test cases.

Public Health Relevance

The biology and clinical course of cancers are often predicted by the presence and distribution of proteins within cells that are involved in cell division, invasion, metastasis and evasion of the host's immune system. We describe a new, non-optical microscopic technology that can visualize as many as 100 proteins simultaneously. We think this may prove to be a remarkable engine for research and clinical advances in the understanding and treatment of cancer.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants Phase II (R33)
Project #
5R33CA183654-02
Application #
8860156
Study Section
Special Emphasis Panel (ZCA1)
Program Officer
Knowlton, John R
Project Start
2014-07-01
Project End
2017-06-30
Budget Start
2015-07-01
Budget End
2016-06-30
Support Year
2
Fiscal Year
2015
Total Cost
Indirect Cost
Name
Stanford University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94304
Good, Zinaida; Sarno, Jolanda; Jager, Astraea et al. (2018) Single-cell developmental classification of B cell precursor acute lymphoblastic leukemia at diagnosis reveals predictors of relapse. Nat Med 24:474-483
Goltsev, Yury; Samusik, Nikolay; Kennedy-Darling, Julia et al. (2018) Deep Profiling of Mouse Splenic Architecture with CODEX Multiplexed Imaging. Cell 174:968-981.e15
O'Gorman, W E; Kong, D S; Balboni, I M et al. (2017) Mass cytometry identifies a distinct monocyte cytokine signature shared by clinically heterogeneous pediatric SLE patients. J Autoimmun :
Burns, Tyler J; Frei, Andreas P; Gherardini, Pier F et al. (2017) High-throughput precision measurement of subcellular localization in single cells. Cytometry A 91:180-189
Han, Guojun; Chen, Shih-Yu; Gonzalez, Veronica D et al. (2017) Atomic mass tag of bismuth-209 for increasing the immunoassay multiplexing capacity of mass cytometry. Cytometry A 91:1150-1163
Jensen, Helle; Chen, Shih-Yu; Folkersen, Lasse et al. (2017) EBI3 regulates the NK cell response to mouse cytomegalovirus infection. Proc Natl Acad Sci U S A 114:1625-1630
Mukherjee, Sayak; Jensen, Helle; Stewart, William et al. (2017) In silico modeling identifies CD45 as a regulator of IL-2 synergy in the NKG2D-mediated activation of immature human NK cells. Sci Signal 10:
Kleppe, Maria; Spitzer, Matthew H; Li, Sheng et al. (2017) Jak1 Integrates Cytokine Sensing to Regulate Hematopoietic Stem Cell Function and Stress Hematopoiesis. Cell Stem Cell 21:489-501.e7
Spitzer, Matthew H; Carmi, Yaron; Reticker-Flynn, Nathan E et al. (2017) Systemic Immunity Is Required for Effective Cancer Immunotherapy. Cell 168:487-502.e15
Dodd, Dylan; Spitzer, Matthew H; Van Treuren, William et al. (2017) A gut bacterial pathway metabolizes aromatic amino acids into nine circulating metabolites. Nature 551:648-652

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