Single-cell proteomic chip for profiling intracellular signaling pathways in single tumor cells

We describe a microchip designed to quantify the levels of a dozen cytoplasmic and membrane proteins from single cells. We use the platform to assess protein–protein interactions associated with the EGF-receptor-mediated PI3K signaling pathway. Single-cell sensitivity is achieved by isolating a defi...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2012-01, Vol.109 (2), p.419-424
Main Authors: Shi, Qihui, Qin, Lidong, Wei, Wei, Geng, Feng, Fan, Rong, Shik Shin, Young, Guo, Deliang, Hood, Leroy, Mischel, Paul S, Heath, James R
Format: Article
Language:English
Subjects:
Antibodies
Bar codes
Biological Sciences
Cell Line, Tumor
Cell lines
cell membranes
Cells
DNA
Erlotinib Hydrochloride
Fluorescence
Glioblastoma
Humans
Intracellular Signaling Peptides and Proteins - genetics
Intracellular Signaling Peptides and Proteins - metabolism
membrane proteins
phosphatidylinositol 3-kinase
Phosphorylation
Physical Sciences
Protein Array Analysis - methods
Proteins
Proteomics
Proteomics - methods
Quinazolines
Signal transduction
Signal Transduction - genetics
T cell receptors
T lymphocytes
Tumor cell line
Tumors
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Description
Summary:We describe a microchip designed to quantify the levels of a dozen cytoplasmic and membrane proteins from single cells. We use the platform to assess protein–protein interactions associated with the EGF-receptor-mediated PI3K signaling pathway. Single-cell sensitivity is achieved by isolating a defined number of cells (n = 0–5) in 2 nL volume chambers, each of which is patterned with two copies of a miniature antibody array. The cells are lysed on-chip, and the levels of released proteins are assayed using the antibody arrays. We investigate three isogenic cell lines representing the cancer glioblastoma multiforme, at the basal level, under EGF stimulation, and under erlotinib inhibition plus EGF stimulation. The measured protein abundances are consistent with previous work, and single-cell analysis uniquely reveals single-cell heterogeneity, and different types and strengths of protein–protein interactions. This platform helps provide a comprehensive picture of altered signal transduction networks in tumor cells and provides insight into the effect of targeted therapies on protein signaling networks.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1110865109