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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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 2012-01, Vol.109 (2), p.419-424 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
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| creator | Shi, Qihui Qin, Lidong Wei, Wei Geng, Feng Fan, Rong Shik Shin, Young Guo, Deliang Hood, Leroy Mischel, Paul S Heath, James R |
| description | 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. |
| doi_str_mv | 10.1073/pnas.1110865109 |
| format | Article |
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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.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.1110865109</identifier><identifier>PMID: 22203961</identifier><language>eng</language><publisher>United States: National Academy of Sciences</publisher><subject>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</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 2012-01, Vol.109 (2), p.419-424</ispartof><rights>copyright © 1993—2008 National Academy of Sciences of the United States of America</rights><rights>Copyright National Academy of Sciences Jan 10, 2012</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c553t-f38bef58388da69495290014e1af4c500e6cd2d0c4e86227767aac5030ac8b8a3</citedby><cites>FETCH-LOGICAL-c553t-f38bef58388da69495290014e1af4c500e6cd2d0c4e86227767aac5030ac8b8a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/109/2.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/23075987$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/23075987$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,315,728,781,785,804,886,27929,27930,53796,53798,58022,58255</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22203961$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Shi, Qihui</creatorcontrib><creatorcontrib>Qin, Lidong</creatorcontrib><creatorcontrib>Wei, Wei</creatorcontrib><creatorcontrib>Geng, Feng</creatorcontrib><creatorcontrib>Fan, Rong</creatorcontrib><creatorcontrib>Shik Shin, Young</creatorcontrib><creatorcontrib>Guo, Deliang</creatorcontrib><creatorcontrib>Hood, Leroy</creatorcontrib><creatorcontrib>Mischel, Paul S</creatorcontrib><creatorcontrib>Heath, James R</creatorcontrib><title>Single-cell proteomic chip for profiling intracellular signaling pathways in single tumor cells</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>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.</description><subject>Antibodies</subject><subject>Bar codes</subject><subject>Biological Sciences</subject><subject>Cell Line, Tumor</subject><subject>Cell lines</subject><subject>cell membranes</subject><subject>Cells</subject><subject>DNA</subject><subject>Erlotinib Hydrochloride</subject><subject>Fluorescence</subject><subject>Glioblastoma</subject><subject>Humans</subject><subject>Intracellular Signaling Peptides and Proteins - genetics</subject><subject>Intracellular Signaling Peptides and Proteins - metabolism</subject><subject>membrane proteins</subject><subject>phosphatidylinositol 3-kinase</subject><subject>Phosphorylation</subject><subject>Physical Sciences</subject><subject>Protein Array Analysis - methods</subject><subject>Proteins</subject><subject>Proteomics</subject><subject>Proteomics - methods</subject><subject>Quinazolines</subject><subject>Signal transduction</subject><subject>Signal Transduction - genetics</subject><subject>T cell receptors</subject><subject>T lymphocytes</subject><subject>Tumor cell line</subject><subject>Tumors</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdkc1v1DAQxS0EokvhzAmIuHBKO7bjrwsSqviSKnEoPVuzXmfXqyQOdgLqf4_TXbrAxZbm_eZ5PI-QlxQuKCh-OQ6YLyiloKWgYB6RVTlpLRsDj8kKgKlaN6w5I89y3gOAERqekjPGGHAj6YrYmzBsO18733XVmOLkYx9c5XZhrNqYllIbusJUYZgSLtjcYapy2A54Xx9x2v3Cu1yAUl3MqmnuS-vC5ufkSYtd9i-O9zm5_fTx-9WX-vrb569XH65rJwSf6pbrtW-F5lpvUJrGCGYAaOMpto0TAF66DduAa7yWjCklFWKpc0Cn1xr5OXl_8B3nde83zi_TdnZMocd0ZyMG-68yhJ3dxp-WM6GFlsXg3dEgxR-zz5PtQ16-gIOPc7aGClneVaaQb_8j93FOZRv3kFG85FGgywPkUsw5-fZhFAp2ic4u0dlTdKXj9d8_eOD_ZFWAV0dg6TzZGctsQ81J3-cpplM_ByWMVkV_c9BbjBa3KWR7e8PKksuiNdNK89-woLNg</recordid><startdate>20120110</startdate><enddate>20120110</enddate><creator>Shi, Qihui</creator><creator>Qin, Lidong</creator><creator>Wei, Wei</creator><creator>Geng, Feng</creator><creator>Fan, Rong</creator><creator>Shik Shin, Young</creator><creator>Guo, Deliang</creator><creator>Hood, Leroy</creator><creator>Mischel, Paul S</creator><creator>Heath, James R</creator><general>National Academy of Sciences</general><general>National Acad Sciences</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QG</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20120110</creationdate><title>Single-cell proteomic chip for profiling intracellular signaling pathways in single tumor cells</title><author>Shi, Qihui ; 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| 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 |
| title | Single-cell proteomic chip for profiling intracellular signaling pathways in single tumor cells |
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