PMID- 20631065 OWN - NLM STAT- MEDLINE DCOM- 20100913 LR - 20211203 IS - 1538-7445 (Electronic) IS - 0008-5472 (Print) IS - 0008-5472 (Linking) VI - 70 IP - 15 DP - 2010 Aug 1 TI - A microfluidic platform for systems pathology: multiparameter single-cell signaling measurements of clinical brain tumor specimens. PG - 6128-38 LID - 10.1158/0008-5472.CAN-10-0076 [doi] AB - The clinical practice of oncology is being transformed by molecular diagnostics that will enable predictive and personalized medicine. Current technologies for quantitation of the cancer proteome are either qualitative (e.g., immunohistochemistry) or require large sample sizes (e.g., flow cytometry). Here, we report a microfluidic platform-microfluidic image cytometry (MIC)-capable of quantitative, single-cell proteomic analysis of multiple signaling molecules using only 1,000 to 2,800 cells. Using cultured cell lines, we show simultaneous measurement of four critical signaling proteins (EGFR, PTEN, phospho-Akt, and phospho-S6) within the oncogenic phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling pathway. To show the clinical application of the MIC platform to solid tumors, we analyzed a panel of 19 human brain tumor biopsies, including glioblastomas. Our MIC measurements were validated by clinical immunohistochemistry and confirmed the striking intertumoral and intratumoral heterogeneity characteristic of glioblastoma. To interpret the multiparameter, single-cell MIC measurements, we adapted bioinformatic methods including self-organizing maps that stratify patients into clusters that predict tumor progression and patient survival. Together with bioinformatic analysis, the MIC platform represents a robust, enabling in vitro molecular diagnostic technology for systems pathology analysis and personalized medicine. FAU - Sun, Jing AU - Sun J AD - Crump Institute for Molecular Imaging, University of California at Los Angeles, Los Angeles, California 90095, USA. FAU - Masterman-Smith, Michael D AU - Masterman-Smith MD FAU - Graham, Nicholas A AU - Graham NA FAU - Jiao, Jing AU - Jiao J FAU - Mottahedeh, Jack AU - Mottahedeh J FAU - Laks, Dan R AU - Laks DR FAU - Ohashi, Minori AU - Ohashi M FAU - DeJesus, Jason AU - DeJesus J FAU - Kamei, Ken-ichiro AU - Kamei K FAU - Lee, Ki-Bum AU - Lee KB FAU - Wang, Hao AU - Wang H FAU - Yu, Zeta T F AU - Yu ZT FAU - Lu, Yi-Tsung AU - Lu YT FAU - Hou, Shuang AU - Hou S FAU - Li, Keyu AU - Li K FAU - Liu, Max AU - Liu M FAU - Zhang, Nangang AU - Zhang N FAU - Wang, Shutao AU - Wang S FAU - Angenieux, Brigitte AU - Angenieux B FAU - Panosyan, Eduard AU - Panosyan E FAU - Samuels, Eric R AU - Samuels ER FAU - Park, Jun AU - Park J FAU - Williams, Dirk AU - Williams D FAU - Konkankit, Vera AU - Konkankit V FAU - Nathanson, David AU - Nathanson D FAU - van Dam, R Michael AU - van Dam RM FAU - Phelps, Michael E AU - Phelps ME FAU - Wu, Hong AU - Wu H FAU - Liau, Linda M AU - Liau LM FAU - Mischel, Paul S AU - Mischel PS FAU - Lazareff, Jorge A AU - Lazareff JA FAU - Kornblum, Harley I AU - Kornblum HI FAU - Yong, William H AU - Yong WH FAU - Graeber, Thomas G AU - Graeber TG FAU - Tseng, Hsian-Rong AU - Tseng HR LA - eng GR - R56 NS052563/NS/NINDS NIH HHS/United States GR - DP2 OD006462/OD/NIH HHS/United States GR - DP2 OD006462-01/OD/NIH HHS/United States GR - R01 NS052563-01A1/NS/NINDS NIH HHS/United States GR - T32 CA009056-35/CA/NCI NIH HHS/United States GR - NS052563/NS/NINDS NIH HHS/United States GR - T32 CA009056/CA/NCI NIH HHS/United States GR - U54 CA119347/CA/NCI NIH HHS/United States GR - R01 NS052563/NS/NINDS NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20100714 PL - United States TA - Cancer Res JT - Cancer research JID - 2984705R RN - 0 (Intracellular Signaling Peptides and Proteins) RN - EC 2.7.1.1 (MTOR protein, human) RN - EC 2.7.10.1 (ErbB Receptors) RN - EC 2.7.11.1 (Protein Serine-Threonine Kinases) RN - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt) RN - EC 2.7.11.1 (Ribosomal Protein S6 Kinases) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - EC 3.1.3.67 (PTEN Phosphohydrolase) RN - EC 3.1.3.67 (PTEN protein, human) SB - IM MH - Brain Neoplasms/metabolism/*pathology MH - Cell Line, Tumor MH - ErbB Receptors/metabolism MH - Glioblastoma/metabolism/*pathology MH - Humans MH - Immunohistochemistry MH - Intracellular Signaling Peptides and Proteins/metabolism MH - Microfluidic Analytical Techniques/instrumentation/*methods MH - PTEN Phosphohydrolase/metabolism MH - Phosphatidylinositol 3-Kinases/metabolism MH - Phosphorylation MH - Protein Serine-Threonine Kinases/metabolism MH - Proto-Oncogene Proteins c-akt/metabolism MH - Reproducibility of Results MH - Ribosomal Protein S6 Kinases/metabolism MH - Signal Transduction MH - TOR Serine-Threonine Kinases PMC - PMC3163840 MID - NIHMS317232 EDAT- 2010/07/16 06:00 MHDA- 2010/09/14 06:00 PMCR- 2011/08/30 CRDT- 2010/07/16 06:00 PHST- 2010/07/16 06:00 [entrez] PHST- 2010/07/16 06:00 [pubmed] PHST- 2010/09/14 06:00 [medline] PHST- 2011/08/30 00:00 [pmc-release] AID - 0008-5472.CAN-10-0076 [pii] AID - 10.1158/0008-5472.CAN-10-0076 [doi] PST - ppublish SO - Cancer Res. 2010 Aug 1;70(15):6128-38. doi: 10.1158/0008-5472.CAN-10-0076. Epub 2010 Jul 14.