title: A microfluidic platform for systems pathology: multiparameter single-cell signaling measurements of clinical brain tumor specimens.
abstract: 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.
authors.affiliation: Crump Institute for Molecular Imaging, University of California at Los Angeles, Los Angeles, California 90095, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Sun
authors.forename: Michael D
authors.initials: MD
authors.lastname: Masterman-Smith
authors.forename: Nicholas A
authors.initials: NA
authors.lastname: Graham
authors.forename: Jing
authors.initials: J
authors.lastname: Jiao
authors.forename: Jack
authors.initials: J
authors.lastname: Mottahedeh
authors.forename: Dan R
authors.initials: DR
authors.lastname: Laks
authors.forename: Minori
authors.initials: M
authors.lastname: Ohashi
authors.forename: Jason
authors.initials: J
authors.lastname: DeJesus
authors.forename: Ken-ichiro
authors.initials: K
authors.lastname: Kamei
authors.forename: Ki-Bum
authors.initials: KB
authors.lastname: Lee
authors.forename: Hao
authors.initials: H
authors.lastname: Wang
authors.forename: Zeta T F
authors.initials: ZT
authors.lastname: Yu
authors.forename: Yi-Tsung
authors.initials: YT
authors.lastname: Lu
authors.forename: Shuang
authors.initials: S
authors.lastname: Hou
authors.forename: Keyu
authors.initials: K
authors.lastname: Li
authors.forename: Max
authors.initials: M
authors.lastname: Liu
authors.forename: Nangang
authors.initials: N
authors.lastname: Zhang
authors.forename: Shutao
authors.initials: S
authors.lastname: Wang
authors.forename: Brigitte
authors.initials: B
authors.lastname: Angenieux
authors.forename: Eduard
authors.initials: E
authors.lastname: Panosyan
authors.forename: Eric R
authors.initials: ER
authors.lastname: Samuels
authors.forename: Jun
authors.initials: J
authors.lastname: Park
authors.forename: Dirk
authors.initials: D
authors.lastname: Williams
authors.forename: Vera
authors.initials: V
authors.lastname: Konkankit
authors.forename: David
authors.initials: D
authors.lastname: Nathanson
authors.forename: R Michael
authors.initials: RM
authors.lastname: van Dam
authors.forename: Michael E
authors.initials: ME
authors.lastname: Phelps
authors.forename: Hong
authors.initials: H
authors.lastname: Wu
authors.forename: Linda M
authors.initials: LM
authors.lastname: Liau
authors.forename: Paul S
authors.initials: PS
authors.lastname: Mischel
authors.forename: Jorge A
authors.initials: JA
authors.lastname: Lazareff
authors.forename: Harley I
authors.initials: HI
authors.lastname: Kornblum
authors.forename: William H
authors.initials: WH
authors.lastname: Yong
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Graeber
authors.forename: Hsian-Rong
authors.initials: HR
authors.lastname: Tseng
mesh_terms: D001932:Brain Neoplasms / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D066246:ErbB Receptors / Q000378:metabolism; D005909:Glioblastoma / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D007150:Immunohistochemistry; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D051059:PTEN Phosphohydrolase / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015203:Reproducibility of Results; D019893:Ribosomal Protein S6 Kinases / Q000378:metabolism; D015398:Signal Transduction; D058570:TOR Serine-Threonine Kinases
chemical_list: D066246:ErbB Receptors; D047908:Intracellular Signaling Peptides and Proteins; D051059:PTEN Phosphohydrolase; D019869:Phosphatidylinositol 3-Kinases; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D019893:Ribosomal Protein S6 Kinases; D058570:TOR Serine-Threonine Kinases; C546842:MTOR protein, human; C494929:PTEN protein, human