A microfluidic platform for systems pathology: multiparameter single-cell signaling measurements of clinical brain tumor specimens.
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.
Sun
Jing
J
Crump Institute for Molecular Imaging, University of California at Los Angeles, Los Angeles, California 90095, USA.
Masterman-Smith
Michael D
MD
Graham
Nicholas A
NA
Jiao
Jing
J
Mottahedeh
Jack
J
Laks
Dan R
DR
Ohashi
Minori
M
DeJesus
Jason
J
Kamei
Ken-ichiro
K
Lee
Ki-Bum
KB
Wang
Hao
H
Yu
Zeta T F
ZT
Lu
Yi-Tsung
YT
Hou
Shuang
S
Li
Keyu
K
Liu
Max
M
Zhang
Nangang
N
Wang
Shutao
S
Angenieux
Brigitte
B
Panosyan
Eduard
E
Samuels
Eric R
ER
Park
Jun
J
Williams
Dirk
D
Konkankit
Vera
V
Nathanson
David
D
van Dam
R Michael
RM
Phelps
Michael E
ME
Wu
Hong
H
Liau
Linda M
LM
Mischel
Paul S
PS
Lazareff
Jorge A
JA
Kornblum
Harley I
HI
Yong
William H
WH
Graeber
Thomas G
TG
Tseng
Hsian-Rong
HR
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
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