Nanofluidic proteomic assay for serial analysis of oncoprotein activation in clinical specimens.
Current methods of protein detection are insensitive to detecting subtle changes in oncoprotein activation that underlie key cancer signaling processes. The requirement for large numbers of cells precludes serial tumor sampling for assessing a response to therapeutics. Therefore, we have developed a nanofluidic proteomic immunoassay (NIA) to quantify total and low-abundance protein isoforms in nanoliter volumes. Our method can quantify amounts of MYC oncoprotein and B cell lymphoma protein-2 (BCL2) in Burkitt's and follicular lymphoma; identify changes in activation of extracellular signal-related kinases-1 (ERK1) and ERK2, mitogen-activated kinase-1 (MEK), signal transducer and activator of transcription protein-3 (STAT3) and STAT5, c-Jun N-terminal kinase (JNK) and caspase-3 in imatinib-treated chronic myelogeneous leukemia (CML) cells; measure an unanticipated change in the phosphorylation of an ERK2 isomer in individuals with CML who responded to imatinib; and detect a decrease in STAT3 and STAT5 phosphorylation in individuals with lymphoma who were treated with atorvastatin. Therefore, we have described a new and highly sensitive method for determining oncoprotein expression and phosphorylation in clinical specimens for the development of new therapeutics for cancer.
Stanford University, Departments of Medicine and Pathology, California, USA.
Alice C
AC
Fan
Debabrita
D
Deb-Basu
Mathias W
MW
Orban
Jason R
JR
Gotlib
Yasodha
Y
Natkunam
Roger
R
O'Neill
Rose-Ann
RA
Padua
Liwen
L
Xu
Daryl
D
Taketa
Amy E
AE
Shirer
Shelly
S
Beer
Ada X
AX
Yee
David W
DW
Voehringer
Dean W
DW
Felsher
D002051:Burkitt Lymphoma / Q000235:genetics / Q000628:therapy; D004789:Enzyme Activation; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D016259:Genes, myc; D006801:Humans; D007118:Immunoassay / Q000379:methods; D016393:Lymphoma, B-Cell / Q000235:genetics / Q000628:therapy; D008224:Lymphoma, Follicular / Q000235:genetics / Q000628:therapy; D009369:Neoplasms / Q000235:genetics* / Q000628:therapy; D015513:Oncogene Proteins / Q000235:genetics*; D009857:Oncogenes; D010750:Phosphoproteins / Q000235:genetics; D020033:Protein Isoforms / Q000032:analysis / Q000235:genetics; D040901:Proteomics / Q000379:methods*
D015513:Oncogene Proteins; D010750:Phosphoproteins; D020033:Protein Isoforms; D048049:Extracellular Signal-Regulated MAP Kinases