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PMID: 19363496 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Nanofluidic proteomic assay for serial analysis of oncoprotein activation in clinical specimens.

Nature medicine ·Vol. 15 ·No. 5 ·2009-05-00 ·Pages 566-71

Fan AC, Deb-Basu D, Orban MW, Gotlib JR, Natkunam Y, O'Neill R, Padua RA, Xu L, Taketa D, Shirer AE, Beer S, Yee AX, Voehringer DW, Felsher DW

Abstract

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.

MeSH Terms
Burkitt Lymphoma/genetics,therapy Enzyme Activation Extracellular Signal-Regulated MAP Kinases/genetics Gene Expression Regulation, Neoplastic Genes, myc Humans Immunoassay/methods Lymphoma, B-Cell/genetics,therapy Lymphoma, Follicular/genetics,therapy Neoplasms/genetics,therapy Oncogene Proteins/genetics Oncogenes Phosphoproteins/genetics Protein Isoforms/analysis,genetics Proteomics/methods
Chemicals
Oncogene Proteins Phosphoproteins Protein Isoforms Extracellular Signal-Regulated MAP Kinases
Authors & Affiliations
14 authors, click to expand affiliations / ORCID
Fan Alice C
Stanford University, Departments of Medicine and Pathology, California, USA.
Deb-Basu Debabrita
Orban Mathias W
Gotlib Jason R
Natkunam Yasodha
O'Neill Roger
Padua Rose-Ann
Xu Liwen
Taketa Daryl
Shirer Amy E
Beer Shelly
Yee Ada X
Voehringer David W
Felsher Dean W
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Article Info
Journal
Nature medicine
Abbr.
Nat Med
ISSN
1546-170X
DOI
10.1038/nm.1903
Published
2009-05-00
Epub
2009-00-12
Pages
566-71
Language
English
Region
United States
NLM ID
9502015
PMCID
PMC4006986
Subset
IM
Grants
NCI NIH HHS · CA89305 · United States
NCI NIH HHS · CA034233 · United States
NCI NIH HHS · R01 CA089305 · United States
NCI NIH HHS · P01 CA034233 · United States
NCI NIH HHS · T32 CA009151 · United States
Corrections
CommentIn
PMID: 19792963
External Links
PubMed source DOI: 10.1038/nm.1903 PMC Full Text View journal details
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