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Impact of upfront cellular enrichment by laser capture microdissection on protein and phosphoprotein drug target signaling activation measurements in human lung cancer: Implications for personalized medicine.

AbstractPURPOSE:
The aim of this study was to evaluate whether upfront cellular enrichment via laser capture microdissection (LCM) is necessary for accurately quantifying predictive biomarkers in nonsmall cell lung cancer tumors.
EXPERIMENTAL DESIGN:
Fifteen snap frozen surgical biopsies were analyzed. Whole tissue lysate and matched highly enriched tumor epithelium via LCM were obtained for each patient. The expression and activation/phosphorylation levels of 26 proteins were measured by reverse phase protein microarray. Differences in signaling architecture of dissected and undissected matched pairs were visualized using unsupervised clustering analysis, bar graphs, and scatter plots.
RESULTS:
Overall patient matched LCM and undissected material displayed very distinct and differing signaling architectures with 93% of the matched pairs clustering separately. These differences were seen regardless of the amount of starting tumor epithelial content present in the specimen.
CONCLUSIONS AND CLINICAL RELEVANCE:
These results indicate that LCM driven upfront cellular enrichment is necessary to accurately determine the expression/activation levels of predictive protein signaling markers although results should be evaluated in larger clinical settings. Upfront cellular enrichment of the target cell appears to be an important part of the workflow needed for the accurate quantification of predictive protein signaling biomarkers. Larger independent studies are warranted.
AuthorsElisa Baldelli, Eric B Haura, Lucio Crinò, Douglas W Cress, Vienna Ludovini, Matthew B Schabath, Lance A Liotta, Emanuel F Petricoin, Mariaelena Pierobon
JournalProteomics. Clinical applications (Proteomics Clin Appl) Vol. 9 Issue 9-10 Pg. 928-37 (Oct 2015) ISSN: 1862-8354 [Electronic] Germany
PMID25676683 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Copyright© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Chemical References
  • Phosphoproteins
Topics
  • Carcinoma, Non-Small-Cell Lung (metabolism)
  • Humans
  • Laser Capture Microdissection (methods)
  • Lung Neoplasms (metabolism)
  • Phosphoproteins (metabolism)
  • Precision Medicine (methods)
  • Protein Array Analysis

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