title: 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.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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
  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.
authors: Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA, USA.|Medical Oncology Division, S. Maria della Misericordia Hospital, Perugia, Italy.
authors: Elisa
authors: E
authors: Baldelli
authors: Department of Thoracic Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA.
authors: Eric B
authors: EB
authors: Haura
authors: Medical Oncology Division, S. Maria della Misericordia Hospital, Perugia, Italy.
authors: Lucio
authors: L
authors: Crinò
authors: Department of Thoracic Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA.
authors: Douglas W
authors: DW
authors: Cress
authors: Medical Oncology Division, S. Maria della Misericordia Hospital, Perugia, Italy.
authors: Vienna
authors: V
authors: Ludovini
authors: Department of Thoracic Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA.
authors: Matthew B
authors: MB
authors: Schabath
authors: Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA, USA.
authors: Lance A
authors: LA
authors: Liotta
authors: Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA, USA.
authors: Emanuel F
authors: EF
authors: Petricoin
authors: Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA, USA.
authors: Mariaelena
authors: M
authors: Pierobon
mesh_terms: D002289:Carcinoma, Non-Small-Cell Lung / Q000378:metabolism; D006801:Humans; D061346:Laser Capture Microdissection / Q000379:methods*; D008175:Lung Neoplasms / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D057285:Precision Medicine / Q000379:methods*; D040081:Protein Array Analysis
chemical_list: D010750:Phosphoproteins
keywords: Individualized therapy; Laser capture microdissection; Nonsmall cell lung cancer; Pathway activation mapping; Reverse phase protein microarray