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Title: Residual tissue repositories as a resource for population-based cancer proteomic studies

Abstract

Background: Mass spectrometry-based proteomics has become a powerful tool for the identification and quantification of proteins from a wide variety of biological specimens. To date, the majority of studies utilizing tissue samples have been carried out on prospectively collected fresh frozen or optimal cutting temperature (OCT) embedded specimens. However, such specimens are often difficult to obtain, in limited in supply, and clinical information and outcomes on patients are inherently delayed as compared to banked samples. Annotated formalin fixed, paraffin embedded (FFPE) tumor tissue specimens are available for research use from a variety of tissue banks, such as from the surveillance, epidemiology and end results (SEER) registries’ residual tissue repositories. Given the wealth of outcomes information associated with such samples, the reuse of archived FFPE blocks for deep proteomic characterization with mass spectrometry technologies would provide a valuable resource for population-based cancer studies. Further, due to the widespread availability of FFPE specimens, validation of specimen integrity opens the possibility for thousands of studies that can be conducted worldwide. Methods: To examine the suitability of the SEER repository tissues for proteomic and phosphoproteomic analysis, we analyzed 60 SEER patient samples, with time in storage ranging from 7 to 32 years; 60 samplesmore » with expression proteomics and 18 with phosphoproteomics, using isobaric labeling. Linear modeling and gene set enrichment analysis was used to evaluate the impacts of collection site and storage time. Results: All samples, regardless of age, yielded suitable protein mass after extraction for expression analysis and 18 samples yielded sufficient mass for phosphopeptide analysis. Although peptide, protein, and phosphopeptide identifications were reduced by 50, 20 and 76% respectively, from comparable OCT specimens, we found no statistically significant differences in protein quantitation correlating with collection site or specimen age. GSEA analysis of GOterm level measurements of protein abundance differences between FFPE and OCT embedded specimens suggest that the formalin fixation process may alter representation of protein categories in the resulting dataset. Conclusions: These studies demonstrate that residual FFPE tissue specimens, of varying age and collection site, are a promising source of protein for proteomic investigations if paired with rigorously verified mass spectrometry workflows.« less

Authors:
 [1];  [1];  [1];  [1];  [1];  [1];  [1];  [2];  [2];  [2];  [2];  [2];  [1];  [2];  [1]; ORCiD logo [1]
  1. Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
  2. National Cancer Inst., Rockville, MD (United States)
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
Sponsoring Org.:
USDOE; National Institutes of Health (NIH); National Cancer Institute Clinical Proteomic Tumor Analysis Consortium (CPTAC)
OSTI Identifier:
1503586
Report Number(s):
PNNL-SA-130920
Journal ID: ISSN 1542-6416
Grant/Contract Number:  
AC05-76RL01830; U24CA160019; U24CA210955
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Clinical Proteomics
Additional Journal Information:
Journal Volume: 15; Journal Issue: 1; Journal ID: ISSN 1542-6416
Country of Publication:
United States
Language:
English
Subject:
Proteomics; Phosphoproteomics; Tandem mass tags; Formalin fixed paraffin embedded; Surveillance, epidemiology and end results

Citation Formats

Piehowski, Paul D., Petyuk, Vladislav A., Sontag, Ryan L., Gritsenko, Marina A., Weitz, Karl K., Fillmore, Thomas L., Moon, Jamie, Makhlouf, Hala, Chuaqui, Rodrigo F., Boja, Emily S., Rodriguez, Henry, Lee, Jerry S. H., Smith, Richard D., Carrick, Danielle M., Liu, Tao, and Rodland, Karin D. Residual tissue repositories as a resource for population-based cancer proteomic studies. United States: N. p., 2018. Web. doi:10.1186/s12014-018-9202-4.
Piehowski, Paul D., Petyuk, Vladislav A., Sontag, Ryan L., Gritsenko, Marina A., Weitz, Karl K., Fillmore, Thomas L., Moon, Jamie, Makhlouf, Hala, Chuaqui, Rodrigo F., Boja, Emily S., Rodriguez, Henry, Lee, Jerry S. H., Smith, Richard D., Carrick, Danielle M., Liu, Tao, & Rodland, Karin D. Residual tissue repositories as a resource for population-based cancer proteomic studies. United States. doi:10.1186/s12014-018-9202-4.
Piehowski, Paul D., Petyuk, Vladislav A., Sontag, Ryan L., Gritsenko, Marina A., Weitz, Karl K., Fillmore, Thomas L., Moon, Jamie, Makhlouf, Hala, Chuaqui, Rodrigo F., Boja, Emily S., Rodriguez, Henry, Lee, Jerry S. H., Smith, Richard D., Carrick, Danielle M., Liu, Tao, and Rodland, Karin D. Fri . "Residual tissue repositories as a resource for population-based cancer proteomic studies". United States. doi:10.1186/s12014-018-9202-4. https://www.osti.gov/servlets/purl/1503586.
@article{osti_1503586,
title = {Residual tissue repositories as a resource for population-based cancer proteomic studies},
author = {Piehowski, Paul D. and Petyuk, Vladislav A. and Sontag, Ryan L. and Gritsenko, Marina A. and Weitz, Karl K. and Fillmore, Thomas L. and Moon, Jamie and Makhlouf, Hala and Chuaqui, Rodrigo F. and Boja, Emily S. and Rodriguez, Henry and Lee, Jerry S. H. and Smith, Richard D. and Carrick, Danielle M. and Liu, Tao and Rodland, Karin D.},
abstractNote = {Background: Mass spectrometry-based proteomics has become a powerful tool for the identification and quantification of proteins from a wide variety of biological specimens. To date, the majority of studies utilizing tissue samples have been carried out on prospectively collected fresh frozen or optimal cutting temperature (OCT) embedded specimens. However, such specimens are often difficult to obtain, in limited in supply, and clinical information and outcomes on patients are inherently delayed as compared to banked samples. Annotated formalin fixed, paraffin embedded (FFPE) tumor tissue specimens are available for research use from a variety of tissue banks, such as from the surveillance, epidemiology and end results (SEER) registries’ residual tissue repositories. Given the wealth of outcomes information associated with such samples, the reuse of archived FFPE blocks for deep proteomic characterization with mass spectrometry technologies would provide a valuable resource for population-based cancer studies. Further, due to the widespread availability of FFPE specimens, validation of specimen integrity opens the possibility for thousands of studies that can be conducted worldwide. Methods: To examine the suitability of the SEER repository tissues for proteomic and phosphoproteomic analysis, we analyzed 60 SEER patient samples, with time in storage ranging from 7 to 32 years; 60 samples with expression proteomics and 18 with phosphoproteomics, using isobaric labeling. Linear modeling and gene set enrichment analysis was used to evaluate the impacts of collection site and storage time. Results: All samples, regardless of age, yielded suitable protein mass after extraction for expression analysis and 18 samples yielded sufficient mass for phosphopeptide analysis. Although peptide, protein, and phosphopeptide identifications were reduced by 50, 20 and 76% respectively, from comparable OCT specimens, we found no statistically significant differences in protein quantitation correlating with collection site or specimen age. GSEA analysis of GOterm level measurements of protein abundance differences between FFPE and OCT embedded specimens suggest that the formalin fixation process may alter representation of protein categories in the resulting dataset. Conclusions: These studies demonstrate that residual FFPE tissue specimens, of varying age and collection site, are a promising source of protein for proteomic investigations if paired with rigorously verified mass spectrometry workflows.},
doi = {10.1186/s12014-018-9202-4},
journal = {Clinical Proteomics},
issn = {1542-6416},
number = 1,
volume = 15,
place = {United States},
year = {2018},
month = {8}
}

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