Proteomic surfaceome analysis of mesothelioma

Our plain-English summary of Proteomic surfaceome analysis of mesothelioma, by A. Ziegler, F. Cerciello, C. Bigosch, et al (Lung Cancer, 2012).

The study

• Authors: A. Ziegler, F. Cerciello, C. Bigosch, D. Bausch-Fluck, E. Felley-Bosco, R. Ossola, A. Soltermann, R.A. Stahel, B. Wollscheid
• Journal: Lung Cancer
• Published: 2012;75(2):189-96
• Cited by: 26 articles, as counted by NIH iCite
• Read it: PubMed · Publisher

Abstract

Identification of new markers for malignant pleural mesothelioma (MPM) is a challenging clinical need. Here, we propose a quantitative proteomics primary screen of the cell surface exposed MPM N-glycoproteins, which provides the basis for the development of new protein-based diagnostic assays. Using the antibody-independent mass-spectrometry based cell surface capturing (CSC) technology, we specifically investigated the N-glycosylated surfaceome of MPM towards the identification of protein-marker candidates discriminatory between MPM and lung adenocarcinoma (ADCA). Relative quantitative CSC analysis of MPM cell line ZL55 in comparison with ADCA cell line Calu-3 revealed a bird's eye view of their respective surfaceomes. In a secondary screen of fifteen MPM and six ADCA, we used high throughput low density microarrays (LDAs) to verify specificity and sensitivity of nineteen N-glycoproteins overregulated in the surfaceome of MPM. This proteo-transcriptomic approach revealed thy-1/CD90 (THY1) and teneurin-2 (ODZ2) as protein-marker candidates for the discrimination of MPM from ADCA. Thy-1/CD90 was further validated by immunohistochemistry on frozen tissue sections of MPM and ADCA samples. Together, we present a combined proteomic and transcriptomic approach enabling the relative quantitative identification and pre-clinical selection of new MPM marker candidates.

Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

In plain English

This laboratory study looked for cell surface proteins that might help tell malignant pleural mesothelioma (MPM) apart from lung adenocarcinoma (ADCA). It did not study asbestos exposure or patient outcomes. The authors first used mass spectrometry to compare one mesothelioma cell line with one adenocarcinoma cell line, and identified 100 cell surface N-glycoproteins. They then measured mRNA for 19 selected proteins in fifteen MPM lines and six comparison lines (five adenocarcinoma, one adenosquamous). Thy-1/CD90 mRNA was present in the mesothelioma lines and absent from the comparison lines. Teneurin-2 was found in eleven MPM lines and, at low levels, in two comparison lines. An antibody the authors made against teneurin-2 gave no specific signals. In frozen tissue from six MPM and eight ADCA tumors, three MPM cases had limited numbers of tumor cells with moderate membrane staining for thy-1/CD90. ADCA tumor cells stained only modestly, if at all, and that staining was inside the cells rather than on the membrane. Supporting (stromal) cells in both tumor types also stained. The authors concluded that teneurin-2, and especially thy-1/CD90, are marker candidates. They called the tissue findings preliminary.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: Laboratory study combining a mass-spectrometry screen of cell surface proteins, mRNA testing on cell lines, and antibody tests on cell lines and frozen tumor tissue
• Who was studied: Fifteen malignant pleural mesothelioma (MPM) cell lines and six lung cancer comparison lines, studied in Zürich, Switzerland. The comparison lines were five adenocarcinoma lines and one adenosquamous line, H596. The initial screen compared one line of each type (ZL55 and Calu-3). Frozen tumor sections came from four epithelioid and two biphasic MPM tumors and eight ADCA tumors.
• Exposure: No exposure was studied. The study compared mesothelioma cells with lung adenocarcinoma cells. Asbestos is mentioned only as background in the introduction, as the main cause of MPM.
• Main results:
  • The surface protein screen of ZL55 (MPM) and Calu-3 (ADCA) cells identified 100 cell surface N-glycoproteins, including 37 CD annotated proteins
  • The known mesothelioma markers mesothelin and N-cadherin were higher in the MPM cell line. The ADCA-over-MPM abundance ratio was 0.04 plus or minus 0.00 for mesothelin and 0.05 plus or minus 0.04 for N-cadherin.
  • Mesothelin mRNA was found in 10 of 15 (67%) MPM cell lines and in the adeno-squamous line H596
  • N-cadherin mRNA was found in all (100%) MPM cell lines and in three (50%) of the comparison lines
  • Thy-1/CD90 mRNA was present in the MPM cell lines and absent from the comparison lines
  • Teneurin-2 mRNA was found in eleven MPM cell lines and only at low levels in one ADCA line (SK-LU-1) and one adeno-squamous line (H596)
  • In protein tests (Western blot), deglycosylated thy-1/CD90 was detected in the MPM cell lines tested but not in the ADCA cell lines
  • A rabbit antiserum the authors made against teneurin-2 gave no specific signals
  • In frozen tumor sections, tumor-stromal cells of both tumor types reacted to the thy-1 antibody. Three MPM cases had discrete numbers of tumor cells with moderate membrane staining.
  • ADCA tumor cells reacted only modestly, if at all, to the thy-1 antibody. Where they did stain, the staining was weak to moderate, inside the cells (cytoplasmic), and toward the stroma.
• The authors' conclusion: The authors concluded that teneurin-2, and especially thy-1/CD90, are cell surface accessible marker candidates for telling MPM apart from ADCA
• Limitations the authors state:
  • The authors described the tissue findings as preliminary. They said confirming them on a larger scale would require antibodies that work on paraffin-embedded tissue, and none of the antibodies they tested did.
  • The method could not detect the widely accepted MPM markers calretinin, cytokeratins 5/6 or WT-1, and did not observe podoplanin either. The authors explained that these markers are not N-glycosylated, and most are not exposed on the cell surface.
  • The antibody the authors produced against teneurin-2 did not work satisfactorily, so teneurin-2 could not be verified at the protein level

Topics

The National Library of Medicine indexes this study under subjects our site covers: Mesothelioma.

Medical terms in this study, explained in our Lexicon: Mesothelioma, Pleural Mesothelioma, Glycoprotein, Lung Adenocarcinoma, Immunohistochemistry.

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