April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Human tear peptide/protein profiling study of Keratoconus grades by SPE-MALDI-TOF Mass Spectrometry analyses
Author Affiliations & Notes
  • Tatiana Maria Suarez-Cortes
    Biomedical R & D, Bioftalmik, Derio, Spain
  • Javier Soria
    Biomedical R & D, Bioftalmik, Derio, Spain
  • Arantxa Acera
    Biomedical R & D, Bioftalmik, Derio, Spain
  • Nerea Gonzalez
    Biomedical R & D, Bioftalmik, Derio, Spain
  • Ibon Iloro
    Proteomics Platform, CIC bioGUNE, Derio, Spain
  • Felix Elortza
    Proteomics Platform, CIC bioGUNE, Derio, Spain
  • Jesus Merayo-Lloves
    Fundacion de Investigacion Oftalmologica, FIO, Fernández Vega Ophthalmological Institute, Oviedo, Spain
    Cirugia y especialidades Medico-Quirugicas, Universidad de Oviedo, Oviedo, Spain
  • Footnotes
    Commercial Relationships Tatiana Suarez-Cortes, Bioftalmik (E); Javier Soria, Bioftalmik (E); Arantxa Acera, Bioftalmik (E); Nerea Gonzalez, Bioftalmik (E); Ibon Iloro, None; Felix Elortza, None; Jesus Merayo-Lloves, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 2006. doi:
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      Tatiana Maria Suarez-Cortes, Javier Soria, Arantxa Acera, Nerea Gonzalez, Ibon Iloro, Felix Elortza, Jesus Merayo-Lloves, ; Human tear peptide/protein profiling study of Keratoconus grades by SPE-MALDI-TOF Mass Spectrometry analyses. Invest. Ophthalmol. Vis. Sci. 2014;55(13):2006.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract
 
Purpose
 

Changes in tears of Keratoconus (KC) patients have been widely reported, however, peptide/protein profiling has not been widely approached. The goal of this study was to analyze the tear peptidome/protein profiles (below 20kDa) of KC patients and to determinate their relation with clinical grades of the disease. Secondly, this work aimed to evaluate whether the tear profiling allows a categorization of patients which would be potentially useful for early diagnosis and disease grading.

 
Methods
 

A total of 72 KC patients (20 mild, 21 moderate and 31 severe) and 14 healthy subjects were studied. The biomicroscopy exam was performed to detect signs of KC and corneal topography study was used to quantify topographic parameters. The modified Rabinowitz McDonnell test was used to confirm KC diagnosis and KC severity was classified according to the steepest simulated keratometry reading on the keratometric map (K2 <45 D, mild; K2 between 45 -52 D, moderate; K2 >52 D, severe KC). Tear samples were collected by capillary and solid-phase extraction using magnetic beads (MB) with reverse phase affinity (C18), in combination with matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF) was used for human tear peptide/protein profiling analysis. Spectra obtained in the range of 1000-20000 Da were pre-processed and analyzed using multivariate statistical techniques.

 
Results
 

The tear peptide/protein profiling revealed differentially expressed peaks depending on grade severity of KC. From 243 peaks detected, 43 presented intensities significantly changed. Canonical discriminate analysis using these 43 most discriminate variables showed a good spatial separation of groups based on tear profile indicating a high correlation between peptide/profile and disease severity.

 
Conclusions
 

Tear peptide/proteome profiling of KC patients analyzed by SPE-MALDI-TOF allows classifying of patients according to clinical severity with high accuracy. Early diagnosis of KC patients is possible based on tear peptidome/protein profile changes. MALDI-TOF profiling of tears constitutes a sensitive method for grading severity of KC and for identifying outliers. Finally, peptidome/protein patterns may provide opportunities for research into the pathogenesis of the disease and to improve diagnosis, prognosis and monitoring of response to therapies in clinical trials.

 
Keywords: 574 keratoconus • 663 proteomics • 486 cornea: tears/tear film/dry eye  
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