Tear fluid proteomics multimarkers for diabetic retinopathy screening

BackgroundThe aim of the project was to develop a novel method for diabetic retinopathy screening based on the examination of tear fluid biomarker changes. In order to evaluate the usability of protein biomarkers for pre-screening purposes several different approaches were used, including machine learning algorithms.MethodsAll persons involved in the study had diabetes. Diabetic retinopathy (DR) was diagnosed by capturing 7-field fundus images, evaluated by two independent ophthalmologists. 165 eyes were examined (from 119 patients), 55 were diagnosed healthy and 110 images showed signs of DR. Tear samples were taken from all eyes and state-of-the-art nano-HPLC coupled ESI-MS/MS mass spectrometry protein identification was performed on all samples. Applicability of protein biomarkers was evaluated by six different optimally parameterized machine learning algorithms: Support Vector Machine, Recursive Partitioning, Random Forest, Naive Bayes, Logistic Regression, K-Nearest Neighbor.ResultsOut of the six investigated machine learning algorithms the result of Recursive Partitioning proved to be the most accurate. The performance of the system realizing the above algorithm reached 74% sensitivity and 48% specificity.ConclusionsProtein biomarkers selected and classified with machine learning algorithms alone are at present not recommended for screening purposes because of low specificity and sensitivity values. This tool can be potentially used to improve the results of image processing methods as a complementary tool in automatic or semiautomatic systems.

[1]  Pål Gulbrandsen,et al.  Sensitivity and specificity of Norwegian optometrists’ evaluation of diabetic retinopathy in single-field retinal images – a cross-sectional experimental study , 2013, BMC Health Services Research.

[2]  D. Klonoff,et al.  An economic analysis of interventions for diabetes. , 2000, Diabetes care.

[3]  Kurt Hornik,et al.  kernlab - An S4 Package for Kernel Methods in R , 2004 .

[4]  Forest M White,et al.  The Potential Cost of High-Throughput Proteomics , 2011, Science Signaling.

[5]  F. Chew,et al.  Proteomic analysis of human tears: defensin expression after ocular surface surgery. , 2004, Journal of proteome research.

[6]  E. Campos,et al.  Tear proteomics in evaporative dry eye disease , 2010, Eye.

[7]  Nan Wang,et al.  Characterization of human tear proteome using multiple proteomic analysis techniques. , 2005, Journal of proteome research.

[8]  Adrienne Csutak,et al.  Quantitative analysis of proteins in the tear fluid of patients with diabetic retinopathy. , 2012, Journal of proteomics.

[9]  Matthias Mann,et al.  Identification of 491 proteins in the tear fluid proteome reveals a large number of proteases and protease inhibitors , 2006, Genome Biology.

[10]  Kari B. Green-Church,et al.  Investigation of the human tear film proteome using multiple proteomic approaches , 2008, Molecular vision.

[11]  Lloyd Paul Aiello,et al.  Telemedicine and diabetic retinopathy: moving beyond retinal screening. , 2011, Archives of ophthalmology.

[12]  T. Sano,et al.  [Diabetic retinopathy]. , 2001, Nihon rinsho. Japanese journal of clinical medicine.

[13]  A. Gooley,et al.  Establishment of the human reflex tear two‐dimensional polyacrylamide gel electrophoresis reference map: New proteins of potential diagnostic value , 1997, Electrophoresis.

[14]  P. Fort,et al.  Impact of diabetes on alpha-crystallins and other heat shock proteins in the eye , 2011, Journal of ocular biology, diseases, and informatics.

[15]  R. Schlingemann,et al.  Molecular basis of the inner blood-retinal barrier and its breakdown in diabetic macular edema and other pathological conditions , 2013, Progress in Retinal and Eye Research.

[16]  Lei Zhou,et al.  Identification of tear fluid biomarkers in dry eye syndrome using iTRAQ quantitative proteomics. , 2009, Journal of proteome research.

[17]  F. Grus,et al.  Changes in the tear protein patterns of diabetic patients using two-dimensional electrophoresis. , 2000, Advances in experimental medicine and biology.

[18]  D. Singer,et al.  Screening for Diabetic Retinopathy , 1993, Diabetes Care.

[19]  F. Grus,et al.  High Performance Liquid Chromatography Analysis of Tear Protein Patterns in Diabetic and Non-Diabetic Dry-Eye Patients , 2001, European journal of ophthalmology.

[20]  A. Augustin,et al.  Analysis of tear proteins by one- and two-dimensional thin-layer iosoelectric focusing, sodium dodecyl sulfate electrophoresis and lectin blotting. Detection of a new component: cystatin C , 1998, Graefe's Archive for Clinical and Experimental Ophthalmology.

[21]  D. Lamberts,et al.  The preocular tear film : in health, disease, and contact lens wear , 1986 .

[22]  Qin Li,et al.  Retinopathy Online Challenge: Automatic Detection of Microaneurysms in Digital Color Fundus Photographs , 2010, IEEE Transactions on Medical Imaging.

[23]  S. Garg,et al.  Diabetic Retinopathy Screening Update , 2009, Clinical Diabetes.

[24]  Peter Bragge,et al.  Screening for presence or absence of diabetic retinopathy: a meta-analysis. , 2011, Archives of ophthalmology.

[25]  N. Komori,et al.  Proteome profiling of vitreoretinal diseases by cluster analysis , 2008, Proteomics. Clinical applications.

[26]  D. Owens,et al.  Practical application of the European Field Guide in screening for diabetic retinopathy by using ophthalmoscopy and 35 mm retinal slides , 1998, Diabetologia.

[27]  J. Olson,et al.  Automated grading for diabetic retinopathy: a large-scale audit using arbitration by clinical experts , 2010, British Journal of Ophthalmology.

[28]  R. Sack,et al.  Characterization of the in vivo forms of lacrimal‐specific proline‐rich proteins in human tear fluid , 2004, Proteomics.

[29]  M. Larsen,et al.  Automated detection of diabetic retinopathy in a fundus photographic screening population. , 2003, Investigative ophthalmology & visual science.

[30]  J. P. O’Hare,et al.  Adding retinal photography to screening for diabetic retinopathy: a prospective study in primary care , 1996, BMJ.

[31]  Manal Bouhaimed,et al.  Automated detection of diabetic retinopathy: results of a screening study. , 2008, Diabetes technology & therapeutics.

[32]  R. Phillips,et al.  Effectiveness of optometrist screening for diabetic retinopathy using slit-lamp biomicroscopy , 2001, Eye.

[33]  R. Mcdermott,et al.  A simple diabetes vascular severity staging instrument and its application to a Torres Strait Islander and Aboriginal adult cohort of north Australia , 2012, BMC Health Services Research.

[34]  Kai Bruns,et al.  SELDI-TOF-MS ProteinChip array profiling of tears from patients with dry eye. , 2005, Investigative ophthalmology & visual science.

[35]  R Core Team,et al.  R: A language and environment for statistical computing. , 2014 .

[36]  J. Tőzsér,et al.  Plasminogen activator inhibitor in human tears after laser refractive surgery , 2004, Journal of cataract and refractive surgery.