SELDI-TOF plasma profiles distinguish individuals in a protein C-deficient family with thrombotic episodes occuring before age 40

Summary

We tested the hypothesis that differences in the low-molecularweight (500–20,000 Da) proteomic profile of plasma may be detectable between members ofa protein C-deficient family who have suffered thrombotic events before age 40 compared to family members without a history of venous thrombosis. Unfractionated plasma samples from members of a previously described large thrombophilic kindred with type I protein C deficiency were applied to ProteinChip weak cation exchange interaction arrays (WCX2; Ciphergen Biosystems, Fremont, CA, USA) and subjected to SELDI-TOF (surface-enhanced laser desorption/ionization time-of-flight) mass spectrometry using the Ciphergen PBSII ProteinChip System (Ciphergen Biosystems). Profiles were analyzed by a boosted decision-tree algorithm. When individuals who had presented with deep venous thrombosis (DVT) before the age of 40 (n=21) were compared to age-matched, healthy family members (n=50), the proteomic patterns defined by the decision-tree analysis could classify the entity of DVT before age 40 with 67% sensitivity, ata specificity of 86%.Whena small group of cases with history of superficial venous thrombosis (n=6) was added to the case group, the sensitivity was 87.5% at a specificity of 80%.These data support the hypothesis that members of the protein C deficient Vermont kindred II who suffer a thrombotic event before age 40 display significant differences in low-molecular-weight proteomics profile compared to those who remain disease-free.This is the first study to apply SELDI-TOF technology in conjunction with a bioinformatics tool to analyze low-molecular-weight proteomic patterns in patients with venous thrombosis.

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Keywords

Venous thrombosis - SELDI-TOF - mass spectrometry - proteomics - protein C deficiency

^* Present address: Department of Pathology, University of Utah, Salt Lake City, Utah, USA.

• References

• 1 Heit JA. Venous thromboembolism: disease burden, outcomes and risk factors. J Thromb Haemost 2005; 03: 1611-8.
• 2 Decousus H, Leizorovicz A. Superficial thrombophlebitis of the legs: still a lot to learn. J Thromb Haemost 2005; 03: 1149-51.
• 3 Rosendaal FR. Venous thrombosis: a multicausal disease. Lancet 1999; 353: 1167-73.
• 4 Bovill EG, Hasstedt SJ, Leppert MF. et al. Hereditary thrombophilia as a model for multigenic disease. Thromb Haemost 1999; 82: 662-6.
• 5 Hasstedt SJ, Bovill EG, Callas PW. et al. An unknown genetic defect increases venous thrombosis risk, through interaction with protein C deficiency. Am J Hum Genet 1998; 63: 569-76.
• 6 Hasstedt SJ, Scott BT, Callas PW. et al. Genome scan of venous thrombosis in a pedigree with protein C deficiency. J Thromb Haemost 2004; 02: 868-73.
• 7 Scott BT, Bovill EG, Callas PW. et al. Genetic screening of candidate genes for a prothrombotic interaction with type 1 protein C deficiency in a large kindred. Thromb Haemost 2001; 85: 82-7.
• 8 Scott BT, Hasstedt SJ, Bovill EG. et al. Characterization of the human prostaglandin H syntase 1 gene (PTGS1): exclusion by genetic linkage analysis as a second modifier gene in familial thrombosis. Blood Coagul Fibrinol 2002; 13: 519-31.
• 9 Bovill EG, Hasstedt SJ, Callas PW. et al. The G20210A prothrombin polymorphism is not associated with increased thromboembolic risk in a large protein C deficient kindred. Thromb Haemost 2000; 83: 366-70.
• 10 Petricoin EF, Ardekani AM, Hitt BA. et al. Use of proteomic patterns in serum to identify ovarian cancer. Lancet 2002; 359: 572-7.
• 11 Petricoin 3rd EF, Ornstein DK, Paweletz CP. et al. Serum proteomic patterns for detection of prostate cancer. J Natl Cancer Inst 2002; 94: 1576-8.
• 12 Li J, Orlandi R, White CN. et al. Independent validation of candidate breast cancer serum biomarkers identified by mass spectrometry. Clin Chem 2005; 51: 2229-35.
• 13 Rosenblatt KR, Bryant-Greenwood P, Killian JK. et al. Serum proteomics in cancer diagnosis and management. Annu Rev Med 2004; 55: 97-112.
• 14 Lopez MF, Mikulskis A, Kuzdzal S. et al. Highresolution serum proteomic profiling of Alzheimer disease samples reveals disease-specific, carrier-protein-bound mass signatures. Clin Chem 2005; 51: 1946-54.
• 15 Stanley BA, Gundry RL, Cotter RJ, Van Eyk JE. Heart disease, clinical proteomics and mass spectrometry. Dis Markers 2004; 20: 167-78.
• 16 Bovill EG, Bauer KA, Dickerman JD. et al. The clinical spectrum of heterozygous protein C deficiency in a large New England kindred. Blood 1989; 73: 712-7.
• 17 Johann DJ, McGuigan MD, Tomov S. et al. Novel approaches to visualization and data mining reveals diagnostic information in the low amplitude region of serum mass spectra from ovarian cancer patients. Dis Markers 2003-2004 19: 197-207.
• 18 Johann DJ, McGuigan MD, Patel AR. et al. Clinical Proteomics and Biomarker Discovery. Ann NY Acad Sci 2004; 1022: 295-305.
• 19 Tomczak JA, Ando RA, Sobel HG. et al. Genetic analysis of a large kindred exhibiting type I protein C deficiency and associated thrombosis. Thrombosis Res 1994; 74: 243-54.