Comparative Plasma Proteomic Analysis of Patients with Multiple Myeloma Treated with Bortezomib-based Regimens
Authors:
J. Čumová 1,2; L. Jedličková 3; D. Potěšil 3,4; O. Šedo 4; K. Stejskal 3; A. Potáčová 1; Z. Zdráhal 3; R. Hájek 1,5,6
Authors‘ workplace:
Babákova myelomová skupina, Ústav patologické fyziologie, LF MU Brno
1; Oddělení klinických laboratoří, Nemocnice Blansko
2; Oddělení funkční genomiky a proteomiky, Ústav experimentální biologie, PřF MU Brno
3; Výzkumná skupina Proteomika CF, Mendelovo centrum genomiky a proteomiky rostlin, Středoevropský technologický institut, MU Brno
4; Laboratoř experimentální hematologie a buněčné imunoterapie, Oddělení klinické hematologie, FN Brno
5; Interní hematoonkologická klinika, LF MU a FN Brno
6
Published in:
Klin Onkol 2012; 25(1): 17-25
Category:
Original Articles
Overview
Backgrounds:
Recently, the term biomarker has become, especially in connection with the term clinical proteomics, one of the most frequent terms in the field of biomedical research. The aim of this work was to select an appropriate pre-fractionation method of blood plasma prior to a subsequent proteomic analysis of low-abundant fraction of proteins by two dimensional gel electrophoresis (2-DE) and mass spectrometry to improve the resolution of 2-DE maps and protein identification.
Materials and Methods:
First, we compared two prefractionation methods (MARS versus ProteoMiner) preceding 2-DE analysis using 10 blood plasma samples. Based on the results of the comparative experiments, low-abundant plasma protein fractions from 18 multiple myeloma patients treated with bortezomib were analyzed. Patients were divided into two groups: a group resistant to chemotherapy (9 patients – disease progression, stable disease) and a group with positive clinical response (9 patients – complete and partial remission).
Results and Conclusion:
Samples prefractioned by ProteoMiner method yielded 2-DE maps with a significantly increased number of detected protein spots, as compared to immunodepletion method MARS (Multiple Affinity Removal System). Between groups of chemoresistant and sensitive patients treated with bortezomib, 15 differently intense spots were revealed by image analysis. These spots were found to correspond to 10 proteins, as confirmed by mass spectrometry. Seven proteins had significantly lower protein level in the group of chemosensitive patients (serum amyloid P, fibrinogen – gamma chain, retinol-binding protein 4, complement factor C4-A, apolipoprotein E, carboxypeptidase N and complement factor H-related protein 1) and 3 proteins showed significantly higher levels of protein (or were only detected) in the group of chemosensitive patients (serum paraoxonase 1, alpha-1-antitrypsin and complement factor B).
Key words:
multiple myeloma – proteomics – two dimensional gel electrophoresis – plasma – protein – bortezomib
This work was supported by grants of Ministry of Education, Youth and Sports of the CR (LC06027 a MSM0021622434) and IGA MZ (NS9683) and NT12130) and grant of The Czech Science Foundation GAP304/10/1395..
The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study.
The Editorial Board declares that the manuscript met the ICMJE “uniform requirements” for biomedical papers.
Submitted:
3. 6. 2011
Accepted:
8. 9. 2011
Sources
1. Ahn SM, Simpson RJ. Body fluid proteomics: Prospects for biomarker discovery. Proteomics Clin Appl 2007; 1(9): 1004–1015.
2. Apweiler R, Aslanidis C, Deufel T et al. Approaching clinical proteomics: current state and future fields of application in fluid proteomics. Clin Chem Lab Med 2009; 47(6): 724–744.
3. Anderson NL, Anderson NG. The human plasma proteome: history, character, and diagnostic prospects. Mol Cell Proteomics 2002; 1(11): 845–867.
4. Philips AV, Cooper TA. RNA processing and human disease. Cell Mol Life Sci 2000; 57(2): 235–249.
5. Mann M, Jensen ON. Proteomic analysis of post-translational modifications. Nat Biotechnol 2003; 21(3): 255–261.
6. O’Farrell PH. High resolution two-dimensional electrophoresis of proteins. J Biol Chem 1975; 25(10): 4007–4021.
7. Zelená J, Hájek R. Proteomické techniky a jejich aplikace u hematoonkologických onemocnění. Čas Lék Čes 2007; 146(7): 586–592.
8. Potáčová A, Čumová J, Hájek R. Proteomická analýza a její využití ve výzkumu mnohočetného myelomu. Klin Onkol 2008; 21 (Suppl 1): 243–246.
9. Kim HJ, Kim MR, So EJ et al. Comparison of proteomes in various human plasma preparations by two-dimensional gel electrophoresis. J Biochem Biophys Methods 2007; 70(4): 619–625.
10. Kim MR, Kim CW. Human blood plasma preparation for two-dimensional gel electrophoresis. J Chromatogr B Analyt Technol Biomed Life Sci 2007; 849(1–2): 203–210.
11. Cho SY, Lee EY, Lee JS et al. Efficient prefractionation of low-abundance proteins in human plasma and construction of a two-dimensional map. Proteomics 2005; 5(13): 3386–3396.
12. Vasudev NS, Ferguson RE, Cairns DA et al. Serum biomarker discovery in renal cancer using 2-DE and prefractionation by immunodepletion and isoelectric focusing; increasing coverage or more of the same? Proteomics 2008; 8(23–24): 5074–5085.
13. Polaskova V, Kapur A, Khan A et al. High-abundance protein depletion: comparison of methods for human plasma biomarker discovery. Electrophoresis 2010; 31(3): 471–482.
14. Roche S, Tiers L, Provansal M et al. Depletion of one, six, twelve or twenty major blood proteins before proteomic analysis: the more the better? J Proteomics 2009; 72(6): 945–951.
15. Björhall K, Miliotis T, Davidsson P. Comparison of different depletion strategies for improved resolution in proteomic analysis of human serum samples. Proteomics 2005; 5(1): 307–317.
16. Echan LA, Tang HY, Ali-Khan N et al. Depletion of multiple high-abundance proteins improves protein profiling capacities of human serum and plasma. Proteomics 2005; 5(13): 3292–3303.
17. Magagnotti C, Fermo I, Carletti RM et al. Comparison of different depletion strategies for improving resolution of the human urine proteome. Clin Chem Lab Med 2010; 48(4): 531–535.
18. Yocum AK, Yu K, Oe T et al. Effect of immunoaffinity depletion of human serum during proteomic investigations. J Proteome Res 2005; 4(5): 1722–1731.
19. Brand J, Haslberger T, Zolg W et al. Depletion efficiency and recovery of trace markers from a multiparameter immunodepletion column. Proteomics 2006; 6(11): 3236–3242.
20. Bellei E, Bergamini S, Monari E et al. High-abundance proteins depletion for serum proteomic analysis: concomitant removal of non-targeted proteins. Amino Acids 2011; 40(1): 145–156.
21. Tu C, Rudnick PA, Martinez MY et al. Depletion of abundant plasma proteins and limitations of plasma proteomics. J Proteome Res 2010; 9(10): 4982–4991.
22. Thulasiraman V, Lin SH, Gheorghiu L et al. Reduction of the concentration difference of proteins in biological liquids using a library of combinatorial ligands. Electrophoresis 2005; 26(18): 3561–3571.
23. Righetti PG, Boschetti E, Lomas L et al. Protein Equalizer Technology: the quest for a „democratic proteome“. Proteomics 2006; 6(14): 3980–3992.
24. Boschetti E, Righetti PG. The ProteoMiner in the proteomic arena: a non-depleting tool for discovering low-abundance species. J Proteomics 2008; 71(3): 255–264.
25. Keidel EM, Ribitsch D, Lottspeich F. Equalizer technology – Equal rights for disparate beads. Proteomics 2010; 10(11): 2089–2098.
26. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72: 248–254.
27. Havlis J, Thomas H, Sebela M et al. Fast-response proteomics by accelerated in-gel digestion of proteins. Anal Chem 2003; 75(6): 1300–1306.
28. Planeta J, Karasek P, Vejrosta J. Development of packed capillary columns using carbon dioxide slurries. J Sep Sci 2003; 26: 525–530.
29. Apweiler R, Martin MJ, O’Donovan C et al. The universal protein resource (UniProt) in 2010. Nucleic Acids Res 2010; 38: D142–D148.
30. Mi H, Lazareva-Ulitsky B, Loo R et al. The PANTHER database of protein families, subfamilies, functions and pathways. Nucleic Acids Res 2005; 33: D284–D288.
31. Tirumalai RS, Chan KC, Prieto DA et al. Characterization of the low molecular weight human serum proteome. Mol Cell Proteomics 2003; 2(10): 1096–1103.
32. Fang X, Zhang WW. Affinity separation and enrichment methods in proteomic analysis. J Proteomics 2008; 71(3): 284–303.
33. Sihlbom C, Kanmert I, Bahr H et al. Evaluation of the combination of bead technology with SELDI-TOF-MS and 2-D DIGE for detection of plasma proteins. J Proteome Res 2008; 7(9): 4191–4198.
34. Hartwig S, Czibere A, Kotzka J et al. Combinatorial hexapeptide ligand libraries (ProteoMiner™): an innovative fractionation tool for differential quantitative clinical proteomics. Arch Physiol Biochem 2009; 115(3): 155–160.
35. Porcel JM, Ordi J, Castro-Salomo A et al. The value of complement activation products in the assessment of systemic lupus erythematosus flares. Clin Immunol Immunopathol 1995; 74(3): 283–288.
36. American Thoracic Society, European Respiratory Society. American Thoracic Society/European Respiratory Society Statement: standards for the diagnosis and management of individuals with alpha-1 antitrypsin deficiency. Am J Respir Crit Care Med 2003; 168(7): 818–900.
37. Durrington PN, Mackness B, Mackness MI. Paraoxonase and atherosclerosis. Arterioscler Thromb Vasc Biol 2001; 21(4): 473–480.
38. Flekač M, Škrha J, Novotný Z. Faktory ovlivňující aktivitu a koncentraci antioxidačního enzymu paraoxonáza 1. Klin Biochem Metab 2006; 14(35): 33–39.
39. Goswami B, Tayal D, Gupta N et al. Paraoxonase: a multifaceted biomolecule. Clin Chim Acta 2009; 410(1–2): 1–12.
40. Graham TE, Yang Q, Blüher M et al. Retinol-binding protein 4 and insulin resistance in lean, obese, and diabetic subjects. N Engl J Med 2006; 354(24): 2552–2563.
41. Hofman A, Ott A, Breteler MM et al. Atherosclerosis, apolipoprotein E, and prevalence of dementia and Alzheimer’s disease in the Rotterdam Study. Lancet 1997; 349(9046): 151–154.
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