Rapid and deep plasma proteomics workflows for robust identification and quantification of biomarkers of sickle cell anaemia

Springer Science and Business Media LLC - Tập 13 - Trang 205-218 - 2022
Sravani Polepalli1, Richa Singh1, Shoma Naskar2, Pasupuleti SKDB Punyasri2, Kongari Ranjith Kumar3, Kameshwari Yele3, Viswanatha Krishnakumari3, Raman Bakthisaran3, Dipty Jain4, Giriraj Ratan Chandak2,5, Swasti Raychaudhuri1,3,5
1Cellular Biochemistry and Proteomics Group, CSIR-Centre for Cellular and Molecular Biology, Hyderabad, India
2Genomic Research on Complex Diseases (GRC-Group), CSIR-Centre for Cellular and Molecular Biology, Hyderabad, India
3Central Proteomics Facility, CSIR-Centre for Cellular and Molecular Biology, Hyderabad, India
4Consultant Paediatrician, Arihant Multispeciality Hospitals, Nagpur, India
5Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, India

Tóm tắt

Plasma serves as a rich source of protein biomarkers but in-depth proteomic analysis is challenging due to vast dynamic range of protein abundance. Pre-fractionation of plasma proteins is commonly practiced to enhance the proteome coverage but the protocols are time-expensive, suffer from flowchart complexity, and often less reproducible. Here, we explore multiple strategies of shotgun proteomics to optimize biomarker discovery workflows for Sickle Cell Anaemia (SCA) patients from Maharashtra, India. A deep proteomics workflow via off-line reverse phase ultra-high-pressure liquid chromatography-based fractionation of tryptic digested plasma peptides followed by optimized pooling of peptides based on charge and hydrophobicity yielded the best depth of plasma proteome with a trade-off of significantly long experimental time. Alternatively, a rapid analysis of tryptic digested plasma peptides via a shorter gradient mass spectrometry run saves time but quantifies only ~ 50% of the proteins than the deep workflow. Intriguingly, despite the difference in proteome coverage, more than 80% of known FDA and SCA biomarkers quantified in the deep workflow are also quantified in the rapid workflow. Given the practical difficulties of sample collection and plasma preservation in rural India, we propose the deep proteomics workflow for biomarker discovery in smaller cohorts and the rapid workflow for biomarker validations in larger cohorts. Targeted-proteomics-based strategies may be designed for the validation of missing biomarkers in the rapid workflow.

Tài liệu tham khảo

Akinsheye I, Alsultan A, Solovieff N et al (2011) Fetal hemoglobin in sickle cell anemia. Blood 118:19–27. https://doi.org/10.1182/blood-2011-03-325258 Anderson NL (2010) The clinical plasma proteome: a survey of clinical assays for proteins in plasma and serum. Clin Chem 56:177–185. https://doi.org/10.1373/clinchem.2009.126706 Anderson NL, Anderson NG (2002) The human plasma proteome: history, character, and diagnostic prospects. Mol Cell Proteomics 1:845–867. https://doi.org/10.1074/mcp.r200007-mcp200 Ballas SK, Kesen MR, Goldberg MF et al (2012) Beyond the definitions of the phenotypic complications of sickle cell disease: an update on management. Sci World J 2012:949535. https://doi.org/10.1100/2012/949535 Bellei E, Bergamini S, Monari E et al (2011) High-abundance proteins depletion for serum proteomic analysis: concomitant removal of non-targeted proteins. Amino Acids 40:145–156. https://doi.org/10.1007/s00726-010-0628-x Brousse V, Rees DC (2021) Sickle cell disease: More than a century of progress. Where do we stand now? Indian J Med Res 154:4–7. https://doi.org/10.4103/ijmr.ijmr_1435_21 Cao Z, Tang HY, Wang H et al (2012) Systematic comparison of fractionation methods for in-depth analysis of plasma proteomes. J Proteome Res 11:3090–3100. https://doi.org/10.1021/pr201068b Chakravorty S, Williams TN (2015) Sickle cell disease: a neglected chronic disease of increasing global health importance. Arch Dis Child 100:48–53. https://doi.org/10.1136/archdischild-2013-303773 Dieckmann-Schuppert A, Schnittler HJ (1997) A simple assay for quantification of protein in tissue sections, cell cultures, and cell homogenates, and of protein immobilized on solid surfaces. Cell Tissue Res 288:119–126. https://doi.org/10.1007/s004410050799 Echan LA, Tang HY, Ali-Khan N et al (2005) Depletion of multiple high-abundance proteins improves protein profiling capacities of human serum and plasma. Proteomics 5:3292–3303. https://doi.org/10.1002/pmic.200401228 Geyer PE, Kulak NA, Pichler G et al (2016) Plasma proteome profiling to assess human health and disease. Cell Syst 2:185–195. https://doi.org/10.1016/j.cels.2016.02.015 Geyer PE, Voytik E, Treit PV et al (2019) Plasma Proteome Profiling to detect and avoid sample-related biases in biomarker studies. EMBO Mol Med 11:e10427. https://doi.org/10.15252/emmm.201910427 Heikkila K, Ebrahim S, Lawlor DA (2007) A systematic review of the association between circulating concentrations of C reactive protein and cancer. J Epidemiol Community Health 61:824–833. https://doi.org/10.1136/jech.2006.051292 Hortin GL, Sviridov D (2010) The dynamic range problem in the analysis of the plasma proteome. J Proteomics 73:629–636. https://doi.org/10.1016/j.jprot.2009.07.001 Hu S, Loo JA, Wong DT (2006) Human body fluid proteome analysis. Proteomics 6:6326–6353. https://doi.org/10.1002/pmic.200600284 Kakhniashvili DG, Griko NB, Bulla LA Jr et al (2005) The proteomics of sickle cell disease: profiling of erythrocyte membrane proteins by 2D-DIGE and tandem mass spectrometry. Exp Biol Med (maywood) 230:787–792. https://doi.org/10.1177/153537020523001102 Kelstrup CD, Jersie-Christensen RR, Batth TS et al (2014) Rapid and deep proteomes by faster sequencing on a benchtop quadrupole ultra-high-field Orbitrap mass spectrometer. J Proteome Res 13:6187–6195. https://doi.org/10.1021/pr500985w Keshishian H, Addona T, Burgess M et al (2009) Quantification of cardiovascular biomarkers in patient plasma by targeted mass spectrometry and stable isotope dilution. Mol Cell Proteomics 8:2339–2349. https://doi.org/10.1074/mcp.M900140-MCP200 Keshishian H, Burgess MW, Gillette MA et al (2015) Multiplexed, quantitative workflow for sensitive biomarker discovery in plasma yields novel candidates for early myocardial injury. Mol Cell Proteomics 14:2375–2393. https://doi.org/10.1074/mcp.M114.046813 Keshishian H, Burgess MW, Specht H et al (2017) Quantitative, multiplexed workflow for deep analysis of human blood plasma and biomarker discovery by mass spectrometry. Nat Protoc 12:1683–1701. https://doi.org/10.1038/nprot.2017.054 Leon IR, Schwammle V, Jensen ON et al (2013) Quantitative assessment of in-solution digestion efficiency identifies optimal protocols for unbiased protein analysis. Mol Cell Proteomics 12:2992–3005. https://doi.org/10.1074/mcp.M112.025585 Miller SA, Dykes DD, Polesky HF (1988) A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 16:1215. https://doi.org/10.1093/nar/16.3.1215 Niu L, Geyer PE, Wewer Albrechtsen NJ et al (2019) Plasma proteome profiling discovers novel proteins associated with non-alcoholic fatty liver disease. Mol Syst Biol 15:e8793. https://doi.org/10.15252/msb.20188793 Piel FB, Patil AP, Howes RE et al (2013) Global epidemiology of sickle haemoglobin in neonates: a contemporary geostatistical model-based map and population estimates. Lancet 381:142–151. https://doi.org/10.1016/S0140-6736(12)61229-X Rappsilber J, Ishihama Y, Mann M (2003) Stop and go extraction tips for matrix-assisted laser desorption/ionization, nanoelectrospray, and LC/MS sample pretreatment in proteomics. Anal Chem 75:663–670 Shevchenko A, Wilm M, Vorm O et al (1996) Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal Chem 68:850–858 Singh P, Chakraborty R, Marwal R et al (2020) A rapid and sensitive method to detect SARS-CoV-2 virus using targeted-mass spectrometry. J Proteins Proteom 11:159–165. https://doi.org/10.1007/s42485-020-00044-9 Tu C, Rudnick PA, Martinez MY et al (2010) Depletion of abundant plasma proteins and limitations of plasma proteomics. J Proteome Res 9:4982–4991. https://doi.org/10.1021/pr100646w Vizcaino JA, Deutsch EW, Wang R et al (2014) ProteomeXchange provides globally coordinated proteomics data submission and dissemination. Nat Biotechnol 32:223–226 Wu C, Duan J, Liu T et al (2016) Contributions of immunoaffinity chromatography to deep proteome profiling of human biofluids. J Chromatogr B Analyt Technol Biomed Life Sci 1021:57–68. https://doi.org/10.1016/j.jchromb.2016.01.015