Altered erythroid-related miRNA levels as a possible novel biomarker for detection of autologous blood transfusion misuse in sport.
Journal
Transfusion
ISSN: 1537-2995
Titre abrégé: Transfusion
Pays: United States
ID NLM: 0417360
Informations de publication
Date de publication:
08 2019
08 2019
Historique:
received:
09
07
2018
revised:
01
04
2019
accepted:
11
04
2019
pubmed:
31
5
2019
medline:
2
6
2020
entrez:
1
6
2019
Statut:
ppublish
Résumé
Autologous blood transfusion (ABT) is a performance-enhancing method prohibited in sport; its detection is a key issue in the field of anti-doping. Among novel markers enabling ABT detection, microRNAs (miRNAs) might be considered a promising analytical tool. We studied the changes of erythroid-related microRNAs following ABT, to identify novel biomarkers. Fifteen healthy trained males were studied from a population of 24 subjects, enrolled and randomized into a Transfusion (T) and a Control (C) group. Seriated blood samples were obtained in the T group before and after the two ABT procedures (withdrawal, with blood refrigerated or cryopreserved, and reinfusion), and in the C group at the same time points. Traditional hematological parameters were assessed. Samples were tested by microarray analysis of a pre-identified set of erythroid-related miRNAs. Hematological parameters showed moderate changes only in the T group, particularly following blood withdrawal. Among erythroid-related miRNAs tested, following ABT a pool of 7 miRNAs associated with fetal hemoglobin and regulating transcriptional repressors of gamma-globin gene was found stable in C and differently expressed in three out of six T subjects in the completed phase of ABT, independently from blood conservation. Particularly, two or more erythropoiesis-related miRNAs within the shortlist constituted of miR-126-3p, miR-144-3p, miR-191-3p, miR-197-3p, miR-486-3p, miR-486-5p, and miR-92a-3p were significantly upregulated in T subjects after reinfusion, with a person-to-person variability but with congruent changes. This study describes a signature of potential interest for ABT detection in sports, based on the analysis of miRNAs associated with erythroid features.
Sections du résumé
BACKGROUND
Autologous blood transfusion (ABT) is a performance-enhancing method prohibited in sport; its detection is a key issue in the field of anti-doping. Among novel markers enabling ABT detection, microRNAs (miRNAs) might be considered a promising analytical tool.
STUDY DESIGN AND METHODS
We studied the changes of erythroid-related microRNAs following ABT, to identify novel biomarkers. Fifteen healthy trained males were studied from a population of 24 subjects, enrolled and randomized into a Transfusion (T) and a Control (C) group. Seriated blood samples were obtained in the T group before and after the two ABT procedures (withdrawal, with blood refrigerated or cryopreserved, and reinfusion), and in the C group at the same time points. Traditional hematological parameters were assessed. Samples were tested by microarray analysis of a pre-identified set of erythroid-related miRNAs.
RESULTS
Hematological parameters showed moderate changes only in the T group, particularly following blood withdrawal. Among erythroid-related miRNAs tested, following ABT a pool of 7 miRNAs associated with fetal hemoglobin and regulating transcriptional repressors of gamma-globin gene was found stable in C and differently expressed in three out of six T subjects in the completed phase of ABT, independently from blood conservation. Particularly, two or more erythropoiesis-related miRNAs within the shortlist constituted of miR-126-3p, miR-144-3p, miR-191-3p, miR-197-3p, miR-486-3p, miR-486-5p, and miR-92a-3p were significantly upregulated in T subjects after reinfusion, with a person-to-person variability but with congruent changes.
CONCLUSIONS
This study describes a signature of potential interest for ABT detection in sports, based on the analysis of miRNAs associated with erythroid features.
Substances chimiques
Biomarkers
0
MicroRNAs
0
Types de publication
Clinical Trial
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2709-2721Informations de copyright
© 2019 AABB.
Références
World Anti-Doping Agency 2018 list of prohibited substances and methods. [cited 2018 Feb 14]. Available from: https://www.wada-ama.org/en/content/what-is-prohibited.
Jelkmann W, Lundby C. Blood doping and its detection. Blood 2011;118:2395-404.
Mørkeberg J. Detection of autologous blood transfusions in athletes: a historical perspective. Transfus Med Rev 2012;26:199-208.
Segura J, Lundby C. Blood doping: potential of blood and urine sampling to detect autologous transfusion. Br J Sports Med 2014;48:837-41.
Segura J, Monfort N, Ventura R. Detection methods for autologous blood doping. Drug Test Anal 2012;4:876-81.
Gore CJ, Parisotto R, Ashenden MJ, et al. Second-generation blood tests to detect erythropoietin abuse by athletes. Haematologica 2003;88:333-44.
Pottgiesser T, Sottas PE, Echteler T, et al. Detection of autologous blood doping with adaptively evaluated biomarkers of doping: a longitudinal blinded study. Transfusion 2011;51:1707-15.
Schumacher YO, Saugy M, Pottgiesser T, et al. Detection of EPO doping and blood doping: the haematological module of the Athlete Biological Passport. Drug Test Anal 2012;4:846-53.
Salamin O, De Angelis S, Tissot JD, et al. Autologous blood transfusion in sports: emerging biomarkers. Transfus Med Rev 2016;30:109-15.
Malm CB, Khoo NS, Granlund I, et al. Autologous doping with cryopreserved red blood cells - effects on physical performance and detection by multivariate statistics. PLoS One 2016;11:e0156157.
Damsgaard R, Munch T, Mørkeberg J, et al. Effects of blood withdrawal and reinfusion on biomarkers of erythropoiesis in humans: implications for antidoping strategies. Haematologica 2006;91:1006-8.
Leuenberger N, Barras L, Nicoli R, et al. Hepcidin as a new biomarker for detecting autologous blood transfusion. Am J Hematol 2016;91:467-72.
Leuenberger N, Barras L, Nicoli R, et al. Urinary di-(2-ethylhexyl) phthalate metabolites for detecting transfusion of autologous blood stored in plasticizer-free bags. Transfusion 2016;56:571-8.
Nikolovski Z, De La Torre C, Chiva C, et al. Alterations of the erythrocyte membrane proteome and cytoskeleton network during storage-a possible tool to identify autologous blood transfusion. Drug Test Anal 2012;4:882-90.
Reichel C. OMICS-strategies and methods in the fight against doping. Forensic Sci Int 2011;213:20-34.
Donati F, Boccia F, De La Torre X, et al. MicroRNA analysis for the detection of autologous blood transfusion in doping control. Agilent Application Note 2015; 25 March:5991-5717EN.
Leuenberger N, Schumacher YO, Pradervand S, et al. Circulating microRNAs as biomarkers for detection of autologous blood transfusion. PLoS One 2013;8:e66309.
He L, Hannon GJ. MicroRNAs: small RNAs with a big role in gene regulation. Nat Rev Genet 2004;5:522-31.
Gasparello J, Fabbri E, Bianchi N, et al. BCL11A mRNA targeting by miR-210: a possible network regulating γ-globin gene expression. Int J Mol Sci 2017;18:E2530.
Ferrari D, Bianchi N, Eltzschig HK, et al. MicroRNAs modulate the purinergic signaling network. Trends Mol Med 2016;22:905-18.
Brognara E, Fabbri E, Bazzoli E, et al. Uptake by human glioma cell lines and biological effects of a peptide-nucleic acids targeting miR-221. J Neurooncol 2014;11:19-28.
Kroh EM, Parkin RK, Mitchell PS, et al. Analysis of circulating microRNA biomarkers in plasma and serum using quantitative reverse transcription-PCR (qRT-PCR). Methods 2010;50:298-301.
Costa MC, Leitão AL, Enguita FJ. MicroRNA profiling in plasma or serum using quantitative RT-PCR. Methods Mol Biol 2014;1182:121-9.
Watier T, Mj Sanchez A. Micro-RNAs, exercise and cellular plasticity in humans: the impact of dietary factors and hypoxia. Microrna 2017;6:110-24.
Correia CN, Nalpas NC, McLoughlin KE, et al. Circulating microRNAs as potential biomarkers of infectious disease. Front Immunol 2017;8:118.
Zhao Y, Song Y, Yao L, et al. Circulating microRNAs: promising biomarkers involved in several cancers and other diseases. DNA Cell Biol 2017;36:77-94.
Lamberti N, Finotti A, Gasparello J, et al. Changes in hemoglobin profile reflect autologous blood transfusion misuse in sports. Intern Emerg Med 2018;13:517-26.
Finotti A, Bianchi N, Fabbri E, et al. Erythroid induction of K562 cells treated with mithramycin is associated with inhibition of raptor gene transcription and mammalian target of rapamycin complex 1 (mTORC1) functions. Pharmacol Res 2015;91:57-68.
Bianchi N, Finotti A, Ferracin M, et al. Increase of microRNA-210, decrease of raptor gene expression and alteration of mammalian target of rapamycin regulated proteins following mithramycin treatment of human erythroid cells. PLoS One 2015;10:e0121567.
El-Khoury V, Pierson S, Kaoma T, et al. Assessing cellular and circulating miRNA recovery: the impact of the RNA isolation method and the quantity of input material. Sci Rep 2016;6:19529.
Duy J, Koehler JW, Honko AN, et al. Optimized microRNA purification from TRIzol-treated plasma. BMC Genomics 2015;16:95.
Rice J, Roberts H, Burton J, et al. Assay reproducibility in clinical studies of plasma miRNA. PLoS One 2015;10:e0121948.
Brzobohatá K, Drozdová E, Smutný J, et al. Comparison of suitability of the most common ancient DNA quantification methods. Genet Test Mol Biomarkers 2017;21:265-71.
Bolstad BM, Irizarry RA, Astrand M, et al. A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics 2003;19:185-93.
Gasparello J, Allegretti M, Tremante E, et al. Liquid biopsy in mice bearing colorectal carcinoma xenografts: gateways regulating the levels of circulating tumor DNA (ctDNA) and miRNA (ctmiRNA). J Exp Clin Cancer Res 2018;37:124.
Risso A, Fabbro D, Damante G, et al. Expression of fetal hemoglobin in adult humans exposed to high altitude hypoxia. Blood Cells Mol Dis 2012;48:147-53.
Seeley TW, Sternlicht MD, Klaus SJ, et al. Induction of erythropoiesis by hypoxia-inducible factor prolyl hydroxylase inhibitors without promotion of tumor initiation, progression, or metastasis in a VEGF-sensitive model of spontaneous breast cancer. Hypoxia (Auckl) 2017;5:1-9.
Noh SJ, Miller SH, Lee YT, et al. Let-7 microRNAs are developmentally regulated in circulating human erythroid cells. J Transl Med 2009;7:98.
Ruan J, Liu X, Xiong X, et al. miR-107 promotes the erythroid differentiation of leukemia cells via the downregulation of Cacna2d1. Mol Med Rep 2015;11:1334-9.
Xiao S, Liao S, Zhou Y, et al. High expression of octamer transcription factor 1 in cervical cancer. Oncol Lett 2014;7:1889-94.
Gañán-Gómez I, Wei Y, Yang H, et al. Overexpression of miR-125a in myelodysplastic syndrome CD34+ cells modulates NF-κB activation and enhances erythroid differentiation arrest. PLoS One 2014;9:e93404.
Huang X, Gschweng E, Van Handel B, et al. Regulated expression of microRNAs-126/126* inhibits erythropoiesis from human embryonic stem cells. Blood 2011;117:2157-65.
Pase L, Layton JE, Kloosterman WP, et al. miR-451 regulates zebrafish erythroid maturation in vivo via its target gata2. Blood 2009;113:1794-804.
Fu YF, Du TT, Dong M, et al. Mir-144 selectively regulates embryonic alpha-hemoglobin synthesis during primitive erythropoiesis. Blood 2009;113:1340-9.
Zhai PF, Wang F, Su R, et al. The regulatory roles of microRNA-146b-5p and its target platelet-derived growth factor receptor α (PDGFRA) in erythropoiesis and megakaryocytopoiesis. PLoS One 2014;9:e93404.
Sun Z, Wang Y, Han X, et al. miR-150 inhibits terminal erythroid proliferation and differentiation. Blood 2015;125:1302-13.
Xiao C, Calado DP, Galler G, et al. MiR-150 controls B cell differentiation by targeting the transcription factor c-Myb. Cell 2007;131:146-59.
Pule GD, Mowla S, Novitzky N, et al. Hydroxyurea down-regulates BCL11A, KLF-1 and MYB through miRNA-mediated actions to induce γ-globin expression: implications for new therapeutic approaches of sickle cell disease. Clin Transl Med 2016;5:15.
Finotti A, Gambari R. Recent trends for novel options in experimental biological therapy of β-thalassemia. Expert Opin Biol Ther 2014;14:1443-54.
O'Connell RM, Rao DS, Chaudhuri AA, et al. Sustained expression of microRNA-155 in hematopoietic stem cells causes a myeloproliferative disorder. J Exp Med 2008;205:585-94.
Xie Q, Chen X, Lu F, et al. Aberrant expression of microRNA 155 may accelerate cell proliferation by targeting sex-determining region Y box 6 in hepatocellular carcinoma. Cancer 2012;118:2431-42.
Sankaran VG, Menne TF, Šćepanović D, et al. MicroRNA-15a and −16-1 act via MYB to elevate fetal hemoglobin expression in human trisomy 13. Proc Natl Acad Sci U S A 2011;108:1519-24.
Zhang L, Flygare J, Wong P, et al. miR-191 regulates mouse erythroblast enucleation by down-regulating Riok3 and Mxi1. Genes Dev 2011;25:119-24.
Li Y, Bai H, Zhang Z, et al. The up-regulation of miR-199b-5p in erythroid differentiation is associated with GATA-1 and NF-E2. Mol Cells 2014;37:213-9.
Ding J, Chen J, Wang Y, et al. Trbp regulates heart function through microRNA-mediated Sox6 repression. Nat Genet 2015;47:776-83.
Kim JM, Lim KS, Hong JS, et al. A polymorphism in the porcine miR-208b is associated with microRNA biogenesis and expressions of SOX-6 and MYH7 with effects on muscle fibre characteristics and meat quality. Anim Genet 2015;46:73-7.
Bianchi N, Zuccato C, Lampronti I, et al. Expression of miR-210 during erythroid differentiation and induction of gamma-globin gene expression. BMB Rep 2009;42:493-9.
Li Y, Liu S, Sun H, et al. MiR-218 inhibits erythroid differentiation and alters iron metabolism by targeting ALAS2 in K562 cells. Int J Mol Sci 2015;16:28156-68.
Gabbianelli M, Testa U, Morsilli O, et al. Mechanism of human Hb switching: a possible role of the kit receptor/miR 221-222 complex. Haematologica 2010;95:1253-60.
Felli N, Pedini F, Romania P, et al. MicroRNA 223-dependent expression of LMO2 regulates normal erythropoiesis. Int J Biochem Cell Biol 2013;45:2519-29.
Zhu Y, Wang D, Wang F, et al. A comprehensive analysis of GATA-1-regulated miRNAs reveals miR-23a to be a positive modulator of erythropoiesis. Nucleic Acids Res 2013;41:4129-43.
Wang F, Zhu Y, Guo L, et al. A regulatory circuit comprising GATA1/2 switch and microRNA-27a/24 promotes erythropoiesis. Nucleic Acids Res 2014;42:442-57.
Alijani S, Alizadeh S, Kazemi A, et al. Evaluation of the effect of miR-26b up-regulation on HbF expression in erythroleukemic K-562 cell line. Avicenna J Med Biotechnol 2014;6:53-6.
Jiang BY, Zhang XC, Su J, et al. BCL11A overexpression predicts survival and relapse in non-small cell lung cancer and is modulated by microRNA-30a and gene amplification. Mol Cancer 2013;12:61.
Mittal SP, Mathai J, Kulkarni AP, et al. miR-320a regulates erythroid differentiation through MAR binding protein SMAR1. Cell Res 2011;21:1196-209.
Navarro F, Gutman D, Meire E, et al. miR-34a contributes to megakaryocytic differentiation of K562 cells independently of p53. Blood 2009;114:2181-92.
Wang F, Yu J, Yang GH, et al. Regulation of erythroid differentiation by miR-376a and its targets. Cell Res 2011;21:1196-209.
Liu Y, Wang Y, Sun X, et al. miR-449a promotes liver cancer cell apoptosis by downregulation of Calpain 6 and POU2F1. Oncotarget 2016;7:13491-501.
Lulli V, Romania P, Morsilli O, et al. MicroRNA-486-3p regulates γ-globin expression in human erythroid cells by directly modulating BCL11A. PLoS One 2013;8:e60436.
Wang F, Yu J, Yang GH, et al. Regulation of erythroid differentiation by miR-376a and its targets. Blood 2009;113:1794-804.
Hojjati MT, Azarkeivan A, Pourfathollah AA, et al. Comparison of microRNAs mediated in reactivation of the γ-globin in β-thalassemia patients, responders and non-responders to hydroxyurea. Hemoglobin 2017;41:110-5.
Yoo JK, Kim J, Choi SJ, et al. Discovery and characterization of novel microRNAs during endothelial differentiation of human embryonic stem cells. Stem Cells Dev 2012;21:2049-57.
Li YC, Li CF, Chen LB, et al. MicroRNA-766 targeting regulation of SOX6 expression promoted cell proliferation of human colorectal cancer. Onco Targets Ther 2015;8:2981-8.
Li Y, Vecchiarelli-Federico LM, Li YJ, et al. The miR-17-92 cluster expands multipotent hematopoietic progenitors whereas imbalanced expression of its individual oncogenic miRNAs promotes leukemia in mice. Blood 2012;119:4486-98.
Azzouzi I, Moest H, Winkler J, et al. MicroRNA-96 directly inhibits γ-globin expression in human erythropoiesis. PLoS One 2011;6:e22838.
Emmrich S, Rasche M, Schöning J, et al. miR-99a/100~125b tricistrons regulate hematopoietic stem and progenitor cell homeostasis by shifting the balance between TGFβ and Wnt signaling. Genes Dev 2014;28:858-74.
Willinger CM, Rong J, Tanriverdi K, et al. MicroRNA signature of cigarette smoking and evidence for a putative causal role of microRNAs in smoking-related inflammation and target organ damage. Circ Cardiovasc Genet 2017;10:e001678.
Zhao J, Qiao CR, Ding Z, et al. A novel pathway in NSCLC cells: miR 191, targeting NFIA, is induced by chronic hypoxia, and promotes cell proliferation and migration. Mol Med Rep 2017;15:1319-25.
Nagpal N, Ahmad HM, Chameettachal S, et al. HIF-inducible miR-191 promotes migration in breast cancer through complex regulation of TGFβ-signaling in hypoxic microenvironment. Sci Rep 2015;5:9650.
Song Z, Ren H, Gao S, et al. The clinical significance and regulation mechanism of hypoxia-inducible factor-1 and miR-191expression in pancreatic cancer. Tumour Biol 2014;35:11319-28.
Manfredini AF, Malagoni AM, Litmanen H, et al. Performance and blood monitoring in sports: the artificial intelligence evoking target testing in antidoping (AR.I.E.T.T.A.) project. J Sports Med Phys Fitness 2011;51:153-9.