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A Gel-Free Quantitative Proteomics Analysis of Factors Released From Hypoxic-Conditioned Placentae

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Abstract

Characterizing the protein factors released from placentae during pathogenesis remains a key objective toward understanding preeclampsia and related pregnancy disorders. Gel-free proteomics technologies applied to placental explant-conditioned media offers the potential of identifying these factors. Relative quantification mass spectrometry using isobaric tagging for relative and absolute quantification (iTRAQ) labeling was employed to compare the “secretome” between healthy term placental tissue cultured under both normoxic and hypoxic oxygen tensions. Of the 499 proteins identified, 45 were differentially expressed (P < .01 level), including interleukin 8 (IL-8) which was significantly upregulated under hypoxia. Global protein level changes are suggestive of decreased extracellular matrix remodeling under the same conditions. A significant enrichment of soluble liberated placental factors is achieved using this model system. Identifying these changes resulting from hypoxic conditioning is hypothesis generating and may provide new mechanistic insights into preeclampsia.

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References

  1. Crews JK, Herrington JN, Granger JP, Khalil RA. Decreased endothelium-dependent vascular relaxation during reduction of uterine perfusion pressure in pregnant rat. Hypertension. 2000;35(1 pt 2):367–372.

    Article  CAS  Google Scholar 

  2. Roberts JM, Taylor RN, Musci TJ, Rodgers GM, Hubel CA, McLaughlin MK. Preeclampsia: an endothelial cell disorder. Am J Obstet Gynecol. 1989;161(5):1200–1204.

    Article  CAS  Google Scholar 

  3. Thomson NF, Thornton S, Clark JF. The effects of placental extracts from normotensive and preeclamptic women on vasoconstriction and oxidative metabolism. Am J Obstet Gynecol. 2000;183(1):206–210.

    CAS  PubMed  Google Scholar 

  4. Rajakumar A, Brandon HM, Daftary A, Ness R, Conrad KP. Evidence for the functional activity of hypoxia-inducible transcription factors overexpressed in preeclamptic placentae. Placenta. 2004;25(10):763–769.

    Article  CAS  Google Scholar 

  5. Hayman R, Warren A, Brockelsby J, Johnson I, Baker P. Plasma from women with pre-eclampsia induces an in vitro alteration in the endothelium-dependent behaviour of myometrial resistance arteries. BJOG. 2000;107(1):108–15.

    Article  CAS  Google Scholar 

  6. Myers J, Mires G, Macleod M, Baker P. In preeclampsia, the circulating factors capable of altering in vitro endothelial function precede clinical disease. Hypertension. 2005;45(2):258–263.

    Article  CAS  Google Scholar 

  7. Hayman R, Warren A, Johnson I, Baker P. The preliminary characterization of a vasoactive circulating factor(s) in pree-clampsia. Am J Obstet Gynecol. 2001;184(6):1196–1203.

    Article  CAS  Google Scholar 

  8. Myers JE, Hart S, Armstrong S, et al. Evidence for multiple circulating factors in preeclampsia. Am J Obstet Gynecol. 2007;196(3):266 e1–e6.

    Article  Google Scholar 

  9. Chappell L, Bewley S. Pre-eclamptic toxaemia: the role of uterine artery Doppler. Br J Obstet Gynaecol. 1998;105(4):379–382.

    Article  CAS  Google Scholar 

  10. Robinson NJ, Wareing M, Hudson NK, et al. Oxygen and the liberation of placental factors responsible for vascular compromise. Lab Invest. 2008;88(3):293–305.

    Article  CAS  Google Scholar 

  11. Keshamouni VG, Michailidis G, Grasso CS, et al. Differential protein expression profiling by iTRAQ-2DLC-MS/MS of lung cancer cells undergoing epithelial-mesenchymal transition reveals a migratory/invasive phenotype. J Proteome Res. 2006;5(5):1143–1154.

    Article  CAS  Google Scholar 

  12. Pierce A, Unwin RD, Evans CA, et al. Eight-channel iTRAQ enables comparison of the activity of 6 leukaemogenic tyrosine kinases. Mol Cell Proteomics. 2008;7(5):853–863.

    Article  CAS  Google Scholar 

  13. Sui J, Zhang J, Tan TL, Ching CB, Chen WN. Comparative proteomic analysis of vascular smooth muscle cells incubated with S- and R-enantiomers of atenolol using iTRAQ-coupled 2D LC-MS/MS. Mol Cell Proteomics. 2008;7(6):1007–1018.

    Article  CAS  Google Scholar 

  14. Yang Y, Zhang S, Howe K, et al. A comparison of nLC-ESI-MS/MS and nLC-MALDI-MS/MS for GeLC-based protein identification and iTRAQ-based shotgun quantitative proteomics. J Biomol Tech. 2007;18(4):226–237.

    PubMed  PubMed Central  Google Scholar 

  15. Daayana S, Baker P, Crocker I. An image analysis technique for the investigation of variations in placental morphology in pregnancies complicated by preeclampsia with and without intrauterine growth restriction. J Soc Gynecol Investig. 2004; 11(8):545–552.

    Article  Google Scholar 

  16. Shilov IV, Seymour SL, Patel AA, et al. The Paragon Algorithm, a next generation search engine that uses sequence temperature values and feature probabilities to identify peptides from tandem mass spectra. Mol Cell Proteomics. 2007;6(9): 1638–1655.

    Article  CAS  Google Scholar 

  17. Mi H, Lazareva-Ulitsky B, Loo R, et al. The PANTHER database of protein families, subfamilies, functions and pathways. Nucleic Acids Res. 2005;33(Database issue):D284–D288.

    Article  CAS  Google Scholar 

  18. Thomas PD, Campbell MJ, Kejariwal A, et al. PANTHER: a library of protein families and subfamilies indexed by function. Genome Res. 2003;13(9):2129–2141.

    Article  CAS  Google Scholar 

  19. Robinson JM, Ackerman WE 4th, Kniss DA, Takizawa T, Vandre DD. Proteomics of the human placenta: promises and realities. Placenta. 2008;29(2):135–143.

    Article  CAS  Google Scholar 

  20. Chen CP, Aplin JD. Placental extracellular matrix: gene expression, deposition by placental fibroblasts and the effect of oxygen. Placenta. 2003;24(4):316–325.

    Article  CAS  Google Scholar 

  21. Chen CP, Yang YC, Su TH, Chen CY, Aplin JD. Hypoxia and transforming growth factor-beta 1 act independently to increase extracellular matrix production by placental fibroblasts. J Clin Endocrinol Metab. 2005;90(2):1083–1090.

    Article  CAS  Google Scholar 

  22. Rahat MA, Marom B, Bitterman H, Weiss-Cerem L, Kinarty A, Lahat N. Hypoxia reduces the output of matrix metalloproteinase-9 (MMP-9) in monocytes by inhibiting its secretion and elevating membranal association. J Leukoc Biol. 2006;79(4):706–718.

    Article  CAS  Google Scholar 

  23. Zhao W, Darmanin S, Fu Q, et al. Hypoxia suppresses the production of matrix metalloproteinases and the migration of human monocyte-derived dendritic cells. Eur J Immunol. 2005;35(12):3468–3477.

    Article  CAS  Google Scholar 

  24. Fox H. Pathology of the placenta. Clin Obstet Gynaecol. 1986; 13(3):501–519.

    CAS  PubMed  Google Scholar 

  25. Hu R, Jin H, Zhou S, Yang P, Li X. Proteomic analysis of hypoxia-induced responses in the syncytialization of human placental cell line BeWo. Placenta. 2007;28(5–6):399–407.

    Article  CAS  Google Scholar 

  26. Hoang VM, Foulk R, Clauser K, Burlingame A, Gibson BW, Fisher SJ. Functional proteomics: examining the effects of hypoxia on the cytotrophoblast protein repertoire. Biochemistry. 2001;40(13):4077–4086.

    Article  CAS  Google Scholar 

  27. Bowen RS, Gu Y, Zhang Y, Lewis DF, Wang Y. Hypoxia promotes interleukin-6 and -8 but reduces interleukin-10 production by placental trophoblast cells from preeclamptic pregnancies. J Soc Gynecol Investig. 2005;12(6):428–432.

    Article  CAS  Google Scholar 

  28. Jonsson Y, Ruber M, Matthiesen L, et al. Cytokine mapping of sera from women with preeclampsia and normal pregnancies. J Reprod Immunol. 2006;70(1–2):83–91.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Maternal and Fetal Health Research Group, St Mary’s Hospital, University of Manchester, UK

    Richard T. Blankley PhD, Nicola J. Robinson PhD, John D. Aplin PhD, Ian P. Crocker PhD, Philip N. Baker DM & Jenny E. Myers PhD

  2. Manchester Interdisciplinary Biocentre, University of Manchester, 131 Princess Street, M1 7DN, Manchester, UK

    Richard T. Blankley PhD & Simon J. Gaskell PhD

  3. School of Cancer and Imaging Sciences, Christie Hospital, University of Manchester, UK

    Anthony D. Whetton PhD

Authors
  1. Richard T. Blankley PhD
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  2. Nicola J. Robinson PhD
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  3. John D. Aplin PhD
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  4. Ian P. Crocker PhD
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  5. Simon J. Gaskell PhD
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  6. Anthony D. Whetton PhD
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  7. Philip N. Baker DM
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  8. Jenny E. Myers PhD
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Correspondence to Richard T. Blankley PhD.

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Blankley, R.T., Robinson, N.J., Aplin, J.D. et al. A Gel-Free Quantitative Proteomics Analysis of Factors Released From Hypoxic-Conditioned Placentae. Reprod. Sci. 17, 247–257 (2010). https://doi.org/10.1177/1933719109351320

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  • DOI: https://doi.org/10.1177/1933719109351320

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