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Assessing the Circulating Placental-Specific Anti-angiogenic Protein sFLT-1 e15a in Preeclampsia

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Preeclampsia

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1710))

Abstract

Preeclampsia is a common obstetric complication globally responsible for a significant burden of maternal and perinatal morbidity and mortality. The anti-angiogenic protein, sFLT-1, plays a central role in its pathophysiology. sFLT-1 is released from a range of tissues into the circulation, where it antagonizes the activity of vascular endothelial growth factor and placental growth factor leading to endothelial dysfunction. The resulting widespread endothelial dysfunction produces the clinical features of preeclampsia including hypertension and proteinuria. Multiple splice variants of sFLT-1 have been identified, with one, known as sFLT-1 e15a, present only in humans and higher-order primates. This sFLT-1 variant is also the main form of sFLT-1 produced by the placenta. Recent work has shown that sFLT-1 e15a is significantly elevated in the placenta and circulation of women with preeclampsia. It is also biologically active, capable of causing endothelial dysfunction and end-organ dysfunction seen in preeclampsia. Indeed, overexpression of sFLT-1 e15a in mice recapitulates the preeclamptic phenotype in pregnancy. No commercial assay currently exists to analyze sFLT-1 e15a protein levels. Here, a new ELISA method to determine circulating sFLT-1 variant levels is described.

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References

  1. Levine RJ, Maynard SE, Qian C, Lim K-H, England LJ, KF Y, Schisterman EF, Thadhani R, Sachs BP, Epstein FH, Sibai BM, Sukhatme VP, Karumanchi SA (2004) Circulating angiogenic factors and the risk of preeclampsia. N Engl J Med 350(7):672–683. https://doi.org/10.1056/NEJMoa031884

    Article  CAS  PubMed  Google Scholar 

  2. Levine RJ, Qian C, Maynard SE, KF Y, Epstein FH, Karumanchi SA (2006) Serum sFlt1 concentration during preeclampsia and mid trimester blood pressure in healthy nulliparous women. Am J Obstet Gynecol 194(4):1034–1041. https://doi.org/10.1016/j.ajog.2005.10.192

    Article  CAS  PubMed  Google Scholar 

  3. Shibata E, Rajakumar A, Powers RW, Larkin RW, Gilmour C, Bodnar LM, Crombleholme WR, Ness RB, Roberts JM, Hubel CA (2005) Soluble fms-like tyrosine kinase 1 is increased in preeclampsia but not in normotensive pregnancies with small-for-gestational-age neonates: relationship to circulating placental growth factor. J Clin Endocrinol Metab 90(8):4895–4903. https://doi.org/10.1210/jc.2004-1955

    Article  CAS  PubMed  Google Scholar 

  4. Clark DE, Smith SK, He Y, Day KA, Licence DR, Corps AN, Lammoglia R, Charnock-Jones DS (1998) A vascular endothelial growth factor antagonist is produced by the human placenta and released into the maternal circulation. Biol Reprod 59(6):1540–1548

    Article  CAS  PubMed  Google Scholar 

  5. Maynard S, Min J, Merchan J, Lim K, Li J, Mondal S, Libermann T, Morgan J, Sellke F, Stillman I, Epstein F, Sukhatme V, Ananth Karumanchi S (2003) Excess placental soluble fms-like tyrosine kinase 1 (sFlt-1) may contribute to endothelial dysfunction, hypertension, and proteinuria in preeclampsia. J Clin Invest 111(5):649–658

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Christinger HW, Fuh G, de Vos AM, Wiesmann C (2004) The crystal structure of placental growth factor in complex with domain 2 of vascular endothelial growth factor receptor-1. J Biol Chem 279(11):10382–10388. https://doi.org/10.1074/jbc.M313237200

    Article  CAS  PubMed  Google Scholar 

  7. Wiesmann C, Fuh G, Christinger HW, Eigenbrot C, Wells JA, de Vos AM (1997) Crystal structure at 1.7 A resolution of VEGF in complex with domain 2 of the Flt-1 receptor. Cell 91(5):695–704

    Article  CAS  PubMed  Google Scholar 

  8. Ahmad SAA (2004) Elevated placental soluble vascular endothelial growth factor receptor-1 inhibits angiogenesis in preeclampsia. Circ Res 95(9):884–891. https://doi.org/10.1161/01.RES.0000147365.86159.f5

    Article  CAS  PubMed  Google Scholar 

  9. Gilbert JS, Babcock SA, Granger JP (2007) Hypertension produced by reduced uterine perfusion in pregnant rats is associated with increased soluble fms-like tyrosine kinase-1 expression. Hypertension 50(6):1142–1147. https://doi.org/10.1161/HYPERTENSIONAHA.107.096594

    Article  CAS  PubMed  Google Scholar 

  10. Kumasawa K, Ikawa M, Kidoya H, Hasuwa H, Saito-Fujita T, Morioka Y, Takakura N, Kimura T, Okabe M (2011) Pravastatin induces placental growth factor (PGF) and ameliorates preeclampsia in a mouse model. Proc Natl Acad Sci U S A 108:1451. https://doi.org/10.1073/pnas.1011293108

    Article  PubMed  Google Scholar 

  11. Bergmann A, Ahmad S, Cudmore M, Gruber AD, Wittschen P, Lindenmaier W, Christofori G, Gross V, Gonzalves A, Grone HJ, Ahmed A, Weich HA (2010) Reduction of circulating soluble Flt-1 alleviates preeclampsia-like symptoms in a mouse model. J Cell Mol Med 14(6B):1857–1867. https://doi.org/10.1111/j.1582-4934.2009.00820.x

    Article  CAS  PubMed  Google Scholar 

  12. Gilbert JS, Verzwyvelt J, Colson D, Arany M, Karumanchi SA, Granger JP (2010) Recombinant vascular endothelial growth factor 121 infusion lowers blood pressure and improves renal function in rats with placentalischemia-induced hypertension. Hypertension 55(2):380–385. https://doi.org/10.1161/HYPERTENSIONAHA.109.141937

    Article  CAS  PubMed  Google Scholar 

  13. Li Z, Zhang Y, Ying Ma J, Kapoun AM, Shao Q, Kerr I, Lam A, O’Young G, Sannajust F, Stathis P, Schreiner G, Karumanchi SA, Protter AA, Pollitt NS (2007) Recombinant vascular endothelial growth factor 121 attenuates hypertension and improves kidney damage in a rat model of preeclampsia. Hypertension 50(4):686–692. https://doi.org/10.1161/HYPERTENSIONAHA.107.092098

    Article  CAS  PubMed  Google Scholar 

  14. Thadhani R, Hagmann H, Schaarschmidt W, Roth B, Cingoez T, Karumanchi SA, Wenger J, Lucchesi KJ, Tamez H, Lindner T, Fridman A, Thome U, Kribs A, Danner M, Hamacher S, Mallmann P, Stepan H, Benzing T (2016) Removal of soluble Fms-like tyrosine kinase-1 by dextran sulfate apheresis in preeclampsia. J Am Soc Nephrol 27(3):903–913. https://doi.org/10.1681/ASN.2015020157

    Article  CAS  PubMed  Google Scholar 

  15. He Y, Smith SK, Day KA, Clark DE, Licence DR, Charnock-Jones DS (1999) Alternative splicing of vascular endothelial growth factor (VEGF)-R1 (FLT-1) pre-mRNA is important for the regulation of VEGF activity. Mol Endocrinol 13(4):537–545. https://doi.org/10.1210/mend.13.4.0265

    Article  CAS  PubMed  Google Scholar 

  16. Rahimi N, Golde TE, Meyer RD (2009) Identification of ligand-induced proteolytic cleavage and ectodomain shedding of VEGFR-1/FLT1 in leukemic cancer cells. Cancer Res 69(6):2607–2614. https://doi.org/10.1158/0008-5472.CAN-08-2905

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Heydarian M, McCaffrey T, Florea L, Yang Z, Ross MM, Zhou W, Maynard SE (2009) Novel splice variants of sFlt1 are upregulated in preeclampsia. Placenta 30(3):250–255. https://doi.org/10.1016/j.placenta.2008.12.010

    Article  CAS  PubMed  Google Scholar 

  18. Sela S, Itin A, Natanson-Yaron S, Greenfield C, Goldman-Wohl D, Yagel S, Keshet E (2008) A novel human-specific soluble vascular endothelial growth factor receptor 1: cell type-specific splicing and implications to vascular endothelial growth factor homeostasis and preeclampsia. Circ Res 102(12):1566–1574. https://doi.org/10.1161/circresaha.108.171504

    Article  CAS  PubMed  Google Scholar 

  19. Thomas CP, Andrews JI, Raikwar NS, Kelley EA, Herse F, Dechend R, Golos TG, Liu KZ (2009) A recently evolved novel trophoblast-enriched secreted form of fms-like tyrosine kinase-1 variant is up-regulated in hypoxia and preeclampsia. J Clin Endocrinol Metab 94(7):2524–2530. https://doi.org/10.1210/jc.2009-0017

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Jebbink J, Keijser R, Veenboer G, van der Post J, Ris-Stalpers C, Afink G (2011) Expression of placental FLT1 transcript variants relates to both gestational hypertensive disease and fetal growth. Hypertension 58(1):70–76. https://doi.org/10.1161/HYPERTENSIONAHA.110.164079

    Article  CAS  PubMed  Google Scholar 

  21. Whitehead CL, Palmer KR, Nilsson U, Gao Y, Saglam B, Lappas M, Tong S (2011) Placental expression of a novel primate-specific splice variant of sFlt-1 is upregulated in pregnancies complicated by severe early onset pre-eclampsia. BJOG 118(10):1268–1271. https://doi.org/10.1111/j.1471-0528.2011.02962.x

    Article  CAS  PubMed  Google Scholar 

  22. Palmer KR, Kaitu’u-Lino TJ, Hastie R, Hannan NJ, Ye L, Binder N, Cannon P, Tuohey L, Johns TG, Shub A, Tong S (2015) Placental-specific sFLT-1 e15a protein is increased in preeclampsia, antagonizes vascular endothelial growth factor signaling, and has antiangiogenic activity. Hypertension 66(6):1251–1259. https://doi.org/10.1161/HYPERTENSIONAHA.115.05883

    Article  CAS  PubMed  Google Scholar 

  23. Souders CA, Maynard SE, Yan J, Wang Y, Boatright NK, Sedan J, Balyozian D, Cheslock PS, Molrine DC, Simas TA (2015) Circulating levels of sFlt1 splice variants as predictive markers for the development of preeclampsia. Int J Mol Sci 16(6):12436–12453. https://doi.org/10.3390/ijms160612436

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Palmer KR, Kaitu’u-Lino TJ, Cannon P, Tuohey L, De Silva MS, Varas-Godoy M, Acuna S, Galaz J, Tong S, Illanes SE (2017) Maternal plasma concentrations of the placental specific sFLT-1 variant, sFLT-1 e15a, in fetal growth restriction and preeclampsia. J Matern Fetal Neonatal Med 30:635–639. https://doi.org/10.1080/14767058.2016.1182975

    Article  CAS  PubMed  Google Scholar 

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Acknowledgment

I would like to acknowledge Professor Terrence Johns for his assistance with peptide design and ELISA optimization. I would also like to acknowledge Dr. Tu’uhevaha Kaitu’u-Lino for her advice with ELISA development and Dr. Sandra Nicholson for her assistance with optimizing sFLT-1 e15a protein production. This work received funding support from the Royal Australian and New Zealand College of Obstetricians and Gynaecologists Arthur Wilson Memorial scholarship and Keith Fitzmaurice Bursary, as well as the National Health and Medical Research Council (#607219).

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, School of Clinical Sciences, Monash University, Level 5, 246 Clayton Rd, Clayton, 3168, VIC, Australia

    Kirsten Palmer

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  1. Kirsten Palmer
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Correspondence to Kirsten Palmer .

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Editors and Affiliations

  1. Monash Medical Centre, Monash University, Clayton, Victoria, Australia

    Padma Murthi

  2. INRS-Institut Armand-Frappier, Laval, Québec, Canada

    Cathy Vaillancourt

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Palmer, K. (2018). Assessing the Circulating Placental-Specific Anti-angiogenic Protein sFLT-1 e15a in Preeclampsia. In: Murthi, P., Vaillancourt, C. (eds) Preeclampsia . Methods in Molecular Biology, vol 1710. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7498-6_3

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  • DOI: https://doi.org/10.1007/978-1-4939-7498-6_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7497-9

  • Online ISBN: 978-1-4939-7498-6

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