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Published in: Journal of Assisted Reproduction and Genetics 6/2019

Open Access 01-06-2019 | Laser | Embryo Biology

Secretome profile selection of optimal IVF embryos by matrix-assisted laser desorption ionization time-of-flight mass spectrometry

Authors: Ray K. Iles, Fady I. Sharara, Raminta Zmuidinaite, Galal Abdo, Sholeh Keshavarz, Stephen A. Butler

Published in: Journal of Assisted Reproduction and Genetics | Issue 6/2019

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Abstract

Purpose

Selecting an embryo at the transfer stage with the best chance of a successful pregnancy is still largely dependent on preceding subjective evaluation of morphokinetics. Expensive prenatal genomic profiling has been so far proved ineffective. Proteomics and metabolomics are promising new approaches to assess embryo viability, but methodologies are often complex and do not lend themselves to rapid analysis in the critical time between blastocyst formation and embryo transfer. Here, we used matrix-assisted laser desorption ionization time-of-flight (MALDI ToF) mass spectrometry to assess the secretome of blastocysts in the minutes prior to embryo transfer and correlated spectral features with pregnancy outcome.

Methods

Four hundred one samples of spent blastocyst culture media were collected from embryo cultures at the time of embryo transfer, of which 136 were used to construct the predictive model. The media samples were frozen at − 20 °C and stored for analysis. Sample analysis was conducted in batches using 1 μl of spent embryo in direct MALDI ToF mass spectral analysis. Quantitative characteristics within this mass range (2000–17,000 m/z) were used to generate a score for selected mass regions (bins) in order to predict pregnancy outcome for each sample.

Results

With a simple algorithm based on nine mass bins within the 2000–10,000 m/z region, it was possible to identify samples with the best chance of becoming an ongoing pregnancy (positive predictive value of 82.9%, p = 0.0018).

Conclusion

A simple, direct and rapid analysis of spent culture fluid from blastocysts at the point of embryo transfer can quickly identify optimal embryos with the best chance of achieving ongoing pregnancy. Methods like this, which take less than 20 min to perform, could dramatically improve the approach to embryo selection and live births.
Literature
1.
go back to reference Gardner DK, Meseguer M, Rubio C, Treff NR. Diagnosis of human preimplantation embryo viability. Hum Reprod Update. 2015;21(6):727–47.CrossRefPubMed Gardner DK, Meseguer M, Rubio C, Treff NR. Diagnosis of human preimplantation embryo viability. Hum Reprod Update. 2015;21(6):727–47.CrossRefPubMed
2.
go back to reference Kupka MS, D'Hooghe T, Ferraretti AP, de Mouzon J, Erb K, Castilla JA, et al. European IVF-Monitoring Consortium (EIM); European Society of Human Reproduction and Embryology (ESHRE). Assisted reproductive technology in Europe, 2011: results generated from European registers by ESHRE. Hum Reprod. 2016;31(2):233–48.PubMed Kupka MS, D'Hooghe T, Ferraretti AP, de Mouzon J, Erb K, Castilla JA, et al. European IVF-Monitoring Consortium (EIM); European Society of Human Reproduction and Embryology (ESHRE). Assisted reproductive technology in Europe, 2011: results generated from European registers by ESHRE. Hum Reprod. 2016;31(2):233–48.PubMed
7.
go back to reference Krisher RL, Schoolcraft WB, Katz-Jaffe MG. Omics as a window to view embryo viability. Fertil Steril. 2015;103(2):333–41.CrossRefPubMed Krisher RL, Schoolcraft WB, Katz-Jaffe MG. Omics as a window to view embryo viability. Fertil Steril. 2015;103(2):333–41.CrossRefPubMed
8.
go back to reference Rødgaard T, Heegaard PM, Callesen H. Non-invasive assessment of in-vitro embryo quality to improve transfer success. Reprod BioMed Online. 2015;31(5):585–92.CrossRefPubMed Rødgaard T, Heegaard PM, Callesen H. Non-invasive assessment of in-vitro embryo quality to improve transfer success. Reprod BioMed Online. 2015;31(5):585–92.CrossRefPubMed
9.
go back to reference Katz-Jaffe MG, Schoolcraft WB, Gardner DK. Analysis of protein expression (secretome) by human and mouse preimplantation embryos. Fertil Steril. 2006;86(3):678–85.CrossRefPubMed Katz-Jaffe MG, Schoolcraft WB, Gardner DK. Analysis of protein expression (secretome) by human and mouse preimplantation embryos. Fertil Steril. 2006;86(3):678–85.CrossRefPubMed
10.
go back to reference Brison DR, Houghton FD, Falconer D, Roberts SA, Hawkhead J, Humpherson PG, et al. Identification of viable embryos in IVF by non-invasive measurement of amino acid turnover. Hum Reprod. 2004;19:2319–24.CrossRefPubMed Brison DR, Houghton FD, Falconer D, Roberts SA, Hawkhead J, Humpherson PG, et al. Identification of viable embryos in IVF by non-invasive measurement of amino acid turnover. Hum Reprod. 2004;19:2319–24.CrossRefPubMed
11.
go back to reference Seli E, Sakkas D, Scott R, Kwok SC, Rosendahl SM, Burns DH. Non-invasive metabolomic profiling of embryo culture media using Raman and near-infrared spectroscopy correlates with reproductive potential of embryos in women undergoing in vitro fertilization. Fertil Steril. 2007;88:1350–7.CrossRefPubMed Seli E, Sakkas D, Scott R, Kwok SC, Rosendahl SM, Burns DH. Non-invasive metabolomic profiling of embryo culture media using Raman and near-infrared spectroscopy correlates with reproductive potential of embryos in women undergoing in vitro fertilization. Fertil Steril. 2007;88:1350–7.CrossRefPubMed
12.
go back to reference Poli M, Ori A, Child T, Jaroudi S, Spath K, Beck M, et al. Characterization and quantification of proteins secreted by single human embryos prior to implantation. EMBO Mol Med. 2015;7(11):1465–79.CrossRefPubMedPubMedCentral Poli M, Ori A, Child T, Jaroudi S, Spath K, Beck M, et al. Characterization and quantification of proteins secreted by single human embryos prior to implantation. EMBO Mol Med. 2015;7(11):1465–79.CrossRefPubMedPubMedCentral
13.
go back to reference Greening DW, Nguyen HP, Evans J, Simpson RJ, Salamonsen LA. Modulating the endometrial epithelial proteome and secretome in preparation for pregnancy: the role of ovarian steroid and pregnancy hormones. J Proteome. 2016;144:99–112.CrossRef Greening DW, Nguyen HP, Evans J, Simpson RJ, Salamonsen LA. Modulating the endometrial epithelial proteome and secretome in preparation for pregnancy: the role of ovarian steroid and pregnancy hormones. J Proteome. 2016;144:99–112.CrossRef
14.
go back to reference Dominguez F, Meseguer M, Aparicio-Ruiz B, Piqueras P, Quiñonero A, Simón C. New strategy for diagnosing embryo implantation potential by combining proteomics and time-lapse technologies. Fertil Steril. 2015;104(4):908–14.CrossRefPubMed Dominguez F, Meseguer M, Aparicio-Ruiz B, Piqueras P, Quiñonero A, Simón C. New strategy for diagnosing embryo implantation potential by combining proteomics and time-lapse technologies. Fertil Steril. 2015;104(4):908–14.CrossRefPubMed
15.
go back to reference Li X, Xu Y, Fu J, Zhang WB, Liu SY, Sun XX. Non-invasive metabolomic profiling of embryo culture media and morphology grading to predict implantation outcome in frozen-thawed embryo transfer cycles. J Assist Reprod Genet. 2015;32(11):1597–605.CrossRefPubMedPubMedCentral Li X, Xu Y, Fu J, Zhang WB, Liu SY, Sun XX. Non-invasive metabolomic profiling of embryo culture media and morphology grading to predict implantation outcome in frozen-thawed embryo transfer cycles. J Assist Reprod Genet. 2015;32(11):1597–605.CrossRefPubMedPubMedCentral
16.
go back to reference Cortezzi SS, Garcia JS, Ferreira CR, Braga DP, Figueira RC, Iaconelli A Jr, et al. Secretome of the preimplantation human embryo by bottom-up label-free proteomics. Anal Bioanal Chem. 2011;401(4):1331–9.CrossRefPubMed Cortezzi SS, Garcia JS, Ferreira CR, Braga DP, Figueira RC, Iaconelli A Jr, et al. Secretome of the preimplantation human embryo by bottom-up label-free proteomics. Anal Bioanal Chem. 2011;401(4):1331–9.CrossRefPubMed
17.
go back to reference Cortezzi SS, Cabral EC, Trevisan MG, Ferreira CR, Setti AS, Braga DP, Figueira Rde C, Iaconelli A Jr, Eberlin MN, Borges E Jr. Prediction of embryo implantation potential by mass spectrometry fingerprinting of the culture medium. Reproduction 2013;145(5):453–462. Cortezzi SS, Cabral EC, Trevisan MG, Ferreira CR, Setti AS, Braga DP, Figueira Rde C, Iaconelli A Jr, Eberlin MN, Borges E Jr. Prediction of embryo implantation potential by mass spectrometry fingerprinting of the culture medium. Reproduction 2013;145(5):453–462.
18.
go back to reference Butler SA, Luttoo J, Freire MO, Abban TK, Borrelli PT, Iles RK. Human chorionic gonadotropin (hCG) in the secretome of cultured embryos: hyperglycosylated hCG and hCG-free beta subunit are potential markers for infertility management and treatment. Reprod Sci. 2013;20(9):1038–45.CrossRefPubMed Butler SA, Luttoo J, Freire MO, Abban TK, Borrelli PT, Iles RK. Human chorionic gonadotropin (hCG) in the secretome of cultured embryos: hyperglycosylated hCG and hCG-free beta subunit are potential markers for infertility management and treatment. Reprod Sci. 2013;20(9):1038–45.CrossRefPubMed
19.
go back to reference Iles RK, Shahpari ME, Cuckle H, Butler SA. Direct and rapid mass spectral fingerprinting of maternal urine for the detection of down syndrome pregnancy. Clin Proteomics. 2015;(12, 1):9. Iles RK, Shahpari ME, Cuckle H, Butler SA. Direct and rapid mass spectral fingerprinting of maternal urine for the detection of down syndrome pregnancy. Clin Proteomics. 2015;(12, 1):9.
20.
go back to reference Sharara FI, Abdo GA. Incidence of monozygotic twins in blastocyst and cleavage stage assisted reproductive technology cycles. Fertil Steril. 2010;93:642–5.CrossRefPubMed Sharara FI, Abdo GA. Incidence of monozygotic twins in blastocyst and cleavage stage assisted reproductive technology cycles. Fertil Steril. 2010;93:642–5.CrossRefPubMed
21.
go back to reference R Development Core Team (2008). R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. ISBN 3–900051–07-0, http://www.R-project.org. R Development Core Team (2008). R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. ISBN 3–900051–07-0, http://​www.​R-project.​org.
23.
go back to reference Rodríguez-Sánchez B, Alcalá L, Marín M, Ruiz A, Alonso E, Bouza E. Evaluation of MALDI-TOF MS (matrix-assisted laser desorption-ionization time-of-flight mass spectrometry) for routine identification of anaerobic bacteria. Anaerobe. 2016;42:101–7.CrossRefPubMed Rodríguez-Sánchez B, Alcalá L, Marín M, Ruiz A, Alonso E, Bouza E. Evaluation of MALDI-TOF MS (matrix-assisted laser desorption-ionization time-of-flight mass spectrometry) for routine identification of anaerobic bacteria. Anaerobe. 2016;42:101–7.CrossRefPubMed
24.
go back to reference Coombes KR, Koomen JM, Baggerly KA, Morris JS, Kobayashi R. Understanding the characteristics of mass spectrometry data through the use of simulation. Cancer Inform. 2005;1:41–52.CrossRefPubMed Coombes KR, Koomen JM, Baggerly KA, Morris JS, Kobayashi R. Understanding the characteristics of mass spectrometry data through the use of simulation. Cancer Inform. 2005;1:41–52.CrossRefPubMed
25.
go back to reference Salamonsen LA, Edgell T, Rombauts LJ, Stephens AN, Robertson DM, Rainczuk A, et al. Proteomics of the human endometrium and uterine fluid: a pathway to biomarker discovery. Fertil Steril. 2013;99(4):1086–92.CrossRefPubMed Salamonsen LA, Edgell T, Rombauts LJ, Stephens AN, Robertson DM, Rainczuk A, et al. Proteomics of the human endometrium and uterine fluid: a pathway to biomarker discovery. Fertil Steril. 2013;99(4):1086–92.CrossRefPubMed
Metadata
Title
Secretome profile selection of optimal IVF embryos by matrix-assisted laser desorption ionization time-of-flight mass spectrometry
Authors
Ray K. Iles
Fady I. Sharara
Raminta Zmuidinaite
Galal Abdo
Sholeh Keshavarz
Stephen A. Butler
Publication date
01-06-2019
Publisher
Springer US
Keyword
Laser
Published in
Journal of Assisted Reproduction and Genetics / Issue 6/2019
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI
https://doi.org/10.1007/s10815-019-01444-7

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