Top

Published in:

Open Access 01-12-2014 | Research

The transcriptome of corona radiata cells from individual MІІ oocytes that after ICSI developed to embryos selected for transfer: PCOS women compared to healthy women

Authors: Marie Louise Wissing, Si Brask Sonne, David Westergaard, Kho do Nguyen, Kirstine Belling, Thomas Høst, Anne Lis Mikkelsen

Published in: Journal of Ovarian Research | Issue 1/2014

Abstract

Background

Corona radiata cells (CRCs) refer to the fraction of cumulus cells just adjacent to the oocyte. The CRCs are closely connected to the oocyte throughout maturation and their gene expression profiles might reflect oocyte quality. Polycystic ovary syndrome (PCOS) is a common cause of infertility. It is controversial whether PCOS associate with diminished oocyte quality. The purpose of this study was to compare individual human CRC samples between PCOS patients and controls.

Methods

All patients were stimulated by the long gonadotropin-releasing hormone (GnRH) agonist protocol. The CRC samples originated from individual oocytes developing into embryos selected for transfer. CRCs were isolated in a two-step denudation procedure, separating outer cumulus cells from the inner CRCs. Extracted RNA was amplified and transcriptome profiling was performed with Human Agilent® arrays.

Results

The transcriptomes of CRCs showed no individual genes with significant differential expression between PCOS and controls, but gene set enrichment analysis identified several cell cycle- and DNA replication pathways overexpressed in PCOS CRCs (FDR < 0.05). Five of the genes contributing to the up-regulated cell cycle pathways in the PCOS CRCs were selected for qRT-PCR validation in ten PCOS and ten control CRC samples. qRT-PCR confirmed significant up-regulation in PCOS CRCs of cell cycle progression genes HIST1H4C (FC = 2.7), UBE2C (FC = 2.6) and cell cycle related transcription factor E2F4 (FC = 2.5).

Conclusion

The overexpression of cell cycle-related genes and cell cycle pathways in PCOS CRCs could indicate a disturbed or delayed final maturation and differentiation of the CRCs in response to the human chorionic gonadotropin (hCG) surge. However, this had no effect on the in vitro development of the corresponding embryos. Future studies are needed to clarify whether the up-regulated cell cycle pathways in PCOS CRCs have any clinical implications.

Available only for authorised users

Rotterdam ESHRE/ASRM-Sponsored PCOS Consensus Workshop Group: Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome Fertil Steril 2004, 81: 19–25.

Desforges-Bullet V, Gallo C, Lefebvre C, Pigny P, Dewailly D, Catteau-Jonard S: Increased anti-Müllerian hormone and decreased FSH levels in follicular fluid obtained in women with polycystic ovaries at the time of follicle puncture for in vitro fertilization. Fertil Steril 2010, 94: 198–204. 10.1016/j.fertnstert.2009.03.004CrossRefPubMed

Stubbs SA, Stark J, Dilworth SM, Franks S, Hardy K: Abnormal preantral folliculogenesis in polycystic ovaries is associated with increased granulosa cell division. J Clin Endocrinol Metab 2007, 92: 4418–4426. 10.1210/jc.2007-0729CrossRefPubMed

Wood JR, Dumesic DA, Abbott DH, Strauss JF: Molecular abnormalities in oocytes from women with polycystic ovary syndrome revealed by microarray analysis. J Clin Endocrinol Metab 2007, 92: 705–713. 10.1210/jc.2006-2123CrossRefPubMed

Haouzi D, Assou S, Monzo C, Vincens C, Dechaud H, Hamamah S: Altered gene expression profile in cumulus cells of mature MII oocytes from patients with polycystic ovary syndrome. Hum Reprod 2012, 27: 3523–3530. 10.1093/humrep/des325CrossRefPubMed

Kenigsberg S, Bentov Y, Chalifa-Caspi V, Potashnik G, Ofir R, Birk OS: Gene expression microarray profiles of cumulus cells in lean and overweight-obese polycystic ovary syndrome patients. Mol Hum Reprod 2009, 15: 89–103. 10.1093/molehr/gan082CrossRefPubMed

Polzikov M, Yakovenko S, Voznesenskaya J, Troshina M, Zatsepina O: Overexpression of ribosomal RNA in cumulus cells of patients with polycystic ovary syndrome. J Assist Reprod Genet 2012, 29: 1141–1145. 10.1007/s10815-012-9827-6PubMedCentralCrossRefPubMed

Heijnen EMEW, Eijkemans MJC, Hughes EG, Laven JSE, Macklon NS, Fauser BCJM: A meta-analysis of outcomes of conventional IVF in women with polycystic ovary syndrome. Hum Reprod Update 2006, 12: 13–21. 10.1093/humupd/dmi036CrossRefPubMed

Albertini DF, Combelles CM, Benecchi E, Carabatsos MJ: Cellular basis for paracrine regulation of ovarian follicle development. Reproduction (Cambridge, England) 2001, 121: 647–653. 10.1530/rep.0.1210647CrossRef

10.

Vermeulen A, Verdonck L, Kaufman JM: A critical evaluation of simple methods for the estimation of free testosterone in serum. J Clin Endocrinol Metab 1999, 84: 3666–3672. 10.1210/jcem.84.10.6079CrossRefPubMed

11.

Alpha Scientists in Reproductive Medicine and ESHRE Special Interest Group of Embryology: The Istanbul consensus workshop on embryo assessment: proceedings of an expert meeting Hum Reprod 2011, 26: 1270–1283. 10.1093/humrep/der037CrossRef

12.

Gardner DK, Schoolcraft WB: Culture and transfer of human blastocysts. Curr Opin Obstet Gynecol 1999, 11: 307–311. 10.1097/00001703-199906000-00013CrossRefPubMed

13.

Teixeira Filho FL, Baracat EC, Lee TH, Suh CS, Matsui M, Chang RJ, Shimasaki S, Erickson GF: Aberrant expression of growth differentiation factor-9 in oocytes of women with polycystic ovary syndrome. J Clin Endocrinol Metab 2002, 87: 1337–1344. 10.1210/jcem.87.3.8316CrossRefPubMed

14.

Kauffmann A, Gentleman R, Huber W: ArrayQualityMetrics--a bioconductor package for quality assessment of microarray data. Bioinformatics (Oxford, England) 2009, 25: 415–416. 10.1093/bioinformatics/btn647CrossRef

15.

Bolstad BM, Irizarry RA, Astrand M, Speed TP: A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics (Oxford, England) 2003, 19: 185–193. 10.1093/bioinformatics/19.2.185CrossRef

16.

Smyth GK: Linear models and empirical bayes methods for assessing differential expression in microarray experiments. Stat Appl Genet Mol Biol 2004, 3: Article3.PubMed

17.

Gentleman RC, Carey VJ, Bates DM, Bolstad B, Dettling M, Dudoit S, Ellis B, Gautier L, Ge Y, Gentry J, Hornik K, Hothorn T, Huber W, Iacus S, Irizarry R, Leisch F, Li C, Maechler M, Rossini AJ, Sawitzki G, Smith C, Smyth G, Tierney L, Yang JYH, Zhang J: Bioconductor: open software development for computational biology and bioinformatics. Genome Biol 2004, 5: R80. 10.1186/gb-2004-5-10-r80PubMedCentralCrossRefPubMed

18.

Smyth GK, Speed T: Normalization of cDNA microarray data. Methods (San Diego, Calif) 2003, 31: 265–273. 10.1016/S1046-2023(03)00155-5CrossRef

19.

Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA, Paulovich A, Pomeroy SL, Golub TR, Lander ES, Mesirov JP: Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci U S A 2005, 102: 15545–15550. 10.1073/pnas.0506580102PubMedCentralCrossRefPubMed

20.

Mootha VK, Lindgren CM, Eriksson K-F, Subramanian A, Sihag S, Lehar J, Puigserver P, Carlsson E, Ridderstråle M, Laurila E, Houstis N, Daly MJ, Patterson N, Mesirov JP, Golub TR, Tamayo P, Spiegelman B, Lander ES, Hirschhorn JN, Altshuler D, Groop LC: PGC-1alpha-responsive genes involved in oxidative phosphorylation are coordinately downregulated in human diabetes. Nat Genet 2003, 34: 267–273. 10.1038/ng1180CrossRefPubMed

21.

Kanehisa M, Goto S, Sato Y, Furumichi M, Tanabe M: KEGG for integration and interpretation of large-scale molecular data sets. Nucleic Acids Res 2012,40(Database issue):D109-D114. 10.1093/nar/gkr988PubMedCentralCrossRefPubMed

22.

Matthews L, Gopinath G, Gillespie M, Caudy M, Croft D, de Bono B, Garapati P, Hemish J, Hermjakob H, Jassal B, Kanapin A, Lewis S, Mahajan S, May B, Schmidt E, Vastrik I, Wu G, Birney E, Stein L, D’Eustachio P: Reactome knowledgebase of human biological pathways and processes. Nucleic Acids Res 2009,37(Database issue):D619-D622. 10.1093/nar/gkn863PubMedCentralCrossRefPubMed

23.

Kruschke JK: Bayesian estimation supersedes the t test. J Exp Psychol Gen 2013, 142: 573–603. 10.1037/a0029146CrossRefPubMed

24.

Anderson RA, Sciorio R, Kinnell H, Bayne RAL, Thong KJ, de Sousa PA, Pickering S: Cumulus gene expression as a predictor of human oocyte fertilisation, embryo development and competence to establish a pregnancy. Reproduction (Cambridge, England) 2009, 138: 629–637. 10.1530/REP-09-0144CrossRef

25.

Adriaenssens T, Segers I, Wathlet S, Smitz J: The cumulus cell gene expression profile of oocytes with different nuclear maturity and potential for blastocyst formation. J Assist Reprod Genet 2011, 28: 31–40. 10.1007/s10815-010-9481-9PubMedCentralCrossRefPubMed

26.

Feuerstein P, Puard V, Chevalier C, Teusan R, Cadoret V, Guerif F, Houlgatte R, Royere D: Genomic assessment of human cumulus cell marker genes as predictors of oocyte developmental competence: impact of various experimental factors. PLoS One 2012, 7: e40449. 10.1371/journal.pone.0040449PubMedCentralCrossRefPubMed

27.

Wathlet S, Adriaenssens T, Segers I, Verheyen G, Janssens R, Coucke W, Devroey P, Smitz J: New candidate genes to predict pregnancy outcome in single embryo transfer cycles when using cumulus cell gene expression. Fertil Steril 2012,98(2):432–439. 10.1016/j.fertnstert.2012.05.007CrossRefPubMed

28.

Ekart J, McNatty K, Hutton J, Pitman J: Ranking and selection of MII oocytes in human ICSI cycles using gene expression levels from associated cumulus cells. Hum Reprod 2013, 28: 2930–2942. 10.1093/humrep/det357CrossRefPubMed

29.

Wathlet S, Adriaenssens T, Segers I, Verheyen G, Van Landuyt L, Coucke W, Devroey P, Smitz J: Pregnancy prediction in single embryo transfer cycles after ICSI using QPCR: validation in oocytes from the same cohort. PLoS One 2013, 8: e54226. 10.1371/journal.pone.0054226PubMedCentralCrossRefPubMed

30.

May-Panloup P, Ferré-L’Hôtellier V, Morinière C, Marcaillou C, Lemerle S, Malinge M-C, Coutolleau A, Lucas N, Reynier P, Descamps P, Guardiola P: Molecular characterization of corona radiata cells from patients with diminished ovarian reserve using microarray and microfluidic-based gene expression profiling. Hum Reprod 2012, 27: 829–843. 10.1093/humrep/der431CrossRefPubMed

31.

Fragouli E, Wells D, Iager AE, Kayisli UA, Patrizio P: Alteration of gene expression in human cumulus cells as a potential indicator of oocyte aneuploidy. Hum Reprod 2012, 27: 2559–2568. 10.1093/humrep/des170CrossRefPubMed

32.

Ouandaogo ZG, Haouzi D, Assou S, Dechaud H, Kadoch IJ, De Vos J, Hamamah S: Human cumulus cells molecular signature in relation to oocyte nuclear maturity stage. PLoS One 2011, 6: e27179. 10.1371/journal.pone.0027179PubMedCentralCrossRefPubMed

33.

Van Montfoort APA, Dumoulin JCM, Kester ADM, Evers JLH: Early cleavage is a valuable addition to existing embryo selection parameters: a study using single embryo transfers. Hum Reprod 2004, 19: 2103–2108. 10.1093/humrep/deh385CrossRefPubMed

34.

Gebhardt KM, Feil DK, Dunning KR, Lane M, Russell DL: Human cumulus cell gene expression as a biomarker of pregnancy outcome after single embryo transfer. Fertil Steril 2011, 96: 47–52. e2 10.1016/j.fertnstert.2011.04.033CrossRefPubMed

35.

Assou S, Haouzi D, De Vos J, Hamamah S: Human cumulus cells as biomarkers for embryo and pregnancy outcomes. Mol Hum Reprod 2010, 16: 531–538. 10.1093/molehr/gaq032CrossRefPubMed

36.

Iager AE, Kocabas AM, Otu HH, Ruppel P, Langerveld A, Schnarr P, Suarez M, Jarrett JC, Conaghan J, Rosa GJM, Fernández E, Rawlins RG, Cibelli JB, Crosby JA: Identification of a novel gene set in human cumulus cells predictive of an oocyte’s pregnancy potential. Fertil Steril 2013, 99: 745–752. e6 10.1016/j.fertnstert.2012.10.041CrossRefPubMed

37.

Wissing ML, Kristensen SG, Andersen CY, Mikkelsen AL, Høst T, Borup R, Grøndahl ML: Identification of new ovulation-related genes in humans by comparing the transcriptome of granulosa cells before and after ovulation triggering in the same controlled ovarian stimulation cycle. Hum Reprod 2014, 29: 997–1010. 10.1093/humrep/deu008CrossRefPubMed

38.

Agca C, Yakan A, Agca Y: Estrus synchronization and ovarian hyper-stimulation treatments have negligible effects on cumulus oocyte complex gene expression whereas induction of ovulation causes major expression changes. Mol Reprod Dev 2012, 80: 102–117. 10.1002/mrd.22141CrossRef

39.

Devjak R, Fon Tacer K, Juvan P, Virant Klun I, Rozman D, Vrtačnik Bokal E: Cumulus cells gene expression profiling in terms of oocyte maturity in controlled ovarian hyperstimulation using GnRH agonist or GnRH antagonist. PLoS One 2012, 7: e47106. 10.1371/journal.pone.0047106PubMedCentralCrossRefPubMed

40.

Harshman SW, Young NL, Parthun MR, Freitas MA: H1 histones: current perspectives and challenges. Nucleic Acids Res 2013, 41: 9593–9609. 10.1093/nar/gkt700PubMedCentralCrossRefPubMed

41.

Plumb M, Marashi F, Green L, Zimmerman A, Zimmerman S, Stein J, Stein G: Cell cycle regulation of human histone H1 mRNA. Proc Natl Acad Sci U S A 1984, 81: 434–438. 10.1073/pnas.81.2.434PubMedCentralCrossRefPubMed

42.

Hao Z, Zhang H, Cowell J: Ubiquitin-conjugating enzyme UBE2C: molecular biology, role in tumorigenesis, and potential as a biomarker. Tumour Biol 2012, 33: 723–730. 10.1007/s13277-011-0291-1CrossRefPubMed

43.

Lee B-K, Bhinge AA, Iyer VR: Wide-ranging functions of E2F4 in transcriptional activation and repression revealed by genome-wide analysis. Nucleic Acids Res 2011, 39: 3558–3573. 10.1093/nar/gkq1313PubMedCentralCrossRefPubMed

Title: The transcriptome of corona radiata cells from individual MІІ oocytes that after ICSI developed to embryos selected for transfer: PCOS women compared to healthy women
Authors: Marie Louise Wissing
Si Brask Sonne
David Westergaard
Kho do Nguyen
Kirstine Belling
Thomas Høst
Anne Lis Mikkelsen
Publication date: 01-12-2014
Publisher: BioMed Central
Published in: Journal of Ovarian Research / Issue 1/2014
Electronic ISSN: 1757-2215
DOI: https://doi.org/10.1186/s13048-014-0110-6

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

The transcriptome of corona radiata cells from individual MІІ oocytes that after ICSI developed to embryos selected for transfer: PCOS women compared to healthy women

Abstract

Background

Methods

Results

Conclusion

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Background

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2014

Outcomes of ovarian transposition in gynaecological cancers; a systematic review and meta-analysis

Role of nerve growth factor and its TRKA receptor in normal ovarian and epithelial ovarian cancer angiogenesis

Brachial-to-ankle pulse wave velocity as an independent prognostic factor for ovulatory response to clomiphene citrate in women with polycystic ovary syndrome

Anti Müllerian Hormone (AMH) level and expression in mural and cumulus cells in relation to age

The expression of the Sprouty 1 protein inversely correlates with growth, proliferation, migration and invasion of ovarian cancer cells

Gemcitabine as a molecular targeting agent that blocks the Akt cascade in platinum-resistant ovarian cancer