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Cumulus cell pappalysin-1, luteinizing hormone/choriogonadotropin receptor, amphiregulin and hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 mRNA levels associate with oocyte developmental competence and embryo outcomes

  • Richard J. Kordus
  • Akhtar Hossain
  • Michael C. Corso
  • Hrishikesh Chakraborty
  • Gail F. Whitman-Elia
  • Holly A. LaVoieEmail author
Assisted Reproduction Technologies

Abstract

Purpose

To determine whether a selected set of mRNA biomarkers expressed in individual cumulus granulosa cell (CC) masses show association with oocyte developmental competence, embryo ploidy status, and embryo outcomes.

Methods

This prospective observational cohort pilot study assessed levels of mRNA biomarkers in 163 individual CC samples from 15 women stimulated in antagonist cycles. Nineteen mRNA biomarker levels were measured by real-time PCR and related to the development of their corresponding individually cultured oocytes and subsequent embryos, embryo ploidy status, and live birth outcomes.

Results

PAPPA mRNA levels were significantly higher in CC from oocytes that led to euploid embryos resulting in live births and aneuploid embryos compared to immature oocytes by ANOVA. LHCGR mRNA levels were significantly higher in CC of oocytes resulting in embryos associated with live birth compared to immature oocytes and oocytes resulting in arrested embryos by ANOVA. Using a general linearized mixed model to assess ploidy status, CC HSD3B mRNA levels in oocytes producing euploid embryos were significantly lower than other oocyte outcomes, collectively. When transferred euploid embryos outcomes were analyzed by ANOVA, AREG mRNA levels were significantly lower and PAPPA mRNA levels significantly higher in CC from oocytes that produced live births compared to transferred embryos that did not form a pregnancy.

Conclusions

Collectively, PAPPA, LHCGR, and AREG mRNA levels in CC may be able to identify oocytes with the best odds of resulting in a live birth, and HSD3B1 mRNA levels may be able to identify oocytes capable of producing euploid embryos.

Keywords

Cumulus cells Real-time PCR mRNA levels Oocyte developmental competence Euploid embryo 

Notes

Funding

This study was supported by an ASPIRE-I grant from the University of South Carolina. MCC was supported by the University of South Carolina School of Medicine Research Program for Medical Students.

Compliance with ethical standards

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and in its later amendments or comparable ethical standards. This study was approved by the University of South Carolina Institution Review Board (IRB registration number: 00000240).

Informed consent

Informed consent was obtained from all individual participants included in the study.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10815_2019_1489_Fig6_ESM.png (733 kb)
Supplemental Fig. 1

Biomarkers for CC mRNA expression not associated with mature oocyte competence and embryo outcomes. To determine the differences in CC mRNA between groups where oocytes had different developmental and embryo outcomes target mRNA levels were compared using repeated measures ANOVA followed by Tukey’s post hoc test (adjusted P-values) for pairwise comparisons. Groups represent CC mRNA from oocytes with the following descriptions: Aneuploid = mature oocytes resulting in aneuploid embryos; Arrested = mature oocytes resulting in embryos that did not reach the blastocyst stage; Failed Fert = mature oocytes that did not fertilize; Immature = immature oocytes that were not fertilized; Live Birth = oocytes that resulted in transferred euploid embryos that resulted in live births; No Pregnancy = oocytes that resulted in transferred euploid embryos that did not result in a pregnancy. No differences were seen in these biomarkers (P > 0.05). Data are presented as stated in fig. 3 legend (PNG 732 kb)

10815_2019_1489_MOESM1_ESM.tif (911 kb)
High resolution image (TIF 911 kb)
10815_2019_1489_Fig7_ESM.png (262 kb)
Supplemental Fig. 2

GLMM model distinguishing CC from oocytes giving rise to live births and oocytes with outcomes not resulting in live birth, collectively. To evaluate the potential of CC biomarker mRNA levels to identify oocytes capable of producing euploid embryos resulting in a live birth versus groups not producing live births, the best fitting model included 110 CC samples from 14 patients and included biomarkers: CYP11A1, CYP19A1, IGFBP5, PAPPA, PGRMC1, and STARD1. Increased PAPPA mRNA expression (P < 0.05) significantly increased the odds of an oocytes producing an embryo resulting in a live birth (OR = 4.591, 95% CI: 1.098, to 19.201). All previously stated categories were included in this model except oocytes yielding euploid embryos that were not transferred. Groups represent CC mRNA from oocytes with the following descriptions: Live Birth = mature oocytes that resulted in euploid embryos that lead to live births; Non-Viable = immature oocytes, mature oocytes resulting in aneuploid blastocysts, oocytes producing embryos that did not reach the blastocyst stage, and mature oocytes that failed to fertilize, collectively. Data are presented as in fig. 3 legend (PNG 262 kb)

10815_2019_1489_MOESM2_ESM.tif (428 kb)
High resolution image (TIF 428 kb)
10815_2019_1489_MOESM3_ESM.docx (33 kb)
ESM 1 (DOCX 32 kb)

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Authors and Affiliations

  1. 1.Department of Cell Biology and AnatomyUniversity of South Carolina School of MedicineColumbiaUSA
  2. 2.Fertility Center of the Carolinas, Department of Obstetrics and GynecologyPrisma Health – UpstateGreenvilleUSA
  3. 3.Department of Epidemiology and BiostatisticsUniversity of South CarolinaColumbiaUSA
  4. 4.Duke Clinical Research InstituteDuke UniversityDurhamUSA
  5. 5.Advanced Fertility and Reproductive Endocrinology Institute, LLCColumbiaUSA
  6. 6.Piedmont Reproductive Endocrinology GroupColumbiaUSA

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