Identifying Expression Markers of Cumulus Cells to Identify Oocyte Developmental Potential Through a Maturation Analysis

Completed

• Conditions: Infertility

• Registration Number: NCT02230449
• Lead Sponsor: Memorial Sisli Hospital, Istanbul

Brief Summary

The aim of the study is to analyze the expression of genes and microRNAs potentially involved in oocyte competence in follicles of different sizes (\<17mm and ≥17mm) at the time of oocyte pick-up.

Detailed Description

In ART, one of the major obstacles encountered in some patients is the wide variation in the follicular size at the oocyte pick-up day and consequently, the number of available mature oocytes and the number of competent embryos for implantation. The follicular size at the day of oocyte pick-up reflects the synchronicity of the individual follicular responsiveness to FSH.

For successful preimplantation development of an embryo, the oocyte must contain all necessary maternal factors. Two of these factors are specific mRNA transcripts (GDF9, PTX3, HAS2, PTGS2, TNFAIP6) and microRNA (miR-224).

For each Cumulus Oocyte Complex, the evaluation of embryo development and morphokinetics will be correlated with the expression and implantation data to identify a quantitative biomarker to predict the embryo with the highest chance of implantation in patients.

Experimental Design:

1. - Collection of Cumulus Oocyte Complex (COC) from patients

2. - Separation of oocytes according to the size of the follicule they have been retrieved from (\<17mm; ≥17mm)

3. - Isolation of mRNA from both groups of cumulus cells

4. - Expression studies, qRT-PCR (GDF9, PTX3, HAS2, PTGS2, TNFAIP6)

5. - Analysis of miR-224, a negative regulator of cumulus expansion via PTX3

6. - Time-lapse culture \& evaluation of morphokinetics

Study Timeline

• Study Start: 2014-08
• Study First Submitted: 2014-08-19
• Study First Submitted QC: 2014-09-02
• Study First Posted: 2014-09-03
• Primary Completion: 2016-06
• Study Completion: 2016-09
• Status Verified: 2017-09
• Last Update Submitted: 2017-09-01
• Last Update Posted: 2017-09-05

Recruitment & Eligibility

• Status: COMPLETED
• Sex: Female
• Target Recruitment: 200

Inclusion Criteria

• No more than 2 previous treatment cycles
• No history of recurrent spontaneous abortions
• Age ≤ 39 years
• BMI < 30 kg / m2
• ≥ 8 cumulus-oocyte-complexes (COC) retrieved
• human Chorionic Gonadotrophin (hCG) triggering

Exclusion Criteria

• Preimplantaion Genetic Diagnosis (PGD) or Preimplantation Genetics Screening (PGS) indication
• Maximum number of 24 COC during pickup
• Failure to culture until Day 5
• Polycystic ovary syndrome
• Severe endometrial factor that can influence implantation
• Uterine malformation or congenital anomaly
• Severe male factor (Azoospermia)

Study & Design

• Study Type: OBSERVATIONAL
• Study Design: Not specified

Primary Outcome Measures

Name	Time	Method
Cumulus cell RNA expression correlation with blastocyst formation potential and quality	6 days	At the day of oocyte pick-up, cumulus cells will be collected from each cumulus oocyte complex. RNA extraction will be performed from those cumulus cells and expression levels of 5 genes and 1 miRNA will be measured (compared with that of a housekeeping gene). Oocytes of those samples will be injected and cultured with routine Assisted Reproduction Techniques. At the day 5 \& 6, after the oocyte pick-up procedure, each blastocyst formed will be noted and the quality of each blastocyst will be recorded. The blastocyst grading will be done according to Gardner's blastocyst morphological criteria (degree of expansion of the blastocoele cavity: 1 to 5, inner cell mass morphology: A to C, trophectoderm morphology: A to C).; The composite primary outcome measure will allow the correlation of the expression levels of 5 genes and 1 miRNA of each cumulus oocyte complex with the corresponding embryo morphology on day 5 (presence or absence of a blastocyst) and blastocyst quality.

Secondary Outcome Measures

Name	Time	Method
Implantation Rate	2 months	Implantation of the embryo(s) transferred will be checked by ultrasound 6 weeks after the pick-up. Patients for whom the number of embryo(s) transferred match the number of sac will allow the correlation of the expression data with implantation.

Trial Locations

Locations (1)

Memorial Sisli Hospital ART Center; 🇹🇷 Istanbul, Turkey