Micro-ribonucleic acids and extracellular vesicles repertoire in the spent culture media is altered in women undergoing In Vitro Fertilization

Masood Abu-Halima,Sigrun Nestel,Andreas Keller,Christina Backes,Irina Nazarenko,Claudia Staib,Tobias Fehlmann,Eckart Meese,Sebastian Häusler

doi:10.1038/s41598-017-13683-8

Masood Abu-Halima, Sigrun Nestel + Show 7 more

Open Access

https://doi.org/10.1038/s41598-017-13683-8

Copy DOI

Abstract

MicroRNAs (miRNAs) are class of small RNA molecules with major impact on gene regulation. We analyzed the potential of miRNAs secreted from pre-implantation embryos into the embryonic culture media as biomarkers to predict successful pregnancy. Using microarray analysis, we profiled the miRNome of the 56 spent culture media (SCM) after embryos transfer and found a total of 621 miRNAs in the SCM. On average, we detected 163 miRNAs in SCM of samples with failed pregnancies, but only 149 SCM miRNAs of embryos leading to pregnancies. MiR-634 predicted an embryo transfer leading to a positive pregnancy with an accuracy of 71% and a sensitivity of 85%. Among the 621 miRNAs, 102 (16.4%) showed a differential expression between positive and negative outcome of pregnancy with miR-29c-3p as the most significantly differentially expressed miRNA. The number of extracellular vehicles was lower in SCM with positive outcomes (3.8 × 109/mL EVs), as compared to a negative outcome (7.35 × 109/mL EVs) possibly explaining the reduced number of miRNAs in the SCM associated with failed pregnancies. The analysis of the miRNome in the SCM of couples undergoing fertility treatment lays the ground towards development of biomarkers to predict successful pregnancy and towards understanding the role of embryonic miRNAs found in the SCM.

Highlights

Worldwide, about 72.4 million couples suffer from subfertility
Seventeen-microliter spent culture media (SCM) from 56 females/single-embryo transfers were used for the miRNA profiling to investigate whether the miRNA(s) correlates with pregnancy outcome and 9 μL SCM from 8 females/single-embryo transfers were used for Extracellular vesicles (EV) concentration measurements as a small ‘Proof-of-Concept’ to investigate whether the EVs concentration correlates with pregnancy outcome
In line with our abovementioned finding that the SCM from embryos leading to a pregnancy have a decreased repertoire of miRNAs, seven of the eight miRNAs presented in Fig. 1b occur significantly less frequent in such samples

Summary

Introduction

About 72.4 million couples suffer from subfertility. To overcome undesired childlessness, about 40 million turns to Assisted Reproduction Techniques (ART) like In Vitro Fertilization (IVF) and/or Intracytoplasmic Sperm Injection (ICSI)[1]. The chances that a specific IVF-cycle (either after fresh stimulation/egg retrieval or after cryopreservation of two-pronuclear zygotes or embryos in earlier treatments) will induce a pregnancy depends on the fitness of the transferred embryo[3]. To reduce the average number of potentially harmful treatment cycles, new diagnostic tools are needed to identify the most capable embryo derived from the retrieved oocytes[6]. It has been shown that RNAs including miRNAs are selectively secreted from pre-implantation embryos into the embryonic culture media and may have a potential as biomarkers of embryo development[17,18,19]. We aimed to identify miRNA expression patterns (miRNomes) in the SCM following embryo transfer and to assess whether specific miRNAs may serve as biomarker for embryo quality and for successful pregnancy. We explore the number and size distribution of extracellular vesicles[21] in the SCM following embryonic transfer

Objectives

Methods

Results

Conclusion