Kouakou, F., Denizot, A. L., L'Hostis, A., Colet, J., Jacques, S., Sallem, A., Ziyyat, A., Vaiman, D., & Wolf, J. P. (2023). Plastic used in in vitro fertilization procedures induces massive placental gene expression alterations. EBioMedicine. https://doi.org/10.1016/j.ebiom.2023.104572
Kouakou F, Denizot AL, L'Hostis A, Colet J, Jacques S, Sallem A, Ziyyat A, Vaiman D, Wolf JP. Plastic used in in vitro fertilization procedures induces massive placental gene expression alterations. EBioMedicine. 2023. doi:10.1016/j.ebiom.2023.104572
Kouakou, F., et al. "Plastic used in in vitro fertilization procedures induces massive placental gene expression alterations." EBioMedicine, 2023.
For EndNote, Zotero or Mendeley:
Open access
No open license is recorded for this paper. Reuse terms are set by the publisher.
Open Access
Free full text on PubMed Central (PMC10149224)
RRM Academy Synopsis
Plastic culture dishes changed placental genes in IVF mouse embryos
In mice, embryos made by IVF in plastic dishes had 1121 altered placental genes, against 200 after glass dishes. Natural mating was the comparison. The changed genes were mostly involved in stress, inflammation and detoxification. Researchers pooled placentas from three groups of pregnant mice.
Key Findings
Placentas from embryos cultured in plastic had 1121 genes with changed activity compared with natural mating (628 up, 493 down). Glass gave 200 (111 up, 89 down).
Plastic and glass placentas differed from each other at 434 genes.
Female placentas had 1627 changed genes after plastic culture and 743 in males. After glass culture the counts were 547 in females and 133 in males.
Fetal brains showed fewer than 50 differently expressed genes in every comparison, a count the authors say is not significantly different from chance.
Implantation rates were 35.6% (plastic), 37.3% (glass) and 56.6% (natural mating), p < 0.004.
Interpretation
This laboratory study used one hybrid mouse strain and measured gene activity in placentas at day 16.5. The study shows no health outcomes in people. Microarray pools were checked by RT-qPCR on separate placentas for 22 genes. The paper reports no placental function or fetal weight data. The authors report more altered genes in female placentas than in male placentas. They call their explanation for that difference, a protective placental response, speculative, and say extending results to humans needs further investigation. They raise the possibility that the placental changes partly buffer the fetus. The brain showed no clear effect.
RRM Context
Restorative reproductive medicine works toward conception in the body. Here the naturally mated group set the reference. Plastic-cultured embryos sat furthest from it and glass-cultured embryos closer. Even in glass, embryos implanted less often (37.3% against 56.6%). The authors cite higher rates of birth defects and pregnancy disorders after ART. IVF bypasses the reason conception fails, and placental gene activity differed by culture material in this mouse model.
Our editorial summary of this paper, not the article's abstract.
Abstract
Background
The exposure to plastic derivatives during human life is deleterious. Infants conceived using ART (IVF or ICSI) have twice as many risks of major birth defects compared to naturally conceived infants. Could plastic ware used during ART trigger defects in the fetal development?
Methods
Three groups of blastocysts were transferred to pseudopregnant mice. One was obtained after IVF and embryo development in plastic ware, the second in glass ware. The third, was obtained in vivo by natural mating. On day 16.5 of pregnancy, females were sacrificed and fetal organs collected for gene expression analysis. Fetal sex was determined by RT-PCR. RNA was extracted from a pool of five placental or brain samples coming from at least two litters from the same group and analyzed by hybridisation onto the mouse Affymetrix 430.2.0 GeneChips, confirmed by RT-qPCR for 22 genes.
Findings
This study highlights a major impact of plastic ware on placental gene expression (1121 significantly deregulated genes), while glassware was much closer to in vivo offspring (only 200 significantly deregulated genes). Gene Ontology indicated that the modified placental genes were mostly involved in stress, inflammation and detoxification. A sex specific analysis revealed in addition a more drastic effect on female than male placentas. In the brains, whatever the comparison, less than 50 genes were found deregulated.
Interpretation
Embryos incubated in plastic ware resulted in pregnancy with massive alterations of placental gene expression profile in concerted biological functions. There were no obvious effects on the brains. Besides other effects, this suggests that plastic ware in ART could be a cause of the increased level of pregnancy disorders observed recurrently in ART pregnancies.
Funding
This study was funded by two grants from the Agence de la Biomedecine in 2017 and 2019.
Aouache R et al., 2018·International journal of molecular sciences·Free full text on PubMed Central
Preeclampsia is a persistent hypertensive gestational disease characterized by high blood pressure and proteinuria, which presents from the second trimester of pregnancy. At the cellular level, preeclampsia has largely been associated with the release of free radicals by the placenta. Placenta-borne oxidative and nitrosative stresses are even sometimes considered as the major molecular determinants of the maternal disease. In this review, we present the recent literature evaluating free radical production in both normal and pathological placentas (including preeclampsia and other major pregnancy diseases), in humans and animal models. We then assess the putative effects of these free radicals on the placenta and maternal endothelium. This analysis was conducted with regard to recent papers and possible therapeutic avenues.
de Waal E et al., 2015·Human molecular genetics·Free full text on PubMed Central
Assisted reproductive technologies (ART) are associated with several complications including low birth weight, abnormal placentation and increased risk for rare imprinting disorders. Indeed, experimental studies demonstrate ART procedures independent of existing infertility induce epigenetic perturbations in the embryo and extraembryonic tissues. To test the hypothesis that these epigenetic perturbations persist and result in adverse outcomes at term, we assessed placental morphology and methylation profiles in E18.5 mouse concepti generated by in vitro fertilization (IVF) in two different genetic backgrounds. We also examined embryo transfer (ET) and superovulation procedures to ascertain if they contribute to developmental and epigenetic effects. Increased placental weight and reduced fetal-to-placental weight ratio were observed in all ART groups when compared with naturally conceived controls, demonstrating that non-surgical embryo transfer alone can impact placental development. Furthermore, superovulation further induced overgrowth of the placental junctional zone. Embryo transfer and superovulation defects were limited to these morphological changes, as we did not observe any differences in epigenetic profiles. IVF placentae, however, displayed hypomethylation of imprinting control regions of select imprinted genes and a global reduction in DNA methylation levels. Although we did not detect significant differences in DNA methylation in fetal brain or liver samples, rare IVF concepti displayed very low methylation and abnormal gene expression from the normally repressed allele. Our findings suggest that individual ART procedures cumulatively increase placental morphological abnormalities and epigenetic perturbations, potentially causing adverse neonatal and long-term health outcomes in offspring.
Zhang L et al., 2022·Journal of assisted reproduction and genetics·Free full text on PubMed Central
Oocyte vitrification is an important assisted reproductive technology (ART) that preserves the fertility of unmarried patients with malignant tumors, and promotes the development of the oocyte donation program. In recent years, the effects of ART, including the vitrification of oocytes and embryos on the health of offspring, have attracted much attention; however, it is difficult to conduct long-term follow-up and biochemical evaluation in humans. In this study, we detected the effect of oocyte vitrification on gene expression in the organs of adult mice offspring by RNA sequencing for the first time. Our results showed that only a small amount of gene expression was significantly affected. Seven genes (Tpm3, Hspe1-rs1, Ntrk2, Cyp4a31, Asic5, Cyp4a14, Retsat) were abnormally expressed in the liver, and ten genes (Lbp, Hspe1-rs1, Prxl2b, Pfn3, Gm9008, Bglap3, Col8a1, Hmgcr, Ero1lb, Ifi44l) were abnormal in the kidney. Several genes were related to metabolism and disease occurrence in the liver or kidney. Besides, we paid special attention to the expression of known imprinted genes and DNA methylation-related genes in adult organs, which are susceptible to oocyte cryopreservation in the preimplantation stage. As a result, some of these transcripts were detected in adult organs, but they were not affected by oocyte vitrification. In conclusion, we first report that oocyte vitrification did not significantly change the global gene expression in offspring organs; nonetheless, it can still influence the transcription of a few functional genes. The potential adverse effects caused by oocyte vitrification need attention and further study.
Håberg SE et al., 2022·Nature communications·Free full text on PubMed Central
Assisted reproductive technology (ART) may affect fetal development through epigenetic mechanisms as the timing of ART procedures coincides with the extensive epigenetic remodeling occurring between fertilization and embryo implantation. However, it is unknown to what extent ART procedures alter the fetal epigenome. Underlying parental characteristics and subfertility may also play a role. Here we identify differences in cord blood DNA methylation, measured using the Illumina EPIC platform, between 962 ART conceived and 983 naturally conceived singleton newborns. We show that ART conceived newborns display widespread differences in DNA methylation, and overall less methylation across the genome. There were 607 genome-wide differentially methylated CpGs. We find differences in 176 known genes, including genes related to growth, neurodevelopment, and other health outcomes that have been associated with ART. Both fresh and frozen embryo transfer show DNA methylation differences. Associations persist after controlling for parents' DNA methylation, and are not explained by parental subfertility.
Market-Velker BA et al., 2010·Biology of reproduction
Assisted reproductive technologies (ARTs) are becoming increasingly prevalent and are generally considered to be safe medical procedures. However, evidence indicates that embryo culture may adversely affect the developmental potential and overall health of the embryo. One of the least studied but most important areas in this regard is the effects of embryo culture on epigenetic phenomena, and on genomic imprinting in particular, because assisted reproduction has been linked to development of the human imprinting disorders Angelman and Beckwith-Wiedemann syndromes. In this study, we performed side-by-side comparisons of five commercial embryo culture systems (KSOMaa, Global, Human Tubal Fluid, Preimplantation 1/Multiblast, and G1v5PLUS/G2v5PLUS) in relation to a best-case (in vivo-derived embryos) and a worst-case (Whitten culture) scenario. Imprinted DNA methylation and expression were examined at three well-studied loci, H19, Peg3, and Snrpn, in mouse embryos cultured from the 2-cell to the blastocyst stage. We show that embryo culture in all commercial media systems resulted in imprinted methylation loss compared to in vivo-derived embryos, although some media systems were able to maintain imprinted methylation levels more similar to those of in vivo-derived embryos in comparison to embryos cultured in Whitten medium. However, all media systems exhibited loss of imprinted H19 expression comparable to that using Whitten medium. Combined treatment of superovulation and embryo culture resulted in increased perturbation of genomic imprinting, above that from culture alone, indicating that multiple ART procedures further disrupt genomic imprinting. These results suggest that time in culture and number of ART procedures should be minimized to ensure fidelity of genomic imprinting during preimplantation development.
Lifestyle and Environment › Environmental Exposures › Plastics and Microplastics · Assisted Reproduction › Safety and Risks › Procedure-Attributable Risk
PMID 37094466 37094466 DOI 10.1016/j.ebiom.2023.104572 10.1016/j.ebiom.2023.104572 Kouakou et al. 2023, Kouakou 2023
Cite this article
Kouakou, F., Denizot, A. L., L'Hostis, A., Colet, J., Jacques, S., Sallem, A., Ziyyat, A., Vaiman, D., & Wolf, J. P. (2023). Plastic used in in vitro fertilization procedures induces massive placental gene expression alterations. EBioMedicine. https://doi.org/10.1016/j.ebiom.2023.104572
Kouakou F, Denizot AL, L'Hostis A, Colet J, Jacques S, Sallem A, Ziyyat A, Vaiman D, Wolf JP. Plastic used in in vitro fertilization procedures induces massive placental gene expression alterations. EBioMedicine. 2023. doi:10.1016/j.ebiom.2023.104572
Kouakou, F., et al. "Plastic used in in vitro fertilization procedures induces massive placental gene expression alterations." EBioMedicine, 2023.