Expressed proteins and activated pathways in conditioned embryo culture media from IVF patients are diverse according to infertility factors

Abstract

Objective:Given that the embryo culture medium secretome reflects the embryo development, we hypothesize that protein profiles are affected according to infertility factors, which can be responsible for detrimental embryonic developmental competence. The aim of this study was to screen the protein profile of conditioned embryo culture media in patients presenting deep infiltrating endometriosis (ENDO) and polycystic ovarian syndrome (PCOS) undergoing IVF, by proteomics approaches. The control group was constituted by tubal factor patients.Methods:Patients underwent in vitro fertilization (IVF) treatment as routine and oocytes were fertilized by ICSI. The embryos were group cultured until day 3 of development, and after transfer the culture media were collected. For the proteomics analysis, two pools of samples were prepared for groups CONTROL and PCOS, and 4 pools of samples for group DIE. Samples were prepared to deplete high abundant proteins and followed evaluated by high throughput proteomics approach.Results:The embryonic organ and tissue development were physiological functions activated, based on proteins identified in the 3 study groups of samples. The samples coming from DIE patients presented a high calcium activity and on the other hand, embryos coming from PCOS patients showed a decreased calcium action. Other pathways as grow factors through the EGF signaling pathway overexpressed in ENDO culture medium and protein kinase A in PCOS were also observed.Conclusions:Proteomic embryonic secretome will advance our knowledge of early embryogenesis and additionally could lead to improved selection of embryos for transfer warrants further investigation.

Investigation of the mechanisms and experimental verification of Cuscuta-Salvia in the treatment of polycystic ovary syndrome (PCOS) via network pharmacology

“…PCOS mice have been rescued by regulating the JUN pathway [ 29 ]. EGF overexpression has been measured in PCOS [ 3 ]. CASP3 plays a critical role in initiating apoptosis [ 7 ].…”

MiR-191-5p is upregulated in culture media of implanted human embryo on day fifth of development

“…Even aneuploid embryos can have normal morphology, and some euploid embryos can have aberrant morphology [ 32 ]. Among the various proteins reported as possible biomarkers for embryonic development [ 31 , 33 ], none have yet proven to be good indicators of a successful implantation in the receptive endometrium.…”

Omics insights into extracellular vesicles in embryo implantation and their therapeutic utility

“…Presently, culture of human embryos requires proprietary commercial media and supplements (including key nutrients, amino acids and carbohydrates) [210] as well as serum albumin [172], which acts as a surfactant, limiting embryos/gametes sticking during the culture process, and acts as a chelator and pH buffer to protect embryos from toxicity in the culture environment [211]. However, albumin (with various co‐associated proteins/lipids observed in IVF media [212]) is a contaminant associated with EVs [86] and interferes with their isolation [53] and proteomic analyses [210, 213]. For EVs from biofluid source, often requires an enrichment step (i.e., abundant protein/lipoprotein depletion [214, 215] or microfiltration to reduce albumin [213]), followed by the isolation of EVs (e.g., size exclusion chromatography [216, 217], immunoaffinity‐based isolation [218], tangential flow filtration [219]).…”

“…However, albumin (with various co‐associated proteins/lipids observed in IVF media [212]) is a contaminant associated with EVs [86] and interferes with their isolation [53] and proteomic analyses [210, 213]. For EVs from biofluid source, often requires an enrichment step (i.e., abundant protein/lipoprotein depletion [214, 215] or microfiltration to reduce albumin [213]), followed by the isolation of EVs (e.g., size exclusion chromatography [216, 217], immunoaffinity‐based isolation [218], tangential flow filtration [219]). However, these purification workflows are not amenable to complex biological sources such as serum/protein‐rich media [220] including human embryo spent media, as they deplete lowly abundant soluble/EV‐associated proteins (i.e., embryo‐secreted factors) [213].…”