Dataset Information

Do extracellular vesicles have specific target cells? Extracellular Vesicle mediated embryo maternal communication

ABSTRACT: Extracellular vesicles (EVs) have emerged as key messengers in intercellular communications. However, the precise mechanisms of how recipient cells read EV messages are not clear. In this study, we investigated the roles of EV protein cargo, EV parent cell type, and recipient cell type in EV function using a human embryo implantation model. The JAr cell line served as an analog of human trophoblast cells, and EVs isolated from these cells were introduced into receptive endometrial epithelial cell analog RL95-2 cells to simulate EV-mediated embryo-maternal communication. Downregulation of Zinc-finger protein 81 (ZNF81) gene expression in RL95-2 cells in response to trophoblast EVs were used as a readout of JAr EV functionality. EVs from the non-trophoblastic cell line (HEK-293 cells) were used as a control to assess the specificity of trophoblast EV functionality in endometrial epithelial cells. Our results demonstrated that ZNF81 gene downregulation is a specific response of RL95-2 cells to JAr EVs, as HEK-293 cell EVs did not induce any gene expression changes. Furthermore, we conducted an analysis of the proteomic cargo profiles of JAr and HEK-293 cell EVs. Interestingly, JAr EVs exhibited a unique proteomic cargo profile compared to that of HEK EVs. Some of the proteins uniquely identified in JAr EVs are also known to play roles in embryo implantation. Moreover, JAr EVs were specifically enriched with important transcription factors that regulate ZNF81 gene expression. These findings contribute to our understanding of the specificity of EV functionality and cellular response in the context of embryo implantation.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: SUBHASHINI MUHANDIRAM MUHANDIRAM RALALAGE

LAB HEAD: Alireza Fazeli

PROVIDER: PXD050024 | Pride | 2024-07-03

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	211208_06_Orbi3_MS_SER_G60_T85_Keerthie_HEK_1_1ug_iRT_50pg.raw	Raw
	211208_08_Orbi3_MS_SER_G60_T85_Keerthie_JAR_2_1_1ug_iRT_50pg.raw	Raw
	211208_10_Orbi3_MS_SER_G60_T85_Keerthie_HEK_2_1ug_iRT_50pg.raw	Raw
	211208_12_Orbi3_MS_SER_G60_T85_Keerthie_JAR_3_1ug_iRT_50pg.raw	Raw
	211208_14_Orbi3_MS_SER_G60_T85_Keerthie_HEK_3_1ug_iRT_50pg.raw	Raw

Items per page:

1 - 5 of 8

Similar Datasets

Project description:Study question: Could extracellular vesicles (EVs) secreted by aneuploid embryos a) serve for development of RNA biomarkers for preimplantation genetic testing for aneuploidy (PGT-A) and b) elicit a relevant transcriptomic response in decidualised primary endometrial stromal cells (dESCs)? Summary answer: Aneuploid embryo EVs a) contain genes PPM1J, LINC00561, ANKRD34C and TMED10 in differential abundance from euploid EVs and b) induce up-regulation of MUC1 transcript in dESCs. What is known already: PGT-A is popular as it is thought to facilitate transfer of euploid embryos to increase implantation probability but the technology is controversial, as it requires an invasive embryo biopsy with an elusive long-term biosafety. The development of non-invasive methods to screen out aneuploid embryos is paramount. It is also critical to decode the embryo-endometrial dialog underlying implantation failure. We have previously reported that IVF embryos secrete EVs that can be internalised by ESCs, conceptualising that successful implantation to the endometrium is facilitated by EVs, which may additionally serve as biomarkers of ploidy status. Study design, size, duration: Embryos destined for biopsy on days 5-7 for PGT-A were grown under standard conditions. Spent media (30μl) were collected from euploid (n=175) and aneuploid (n=140)embryos at cleavage (days 1-3) stage and from euploid (n=187) and aneuploid (n=142) embryos at blastocyst (days 3-5) stage. Media samples from n=35 cleavage embryos were pooled in order to obtain five euploid and four aneuploid pools. Similarly, media samples from blastocyst were pooled to create one euploid and one aneuploid pool. ESCs were obtained from five women undergoing diagnostic laparoscopy. Participants/materials, setting, methods: EVs were isolated from pools of media derived from euploid and aneuploid day 3 embryos with differential centrifugation and EV-RNA sequencing was performed following the SMARTer Stranded Total RNA-Seq approach. ESCs were decidualised (E2:10nM, P4:1uM, cAMP:0.5 mM twice every 48 hours) and treated for 24 hours with 50 ng/ml euploid or aneuploid EVs extracted from blastocyst media. RNA sequencing was performed on ESCs following the Illumina Truseq RNAseq protocol. Main results and the role of chance: Aneuploid cleavage stage embryos secreted EVs that were less abundant in RNA fragments originating from the genes PPM1J (log2fc=-5.13, p=0.011), LINC00561 (log2fc=-7.87, p=0.010) and ANKRD34C (log2fc=-7.30, p=0.017) and more abundant in TMED10 (log2fc=1.63 p=0.025) compared to EVs from euploid embryos. Decidualisation per se induced downregulation of MUC1 (log2FC=-0.54, p=0.0028) in ESCs as prerequisite for the establishment of receptive endometrium. The expression of MUC1 transcript in decidualised ESCs was significantly increased following treatment with aneuploid compared to euploid embryo-secreted EVs (log2FC=0.85, p=0.0201). Limitations, reasons for caution: The findings of the study may require validation utilising a second cohort of EVs samples. Wider implications of the findings: The discovery that the RNA cargo of EVs secreted from aneuploid cleavage stage embryos is diverse from that of euploid embryos potentiates the development of non-invasive methodology for PGT-A. The upregulation of MUC1 in dESCs following aneuploid embryo EV treatment proposes a new mechanism underlying implantation failure. Funding: The study was supported by a MSCA fellowship awarded to SM by the European Commission (CERVINO grant agreement ID: 79620) and by a BIRTH research grant from Theramex HQ UK Ltd

Dataset Information

Do extracellular vesicles have specific target cells? Extracellular Vesicle mediated embryo maternal communication

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets