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MiR-135a-5p regulates window of implantation by suppressing pinopodes development and decidualization of endometrial stromal cells

  • Reproductive physiology and disease
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

The window of implantation (WOI) is a brief period during which the endometrium is receptive to embryo implantation. This study investigated the relationship between miR-135a-5p and endometrial receptivity.

Methods

Peripheral blood was collected on the day of ovulation and the 5th day after ovulation for high-throughput sequencing from women who achieved clinical pregnancy through natural cycle frozen embryo transfer. RT-qPCR assessed miR-135a-5p expression in the endometrium tissue or cells during the mouse implantation window or decidualization. Scanning electron microscopy was utilized to observe pinopode morphology and quantity in mice overexpressing miR-135a-5p during the WOI. Human endometrial stromal cells (HESC) and artificial induction of mouse uterine decidualization were used to explore whether miR-135a-5p overexpression inhibits decidualization by regulating HOXA10 and BMPR2. Furthermore, the impact of miR-135a-5p on HESC proliferation and HTR8/SVneo invasion was explored.

Results

A total of 54 women were enrolled in the study. bioinformatics analysis and animal models demonstrated that miR-135a-5p was significantly downregulated during the WOI, and its high expression can lead to abnormal pregnancy outcomes. Overexpression of miR-135a-5p resulted in the absence of pinopode in mouse endometrial tissue during the WOI. High miR-135a-5p levels were found to potentially inhibit endometrial tissue decidualization by downregulating HOXA10 and BMPR2 expression. Finally, CEBPD was identified as a potential regulator of miR-135a-5p, which would explain the decreased miR-135a-5p expression during the WOI.

Conclusion

MiR-135a-5p expression is significantly downregulated during the WOI. High miR-135a-5p levels suppress pinopode development and endometrial tissue decidualization through HOXA10 and BMPR2, contributing to inadequate endometrial receptivity.

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Data availability

The data generated in this study are provided in the Supplement or are available from the corresponding author.

Abbreviations

ANOVA:

Analysis of variance

ART:

Assisted reproduction technology

BMPR2:

Bone morphogenetic protein receptor 2

c-AMP:

8-Bromo-cAMP sodium salt

CEBPD:

CCAAT/enhancer-binding protein delta

CS-FBS:

Charcoal-stripped fetal bovine serum

CCK-8:

Cell counting kit-8

DAB:

3,3’-diaminobenzidene tetrahydrochloride

DE:

different expression

DMEM/F12:

Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12

dpc:

days post coitus

Dtprp:

decidual/trophoblast prolactin-related protein

E 0.5:

embryonic day 0.5

EDTA:

ethylenediaminetetraacetic acid

EECs:

endometrial epithelial cells

ESCs:

endometrial stromal cells

FET:

frozen embryo transfer

GEO:

Gene Expression Omnibus

HBBS:

hanks balanced salt solution

HE:

hematoxylin-eosin

HEK-293T:

human embryonic kidney cells

HESCs:

human endometrial stromal cells

HOXA10:

(homeobox A10)

IGFBP1:

insulin like growth factor binding protein 1

ITS:

insulin-transferrin-selenium solution

LH:

luteinizing hormone

miRNA:

microRNA

MPA:

Medroxyprogesterone 17-acetate

NGS:

Next Generation Sequencing

PBS:

phosphate buffered saline

pre-WOI:

previous WOI

pHESCs:

human primary endometrial stromal cells

PRL:

prolactin

RIF:

repeated implantation failures

RPMI:

Roswell Park Memorial Institute

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

SEM:

scanning electron microscopy

Si:

small interfering

WOI:

window of implantation

3’-UTR:

3’ untranslated region

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Acknowledgements

We would like to thank the patients participated in the included studies and all researchers.

Funding

This study was supported by Major Clinical Research Projects of the Second Affiliated Hospital, Air Force Medical University (2021LCYJ004).

Author information

Author notes

  1. Yunan He and Ying Ju contributed equally to this work.

Authors and Affiliations

  1. Reproductive Medicine Center, Department of Gynecology and Obstetrics, Tangdu Hospital, Air Force Medical University, No.1 Xinsi Road, Baqiao District, Xi’an, Shaanxi, China

    Yunan He, Ying Ju, Hui Lei, Jie Dong, Ni Jin, Jie Lu, Shuqiang Chen & Xiaohong Wang

  2. Clinical Research Center for Reproductive Medicine and Gynecological Endocrine Diseases of Shaanxi Province, Xi’an, Shaanxi, China

    Yunan He, Ying Ju, Hui Lei, Jie Dong, Ni Jin, Jie Lu, Shuqiang Chen & Xiaohong Wang

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  1. Yunan He
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Contributions

XHW and SQC conceived and designed the study. YNH, HL, JL, and NJ performed the experiments, analyzed the data. YNH, YJ, and SQC wrote the manuscript. SQC, JD and XHW contributed to the writing of final version of the manuscript. All authors agreed and reviewed the final version of the manuscript.

Corresponding authors

Correspondence to Shuqiang Chen or Xiaohong Wang.

Ethics declarations

Ethics approval and consent to participate

This study received approval from Institutional Review Board, Medical Ethics Committee of the Second Affiliated Hospital, Air Force Medical University (TDLL-202311-19), and all patients provided informed consent. Animal experiments were approved by the Experimental Animal Welfare and Ethics Committee of Air Force Medical University (IACUC-20221130).

Consent for publication

All authors agreed to publish the manuscript. What we have done is not involved in previous studies. This manuscript will not be published elsewhere.

Conflict of interest

The authors declare that they have no competing interests.

Competing interests

The authors declare that they have no competing interests.

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He, Y., Ju, Y., Lei, H. et al. MiR-135a-5p regulates window of implantation by suppressing pinopodes development and decidualization of endometrial stromal cells. J Assist Reprod Genet (2024). https://doi.org/10.1007/s10815-024-03088-8

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