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Single Cell Proteomics Profiling Reveals That Embryo-Secreted TNF-α Plays a Critical Role During Embryo Implantation to the Endometrium

  • Reproductive Endocrinology: Original Article
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A Correction to this article was published on 16 February 2022

This article has been updated

Abstract

It has been long-known that endometrium-secreted cytokines play a critical role during embryo implantation. However, whether cytokines secreted from the embryo are relevant to the process of embryo implantation remains unclear. The concentration of cytokines in embryo culture medium was tested using a newly developed, high-sensitivity single-cell proteomic platform and evaluated in comparison to embryo quality and clinical outcome. The effect of TNF-α on embryo and endometrium Ishikawa cells was investigated using immunofluorescence staining, CCK-8 assay, TUNEL staining, and RT-qPCR. Of the 10 cytokines measured, only TNF-α concentration was significantly higher in the group with embryo implantation failure. Immunofluorescence staining showed that the expression of TNF-α was unevenly distributed in blastocysts, and the expression level was significantly correlated with the blastocyst inner cell mass (ICM) quality score. Gene profiling showed that addition of TNF-α led to increased expression of tumor necrosis factor receptor 1 (TNFR1) and apoptosis-related genes and that this could be inhibited by the TNF-α receptor inhibitor etanercept (ETA). In addition, an increased expression of water and ion channels, including AQP3, CFTR, ENaCA, and CRISP2 was also observed which could also be inhibited by ETA. Our results show that higher embryo-secreted TNF-α levels are associated with implantation failure through activation of TNF-α receptor, and TNF-α may be an independent predictor for pre-transfer assessment of the embryo development potential in IVF patients.

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Abbreviations

TNF-α:

tumor necrosis factor alpha

ICM:

blastocysts inner cell mass

TM:

trophectoderm of the embryo

ETA:

etanercept

TNFR1:

tumor necrosis factor receptor 1

TNFR2:

tumor necrosis factor receptor 2

CFTR:

cystic fibrosis transmembrane conductance regulator

ENaCA:

epithelial Na (+) channel subunit alpha

AQP3:

aquaporin-3

CRISP2:

cysteine-rich secretory protein 2

LIF:

leukemia inhibitory factor

CRYBB2:

beta-crystallin B2

Th1/2:

helper T cells 1/2

IL-1β:

interleukin-1 beta

IL-6:

interleukin-6

IL-8:

interleukin-8

IL-10:

interleukin-10

MCP1:

monocyte chemotactic protein 1

MIP-1α:

macrophage inflammatory protein 1-alpha

MIP-1β:

macrophage inflammatory protein 1-beta

RANTES:

C-C motif chemokine 5, T-cell-specific protein

GM-CSF:

granulocyte-macrophage colony-stimulating factor

NF-κB:

nuclear factor of kappa light polypeptide gene enhancer in B-cells 3

IVF:

in vitro fertilization

ICSI:

intracytoplasmic sperm injection

ART:

assisted reproductive technology

ROC:

receiver operating characteristic curve

PCOS:

polycystic ovary syndrome

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Acknowledgements

We thank AIVFO (Inc., Chengdu, China) for providing technical support. Especially, we grateful for the technical assistance provided by Zhang Jie and Bo Lin in Histology and Imaging Platform, Core Facilities of West China Hospital, Sichuan University.

Funding

This work was supported by the National Key Research and Development Program of China (2018YFC1002804), Scientific and Translational Research Fund of Sichuan Provincial Department of Science and Technology (2017YSZH0032), the Key Research and Development Support Program of Chengdu (2019-YF05-00017-SN), and the Natural Sciences Foundation of China (81901435).

Availability of Supporting Data

The dataset used and/or analyzed during the current study is available from the corresponding author upon reasonable request. Registry data are available publicly (see references).

Author information

Author notes

  1. Jiao Lv and Xudong Shan contributed equally to this work.

Authors and Affiliations

  1. The Joint Laboratory for Reproductive Medicine of SCU-CUHK, West China Second University Hospital, Sichuan University, Chengdu, China

    Jiao Lv, Haoxuan Yang, Yuting Wen, Xueguang Zhang, Hanxiao Chen, Chi Chiu Wang, Ruizhe Zhang, Tin Chiu Li, Xiaohu Zhang, Lang Qin & Wenming Xu

  2. Department of Obstetrics/Gynecology, West China Second University Hospital, Sichuan University, Chengdu, China

    Jiao Lv, Haoxuan Yang, Yuting Wen, Xueguang Zhang, Hanxiao Chen, Xiaohu Zhang & Wenming Xu

  3. The Key Laboratory of Birth Defects and Related Diseases of Women and Children, Ministry of Education, West China Second University Hospital, Sichuan University, Chengdu, China

    Jiao Lv, Haoxuan Yang, Yuting Wen, Xueguang Zhang, Hanxiao Chen, Xiaohu Zhang & Wenming Xu

  4. West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu, China

    Xudong Shan

  5. School of Medical and Life Sciences, Chengdu University of Traditional Chinese Medicine, Chengdu, China

    Xudong Shan, Hengli Li, Dongmei Tian & Minghui Zhu

  6. Assisted Reproductive Technology Unit, Department of Obstetrics and Gynecology, The Chinese University of Hong Kong, Shatin, Hong Kong, SAR, China

    Chi Chiu Wang, Ruizhe Zhang & Tin Chiu Li

  7. Department of Obstetrics and Gynecology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 107 Road Yanjiang West, Guangzhou, 510120, China

    Xiaomiao Zhao

  8. Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China

    Yao Lu

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  1. Jiao Lv
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Contributions

WX and MZ developed the concept of the study; all authors contributed to data accumulation; JL and XS contributed to data analysis; all authors contributed to data interpretation. JL and WX wrote the manuscript. All authors contributed to revisions of the manuscript and approved the final submission. JL had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.

Corresponding authors

Correspondence to Minghui Zhu or Wenming Xu.

Ethics declarations

Ethics Approval and Consent to Participate

The Second Affiliated Hospital of Chengdu University of Traditional Chinese Medicine Ethics Committee; approval number: 2017–01. Participants provided consent to participate in this study.

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Not applicable.

Conflict of Interest

The authors declare no competing interests.

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Lv, J., Shan, X., Yang, H. et al. Single Cell Proteomics Profiling Reveals That Embryo-Secreted TNF-α Plays a Critical Role During Embryo Implantation to the Endometrium. Reprod. Sci. 29, 1608–1617 (2022). https://doi.org/10.1007/s43032-021-00833-7

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