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A Proteomic Analysis of Human Follicular Fluid: Proteomic Profile Associated with Embryo Quality

  • Reproductive Biology: Original Article
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Abstract

Embryo selection is a key point of in vitro fertilization (IVF). The most commonly used method for embryo selection is morphological assessment. However, it is sometimes inaccurate. Follicular fluid (FF) contains a complex mixture of proteins that are essential for follicle development and oocyte maturation. Analyzing human FF proteomic profiles and identifying predictive biomarkers might be helpful for evaluating embryo quality. A total of 22 human FF samples were collected from 19 infertile women who underwent IVF/intracytoplasmic sperm injection (ICSI) treatment between October 2021 and November 2021. FFs were grouped into two categories on the basis of the day 3 embryo quality, grade I or II in the hqFF group and grade III in the nhqFF group. FF was analyzed by liquid chromatography-tandem mass spectrometry (LC/MS/MS). The key differentially expressed proteins (DEPs) were validated by parallel reaction monitoring (PRM) and enzyme-linked immunosorbent assay (ELISA). Differentially expressed proteins were further analyzed using DAVID software. A total of 558 proteins were identified, of which 50 proteins were differentially expressed in the hqFF versus nhqFF group, including 32 upregulated proteins (> 1.20-fold, P < 0.05) and 18 downregulated proteins (< 0.67-fold, P < 0.05). Bioinformatics analyses showed that the upregulated DEPs were enriched in components of the coagulation and complement systems and negative regulation of peptidase activity, while the downregulated DEPs were enriched in molecular function of extracellular matrix structural and constituent collagen binding. Our results suggested that a number of protein biomarkers in FF were associated with embryo quality. It may help develop an effective and noninvasive method for embryo selection.

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Data Availability

The original contributions presented in the study are included in the article/Supplementary Material. Further inquiries can be directed to the corresponding authors.

Code Availability

Not applicable.

Abbreviations

AMH:

Anti-Müllerian hormone

CBG:

Corticosteroid binding globulin

C1R:

Complement C1r subcomponent

C7:

Complement component C7

COCs:

Cumulus-oocyte complexes

DDA:

Data-dependent acquisition

DEP:

Differentially expressed protein

FF:

Follicular fluid

GCs:

Glucocorticoids

GnRH:

Gonadotropin releasing hormone

GO:

Gene Ontology

ICSI:

Intracytoplasmic sperm injection

ITIH1:

Inter-alpha-trypsin inhibitor heavy chain H1

ITIH2:

Inter-alpha-trypsin inhibitor heavy chain H2

IVF:

In vitro fertilization

KEGG:

Kyoto Encyclopedia of Genes and Genomes

LC/MS/MS:

Liquid chromatography–tandem mass spectrometry

PPI:

Protein–protein interaction

PRM:

Parallel reaction monitoring

SERPIN:

Serine protease inhibitor

SERPINA6:

Serine protease inhibitor A6

SERPINC1:

Serpin peptidase inhibitor, clade C

SHAP:

Serum-derived hyaluronan-associated proteins

TMT:

Tandem mass tags

VEGF:

Vascular endothelial growth factor

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Acknowledgements

We thank all the patients for participating in this research.

Funding

This research was supported by the National Key Research and Development Project (2019YFA0802600) and the National Natural Science Foundation of China (82171599 and 81971333).

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Authors and Affiliations

  1. Reproductive and Genetic Hospital, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, 230001, China

    Jingjuan Ji, Xinyi Zhu, Yan Zhang, Lijun Shui, Shun Bai, Lingli Huang, Haoyu Wang, Shiwei Fan, Zelin Zhang, Lihua Luo & Bo Xu

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  1. Jingjuan Ji
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Contributions

BX and JJ conceived of the study. SB, BX, LH, and JJ participated in its design and coordination and reviewed the final manuscript for submission. JJ, XZ, LS, SB, and HW carried out the studies and drafted the manuscript. BX and SB carried out the bioinformatics analysis. LL participated in the study design and performed the HFF collection. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Bo Xu.

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Ethics Approval and Consent to Participate

The Ethics Committee of The First Affiliated Hospital of USTC approved this study protocol (2022-KY-354), and informed consent was obtained from each individual.

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The authors declare no competing interests.

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Ji, J., Zhu, X., Zhang, Y. et al. A Proteomic Analysis of Human Follicular Fluid: Proteomic Profile Associated with Embryo Quality. Reprod. Sci. 31, 199–211 (2024). https://doi.org/10.1007/s43032-023-01293-x

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