Biallelic variants in MAD2L1BP (p31comet) cause female infertility characterized by oocyte maturation arrest

  1. Lingli Huang
  2. Wenqing Li
  3. Xingxing Dai
  4. Shuai Zhao
  5. Bo Xu
  6. Fengsong Wang
  7. Ren-Tao Jin
  8. Lihua Luo
  9. Liming Wu
  10. Xue Jiang
  11. Yu Cheng
  12. Jiaqi Zou
  13. Caoling Xu
  14. Xianhong Tong
  15. Heng-Yu Fan  Is a corresponding author
  16. Han Zhao  Is a corresponding author
  17. Jianqiang Bao  Is a corresponding author
  1. University of Science and Technology of China, China
  2. Zhejiang University, China
  3. Shandong University, China
  4. Anhui Medical University, China

Abstract

Human oocyte maturation arrest represents one of the severe conditions for female patients with primary infertility. However, the genetic factors underlying this human disease remain largely unknown. The spindle assembly checkpoint (SAC) is an intricate surveillance mechanism that ensures accurate segregation of chromosomes throughout cell cycles. Once the kinetochores of chromosomes are correctly attached to bipolar spindles and the SAC is satisfied, the MAD2L1BP, best known as p31comet, binds MAD2 and recruits the AAA+-ATPase TRIP13 to disassemble the mitotic checkpoint complex (MCC), leading to the cell cycle progression. In this study, by whole-exome sequencing (WES), we identified homozygous and compound heterozygous MAD2L1BP variants in three families with female patients diagnosed with primary infertility owing to oocyte metaphase I (MI) arrest. Functional studies revealed that the protein variants resulting from the C-terminal truncation of MAD2L1BP lost their binding ability to MAD2. cRNA microinjection of full-length or truncated MAD2L1BP uncovered their discordant roles in driving the extrusion of polar body 1 (PB1) in mouse oocytes. Furthermore, the patient’s oocytes carrying the mutated MAD2L1BP variants resumed polar body extrusion (PBE) when rescued by microinjection of full-length MAD2L1BP cRNAs. Together, our studies identified and characterized novel biallelic variants in MAD2L1BP responsible for human oocyte maturation arrest at MI, and thus prompted new therapeutic avenues for curing female primary infertility.

Data availability

Source Data files have been provided for Figure 2, Figure 3 and Table 2. Sequencing data have been deposited in GEO under accession code GSE232488.

The following data sets were generated

Article and author information

Author details

  1. Lingli Huang

    Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9748-267X
  2. Wenqing Li

    Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China
    Competing interests
    The authors declare that no competing interests exist.
  3. Xingxing Dai

    Life Sciences Institute, Zhejiang University, Hangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  4. Shuai Zhao

    Center for Reproductive Medicine, Shandong University, Jinan, China
    Competing interests
    The authors declare that no competing interests exist.
  5. Bo Xu

    Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China
    Competing interests
    The authors declare that no competing interests exist.
  6. Fengsong Wang

    School of Life Science, Anhui Medical University, Hefei, China
    Competing interests
    The authors declare that no competing interests exist.
  7. Ren-Tao Jin

    Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China
    Competing interests
    The authors declare that no competing interests exist.
  8. Lihua Luo

    Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China
    Competing interests
    The authors declare that no competing interests exist.
  9. Liming Wu

    Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China
    Competing interests
    The authors declare that no competing interests exist.
  10. Xue Jiang

    Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China
    Competing interests
    The authors declare that no competing interests exist.
  11. Yu Cheng

    Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China
    Competing interests
    The authors declare that no competing interests exist.
  12. Jiaqi Zou

    Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China
    Competing interests
    The authors declare that no competing interests exist.
  13. Caoling Xu

    Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China
    Competing interests
    The authors declare that no competing interests exist.
  14. Xianhong Tong

    Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China
    Competing interests
    The authors declare that no competing interests exist.
  15. Heng-Yu Fan

    Life Sciences Institute, Zhejiang University, Hangzhou, China
    For correspondence
    hyfan@zju.edu.cn
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4544-4724
  16. Han Zhao

    Center for Reproductive Medicine, Shandong University, Jinan, China
    For correspondence
    hanzh80@sdu.edu.cn
    Competing interests
    The authors declare that no competing interests exist.
  17. Jianqiang Bao

    Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China
    For correspondence
    jqbao@ustc.edu.cn
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1248-2687

Funding

National Natural Science Foundation of China (81801440)

  • Lingli Huang

National Natural Science Foundation of China (82192874)

  • Han Zhao

National Natural Science Foundation of China (82171842)

  • Han Zhao

National Natural Science Foundation of China (31970793)

  • Jianqiang Bao

National Natural Science Foundation of China (32170856)

  • Jianqiang Bao

the Ministry of Science and Technology of China (2019YFA0802600)

  • Jianqiang Bao

the Fundamental Research Funds for the Central Universities (WK2070000156)

  • Jianqiang Bao

Startup funding (KY9100000001)

  • Jianqiang Bao

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: Animal care and experimental procedures were conducted in accordance with the Animal Research Committee guidelines of Zhejiang University (approval # ZJU20210252 to H.Y.F) and USTC (approval # 2019-N(A)-299 to J.Q.B).

Human subjects: Peripheral blood samples from all affected individuals and their available family members and ten Metaphase I (MI) arrested oocytes from the patient (F1: II-1) were donated for this study with written informed consent. This study was approved by the biomedical research ethics committees of Anhui Medical University on 1 March 2017(reference number 20170121; the Anhui Provincial Hospital Affiliated to Anhui Medical University, now renamed as the First Affiliated Hospital of USTC after December 2017).

Reviewing Editor

  1. Adèle L Marston, University of Edinburgh, United Kingdom

Version history

  1. Received: December 18, 2022
  2. Preprint posted: January 16, 2023 (view preprint)
  3. Accepted: June 15, 2023
  4. Accepted Manuscript published: June 19, 2023 (version 1)
  5. Version of Record published: July 4, 2023 (version 2)

Copyright

© 2023, Huang et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 594
    Page views
  • 86
    Downloads
  • 0
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Lingli Huang
  2. Wenqing Li
  3. Xingxing Dai
  4. Shuai Zhao
  5. Bo Xu
  6. Fengsong Wang
  7. Ren-Tao Jin
  8. Lihua Luo
  9. Liming Wu
  10. Xue Jiang
  11. Yu Cheng
  12. Jiaqi Zou
  13. Caoling Xu
  14. Xianhong Tong
  15. Heng-Yu Fan
  16. Han Zhao
  17. Jianqiang Bao
(2023)
Biallelic variants in MAD2L1BP (p31comet) cause female infertility characterized by oocyte maturation arrest
eLife 12:e85649.
https://doi.org/10.7554/eLife.85649

Further reading

    1. Chromosomes and Gene Expression
    2. Developmental Biology
    Virginia L Pimmett, Mounia Lagha
    Insight

    Imaging experiments reveal the complex and dynamic nature of the transcriptional hubs associated with Notch signaling.

    1. Cell Biology
    2. Developmental Biology
    Simon Schneider, Andjela Kovacevic ... Hubert Schorle
    Research Article

    Cylicins are testis-specific proteins, which are exclusively expressed during spermiogenesis. In mice and humans, two Cylicins, the gonosomal X-linked Cylicin 1 (Cylc1/CYLC1) and the autosomal Cylicin 2 (Cylc2/CYLC2) genes, have been identified. Cylicins are cytoskeletal proteins with an overall positive charge due to lysine-rich repeats. While Cylicins have been localized in the acrosomal region of round spermatids, they resemble a major component of the calyx within the perinuclear theca at the posterior part of mature sperm nuclei. However, the role of Cylicins during spermiogenesis has not yet been investigated. Here, we applied CRISPR/Cas9-mediated gene editing in zygotes to establish Cylc1- and Cylc2-deficient mouse lines as a model to study the function of these proteins. Cylc1 deficiency resulted in male subfertility, whereas Cylc2-/-, Cylc1-/yCylc2+/-, and Cylc1-/yCylc2-/- males were infertile. Phenotypical characterization revealed that loss of Cylicins prevents proper calyx assembly during spermiogenesis. This results in decreased epididymal sperm counts, impaired shedding of excess cytoplasm, and severe structural malformations, ultimately resulting in impaired sperm motility. Furthermore, exome sequencing identified an infertile man with a hemizygous variant in CYLC1 and a heterozygous variant in CYLC2, displaying morphological abnormalities of the sperm including the absence of the acrosome. Thus, our study highlights the relevance and importance of Cylicins for spermiogenic remodeling and male fertility in human and mouse, and provides the basis for further studies on unraveling the complex molecular interactions between perinuclear theca proteins required during spermiogenesis.