Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Dec 2:12:746082.
doi: 10.3389/fgene.2021.746082. eCollection 2021.

Exome-Sequencing Identifies Novel Genes Associated with Recurrent Pregnancy Loss in a Chinese Cohort

Huifen Xiang  1   2 , Chunyan Wang  3   4 , Hong Pan  3   4 , Qian Hu  1   2 , Ruyi Wang  3   4 , Zuying Xu  1   2 , Tengyan Li  4 , Yezhou Su  1   2 , Xu Ma  3   4 , Yunxia Cao  1   2 , Binbin Wang  3   4
Affiliations

Exome-Sequencing Identifies Novel Genes Associated with Recurrent Pregnancy Loss in a Chinese Cohort

Huifen Xiang et al. Front Genet. .

Abstract

Recurrent pregnancy loss (RPL) is a common reproductive problem affecting around 5% of couples worldwide. At present, about half of RPL cases remained unexplained. Previous studies have suggested an important role for genetic determinants in the etiology of RPL. Here, we performed whole-exome sequencing (WES) analysis on 100 unrelated Han Chinese women with a history of two or more spontaneous abortions. We identified 6736 rare deleterious nonsynonymous variants across all patients. To focus on possible candidate genes, we generated a list of 95 highly relevant genes that were functionally associated with miscarriage according to human and mouse model studies, and found 35 heterozygous variants of 28 RPL-associated genes in 32 patients. Four genes (FOXA2, FGA, F13A1, and KHDC3L) were identified as being strong candidates. The FOXA2 nonsense variant was for the first time reported here in women with RPL. FOXA2 knockdown in HEK-293T cells significantly diminished the mRNA and protein expression levels of LIF, a pivotal factor for maternal receptivity and blastocyst implantation. The other genes, with 29 variants, were involved in angiogenesis, the immune response and inflammation, cell growth and proliferation, which are functionally important processes for implantation and pregnancy. Our study identified several potential causal genetic variants in women with RPL by WES, highlighting the important role of genes controlling coagulation, confirming the pathogenic role of KHDC3L and identifying FOXA2 as a newly identified causal gene in women with RPL.

Keywords: FOXA2; KHDC3L; coagulation; recurrent pregnancy loss; whole-exome sequencing.

PubMed Disclaimer

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Correlation between miscarriage phenotype and variant accumulation. (A) The mean number of total deleterious variants in two groups with miscarriages and more than two miscarriages, respectively. (B) The mean number of deleterious variants of RPL-related genes in two groups with miscarriages and more than two miscarriages, respectively.
FIGURE 2
FIGURE 2
Sanger sequencing of the variants in strong candidate genes. (A) KHDC3L c.436_468del: p.146_156del. (B) FGA c.1906_1908del: p.636_636del. (C) FGA c.C2285T:p.A762V. (D) F13A1 c.C1201T:p.Q401X. (E) F13A1 c.C1834T:p.R612C. (F) FOXA2 c.C1242G:p.Y414X.
FIGURE 3
FIGURE 3
Expression of LIF in HEK 293T cells with FOXA2 knockdown. (A) The mRNA level of LIF was measured by qPCR. (B) The protein level of LIF was detected by western blot. Student’s t-test was used for the p value analysis. *p < 0.05; **p < 0.01.
See this image and copyright information in PMC

References

    1. Amri Y., Toumi N. E. H., Hadj Fredj S., De Moerloose P. (2016). Congenital Afibrinogenemia: Identification and Characterization of Two Novel Homozygous Fibrinogen Aα and Bβ Chain Mutations in Two Tunisian Families. Thromb. Res. 143, 11–16. 10.1016/j.thromres.2016.04.016 - DOI - PubMed
    1. Arias-Sosa L. A., Acosta I. D., Lucena-Quevedo E., Moreno-Ortiz H., Esteban-Pérez C., Forero-Castro M. (2018). Genetic and Epigenetic Variations Associated with Idiopathic Recurrent Pregnancy Loss. J. Assist. Reprod. Genet. 35, 355–366. 10.1007/s10815-017-1108-y - DOI - PMC - PubMed
    1. Biswas A., Ivaskevicius V., Thomas A., Varvenne M., Brand B., Rott H., et al. (2014). Eight Novel F13a1 Gene Missense Mutations in Patients with Mild Fxiii Deficiency: In Silico Analysis Suggests Changes in Fxiii-A Subunit Structure/function. Ann. Hematol. 93, 1665–1676. 10.1007/s00277-014-2102-4 - DOI - PubMed
    1. Casini A., Neerman-Arbez M., Ariëns R. A., De Moerloose P. (2015). Dysfibrinogenemia: From Molecular Anomalies to Clinical Manifestations and Management. J. Thromb. Haemost. 13, 909–919. 10.1111/jth.12916 - DOI - PubMed
    1. Casini A., Neerman-Arbez M., de Moerloose P. (2013). Congenital Fibrinogen Disorders: An Update. Semin. Thromb. Hemost. 39, 585–595. 10.1055/s-0033-1349222 - DOI - PubMed