Heterozygous loss-of-function variants in LHX8 cause female infertility characterized by oocyte maturation arrest

Lin Zhao; Qun Li; Yanping Kuang; Peng Xu; Xiaoxi Sun; Qingxia Meng; Wenjing Wang; Yang Zeng; Biaobang Chen; Jing Fu; Jie Dong; Jiawei Zhu; Yuxi Luo; Hao Gu; Caihong Li; Chunyi Li; Ling Wu; Xiaoyan Mao; Huizhen Fan; Ruyi Liu; Zhihua Zhang; Qiaoli Li; Jing Du; Lin He; Li Jin; Lei Wang; Qing Sang

doi:10.1016/j.gim.2022.07.027

Heterozygous loss-of-function variants in LHX8 cause female infertility characterized by oocyte maturation arrest

Genet Med. 2022 Nov;24(11):2274-2284. doi: 10.1016/j.gim.2022.07.027. Epub 2022 Aug 27.

Authors

Lin Zhao¹, Qun Li², Yanping Kuang³, Peng Xu⁴, Xiaoxi Sun⁵, Qingxia Meng⁶, Wenjing Wang⁷, Yang Zeng⁷, Biaobang Chen⁸, Jing Fu⁵, Jie Dong⁷, Jiawei Zhu⁶, Yuxi Luo⁷, Hao Gu⁷, Caihong Li⁹, Chunyi Li⁹, Ling Wu³, Xiaoyan Mao³, Huizhen Fan⁷, Ruyi Liu⁷, Zhihua Zhang⁷, Qiaoli Li⁷, Jing Du⁸, Lin He¹⁰, Li Jin¹¹, Lei Wang¹², Qing Sang¹³

Affiliations

¹ Institute of Pediatrics, Children's Hospital of Fudan University, the Institutes of Biomedical Sciences, the State Key Laboratory of Genetic Engineering, Fudan University, Shanghai, China; NHC Key Lab of Reproduction Regulation, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Fudan University, Shanghai, China.
² Institute of Pediatrics, Children's Hospital of Fudan University, the Institutes of Biomedical Sciences, the State Key Laboratory of Genetic Engineering, Fudan University, Shanghai, China; Human Phenome Institute, Fudan University, Shanghai, China.
³ Reproductive Medicine Center, Shanghai Ninth Hospital, Shanghai Jiao Tong University, Shanghai, China.
⁴ Hainan Jinghua Hejing Hospital for Reproductive Medicine, Haikou, China.
⁵ Shanghai Ji Ai Genetics and IVF Institute, Obstetrics and Gynecology Hospital, Fudan University, Shanghai, China.
⁶ Center for Reproduction and Genetics, The Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou, China.
⁷ Institute of Pediatrics, Children's Hospital of Fudan University, the Institutes of Biomedical Sciences, the State Key Laboratory of Genetic Engineering, Fudan University, Shanghai, China.
⁸ NHC Key Lab of Reproduction Regulation, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Fudan University, Shanghai, China.
⁹ Shenyang Jinghua Hospital, Liaoning, China.
¹⁰ Bio-X Institutes, Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Ministry of Education, Shanghai Jiao Tong University, Shanghai, China.
¹¹ State Key Laboratory of Genetic Engineering and Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Fudan University, Shanghai, China.
¹² Institute of Pediatrics, Children's Hospital of Fudan University, the Institutes of Biomedical Sciences, the State Key Laboratory of Genetic Engineering, Fudan University, Shanghai, China. Electronic address: wangleiwanglei@fudan.edu.cn.
¹³ Institute of Pediatrics, Children's Hospital of Fudan University, the Institutes of Biomedical Sciences, the State Key Laboratory of Genetic Engineering, Fudan University, Shanghai, China. Electronic address: sangqing@fudan.edu.cn.

PMID: 36029299
DOI: 10.1016/j.gim.2022.07.027

Abstract

Purpose: The genetic causes of oocyte maturation arrest leading to female infertility are largely unknown, and no population-based genetic analysis has been applied in cohorts of patients with infertility. We aimed to identify novel pathogenic genes causing oocyte maturation arrest by using a gene-based burden test.

Methods: Through comparison of exome sequencing data from 716 females with infertility characterized by oocyte maturation arrest and 3539 controls, we performed a gene-based burden test and identified a novel pathogenic gene LHX8. Splicing event was evaluated using a minigene assay, expression of LHX8 protein was assessed in HeLa cells, and nuclear subcellular localization was determined in both HeLa cells and mouse oocytes.

Results: A total of 5 heterozygous loss-of-function LHX8 variants were identified from 6 independent families (c.389+1G>T, c.412C>T [p.Arg138∗], c.282C>A [p.Cys94∗]; c.257dup [p.Tyr86∗]; and c.180del, [p.Ser61Profs∗30]). All the identified variants in LHX8 produced truncated LHX8 protein and resulted in loss of LHX8 nuclear localization in both HeLa cells and mouse oocytes.

Conclusion: By combining genetic evidence and functional evaluations, we identified a novel pathogenic gene LHX8 and established the causative relationship between LHX8 haploinsufficiency and female infertility characterized by oocyte maturation arrest.

Keywords: Female infertility; Gene-based burden test; LHX8; Loss-of-function variants; Oocyte maturation arrest.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Exome Sequencing
Female
HeLa Cells
Humans
Infertility, Female* / genetics
Infertility, Female* / pathology
Mice
Oocytes
Oogenesis / genetics