Novel variations in TENT5D lead to teratozoospermia in infertile patients

Ying-Teng Zhang; Gan Shen; Liang-Chai Zhuo; Xue Yang; Su-Yan Wang; Tie-Chao Ruan; Chuan Jiang; Xiang Wang; Yan Wang; Yi-Hong Yang; Ying Shen

doi:10.1111/andr.13589

Novel variations in TENT5D lead to teratozoospermia in infertile patients

Andrology. 2024 Jan 16. doi: 10.1111/andr.13589. Online ahead of print.

Authors

Ying-Teng Zhang¹, Gan Shen¹, Liang-Chai Zhuo¹, Xue Yang¹, Su-Yan Wang¹, Tie-Chao Ruan^{1

2}, Chuan Jiang¹, Xiang Wang¹, Yan Wang³, Yi-Hong Yang³, Ying Shen^{1

4}

Affiliations

¹ Department of Obstetrics/Gynecology, Key Laboratory of Obstetric, Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education, West China Second University Hospital, Sichuan University, Chengdu, China.
² Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu, China.
³ Reproduction Medical Center of West China Second University Hospital, Key Laboratory of Obstetric, Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education, Sichuan University, Chengdu, China.
⁴ NHC Key Laboratory of Chronobiology, Sichuan University, Chengdu, China.

PMID: 38228861
DOI: 10.1111/andr.13589

Abstract

Purpose: Teratozoospermia is the main pathogenic factor of male infertility. However, the genetic etiology of teratozoospermia is largely unknown. This study aims to clarify the relationship between novel variations in TENT5D and teratozoospermia in infertile patients.

Materials and methods: Two infertile patients were enrolled. Routine semen analysis of patients and normal controls was conducted with the WHO guidelines. Whole-exome sequencing (WES) was conducted to identify pathogenic variants in the two patients. Morphology and ultrastructure analysis of spermatozoa in the two patients was determined by Papanicolaou staining, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The functional effect of the identified variants was analyzed by immunofluorescence staining and western blotting. The expression of TENT5D in different germ cells was detected by immunofluorescence staining.

Results: Two new hemizygous variations, c.101C > T (p.P34L) and c.125A > T (p.D42V), in TENT5D were detected in two patients with male infertility. Morphology analysis showed abnormalities in spermatozoa morphology in the two patients, including multiple heads, headless, multiple tails, coiled, and/or bent flagella. Ultrastructure analysis showed that most of the spermatozoa exhibited missing or irregularly arranged '9+2' structures. Further functional experiments confirmed the abrogated TENT5D protein expression in patients. In addition, both p.P34L and p.D42V substitutions resulted in a conformational change of the TENT5D protein. We precisely analyzed the subcellular localization of TENT5D in germ cells in humans and mice. And we found that TENT5D was predominantly detected in the head and flagellum of elongating spermatids and epididymal spermatozoa.

Conclusions: Our results showed further evidence of a relationship between TENT5D mutation and human male infertility, providing new genetic insight for use in the diagnosis and treatment of male infertility.

Keywords: TENT5D; WES; gene variation; male infertility; teratozoospermia.