Altered zygotic gene expression caused by sperm with Tdrd6 variants disrupts early embryonic development

Abstract

The precise mechanisms behind early embryonic arrest due to sperm-related factors and the most effective strategies are not yet fully understood. Here, we present two cases of male infertility linked to novel TDRD6 variants, associated with oligoasthenoteratozoospermia (OAT) and early embryonic arrest. To investigate the underlying mechanisms and promising therapeutic approaches, Tdrd6 knock-in and knock-out mice were generated. The Tdrd6 variant male mice demonstrated OAT and embryonic arrest, mirroring the clinical observations of our patients. Sperm from both affected individuals and mice exhibited aberrant localization of phospholipase C zeta and oocyte activation deficiency (OAD). The application of artificial oocyte activation (AOA) effectively overcame the infertility caused by the variants, facilitating successful pregnancies and live births in both human and mice. Additionally, our research revealed that OAD influences the expression of multitude of genes at the 2-pronuclear (2PN) stage, with the Mos gene playing a pivotal role in early embryonic arrest. The injection of Mos mRNA can mitigate this arrest. To our knowledge, this is the first study to show that sperm-related OAD affects gene expression at the 2PN stage and elucidate how AOA overcomes male factor-derived embryonic arrest, enabling successful pregnancies and live births.

Keywords

artificial oocyte activation / assisted reproductive technology / early embryonic arrest / infertility / TDRD6

References

[1] Wei Y, Wang J, Qu R, et al. Genetic mechanisms of fertilization failure and early embryonic arrest: a comprehensive review. Hum Reprod Update. 2024; 30(1): 48-80.

[2] Vallet-Buisan M, Mecca R, Jones C, Coward K, Yeste M. Contribution of semen to early embryo development: fertilization and beyond. Hum Reprod Update. 2023; 29(4): 395-433.

[3] Colombero LT, Moomjy M, Sills ES, Rosenwaks Z, Palermo GD. The role of structural integrity of the fertilising spermatozoon in early human embryogenesis. Zygote. 1999; 7(2): 157-163.

[4] Yoon SY, Jellerette T, Salicioni AM, et al. Human sperm devoid of PLC, zeta 1 fail to induce Ca2+ release and are unable to initiate the first step of embryo development. J Clin Invest. 2008; 118(11): 3671-3681.

[5] Cui L, Fang L, Zhuang L, Shi B, Lin CP, Ye Y. Sperm-borne microRNA-34c regulates maternal mRNA degradation and preimplantation embryonic development in mice. Reprod Biol Endocrinol. 2023; 21(1): 40.

[6] Yuan S, Schuster A, Tang C, et al. Sperm-borne miRNAs and endo-siRNAs are important for fertilization and preimplantation embryonic development. Development. 2015:dev131755.

[7] Castillo J, Jodar M, Oliva R. The contribution of human sperm proteins to the development and epigenome of the preimplantation embryo. Hum Reprod Update. 2018; 24(5): 535-555.

[8] Guo R, Wu H, Zhu X, et al. Bi-allelic variants in chromatoid body protein TDRD6 cause spermiogenesis defects and severe oligoasthenoteratozoospermia in humans. J Med Genet. 2024:jmg-2023-109766.

[9] Sha YW, Wang X, Su ZY, et al. TDRD6 is associated with oligoasthenoteratozoospermia by sequencing the patient from a consanguineous family. Gene. 2018; 659: 84-88.

[10] Vasileva A, Tiedau D, Firooznia A, Müller-Reichert T, Jessberger R. Tdrd6 is required for spermiogenesis, chromatoid body architecture, and regulation of miRNA expression. Curr Biol. 2009; 19(8): 630-639.

[11] Fanourgakis G, Lesche M, Akpinar M, Dahl A, Jessberger R. Chromatoid body protein TDRD6 supports long 3’ UTR triggered nonsense mediated mRNA decay. PLoS Genet. 2016; 12(5): e1005857.

[12] Campos G, Sciorio R, Esteves SC. Total fertilization failure after ICSI: insights into pathophysiology, diagnosis, and management through artificial oocyte activation. Hum Reprod Update. 2023; 29(4): 369-394.

[13] Jin XL, O’Neill C. The presence and activation of two essential transcription factors (cAMP response element-binding protein and cAMP-dependent transcription factor ATF1) in the two-cell mouse embryo. Biol Reprod. 2010; 82(2): 459-468.

[14] Zhang YL, Zheng W, Ren P, et al. Biallelic variants in MOS cause large polar body in oocyte and human female infertility. Hum Reprod. 2022; 37(8): 1932-1944.

[15] Zhang YL, Zheng W, Ren P, et al. Biallelic mutations in MOS cause female infertility characterized by human early embryonic arrest and fragmentation. EMBO Mol Med. 2021; 13(12): e14887.

[16] Chen C, Sun T, Yin M, et al. Ionomycin-induced mouse oocyte activation can disrupt preimplantation embryo development through increased reactive oxygen species reaction and DNA damage. Mol Hum Reprod. 2020; 26(10): 773-783.

[17] Han Y, Liang C, Manthari RK, et al. Arsenic influences spermatogenesis by disorganizing the elongation of spermatids in adult male mice. Chemosphere. 2020; 238: 124650.

[18] He J, Liu X, Zhu C, et al. ASD2023: towards the integrating landscapes of allosteric knowledgebase. Nucleic Acids Res. 2024; 52(D1): D376-D383.

[19] Li Q, Hu Z, He J, et al. Deciphering the comprehensive knowledgebase landscape featuring infertility with IDDB Xtra. Comput Biol Med. 2024; 170: 108105.

[20] Herbst R, Shearman MS, Obermeier A, Schlessinger J, Ullrich A. Differential effects of W mutations on p145c-kit tyrosine kinase activity and substrate interaction. J Biol Chem. 1992; 267(19): 13210-13216.

[21] Lin Y, Huang Y, Li B, et al. Novel mutations in PLCZ1 lead to early embryonic arrest as a male factor. Front Cell Dev Biol. 2023; 11: 1193248.

[22] Xin A, Qu R, Chen G, et al. Disruption in ACTL7A causes acrosomal ultrastructural defects in human and mouse sperm as a novel male factor inducing early embryonic arrest. Sci Adv. 2020; 6(35): eaaz4796.

[23] Chaigne A, Campillo C, Gov NS, et al. A soft cortex is essential for asymmetric spindle positioning in mouse oocytes. Nat Cell Biol. 2013; 15(8): 958-966.

[24] Colledge WH, Carlton MB, Udy GB, Evans MJ. Disruption of c-mos causes parthenogenetic development of unfertilized mouse eggs. Nature. 1994; 370(6484): 65-68.

[25] Zeng Y, Shi J, Xu S, et al. Bi-allelic mutations in MOS cause female infertility characterized by preimplantation embryonic arrest. Hum Reprod. 2022; 37(3): 612-620.

[26] Lyu QF, Deng L, Xue SG, et al. New technique for mouse oocyte injection via a modified holding pipette. Reprod Biomed Online. 2010; 21(5): 663-666.

[27] Cummins JM, Breen TM, Harrison KL, Shaw JM, Wilson LM, Hennessey JF. A formula for scoring human embryo growth rates in in vitro fertilization: its value in predicting pregnancy and in comparison with visual estimates of embryo quality. J In Vitro Fert Embryo Transf. 1986; 3(5): 284-295.

[28] Heindryckx B, De Gheselle S, Gerris J, Dhont M, De Sutter P. Efficiency of assisted oocyte activation as a solution for failed intracytoplasmic sperm injection. Reprod Biomed Online. 2008; 17(5): 662-668.

[29] Heindryckx B, Van der Elst J, De Sutter P, Dhont M. Treatment option for sperm-or oocyte-related fertilization failure: assisted oocyte activation following diagnostic heterologous ICSI. Hum Reprod. 2005; 20(8): 2237-2241.

[30] Ferrer-Buitrago M, Dhaenens L, Lu Y, et al. Human oocyte calcium analysis predicts the response to assisted oocyte activation in patients experiencing fertilization failure after ICSI. Hum Reprod. 2018; 33(3): 416-425.

[31] Vanden Meerschaut F, Leybaert L, Nikiforaki D, Qian C, Heindryckx B, De Sutter P. Diagnostic and prognostic value of calcium oscillatory pattern analysis for patients with ICSI fertilization failure. Hum Reprod. 2013; 28(1): 87-98.

[32] Santos F, Hendrich B, Reik W, Dean W. Dynamic reprogramming of DNA methylation in the early mouse embryo. Dev Biol. 2002; 241(1): 172-182.

[33] Wossidlo M, Nakamura T, Lepikhov K, et al. 5-Hydroxymethylcytosine in the mammalian zygote is linked with epigenetic reprogramming. Nat Commun. 2011; 2: 241.

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