O-107 Sperm chromosome chaos induced by MEIKIN gene mutations leads to blastocyst formation failure

Abstract

Abstract Study question What are the effect and corresponding mechanisms of chromosome chaos of sperm caused by mutation of the MEIKIN gene on preimplantation embryo development? Summary answer Sperm chromosome chaos induced by MEIKIN gene mutations directly causes blastocyst formation failure by affecting the activation of the embryonic genome and lineage differentiation. What is known already In mice, Meikin is a meiosis-specific kinetochore factor that plays a crucial role in mono-orientation and protection of centromeric cohesion in meiosis I and the regulation of chromosome alignment during meiosis II. Both male and female Meikin gene knockout mice are completely infertile. Infertility in male Meikin-/- mice originates from defects in meiotic chromosome segregation and subsequent spermatogenesis. To our knowledge, no mutation of the MEIKIN gene has been reported in humans. Study design, size, duration In this study, semen from three patients with MEIKIN mutations and arrested embryos from one of them were analyzed to determine the effect of MEIKIN mutations on sperm and early embryonic development. Participants/materials, setting, methods Three male patients with MEIKIN mutations with a specific clinical phenotype were recruited. We assessed mutant sperm morphology using Papanicolaou staining. Sperm FISH and single sperm chromosome aneuploidy analysis（CNV-seq） were applied to identify mutant sperm Chromosomal abnormality. We also investigated the chromosome constitution and gene expression of the arrested embryos from one patient by parallel single-cell genome and transcriptome sequencing. Main results and the role of chance Novel mutations in the MEIKIN gene were identified in three infertility males with a specific clinical phenotype（They had good-quality embryos on day 3, but these embryos repeatedly failed to implant or develop to the blastocyst stage）. Routine semen assessments were conducted based on the WHO’s guidelines, and all these patients were diagnosed with oligoasthenoteratozoospermia. Sperm FISH showed that the frequencies of aneuploidies in spermatozoa were significantly higher than the normal control（89.05%,70.9%,93.24% vs. 5.03%）, and the CNV-seq analysis of single spermatozoa confirmed mutant sperm had severe chromosome chaos. Each cell of the arrested embryos from one patient had complicated chromosomal abnormalities, and the abnormal chromosomes were mainly derived from sperm. At the gene expression level, these embryos were arrested at the 8-cell to morula stage transition, and defects activation of a large ZGA and lineage specification genes led to blastocyst formation failure. Limitations, reasons for caution In this study, we discovered that mutations in MEIKIN caused male infertility manifesting as sperm chromosome chaos and blastocyst formation failure. However, the molecular mechanism of the sperm chromosome chaos involved in blastocyst formation needs to be further illuminated by using Meikin knockout mouse models. Wider implications of the findings We first reported that male with MEIKIN mutations is related to severe sperm chromosome abnormalities and blastocyst formation failure. Therefore, In the process of ICSI, if there are good-quality cleavage embryos that repeatedly fail to implant or develop to the blastocyst stage, it is recommended to examine male factors concurrently Trial registration number not applicable