A copy number variation genotyping method for aneuploidy detection in spontaneous abortion specimens

Chen, Songchang; Liu, Deyuan; Zhang, Junyu; Li, Shuyuan; Zhang, Lanlan; Fan, Jianxia; Luo, Yuqin; Qian, Yeqing; Huang, He‐Feng; Liu, Chao; Zhu, Huanhuan; Jiang, Zhengwen; Xu, Chenming

doi:10.1002/pd.4986

Cited by 21 publications

(39 citation statements)

References 25 publications

(42 reference statements)

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“…An abundance of reports into human miscarriages cite aneuploidy as the greatest cause of the pregnancy failure 17,39 . In contrast, reports of aneuploidy associated with pregnancy loss in domesticated animals is rarely reported.…”

Section: Discussionmentioning

confidence: 99%

Whole genome analysis reveals aneuploidies in early pregnancy loss in the horse

Shilton

Kahler

Davis

et al. 2020

Sci Rep

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The first 8 weeks of pregnancy is a critical time, with the majority of pregnancy losses occurring during this period. Abnormal chromosome number (aneuploidy) is a common finding in human miscarriage, yet is rarely reported in domestic animals. equine early pregnancy loss (epL) has no diagnosis in over 80% of cases. The aim of this study was to characterise aneuploidies associated with equine EPL. Genomic DNA from clinical cases of spontaneous miscarriage (EPLs; 14-65 days of gestation) and healthy control placentae (various gestational ages) were assessed using a high density genotyping array. Aneuploidy was detected in 12/55 EPLs (21.8%), and 0/15 healthy control placentae. Whole genome sequencing (30X) and digital droplet PCR (ddPCR) validated results. The majority of these aneuploidies have never been reported in live born equines, supporting their embryonic/fetal lethality. Aneuploidies were detected in both placental and fetal compartments. Rodents are currently used to study how maternal ageing impacts aneuploidy risk, however the differences in reproductive biology is a limitation of this model. We present the first evidence of aneuploidy in naturally occurring equine EPLs at a similar rate to human miscarriage. We therefore suggest the horse as an alternative to rodent models to study mechanisms resulting in aneuploid pregnancies.

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Section: Discussionmentioning

confidence: 99%

Whole genome analysis reveals aneuploidies in early pregnancy loss in the horse

Shilton

Kahler

Davis

et al. 2020

Sci Rep

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“…An abundance of reports into human miscarriages, cite aneuploidy as the greatest cause of the pregnancy failure [20, 21]. In contrast, reports of aneuploidy associated with pregnancy loss in domesticated animals is rarely reported, and never in the horse.…”

Section: Discussionmentioning

confidence: 99%

“…Aneuploidy (gain or loss of a whole chromosome) is well documented in human spontaneous abortion [20, 21] and is associated with advancing maternal age. Some human autosomal trisomies (13, 18, and 21) have been identified in infants following live births, although these usually present with significant developmental pathologies [22] with only trisomy 21 individuals surviving to adulthood [23], and no documented surviving monosomies.…”

Section: Introductionmentioning

confidence: 99%

Whole genome analysis reveals aneuploidies in early pregnancy loss in the horse

Shilton

Kahler

Davis

et al. 2020

Preprint

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Chromosome abnormalities are well documented in human spontaneous abortion studies, yet rarely reported in domesticated animals. Rodent models have previously been used to study the effects of maternal ageing on oocyte quality and ultimately aneuploidy, however the differing endocrine profiles, oocyte characteristics and the polytocous nature of rodents are limitations for translation into human medicine. Early Pregnancy Loss (EPL) occurs in 5-10% of confirmed equine pregnancies and has no diagnosis in over 80% of cases. Aneuploidy has never been described in equine pregnancy loss, thus the objectives of this study were to quantify the frequency and characteristics of aneuploidy associated with equine EPL. EPL conceptuses were submitted from clinical cases of spontaneous pregnancy loss (14-65 days of gestation) between 2013 and 2018. Age matched control conceptuses were obtained from terminated clinically normal pregnancies (CNP). Aneuploidy was detected in 12/55 EPLs (21.8%), 0/10 CNP, 0/5 healthy term chorioallantois, and 0/5 healthy adult mares via genotyping. Whole genome sequencing (30X) and ddPCR validated results. Aneuploidies involved 10/32 equine chromosomes, consisting of nine trisomies and three monosomies. Autosomal aneuploidies were detected in both placental and fetal compartments in all samples tested. Aneuploid types (7/9) were mostly unique to EPL, supporting their embryonic/fetal lethality. Presenting the first evidence of aneuploidies in failed equine pregnancies not only provides the initial step in identifying genetic causes for these early losses, but also offers the horse as a new model for studying naturally occurring aneuploidy. We also demonstrate that SNP arrays provide a simple, cost effective way to screen aneuploidies across a large population.Author SummaryThe first 8 weeks of pregnancy is a critical time in both humans and horses, as the majority of pregnancy losses occur during this period. Despite such high prevalence, many cases do not have a known cause. Abnormal chromosome number (aneuploidy) is the most common finding in human pregnancy loss studies, but to date no equivalent study has been performed in domesticated animals, including the horse. We studied the genetics of naturally occurring pregnancy losses from Thoroughbred horses and found a similar level of aneuploidy to that observed in women. As humans and horses share similarities in their reproductive biology (ageing eggs, increased pregnancy loss in older mothers, similar key hormones), we suggest that by comparing the genetics of these two species, greater advances in identifying causes of aneuploidy pregnancy can be reached. Thoroughbred horses also tend to be more inbred than humans, facilitating the identification of mutations that increase the chance of aneuploidy, and this knowledge could potentially be applied in human medicine, as well as in species conservation.

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“…Then, universal primers labeled with uorescent markers were used to amplify the concatenate products by PCR. After ampli cation, the PCR products were separated and detected by uorescence capillary electrophoresis, then the copy number of target regions were calculated by analyzing the peak height of electrophoretogram, the detailed work ow refer to the study Chen, S et al reported 14 . There were totally 170 (for Chr1-12 and Chr16-17, there 8 probes for each chromosome; for Chr13-15, Chr21-22 and ChrY, there 5 probes for each chromosome; for Chr18-Chr20 and ChrX, there 7 probes for each chromosome) pairs of probes targeting 24 chromosomes were designed for the aneuploidy detection.…”

Section: Aneuploidy Detection By Hlpamentioning

confidence: 99%

Aneuploid Abortion Positively Correlate With MAD1 Overexpression and mir125b Depression

Zhao¹,

Li²,

Li³

et al. 2020

Preprint

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Background: Aneuploid is the most frequent cause of early embryo abortion, and any defect in chromosome segregation would fail to satisfy spindle assembly checkpoint (SAC) during mitosis, which could lead to the halted metaphase and aneuploid occurrence. Mitotic checkpoint complex(MCC), a complex compound of MAD1、MAD2、Cdc20、BUBR1 and BUB3, plays an important role in SAC activation. Studies have confirmed that the overexpression of MAD2 and BUBR1 can facilitate the correct chromosome segregation and embryo stability. Research identifications also proved that miR-125b negatively regulated MAD1 expression by binding to its 3’UTR. However, the expression of mir125b, MAD1 and BUB3 genes in aneuploidy embryos of spontaneous abortion has not been reported.Methods: In this study, embryonic villi from miscarriage pregnant women were collected and divided into two groups (aneuploidy and euploidy) by HLPA and FISH analysis. The RNA levels of mir125b, MAD1 and BUB3 were detected through QRT-PCR, while Western blot was further used to analyze the protein levels of MAD1 and BUB3.Results: SPSS 17.0 statistical analysis(P<0.05) showed that mir125b and BUB3 were significantly down-regulated in aneuploidy group compared to the control group, MAD1 was significantly up-regulated in RNA level; Additionally, MAD1 protein level was also significantly higher while BUB3 was mildly increased in aneuploidy abortion villus. Correlation analysis revealed that the expression of MAD1 was negatively correlated with Mir125b.Conclusion: these results suggested that aneuploid abortion was positively correlated with MAD1 overexpression which might be caused by insufficient mir125b.

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A copy number variation genotyping method for aneuploidy detection in spontaneous abortion specimens

Abstract: High-throughput ligation-dependent probe amplification is a rapid and accurate method for aneuploidy detection. It can be used as a cost-effective screening procedure in clinical spontaneous abortions. © 2016 John Wiley & Sons, Ltd.

Cited by 21 publications

References 25 publications

Whole genome analysis reveals aneuploidies in early pregnancy loss in the horse

Whole genome analysis reveals aneuploidies in early pregnancy loss in the horse

Whole genome analysis reveals aneuploidies in early pregnancy loss in the horse

Aneuploid Abortion Positively Correlate With MAD1 Overexpression and mir125b Depression

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