Single-cell DNA sequencing reveals a high incidence of chromosomal abnormalities in human blastocysts

Journal of Clinical Investigation, Journal Year: 2024, Volume and Issue: 134(6)

Published: Jan. 4, 2024

Aneuploidy, a deviation from the normal chromosome copy number, is common in human embryos and considered primary cause of implantation failure early pregnancy loss. Meiotic errors lead to uniformly abnormal karyotypes, while mitotic chromosomal mosaicism: presence cells with at least two different karyotypes within an embryo. Knowledge about mosaicism blastocysts mainly derives bulk DNA sequencing multicellular trophectoderm (TE) and/or inner cell mass (ICM) samples. However, this can only detect average net gain or loss above detection threshold 20-30%. To accurately assess mosaicism, we separated TE ICM 55 good quality surplus successfully applied single-cell whole genome (scKaryo-seq) on 1057 cells. Mosaicism involving numerical structural abnormalities was detected 82% embryos, where most affected less than 20% Structural abnormalities, potentially caused by replication stress damage, were observed 69% embryos. In conclusion, our findings indicated that prevalent good-quality blastocysts, these would likely be identified as current techniques used for preimplantation genetic testing aneuploidy (PGT-A).

Language: Английский

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On the reproductive capabilities of aneuploid human preimplantation embryos

The American Journal of Human Genetics, Journal Year: 2022, Volume and Issue: 109(9), P. 1572 - 1581

Published: Sept. 1, 2022

Language: Английский

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APCAD Part 2: A Novel Method for Detection of Meiotic Aneuploidy in Preimplantation Embryos

Genes, Journal Year: 2025, Volume and Issue: 16(2), P. 115 - 115

Published: Jan. 21, 2025

Background/Objectives: Preimplantation genetic testing methods to detect aneuploidy (PGT-A) based on genomewide single nucleotide polymorphism (SNP) data were scarce and did not meet our needs. Methods: Hence, we developed a novel method for this purpose. After the raw B-allele frequency (rBAF) values of Single Nucleotide Polymorfisms (SNPs) are obtained from sample interest with SNP array, BAF specific categories SNPs (cBAF) visualized separately. Results: The analysis cBAF, rBAF Log2R profiles enables distinguish all common types chromosomal abnormalities without haplotyping. This was demonstrated by reanalyzing 359 embryos which had previously been analyzed Karyomapping. We identified additional underrepresented maternal haplotypes in five samples that could In addition, chromosomes meiotic-origin copy number gains (both parental homolog (BPH)) (n = 70) non-mosaic loss larger than 5 Mb 93) detected Conclusions: conclude proposed can be used reliably haplotyping and, hence need phasing reference.

Language: Английский

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Metabolic imaging of human embryos is predictive of ploidy status but is not associated with clinical pregnancy outcomes: a pilot trial

Human Reproduction, Journal Year: 2024, Volume and Issue: 39(3), P. 516 - 525

Published: Jan. 9, 2024

Abstract STUDY QUESTION Does fluorescence lifetime imaging microscopy (FLIM)-based metabolic assessment of human blastocysts prior to frozen transfer correlate with pregnancy outcomes? SUMMARY ANSWER FLIM failed distinguish consistent patterns in mitochondrial metabolism between leading compared those that did not. WHAT IS KNOWN ALREADY measurements provide quantitative information on NAD(P)H and flavin adenine dinucleotide (FAD+) concentrations. The embryos has long been linked their viability, suggesting the potential utility aid selection. DESIGN, SIZE, DURATION This was a pilot trial enrolling 121 IVF couples who consented have blastocyst measured using non-invasive imaging. After being warmed, 105 couples’ good-quality underwent 6-min scan controlled temperature gas environment. FLIM-assessed were then transferred without any intervention management. PARTICIPANTS/MATERIALS, SETTING, METHODS Eight parameters obtained from each (4 for 4 FAD): short lifetime, intensity, fraction molecule engaged enzyme. redox ratio (intensity NAD(P)H)/(intensity FAD) also calculated. data combined known metadata analyzed quantify ability differentiate resulted De-identified discarded aneuploid (n = 158) correlations ploidy status other factors. Statistical comparisons performed logistic regression receiver operating characteristic (ROC) curves 5-fold cross-validation averaged over 100 repeats random sampling. AUC values used classes. MAIN RESULTS AND THE ROLE OF CHANCE No showed significant differences resulting versus A produced an ROC 0.58. In contrast, robust AUCs when classifying factors such as comparison Day 5 64) 6 41) (AUC 0.78), inner cell mass trophectoderm 105: 0.88) euploid positive 108) 0.82). LIMITATIONS, REASONS FOR CAUTION study protocol not select which embryo cohort included all high quality. limited number participants sites. Increased power performing more sites may provided stronger conclusion regarding merits use clinically. WIDER IMPLICATIONS FINDINGS Blastocyst was, however, highly distinguishable. addition, regions day consistently revealed by FLIM. While detects features blastocysts, this indicates it does serve effective viability detection tool. be because plays alternative role post-implantation. FUNDING/COMPETING INTEREST(S) sponsored Optiva Fertility, Inc. Boston contributed clinical site services. Becker Hickl, GmbH, system loan. T.S. founder held stock Inc., D.S. E.S. had options during study. TRIAL REGISTRATION NUMBER approved WCG Connexus IRB (Study Number 1298156).

Language: Английский

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Previously reported and here added cases demonstrate euploid pregnancies followed by PGT-A as “mosaic” as well as “aneuploid” designated embryos

Reproductive Biology and Endocrinology, Journal Year: 2023, Volume and Issue: 21(1)

Published: March 8, 2023

Abstract Background After the longest time opposing all transfers of embryos by preimplantation genetic testing for aneuploidy (PGT-A) diagnosed as “chromosomal-abnormal,” field has over recent years slowly been moving toward selective PGT-A “mosaic” embryos, but is still rejecting defined “aneuploid.” Methods Upon review literature, we report published cases euploid pregnancies following “aneuploid” and add several additional, ongoing at our center. Results Among from center, identified seven four which preceded industry’s 2016 switch binary “euploid” – reporting to “euploid,” “mosaic,” reporting. That those post definition involving therefore, cannot be ruled out. Since then, recently established three additional await confirmation euploidy after delivery. A fourth pregnancy transfer a trisomy 9 embryo miscarried before fetal heart. Outside own center’s experience, literature revealed only one such transfer, “chaotic-aneuploid” with six abnormalities, leading normal In reviewing furthermore demonstrate why current that differentiates between based on relative percentages aneuploid DNA in single trophectoderm biopsy average 5-6 cells, biologically non-sensical. Conclusion Basic biological evidence clinically very limited experience labeled beyond reasonable doubt least some can lead healthy births. Therefore, this observation establishes rejection reduces live birth chances IVF patients. Whether (and what possible degree) differ remains determined. The answer will likely depend aneuploidy(ies) an degree “mosaicism” single, 5/6-cell reflect ploidy-status complete embryo.

Language: Английский

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Implicit bias in diagnosing mosaicism amongst preimplantation genetic testing providers: results from a multicenter study of 36 395 blastocysts

Human Reproduction, Journal Year: 2023, Volume and Issue: 39(1), P. 258 - 274

Published: Oct. 23, 2023

Abstract STUDY QUESTION Does the diagnosis of mosaicism affect ploidy rates across different providers offering preimplantation genetic testing for aneuploidies (PGT-A)? SUMMARY ANSWER Our analysis 36 395 blastocyst biopsies eight laboratories revealed that euploidy were significantly higher in reporting low mosaicism. WHAT IS KNOWN ALREADY Diagnoses consistent with chromosomal have emerged as a third category possible embryo outcomes following PGT-A. However, era mosaicism, selection has become increasingly complex. Biological, technical, analytical, and clinical complexities interpreting such results led to substantial variability PGT-A clinics. Critically, it remains unknown whether these differences impact number euploid embryos available transfer. Ultimately, this may outcomes, important implications patients. DESIGN, SIZE, DURATION In international, multicenter cohort study, we reviewed consecutive results, obtained from 10 035 patients 11 867 treatment cycles, conducted between October 2015 2021. A total 17 IVF centers, providers, five countries three continents participated study. All blastocysts tested using trophectoderm biopsy next-generation sequencing. Both autologous donation cycles assessed. Cycles structural rearrangements excluded analysis. PARTICIPANTS/MATERIALS, SETTING, METHODS The randomly categorized (A H). Providers B, C, D, E, F, G, H all reported whereas Provider either or aneuploid. Ploidy analyzed multilevel mixed linear regression. Analyses adjusted maternal age, paternal oocyte source, biopsied, day biopsy, provider, appropriate. We compared associations including chromosomally normal (euploid) determined be suitable MAIN RESULTS AND THE ROLE OF CHANCE mean age (±SD) was 36.2 (±5.2). findings reveal strong association provider Amongst seven varied 3.1% 25.0%. After adjusting confounders, observed significant difference likelihood diagnosing (P < 0.001), ranging 6.5% (95% CI: 5.2–7.4%) B 35.6% 32.6–38.7%) E. Notably, highest lowest (Provider B: euploidy, 55.7% 54.1–57.4%), 5.2–7.4%); H: 44.5% 43.6–45.4%), 9.9% 9.2–10.6%)); D: 43.8% 39.2–48.4%), 11.0% 7.5–14.5%)). Moreover, overall chance having at least one transfer when not reported, other (OR = 1.30, 95% 1.13–1.50). Differences mosaic raise further concerns regarding accuracy relevance predictions. While confirmed equivalent blastocysts, found proportion are used treatment. LIMITATIONS, REASONS FOR CAUTION Due retrospective nature can ascertained, however, causality cannot established. Certain parameters grade dataset. Furthermore, certain platform-related clinic-specific factors readily quantifiable explicitly captured our As such, full elucidation potential confounders accounting possible. WIDER IMPLICATIONS FINDINGS highlight need standardization quality assurance industry. decision ultimately reduce success cycle by limiting pool embryos. Until diagnoses accurately reflect biological variability, warrants utmost caution. prudent approach is imperative, determine failure some FUNDING/COMPETING INTEREST(S) This work supported Torres Quevedo Grant, awarded M.P. (PTQ2019-010494) Spanish State Research Agency, Ministry Science Innovation, Spain. M.P., L.B., A.R.L., A.L.R.d.C.L., N.P.P., D.S., F.A., A.P., B.M., L.D., F.V.M., M.R., E.P.d.l.B., A.R., R.V. no competing interests declare. B.L., R.M., J.A.O. time employees IB Biotech, genetics company Instituto Bernabeu group, which performs testing. M.G. employee Novagen, Cegyr, TRIAL REGISTRATION NUMBER N/A.

Language: Английский

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A multi-omics genome-and-transcriptome single-cell atlas of human preimplantation embryogenesis reveals the cellular and molecular impact of chromosome instability

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: March 10, 2023

ABSTRACT The frequent acquisition of genomic abnormalities in human preimplantation embryos is a leading cause pregnancy loss, but does not necessarily prohibit healthy offspring. However, the impact on cellular states and development early embryo remains largely unclear. Here, we characterise aneuploidy reconstruct gene regulatory networks embryos, investigate expression developmental perturbations instigated by using single-cell genome-and-transcriptome sequencing (G&T-seq). At level, show that acquired numerical structural chromosomal aberrations are across all stages embryogenesis cell lineages. transcriptome identify regulators identity uncover network 248 transcription factors from 10 major modules distinct lineages embryos. By integrating DNA-with RNA-information, unveil how levels affected losses or gains corresponding genes embryonic cells development, as well copy-number aberrant factor perturb their cognate target euploid regions. Furthermore, reveal majority aneuploid delay reduced fitness, indicating competition within mosaic diploid-aneuploid embryo, which may contribute to selection against birth offspring In summary, our multi-modal analyses provide unprecedented insights into development.

Language: Английский

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Diagnostic or prognostic? Decoding the role of embryo selection on in vitro fertilization treatment outcomes

Fertility and Sterility, Journal Year: 2024, Volume and Issue: 121(5), P. 730 - 736

Published: Jan. 5, 2024

Language: Английский

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Double strand DNA breaks in sperm: the bad guy in the crowd

Journal of Assisted Reproduction and Genetics, Journal Year: 2023, Volume and Issue: 40(4), P. 745 - 751

Published: Feb. 24, 2023

Language: Английский

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Effectiveness and safety of consecutive single embryo transfer compared to double embryo transfer: results from the UK HFEA registry

Human Reproduction, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 14, 2025

Abstract STUDY QUESTION How does two-consecutive single embryo transfer (2xSET) affect reproductive outcomes of IVF and ICSI compared to double (DET)? SUMMARY ANSWER Two-consecutive SET may provide greater or comparable live birth rate (LBR); with lower multiple birth, preterm pregnancy loss neonatal death rates DET. WHAT IS KNOWN ALREADY Elective in IVF/ICSI is widely encouraged over DET minimize the risk births associated morbidities. Despite this, following remain higher than 10% target across Europe USA. Currently, majority evidence regarding based on various studies assessing such as LBR per treatment cycle, opposed oocyte retrieval. As such, representation mostly unfavourable. Analysis cumulative two embryos consecutive cycles, rather one event (DET) more effective at distinguishing methods will therefore valuable information relevant clinical practice. DESIGN, SIZE, DURATION This retrospective cohort study was conducted using Human Fertilisation Embryology Authority (HFEA) register data, which encompasses national data from all clinics UK. All women who underwent their first retrieval cycle subsequent SET, DET, 2xSET between 2010 2019 blastocyst were included (N = 71 807). PARTICIPANTS/MATERIALS, SETTING, METHODS The liveborn baby rate, pregnancies blastocyst-stage embryos, where stratified by maternal age. Data analysis RStudio v4.2, alpha equals 0.05. MAIN RESULTS AND THE ROLE OF CHANCE Blastocyst-stage achieved a median 0.47 (interquartile range [IQR] 0.13) 0.41 (IQR 0.13), 0.38 (P < 0.05). Using reference standard, significantly odds ((odds ratio [OR] 6.87, 95% CI 6.14–7.68) vs 28.20, 25.20–31.57). also (OR 1.11, 1.06–1.15) 2.80, 2.67–2.94). Similarly, 1.14, 1.08–1.21) 2.11, 1.98–2.24). consistently aged 39 years under However, results > Across age groups, had highest In under, 0.05), whereas cohorts Moreover, 37 LIMITATIONS, REASONS FOR CAUTION Certain confounders are not recorded within HFEA registry including patient BMI, evaluation quality, endometrial thickness transfer. Consequently, while our identifies broad trends success morbidity, differ certain populations. WIDER IMPLICATIONS FINDINGS old, overall morbidity should be considered first-line among cohorts, advanced improve reduce ART. FUNDING/COMPETING INTEREST(S) No external funding used for this study. None authors has any conflicts interest. TRIAL REGISTRATION NUMBER did require registration. Following consultation Institutional Review Board Imperial College London, ethical approval deemed necessary.

Language: Английский

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