Preimplantation Genetic Testing For Aneuploidy Results Research Articles

The Association between Embryo Development and Chromosomal Results from PGT-A in Women of Advanced Age: A Prospective Cohort Study.

Embryo morphology and morphokinetics have been studied for their association with euploid embryos. However, the results are controversial, especially in the advanced-aged women group, when the risk of aneuploidy increases significantly. This prospective cohort study evaluated the association between embryo development between day-3 cleavage and day-5 blastocyst stages and euploidy rates, determined using preimplantation genetic testing for aneuploidy (PGT-A). Embryos from women aged 35 years and above who underwent intracytoplasmic sperm injections and PGT-A were studied. Day-3 cleavage-stage embryos were evaluated for their cell number, and day-5 blastocyst-stage embryos were evaluated for their morphological grade. Embryo development from day 3 to day 5 was categorized as either good or poor development and evaluated for its association with the PGT-A results. We evaluated 325 embryos from 101 infertile couples. It was found that 55.17% of blastocysts with good development and 29.83% with poor development were euploid. A significant association was found between embryo development and euploidy rates in advanced-aged women (p < 0.001). Also, there were significantly higher rates of euploid embryos with good blastocyst morphological grades, especially blastocyst expansion grades and trophectoderm grades. In conclusion, embryo morphokinetics shows promising results in predicting euploidy in advanced female age.

P-132 Does the automated blastocyst assessment system, iDAScore®, provide valuable data for predicting the results of preimplantation genetic testing for aneuploidy (PGT-A)?

Abstract Study question What iDAScore® could indicate the presence of euploidy in PGT-A? Is it a valid predictor of the result of PGT-A? Summary answer PGT-A results with an iDAScore® of 8.3 or higher can indicate euploidy. However, patient age and trophectoderm (TE) grade can be effective predictors as well. What is known already PGT-A was approved as a general test in Japan in 2022. Currently, only patients who meet the criteria of Japan Society of Obstetrics and Gynecology can undergo the test. It is expected that the test will improve the live birth rate and reduce the miscarriage rate. The iDAScore® uses artificial intelligence and deep learning to score blastocysts. It is anticipated to be more objective than is morphological evaluation, which varies among individuals. Unlike PGT-A, the iDAScore® can assess embryos without cellular invasion. However, only a few reports have investigated the association between PGT-A and iDAScore® results. Study design, size, duration The study was performed retrospectively on 3,781 cycles of in vitro fertilization (IVF) conducted at our clinic between August 2020 and October 2022. Of the total of 5,684 embryos derived from conventional IVF or intracytoplasmic sperm injection that were cultured in a time lapse incubator, 293 blastocysts were used in the study. Participants/materials, setting, methods On the basis of their PGT-A results, the blastocysts were divided into euploidy and aneuploidy groups. The mean age of patients in each group, the methods of insemination, and the median iDAScore® were evaluated. The area under the curve (AUC) and the cut-off value were calculated from the PGT-A and iDAScore® results. A logistic regression analysis of the predictive parameters of PGT-A results, including the iDAScore®, was also performed. Main results and the role of chance Patients’ mean ages in the euploidy and aneuploidy groups were 38.5 and 41.5 years, respectively. This was significantly higher in the aneuploidy group (P&amp;lt;0.001). There was no significant difference in the stage of maturity at oocyte retrieval (P = 0.302). Similarly, no significant differences were associated with method of insemination (P = 0.961). The median iDAScore® for the euploidy group was 8.7; for the aneuploidy group, it was 7.8. This result was significantly higher in the euploidy group (P&amp;lt;0.001). The AUC, calculated from PGT-A results and the iDAScore® with receiver operating characteristic curves, was 0.685 (95% CI [0.617−0.754]). The cut-off value was 8.3. The logistic regression analysis of age, stage of maturity, insemination method, timing of blastocyst freezing, diameter before blastocyst freezing, inner cell mass, TE grade in the Gardner classification, and iDAScore® as parameters for the PGT-A results indicated significant differences for age and TE grade (age: P&amp;lt;0.001, odds ratio [95%CI]: 1.320 [1.19-1.45]; TE grade 2: P = 0.03, odds ratio [95%CI]: 2.480 [1.11-5.57]; TE grade 3: P = 0.02, odds ratio [95%CI]: 3.740 [1.27-11.00]), while no significant differences were associated with the iDAScore®. Limitations, reasons for caution The iDAScore® is likely to depend on the timing of blastocyst freezing. Furthermore, the iDAScore® utilised in this study was the score of blastocysts that fulfilled our freezing criteria; the blastocyst that reaches a size of at least 150-160 µm in diameter on Day 5,6, or 7 is frozen. Wider implications of the findings An iDAScore® of 8.3 or above could indicate a euploidy PGT-A result. However, the logistic regression analysis indicated that age and TE grade were more significant predictors than the iDAScore® was. Therefore, it is difficult to predict PGT-A results from the iDAScore® itself. Trial registration number Not applicable

P-264 Correlations between the artificial intelligence scoring system (iDAScore v1.0) and live birth outcomes in preimplantation genetic testing for aneuploidy cycles

Abstract Study question Is the iDAScore v1.0 score correlated with pregnancy outcomes in single embryo transfer (SET) cycles following preimplantation genetic testing for aneuploidy (PGT-A)? Summary answer We demonstrated that elevated iDAScore v1.0 scores are positively correlated with the probabilities of pregnancy and live birth (LB) in SETs following PGT-A. What is known already To select the blastocysts with high development potential in advance, several groups have proposed the implementation of artificial intelligence (AI)-based software is capable of predicting implantation in women undergoing in vitro fertilization (IVF). It has also been known that using high resolution next generation sequencing preimplantation genetic screening (hr-NGS) for PGT-A can enable the exclusion of aneuploid embryos before embryo transfers and thus improve pregnancy outcomes of SETs. However, the capacity of AI-based assessments has remained unclear in IVF cycles with PGT-A. Study design, size, duration This retrospective study, approved by the Institutional Review Board of Chung Sun Medical University, was performed to assess the dataset of 317 SET cycles after PGT-A from January 2017 to September 2019. A single euploid or mosaic blastocyst with an AI score calculated using the iDAScore software (v1.0) was selected for frozen embryo transfer according to the given priority of blastocyst morphology. The iDAScore groups were categorized by quartiles of AI scores. Participants/materials, setting, methods Embryos were cultured in a time-lapse incubator and qualified blastocysts underwent next generation sequencing (NGS)-based PGT-A. The confounding factors associated with LB were evaluated using logistic regression analysis in generalized estimating equations (GEEs). The differences between iDAScore groups were assessed using the Mann–Whitney U test or Fisher’s exact test, as applicable. The receiver operating characteristic (ROC) curve analysis was used to estimate the predictive powers. A P value &amp;lt; 0.05 was considered statistically significant. Main results and the role of chance The results revealed the patient age, anti-Müllerian hormone, body mass index, oocyte sources (autologous or donor), PGT-A results (euploidy, low-level mosaicism, or high-level mosaicism), aberrant chromosome types (none, whole chromosome, segmental chromosome, or whole with segmental chromosome), and aberrant chromosome sites (0, 1, 2, or &amp;gt; 2) were not correlated with LB. However, embryo biopsy days (day 5 vs. day 6, OR = 2.271, 95% CI = 1.534–14.824), blastocyst morphology scores (OR = 1.185, 95% CI = 1.034–1.359), and AI scores (OR = 1.640, 95% CI = 1.231–2.183) were significantly correlated with LB. The AI scores were then divided into quartiles (group 1: 5.0–7.9; group 2: 8.0–8.5; group 3: 8.6–8.9; and group 4: 9.0–9.5). The ongoing pregnancy (29.1% vs. 54.2%–57.5%) and live birth (27.8% vs. 52.8%–56.3%) rates of group 1 were significantly lower than other groups. The blastocyst morphology score (5.3 ± 1.2), and KIDScore D5 scores (v1 = 3.0 ± 1.7; v3 = 4.6 ± 1.5) were also lowest in group 1. The ROC curve analysis confirmed a significant but limited LB prediction capability for iDAScore v1.0 (AUC = 0.6), which was similar to the KIDScore D5 (AUC = 0.61–0.62). Limitations, reasons for caution Because of the retrospective nature, the major limitation was the lack of randomization, which may present a risk of selection bias. According to the criteria of blastocyst selection for SETs, the embryos with low (&amp;lt; 5.0) or high (&amp;gt; 9.5) AI scores were not present in this dataset. Wider implications of the findings Although prediction capability of iDAScore remains perfectible, the AI score is still significantly associated with LB probability. Euploid or mosaic blastocysts with low AI scores (&amp;lt;8.0) processed a decreased LB rate, indicating the potential of annotation-free iDAScore system as a decision support tool for deselecting embryos with poor post-implantation development. Trial registration number not applicable

Comparison of clinical outcome between day 5 and day 6 single blastocyst transfers in cycles undergoing preimplantation genetic testing for aneuploidy

ObjectiveIn vitro fertilization (IVF) treatment has gradually adopted the practice of culturing embryos until the blastocyst stage on the D5 or D6 as the standard approach. PGT-A is commonly used in vitro fertilization (IVF). This study aimed to evaluate the clinical outcomes of frozen embryo transfers (FETs) using single blastocyst transfers (SBTs) on the fifth (D5) or sixth (D6) day of development in cycles that underwent preimplantation genetic testing for aneuploidy (PGT-A). Materials and methodsThe patients who had at least one euploid or mosaic blastocyst of good quality determined by PGT-A results and received single embryo transfer (SET) cycles were included in the study. In this study, the live birth rate (LBR) and neonatal outcomes were compared after the transfer of single biopsied D5 and D6 blastocysts in frozen embryo transfer (FET) cycles. ResultsA total of 527 frozen-thawed blastocyst transfer (FET) cycles (8449 biopsied embryos were analyzed). No significant difference in the implantation rate (IR), clinical pregnancy rate (CPR), and live birth rate (LBR) between the transfers of D5 and D6 blastocysts. Birth weight was the only perinatal outcome that showed a significant difference between the D5 and D6 groups. ConclusionThe study confirmed that the transfer of a single euploid or mosaic blastocyst, regardless of whether it was on the fifth (D5) or sixth (D6) day of development, can lead to promising clinical results.

A non-invasive artificial intelligence approach for the prediction of human blastocyst ploidy: a retrospective model development and validation study

One challenge in the field of in-vitro fertilisation is the selection of the most viable embryos for transfer. Morphological quality assessment and morphokinetic analysis both have the disadvantage of intra-observer and inter-observer variability. A third method, preimplantation genetic testing for aneuploidy (PGT-A), has limitations too, including its invasiveness and cost. We hypothesised that differences in aneuploid and euploid embryos that allow for model-based classification are reflected in morphology, morphokinetics, and associated clinical information. In this retrospective study, we used machine-learning and deep-learning approaches to develop STORK-A, a non-invasive and automated method of embryo evaluation that uses artificial intelligence to predict embryo ploidy status. Our method used a dataset of 10 378 embryos that consisted of static images captured at 110 h after intracytoplasmic sperm injection, morphokinetic parameters, blastocyst morphological assessments, maternal age, and ploidy status. Independent and external datasets, Weill Cornell Medicine EmbryoScope+ (WCM-ES+; Weill Cornell Medicine Center of Reproductive Medicine, NY, USA) and IVI Valencia (IVI Valencia, Health Research Institute la Fe, Valencia, Spain) were used to test the generalisability of STORK-A and were compared measuring accuracy and area under the receiver operating characteristic curve (AUC). Analysis and model development included the use of 10 378 embryos, all with PGT-A results, from 1385 patients (maternal age range 21-48 years; mean age 36·98 years [SD 4·62]). STORK-A predicted aneuploid versus euploid embryos with an accuracy of 69·3% (95% CI 66·9-71·5; AUC 0·761; positive predictive value [PPV] 76·1%; negative predictive value [NPV] 62·1%) when using images, maternal age, morphokinetics, and blastocyst score. A second classification task trained to predict complex aneuploidy versus euploidy and single aneuploidy produced an accuracy of 74·0% (95% CI 71·7-76·1; AUC 0·760; PPV 54·9%; NPV 87·6%) using an image, maternal age, morphokinetic parameters, and blastocyst grade. A third classification task trained to predict complex aneuploidy versus euploidy had an accuracy of 77·6% (95% CI 75·0-80·0; AUC 0·847; PPV 76·7%; NPV 78·0%). STORK-A reported accuracies of 63·4% (AUC 0·702) on the WCM-ES+ dataset and 65·7% (AUC 0·715) on the IVI Valencia dataset, when using an image, maternal age, and morphokinetic parameters, similar to the STORK-A test dataset accuracy of 67·8% (AUC 0·737), showing generalisability. As a proof of concept, STORK-A shows an ability to predict embryo ploidy in a non-invasive manner and shows future potential as a standardised supplementation to traditional methods of embryo selection and prioritisation for implantation or recommendation for PGT-A. US National Institutes of Health.

O-176 End-to-end deep learning system for recognition of euploid and aneuploid embryos using time-lapse videos

Abstract Study question Can an Artificial Intelligence (AI) system based on a deep learning algorithm analyze time-lapse videos for ploidy status prediction? Summary answer Our spatiotemporal model can distinguish aneuploid embryos from euploid embryos using time-lapse videos from 10 to 115 hours post-insemination (hpi) with an accuracy of 71,28%. What is known already As the maternal age advances chances of aneuploidy in oocyte also increases and there is a high chance of early termination of pregnancy. Pre-implantation genetic testing for aneuploidy (PGT-A) is a reliable tool for detecting chromosomal status. However, PGT-A is an invasive technique in which protocol requires an embryo biopsy. Continuous monitoring of embryo development led to AI models for the prediction of ploidy based on blastocyst images or morphokinetic parameters. Previous publications showed that euploid embryos reach blastulation earlier than non-euploid embryos. This is the first attempt to predict ploidy by analyzing continuous embryo development through captured time-lapse images. Study design, size, duration The present study consisted of a single-center retrospective analysis for the evaluation of ploidy status with a non-invasive method. We developed our models based on a balanced dataset of 940 videos (from 10 to 115 hpi) extracted from the EmbryoScope time-lapse system. All the videos were divided into 90% for training and validating and 10% for testing. The target class for the predicted models was the results of PGT-A on blastocyst by next-generation sequencing. Participants/materials, setting, methods We used an end-to-end approach to develop an automated AI system capable of extracting features from images and classifying them considering temporal dependencies. First, a convolutional neural network (CNN) extracted the most relevant features from each frame. We used a deep architecture known as ResNet50. Second, a bidirectional long short-term memory (LSTM) layer received this information and analyzed temporal dependencies, obtaining a low-dimensional feature vector that defined each video. Finally, a multilayer perceptron classified them. Main results and the role of chance Euploid and aneuploid precision was 69% and 75%, respectively. Euploid and aneuploid sensitivity was 79% and 64%, respectively. Euploid and aneuploid F1 score was 73% and 69% respectively. The global accuracy associated with our spatiotemporal model to differentiate between the two classes achieved 71,28% on this dataset. Additionally, we trained models with external information such as maternal age (38,3±3,9 versus 39,1±3,1), but the performance did not improve. Note that we did not apply a prior selection of good quality videos to study more reliably the possible inclusion of an AI model for chromosomal status analysis in clinical practice. Limitations, reasons for caution The main limitation of this study is the single-center retrospective approach and the reduced size of the database, therefore future prospective research would improve model performance. However, the preliminary results showed the high potential of the methods. Wider implications of the findings Our results showed potential automation of chromosomic status evaluation. Our findings led to a possible non-invasive method and the research of new unknown key factors for determining ploidy. Further studies with a large number of time-lapse videos could result in a potential translation to clinical use. Trial registration number not applicable

Development of an artificial intelligence model for predicting the likelihood of human embryo euploidy based on blastocyst images from multiple imaging systems during IVF.

Development of an artificial intelligence model for predicting the likelihood of human embryo euploidy based on blastocyst images from multiple imaging systems during IVF.

Comparison of day 5 blastocyst with day 6 blastocyst: Evidence from NGS-based PGT-A results.

Aneuploidy is one genetic factor leading to the failure of embryo implantation. This study aimed to investigate the relationship between the day of embryo blastulation and the ploidy status of embryo, to aid in selecting embryos with the most likelihood of of beingeuploid in anoninvasive way. This retrospective study recruited women undergoing preimplantation genetic testing (PGT) for aneuploidy (PGT-A) with trophectoderm biopsy from January 2019 to December 2020. The ploidy status of embryos was determined by next-generation sequencing (NGS). Altogether, 2531 blastocysts from 839 PGT-A cycles were evaluated. The euploid rate of day 5 blastocysts was significantly higher than that of day 6 blastocysts, either from the same ovarian stimulation (OS) cycles (49.9% vs 35.7%, P < 0.001) or from different OS cycles (48.2% vs 27.8%, P < 0.001). This effect of increasing time to embryo blastulation significantly reducing the prevalence of euploidy was not seen in women at age 38 or older. However, after single euploid embryo transfer, the clinical outcomes of day 5 blastocysts were comparable to those of day 6 blastocysts. In non-PGT cycles, our data support the selection of day 5 blastocysts for transfer over day 6 blastocysts. Further, women with poor ovarian reserve incline to obtain only day 6 blastocysts, and PGT-A is valuable for identifying the ploidy status of embryos, especially for those with advanced age. However, the stage of blastocysts will not affect the clinical outcome after transfer when they are identified as euploid.

THE ASSOCIATION BETWEEN SELF-REPORTED RACE AND EMBRYONIC MOSAICISM

There is evidence to suggest that race is associated with IVF outcomes; yet, little is known about its relationship with embryonic mosaicism. Our objective is to explore the relationship between self-reported race and embryonic aneuploidy and mosaicism for patients undergoing in vitro fertilization (IVF) with preimplantation genetic testing for aneuploidy (PGT-A). This was a retrospective, chart review study of patients at a tertiary care academic medical center with self-reported race data who underwent IVF with PGT-A from 2019-2020. Race was classified as Caucasian (C), Asian (A), African American (AA), or Multiracial/Other (M). PGT-A was performed at an external laboratory and the following numbers were reported for each cycle: embryos diagnosed, euploid embryos, aneuploid embryos, high (HL)/low (LL) level mosaic embryos, and whole or segmental mosaic embryos. After adjusting for age, the association between self-reported race and PGT-A results were analyzed using chi-square tests and regression modeling by Poisson distributions with p<0.05 to be considered significant Of the 493 patients with documented self-reported race who underwent IVF with PGT-A, 309 (62.7%) classified as C, 57 (11.6%) as A, 36 (7.3%) as AA, and 82 (16.6%) as M. A total of 2,470 embryos analyzed by PGT-A were included, with a mean number of 5.15±3.50 embryos diagnosed per patient. After adjusting for age, there was no statistically significant association between race and HL (p=0.27) or LL (p=0.17) mosaicism and whole (HL, p=0.48, LL, p=0.63) or segmental (HL, p=0.75, LL, p=0.08) mosaicism. Additionally, there was no difference in number of embryos biopsied, number of euploid embryos, or number of aneuploid embryos by race (Table 1). This is the first study to explore the association between self-reported race and embryonic mosaicism. Our findings do not support a significant relationship between race and embryonic mosaicism. Future research should employ more diverse patient populations to clarify potential relationships.

RECURRING, UNEXPLAINED SEGMENTAL ANEUPLOIDIES OBSERVED ON PGT-A WITH BREAKPOINTS THAT CORRESPOND TO KNOWN FRAGILE SITES

To present a collection of cases involving recurring, terminal, segmental aneuploidies (RTSAs) observed in preimplantation genetic testing for aneuploidy (PGT-A) that cannot be explained by inheritance or a balanced translocation in a carrier parent. Interestingly, the breakpoints of these unexplained RTSAs correspond to known fragile sites. Our objective is to open discussion about the potential significance of fragile sites for infertile patients pursuing in vitro fertilization (IVF). PGT-A results were reviewed by Igenomix, a reference laboratory, for potential inherited chromosomal rearrangements. The criteria include observing at least two embryos with a terminal segmental aneuploidy occurring at the same breakpoint (1). Karyotyping was recommended for the reproductive couple. When only a single RTSA was observed, instead of the pair typically observed in translocation cases, we compared to known fragile sites (2). Clinical testing to confirm the presence of a fragile site is not available. Thirteen cases were identified out of approximately 66,000 (Table 1). Eight reproductive couples underwent karyotype analysis and were negative for translocations and inversions. One patient was lost to follow up and the remaining four karyotypes are pending.Tabled 1Fragile SiteEmbryos Affected/TotalPt AgeAdd'l Repro HxKaryotypeFRA10A (10q23.3)2/434Lost3/8N/PEgg donorNeg3/154015 unsuccessful IVF cyclesNegFRA7A (7p11.2)2/1030Hx of testicular cancerNeg3/840Neg2/1039NegFRA13C/FRA13B (13q21)2/2423/831NegFRA9A/FRA9C (9p21.2)2/1538FRA5F (5q21.2)6/1235FRA5A (5p13.3)2/636NegFRA4B (4q12)2/635FRA11I/FRA11C (11p15.5)2/339Neg Open table in a new tab RTSAs occurring at known fragile sites can be observed in embryos. Such RTSAs can be observed as either deletions, duplications, or both, and can be mosaic, suggesting a potential mitotic event. When such RTSAs corresponding to fragile sites are observed, karyotypes are expected to be negative for a chromosomal rearrangement.

FACTORS AFFECTING EMBRYO ANEUPLOIDY

To identify maternal and embryonic factors that correlate with and potentially influence embryo aneuploidy. In this prospective observational study, patients underwent routine ovarian stimulation with exogenous gonadotropins. During oocyte collection, each punctured follicle was sonographically measured. Embryos were group-cultured to the blastocyst stage according to follicle diameter. Trophectoderm biopsies were followed by pre-implantation genetic testing for aneuploidy (PGT-A). Multiple logistic regression was used to determine if patient age, the ovulatory trigger agent (agonist, agonist+hCG, or hCG alone), follicle diameter at retrieval, the total number of eggs collected, and the day of blastocyst formation (5, 6, or 7) correlated with abnormal PGT-A results. P<0.05 was considered significant. A total of 344 blastocysts were biopsied in the study period. Patient age at retrieval ranged from 21.0 to 43.6 years. A logistic regression model was developed predicting abnormal PGT-A results. The only significant predictor of an abnormal (aneuploid) test result in this model was patient age at retrieval (P=0.0378). Increasing patient age corresponded with increased risk of an abnormal PGT-A result. The ovulatory trigger agent, follicle diameter at retrieval, the total number of eggs collected, and the day of blastocyst formation were not significantly correlated with embryo ploidy. Only patient age significantly correlated with embryo ploidy. The ovulatory trigger agent, follicle diameter at retrieval, the total number of eggs collected, and the day of blastocyst formation were not significantly correlated with embryo ploidy.

Morphokinetic parameters comparison between euploid and aneuploid blastocysts

Background: Pre-implantation genetic testing for aneuploidy (PGT-A) has been using for years in embryoselection. However, this is an invasive method and may cause harm to the embryos. Therefore, time-lapse monitoring has been thought to be an alternative approach for embryo selection due to its efficiency. Up to now, several studies were investigating the relationship between the morphokinetic parameters and the embryo ploidy. However, the results are not consistent. This study aims to evaluate the correlation between morphokinetic parameters and PGT-A results.&#x0D; Methods: This retrospective cohort study was conducted at IVFMD Phu Nhuan, My Duc Phu Nhuan Hospital, between September 2018 and June 2019. Patients undergoing PGT-A due to advanced maternal age, repeated implantation failure or recurrent miscarriage and having embryo cultured under time-lapse monitoring were included. Patients with the re-thawing embryo for PGT-A were not eligible. The time from insemination to the pronuclear appearing (tPN), the onset of two to eight-cell divisions (t2 to t8) and the duration of the second cell cycle (cc2, t3-t2) were observed.&#x0D; Results: There were 39 patients included in the study, with mean age of 36.4 +/- 5.7 years. A total of 110 blastocysts were biopsied. Amongst them, 63 embryos (57.3%) were euploidy (group 1), and 47 embryos (42.7%) were aneuploidy (group 2). There was no significant difference between euploid, and aneuploid embryos regarding all morphokinetic parameters, including tPN, t2, t3, t4, t5, cc2, and t8 (7.2 +/- 1.5 hours vs. 7.4 +/- 1.6 hours; 25.0 +/- 2.8 hours vs. 25.6 +/- 3.2 hours; 35.8 +/- 3.6 hours vs. 36.9 +/- 3.3 hours; 37.5 +/- 4.4 hours vs. 38.3 +/- 4.3 hours; 49.2 +/- 5.52 hours vs. 49.9 +/- 6.2 hours; 10.7 +/- 2.6 hours vs. 11.2 +/- 1.7 hours; and 55.7 +/- 6.4 hours vs. 58.1 +/- 7.4 hours, respectively).&#x0D; Conclusion: In this study, we found no difference in the morphokinetic parameters between euploid and aneuploid embryos.

End-to-end deep learning for recognition of ploidy status using time-lapse videos.

Our retrospective study is to investigate an end-to-end deep learning model in identifying ploidy status through raw time-lapse video. By randomly dividing the dataset of time-lapse videos with known outcome of preimplantation genetic testing for aneuploidy (PGT-A), a deep learning model on raw videos was trained by the 80% dataset, and used to test the remaining 20%, by feeding time-lapse videos as input and the PGT-A prediction as output. The performance was measured by an average area under the curve (AUC) of the receiver operating characteristic curve. With 690 sets of time-lapse video image, combined with PGT-A results, our deep learning model has achieved an AUC of 0.74 from the test dataset (138 videos), in discriminating between aneuploid embryos (group 1) and others (group 2, including euploid and mosaic embryos). Our model demonstrated a proof of concept and potential in recognizing the ploidy status of tested embryos. A larger scale and further optimization on the exclusion criteria would be included in our future investigation, as well as prospective approach.

Consistency in rates of diagnosis of embryonic mosaicism, segmental abnormalities, and “no call” results among experienced embryologists performing preimplantation genetic testing for aneuploidy

ObjectiveTo determine whether differences exist in rates of subchromosomal abnormalities, mosaicism, and “no call” results among embryologists performing and loading trophectoderm biopsies for preimplantation genetic testing for aneuploidy (PGT-A).DesignRetrospective cohort.SettingLarge infertility center.Patient(s)All patients undergoing in vitro fertilization with PGT-A.Intervention(s)The NexCCS next generation sequencing platform was used for PGT-A. The χ2 testing assessed differences in rates of primary outcomes between embryologists. Intraclass correlation coefficients evaluated inter-embryologist reliability in rates of abnormal and no call results. Median absolute performance difference (MAPD) scores, which quantify the impact of technical variation on analytical performance, were averaged for individual embryologists. Analysis of variance assessed differences in mean MAPD scores.Main Outcome Measure(s)Interoperator variability in rates of mosaic, segmental, and no call results.Result(s)Four embryologists performed 30,899 biopsies and 6 embryologists loaded specimens into designated tubes. Among individuals performing trophectoderm sampling, rates of mosaicism were 4.3% to 6.1%, segmental errors were 9.0% to 10.7%, and inconclusive results were 1.1% to 2.9%. For those loading, the incidence of mosaicism was 4.2% to 5.9%, subchromosomal abnormalities was 9.7% to 10.4%, and no call results was 1.2% to 2.2%. The intraclass correlation coefficient was 0.978 for embryologists performing biopsies and 0.981 for those loading. Differences in mean MAPD scores were within 0.6% and 0.2% of each other for doing biopsies and loading embryologists, respectively.Conclusion(s)Rates of mosaicism, segmental, and no call PGT-A results are consistent among experienced embryologists. Due to the large sample size included, differences within 1% of the mean were deemed clinically irrelevant despite statistical significance.

The reproducibility of trophectoderm biopsies in euploid, aneuploid, and mosaic embryos using independently verified next-generation sequencing (NGS): a pilot study.

To assess the accuracy and reliability of comprehensive chromosome screening by next-generation sequencing (NGS) of human trophectoderm (TE) biopsy specimens. The reliability and accuracy of diagnoses made by preimplantation genetic testing for aneuploidy (PGT-A) from TE biopsy were tested. Repeat biopsies of TE and inner cell mass (ICM) samples were obtained from thawed blastocysts previously tested by NGS. To test for the reliability of the NGS assay, biopsy samples were compared with the original PGT-A results. Prior NGS testing classified the TE samples as euploid, aneuploid, or aneuploid-mosaic. The resulting re-biopsied samples underwent SurePlex whole genome amplification followed by NGS via the MiSeq platform, with copy number value (CNV) determined using BlueFuse Multi Software. The primary outcome measure was reliability, defined as concordance between initial TE result and the repeat biopsies. Accuracy was determined by concordance between the TE and ICM samples, and compared between three chromosome types (disomic, aneuploid, and mosaic). Re-biopsies were performed on 32 embryos with prior PGT-A showing euploidy (10 embryos), aneuploidy of one or two chromosomes (4 embryos), or aneuploid-mosaic with one aneuploid chromosome and one mosaic chromosome (18 embryos). One hundred twenty-nine biopsy samples completed NGS (90 TE and 39 ICM biopsies) and 105 biopsy results were included in the analysis. TE biopsies provide a highly accurate test of the future fetus, with the ICM disomic concordance rate of 97.6%. Clinical concordance rates indicate that TE biopsies provide a reliable test when the result is euploid (99.5%) or aneuploid (97.3%), but less reliable when the result is mosaic (35.2%). TE biopsies predict euploidy or aneuploidy in the ICM with a high degree of accuracy. PGT-A with NGS of TE biopsies is shown to be highly reliable, with clinically relevant concordance rates for aneuploidy and euploidy over 95%. TE biopsies indicating mosaicism were less reliable (35.2%), presumably because mitotic non-disjunction events are not uniformly distributed throughout the blastocyst. However, classification of TE biopsy of PGT-A with NGS results as either aneuploid or euploid provides a highly reliable test.

Non-invasive prenatal testing has altered positive predictive value following transfer of a euploid blastocyst

The positive predictive value (PPV) of non-invasive prenatal testing (NIPT) has been reported to range from 91.3-97.2% in the general population (1,2). However, PPV is dependent upon the prevalence of the disease in the population being tested. Patients who undergo in vitro fertilization (IVF) with preimplantation genetic testing for aneuploidy (PGT-A) and transfer a euploid embryo are presumably a lower risk population when compared to the general population. The objective of this study is to explore the PPV for NIPT following transfer of a euploid blastocyst. Retrospective cohort study. All patients at a single IVF center between 2014 and 2018 who pursued pre-implantation genetic testing for aneuploidy (PGT-A) and underwent transfer of a euploid blastocyst between 2014 and 2018 were contacted to request completion of a medical record release form authorizing release of antenatal records. Records were reviewed, and patients who had documentation of an abnormal NIPT were included in this study. Results of any subsequent prenatal or postnatal diagnostic testing were used to classify each positive NIPT as a “true positive” or a “false positive”. The PPV of NIPT was calculated. A total of 1,202 patients eligible for inclusion were contacted for completion of the medical record release form. Five patients with abnormal NIPT following transfer of a euploid blastocyst were identified. Four of these patients (80%) had subsequent definitive prenatal diagnostic testing which revealed a euploid karyotype concordant with their PGT-A results. One patient, who had a PGT-A result indicating 46,XX but a NIPT positive for Turner syndrome, underwent amniocentesis which confirmed Turner mosaicism (45,X karyotype in 80% of cells). Therefore, the PPV of NIPT in this patient cohort was 20%. The PPV of NIPT for patients undergoing transfer of a euploid blastocyst is lower than that for the general population. PGT-A may be more likely to yield inaccurate results in the presence of embryonic mosaicism, as illustrated by the true positive NIPT case in this study cohort. PGT-A is an imperfect screening tool and follow-up antenatal screening is advisable; however, clinicians and patients should recognize that patients undergoing transfer of a euploid blastocyst are at a relatively lower risk for fetal aneuploidy when compared to the general population and, as a result, the PPV of NIPT is altered in this setting.

Do male embryos grow faster than female embryos?

Preimplantation genetic testing for aneuploidy (PGT-A) gives us insight into the chromosomal makeup of an embryo. It has been suggested that male embryos grow faster than female embryos and as a result embryologists are more likely to select a male embryo for transfer. The objective of this study is to assess whether an embryo’s sex and/or chromosomal normalcy may be related to their rate of development. A retrospective study of PGT-A results obtained between 2016 and 2018. Information was derived from PGT-A results of 151 patients (691 embryos; 21 uncounted due to being beyond parameters or missing information). We determined the ratios of day-5 XY to XX, labeling this as group A, day-6 XY to XX as group B, day-5 euploid (N) to aneuploid (AN) as group C, and day-6 N to AN as group D. These ratios were then tested to determine whether embryo growth exhibited any significant patterns. RESULTS: : A chi-square test for independence was performed on the ratios of four groups: A, B, C, and D. The former three groups’ ratios yielded no significant differences, with group B and C possessing ratios very close to an even split 50/50, which is consistent with the average prediction of embryo development. However, the latter group yielded a significant difference with 92:128 euploid to aneuploid ratio (p = 0.015).Tabled 1group Atotal Day 5 XY242Expected Value225total Day 5 XX208Expected Value225Chi-Squre TestP-Value:0.11Group BTotal Day 6 XY108Expected Value110Total Day 6 XX112Expected Value110Chi-Squre Testp-Value:0.79 Open table in a new tab Our results do not support the notion that male embryos grow faster than female embryos. The was a significant difference in the rate of aneuploidy seen when comparing day 6 blastocysts to those that reached the blastocyst stage on day 5.

Previously vitrified embryos can be successfully warmed, biopsied or rebiopsied, and provide results

This study aimed to evaluate whether vitrified embryos could be warmed, biopsied or rebiopsied, and provide meaningful preimplantation genetic testing for aneuploidy (PGT-A) results. The vitrified and warmed embryos include both those biopsied for the first time, as well as those biopsied for a second time (rebiopsied) due to inconclusive results. This was a retrospective study performed at a large, private IVF center which included 242 embryos being warmed and biopsied for the first time and 28 embryos being warmed and rebiopsied. Day 5 and 6 embryos of good and fair quality were vitrified for potential future use. Embryos were warmed, given time to re-expand, and biopsied. Some embryos that initially survived the warming process were unable to be biopsied due to degeneration or developmental arrest prior to re-expansion. Samples were sent to a third-party testing laboratory for PGT-A testing via aCGH, SNP array or NGS. Two hundred forty-two embryos were warmed and biopsied for the first time. One hundred seventy-eight (74%) reached a stage and quality suitable for biopsy. One hundred ten of these embryos (62%) were euploid. When classified by age group, patients 37 years and under yielded a euploid rate of 66%, while patients 38 years and older yielded a euploid rate of 44%. Twenty-eight embryos were warmed for rebiopsy. Twenty-two (79%) reached a stage and quality suitable for biopsy. Twelve of these (55%) were euploid. When classified by age group, patients 37 years and under yielded a euploid rate of 62%, while patients 38 years and older yielded a euploid rate of 50%. Overall, 96% of thawed embryos (258/270) survived the warming process, and 78% of surviving embryos (200/258) were able to be biopsied or rebiopsied.Tabled 11st BiopsyTotal # of patientsAverage AgeWarmedSurvivalBiopsiedEuploidAneuploidOther*4835.3242232178110561296%74%62%31%7%*Other = 9 mosaic, 2 no DNA amplification, 1 no call Open table in a new tab Tabled 1Rebiopsy2436.2282622128293%79%55%36%9%*Other = 1 mosaic, 1 no DNA amplification Open table in a new tab *Other = 9 mosaic, 2 no DNA amplification, 1 no call *Other = 1 mosaic, 1 no DNA amplification Embryo thaw survival rates are high, but approximately one in five embryos (22%) degenerate or arrest prior to re-expansion and cannot be biopsied. The majority of vitrified embryos can be successfully biopsied or rebiopsied and provide meaningful PGT-A results. With informed consent, embryo thaw and biopsy or rebiopsy is a reasonable clinical option.

34. RETROSPECTIVE PGT-A ANALYSIS FOR MULTIPLE DISPLACEMENT AMPLIFICATION PRODUCTS OF EMBRYOS CORRESPONDING TO 104 LIVE BIRTHS BY NEXT-GENERATION SEQUENCING (NGS)

Introduction Preimplantation genetic testing for aneuploidy(PGT-A) has been widely used, but its clinical application value is still inconclusive. The previous PGS 1.0 technology has been proven to have a negative impact on IVF outcomes. PGS 2.0, blastocyst biopsy combined with high-throughput detection technology, has gradually became the mainstream detection method of PGT-A.Many problems still exist in this technology: 1. Biases caused by whole genome amplification (WGA) or high-throughput detection techniques; 2.Embryo mosaicism.Mosaic embryos were used to be considered unsuitable for transfer. However recently healthy live births after transferring mosaic embryo have been widely reported. And many studies also found that some embryos diagnosed as abnormal by PGT-A were turned out to be normal after retested with their inner cell mass. It is therefore suggested that there may be false positive results in PGT-A technology, resulting in the waste of many valuable embryos with normal developmental potential. The interpretation of PGT-A results in clinical practice remains deficient.Since 2015, our center has adopted multiple displacement amplification (MDA) as the first step of all PGT cycles, so we have established a library of MDA products. Material & methods We investigated PGT cycles conducted for monogenic diseases (PGT-M) between January 2015 and December 2017 in our center. Eligible cycles were in which the blastocyst biopsy and single blastocyst transfer were adopted and healthy live birth was obtained. Exclusion criteria: Cycles in which PGT-A had been performed and the embryo was selected based on PGT-A results. MDA used Repli-g MiDi kit(Qiagen, Germany) or Repli-g Single cell kit(Qiagen, Germany) .All MDA products were conducted PGT-A using next generation sequencing (NGS) by Illumina NextSeq® 550 platform. We sequenced the amplified genome of each sample at approximately 0.01x genome depth and the resolution of chromosomal abnormality detection was 4Mb. Results A total of 103 MDA products were tested, of which 11 failed to amplify. Among the 92 successful amplified products, 47 used the Midi-MDA kit while 46 used the Single cell-MDA kit. The average standard deviation(SD) value of NGS data was 3.58 (2.09-9.25).The PGT-A results showed that: 35.9%(33/92) embryos were euploidy; one was trisomy 22; 30.4%(28/92) embryos were mosaic whole chromosomal aneuploidy (the proportion of mosaicism range from 20% to 41%);6.5% (6/92) embryos were segmental chromosomal aneuploidy(chromosomal fragment length of duplication or deletion range from 4.06 Mb to 191.01 Mb), and 8.7% (8/92) embryos were mosaic segmental aneuploidy; 17.4% (16/92) embryos had segmental aneuploidy combined with mosaic aneuploidy. Overall, mosaic chromosomal aneuploidy was detected in 56.5% (52/92) blastocysts which could produce healthy babies.Prenatal diagnosis or karyotype detection of the newborns were performed in 31 cases and all the results were euploidy. Conclusions Our study demonstrated that PGT-A would lead to the waste of viable embryos, so it should be questioned whether all mosaic embryos and segmental aneuploidy embryos should be abandoned. How to correctly interpret the results of PGT-A is an urgent problem.

42. RETROSPECTIVE PGT-A ANALYSIS FOR MULTIPLE DISPLACEMENT AMPLIFICATION (MDA) PRODUCTS OF EMBRYOS CORRESPONDING TO 21 SPONTANEOUS ABORTIONS BY NGS

Introduction Preimplantation genetic testing for aneuploidy (PGT-A) is widely used to identify aneuploidy embryos, thereby reducing miscarriage rate and improving the clinical outcome of in vitro fertilization (IVF). But there are many reasons for miscarriage. Not all miscarriages are caused by embryonic aneuploidy. Moreover, PGT-A has its inherent defects, the results of PGT-A may not fully represent the true karyotype of embryos. Many problems still exist in this technology: 1. Biases caused by whole genome amplification (WGA) or high-throughput detection techniques; 2.Embryo mosaicism. Therefore, it remains unclear what percentage of spontaneous abortion can be prevented by PGT-A. Since 2015, our center has adopted multiple displacement amplification (MDA) as the first step in all PGT cycles. So a complete library of MDA products has been well-established. Material & methods We collected PGT cycles performed for monogenic diseases (PGT-M) at our center from January 2015 to December 2017. Cycles in which the blastocyst biopsy and single blastocyst transplantation were applied, and spontaneous miscarriage appeared during early pregnancy, and MDA products of the embryos were frozen were considered eligible. Exclusion criteria: cycles in which PGT-A had been conducted and the embryo was already selected according to the PGT-A results. All MDA products were performed PGT-A using next generation sequencing (NGS) by Illumina NextSeq ® 550 platform. The software determined the ploidy status with 0.01X read coverage. Results A total of 21 MDA products were detected, and 4 of them were unable to evaluate due to amplification failure or poor test data. The PGT-A results of the remaining 17 embryos showed that 47.1% (8/17) embryos were euploidy, 11.6% (2/17) embryos were segmental chromosomal aneuploidy (46,XN,-(1)(q23.3-q44)(85.03Mb) and 46,XN,+(13)(q11-q34)(95.75Mb)), 27.8% (5/18) of embryos were whole chromosomal aneuploidy mosaicism (the proportion of mosaicism range from 21 to 47%) and 2 cases were trisomy (trisomy 21 and trisomy 22). 6 patients performed karyotype analysis of abortion tissues, among which PGT-A results of 4 cases (3 cases were normal, one was trisomy 22) were consistent with the karyotype of abortion tissues. However, 2 cases showed aneuploidy mosaicism (46,XN,+(mosaic)(19)(30%) and 46,XN,-(mosaic)(6)(44%)) by PGT-A, while the karyotype of their abortion tissues were normal. Conclusions Our study showed that PGT-A could prevent a proportion of spontaneous abortion. However, embryos with low ratio of mosaicism may not be the cause of miscarriage. In fact, spontaneous abortion of these cases was caused by other factors. How high the proportion of mosaicism may lead to miscarriage still requires further research to confirm.