Use Of Preimplantation Genetic Testing For Aneuploidy Research Articles

Preimplantation Genetic Testing for Aneuploidy (PGT-A) in In-Vitro Fertilisation (IVF) Treatment: Study Protocol for Pilot Phase of a Randomised Controlled Trial.

Introduction: Poor outcomes following IVF treatments are speculated to be due to the transfer of aneuploid embryos that cannot be identified based on morphological evaluation alone. This leads to patients requiring numerous embryo transfers and, consequently, a prolonged time interval before live birth. Embryo selection following preimplantation genetic testing for aneuploidy (PGT-A) with next-generation sequencing (NGS) has been suggested as an intervention to shorten time to pregnancy in women undergoing in vitro fertilisation (IVF). Past studies assessing the clinical efficacy of PGT-A in improving clinical outcomes have been conflicting and the associated clinical pregnancy rates and live birth rates following the transfer of a mosaic embryos have yet to be determined. None of the existing studies solely included women of advanced reproductive age (ARA). The pilot study and proposed RCT will determine if, compared to morphological evaluation alone, the use of PGT-A through NGS is a more clinically effective, safer, and more cost-effective way to provide IVF treatment in women of advanced reproductive age. Method and Analysis: The proposed pilot study will aim to randomise 100 patients within a single-centre study to evaluate recruitment, randomisation, and adherence to study protocol and allocated trail arms by participating patients. The results of the pilot study will enable us to determine the sample size for a larger study to establish the effectiveness of PGT-A in ARA women. Ethics and Dissemination: The study (Integrated Research Application System Number 236067) received approval from the Health Research Authority and Health and Care Research Wales (HCRW) and the East Midlands-Leicester South Research Ethics Committee (20/EM/0290). The results will be made available to patients, the funders, the Reproductive Medicine societies, and other researchers. Trial registration: ClinicalTrials.gov Identifier: NCT05009745, n.

Human embryos harbor complex mosaicism with broad presence of aneuploid cells during early development

Pre-implantation genetic testing for aneuploidy (PGT-A) is used in approximately half of in vitro fertilization cycles. Given the limited understanding of the genetics of human embryos, the current use of PGT-A is based on biologically uncertain assumptions and unvalidated guidelines, leading to the possibility of disposing of embryos with pregnancy potential. We isolated and sequenced all single cells (1133) from in vitro cultured 20 human blastocysts. We found that all blastocysts exhibited mosaicism with mitotic-induced aneuploid cells and showed an ~25% aneuploidy rate per embryo. Moreover, 70% (14/20) of blastocysts contained ‘chromosome-complementary’ cells, suggesting genetic mosaicism is underestimated in routine PGT-A. Additionally, the analysis of 20,945 single cells from day 8–14 embryos (in vitro cultured) and embryonic/fetal organs showed that 97% of the analyzed embryos/organs were mosaic. Over 96% of their aneuploid cells harbored ≤ 2 chromosome errors. Our findings have revealed a high prevalence of mosaicism in human embryos.

Success Rates with Preimplantation Genetic Testing for Aneuploidy (PGT-A) in Good Prognosis Patients is Dependent on Age.

To evaluate cumulative live birth following preimplantation genetic testing for aneuploidy (PGT-A) with next generation sequencing (NGS) compared to morphology alone among patients aged 21-40 years undergoing single blastocyst transfer. Retrospective cohort study SUBJECTS: Patients aged 21 to 40 years undergoing first, autologous retrieval cycles resulting in ≥ 5 fertilized oocytes, with subsequent single blastocyst transfer in SART clinics from 2016 to 2019. PGT-A using NGS MAIN OUTCOME MEASURES: The primary outcome was cumulative live birth per retrieval. Secondary outcomes included clinical pregnancy, miscarriage, and live birth per transfer. A total of 56,469 retrieval cycles were included in the analysis. Retrieval cycles were stratified based on age (< 35, 35-37, and 38-40 years) and exposure to PGT-A with NGS. Modified Poisson regression modeling was used to evaluate the association between PGT-A and cumulative live birth per retrieval while controlling for covariates. In this cohort, most cycles did not utilize PGT-A (n=49,608; 88%). After adjusting for covariates, the use of PGT-A was associated with a slightly lower cumulative live birth in individuals aged <35 years (risk ratio [RR] 0.96; 95% CI: 0.93, 0.99) compared with no PGT, but higher cumulative live birth in ages 35-37 years (RR 1.04; 95% CI: 1.00, 1.08), and 38-40 years (RR 1.14; 95% CI: 1.07, 1.20). A subgroup analysis limited to freeze-all cycles (n=29,041) showed that PGT-A was associated with higher cumulative live birth in individuals aged ≥ 35 years and was similar to no PGT in individuals aged < 35 years. Miscarriage was significantly less likely in individuals aged ≥ 35 years utilizing PGT-A compared with no PGT-A. In this large national database study, success rates in cycles utilizing PGT-A were dependent on age. Cumulative live birth was observed to be significantly less likely in PGT-A cycles among individuals aged < 35 years and more likely among individuals aged 38 to 40 years, compared to no PGT-A. In individuals with no fresh transfer, results were similar. Moreover, miscarriage was significantly less likely with PGT-A among individuals aged 35-40 years in a subgroup analysis of freeze-all cycles.

Preimplantation genetic testing for aneuploidy is associated with reduced live birth rates in fresh but not frozen donor oocyte in vitro fertilization cycles: an analysis of 18,562 donor cycles reported to Society for Assisted Reproductive Technology Clinic Outcome Reporting System

ObjectiveTo evaluate the impact of preimplantation genetic testing for aneuploidy (PGT-A) on first transfer live birth rate (LBR) and cumulative LBR (CLBR) in donor oocyte IVF cycles. DesignRetrospective cohort study of the SART CORS database. Subjects11,348 fresh and 7,214 frozen-thawed donor oocyte IVF cycles were analyzed. ExposureThe first reported donor stimulation cycle per patient between January 1, 2014 and December 31, 2015, and all linked embryo transfer cycles between January 1, 2014 and December 31, 2016, were included in the study. Main Outcome MeasuresLBR was compared for patients using fresh and frozen-thawed donor oocytes, with or without PGT-A. Logistic regression models were adjusted for age, body mass index, gravidity, infertility etiology, and prior IVF cycles. ResultsAmong patients who had blastocysts available for transfer or PGT-A, use of PGT-A was associated with a decreased first transfer LBR (46.9 vs 53.2%, p <0.001) and CLBR (58.4 vs 66.6%, p <0.001) in fresh oocyte donor cycles compared with no PGT-A. LBR in frozen-thawed oocyte donor cycles with PGT-A were nominally higher than those without PGTA (48.3% vs. 40.5%) but were not statistically significant in multivariable logistic regression models (p=0.14). Early pregnancy loss was not significantly different with and without PGT-A. Multiple gestation, preterm birth, and low birthweight infants were all reduced with addition of PGT-A in fresh donor oocyte cycles, though these outcomes were not significantly different when comparing single embryo transfers in fresh oocyte cycles and also not significantly different among frozen-thawed donor oocyte cycles. ConclusionPGT-A in fresh oocyte donor cycles was associated with decreased LBR and CLBR, while effects on frozen-thawed oocyte donor cycles were clinically negligible. Obstetrical benefits associated with PGT-A in fresh donor cycles appear linked to increased single embryo transfer.

P-557 The contribution of PGT-A to PGT-M and PGT-SR clinical outcomes – A single centre retrospective analysis

Abstract Study question To determine if the introduction of concurrent testing with PGT-A has improved clinical outcomes for PGT-M/SR patients. Summary answer This study demonstrates that the introduction of concurrent testing may have improved clinical outcomes for this patient demographic irrespective of age. What is known already Preimplantation Genetic Testing (PGT) is utilised to investigate the genetic constitution of embryos. PGT-M (monogenic) and PGT-SR (structural rearrangements) are screening techniques considered in treatment for couples at risk of conceiving offspring with known genetic or structural chromosomal abnormalities respectively. Preimplantation Genetic testing for Aneuploidy (PGT-A) was developed in order to screen embryos for numerical chromosomal abnormalities, a leading cause of failed implantation. Nevertheless, routine use of PGT-A is discouraged due to lack of evidence regarding its efficacy. However, PGT-A is routinely used alongside PGT-M and PGT-SR despite the lack of evidence of its effectiveness in improving patient outcomes. Study design, size, duration The retrospective analysis of 496 PGT-M/SR single embryo transfer patient cycles between December 2013 and December 2021. Participants/materials, setting, methods The retrospective analysis of 496 PGT-M/SR single embryo transfer cycles at CRGH Portland was undertaken (single testing n = 77; concurrent testing n = 419). The clinical outcomes (positive urine pregnancy test, clinical pregnancy, live birth and miscarriage rates) were compared between groups using IBM SPSS Statistics (premium 27). Binary logistic regression was performed for statistical analysis, results with p &amp;lt; 0.05 were determined statistically significant. Main results and the role of chance Overall, concurrent testing was associated with statistically significant improved clinical outcomes compared to the reference group. Statistical significance was seen in positive urine pregnancy test (49% vs 68%, p = 0.012), clinical pregnancy (32.5% vs 63%; p = 0.000013), live birth rate (32.5% vs 59%; p = &amp;lt;0.00016) and miscarriage rates (34.2% vs 13%; p = 0.001). The results imply that the introduction of concurrent testing has improved the probability of obtaining a positive clinical outcome and has reduced the chance of miscarriage in this patient demographic. Further analysis showed concurrent testing improved the chances of obtaining a positive clinical outcome irrespective of the patients age or quality of the embryo. Limitations, reasons for caution This study has produced promising results. However, there are several limitations such as the small sample size, embryo biopsy techniques used, and the different molecular diagnostic techniques and providers utilised. Wider implications of the findings The results imply a justification for the use of PGT-A alongside PGT-M/SR screening techniques to improve embryo selection and clinical outcomes for this cohort of patients. Additionally, they demonstrate the efficacy of PGT-A in a previously underrepresented demographic. However, further research is required to confirm the clinical value of PGT-A. Trial registration number not applicable

P-612 Euploidy rates between cycles triggered with gonadotrophin-releasing hormone agonist (GnRHa), human chorionic gonadotrophin (hCG) and dual trigger

Abstract Study question Is there any difference in euploidy rates between in vitro fertilization (IVF) cycles triggered with GnRHa, hCG or dual trigger? Summary answer There is no difference in the euploidy rate in preimplantation genetic testing for aneuploidy (PGT-A) cycles according to the trigger method used. What is known already The use of PGT-A has shown that the success of meiosis in the IVF laboratory is inversely correlated with maternal age. However, other uninvestigated factors involved in the IVF process may interfere with normal oocyte physiology. Since the final maturation obtained after ovarian stimulation depends on the use of drugs to promote the resumption of meiosis, it is worth investigating whether the triggering method could impact on oocyte and embryo euploidy. This study aims is to evaluate differences in embryo euploidy rates between PGT-A cycles triggered with GnRHa, hCG or dual trigger (GnRHa + hCG). Study design, size, duration This is a single-center retrospective study of 934 patients who underwent 1085 PGT-A cycles by Next-Generation Sequencing (NGS) from January 2017 to December 2023, using one of three possible trigger methods to induce final oocyte maturation: GnRHa, hCG, or dual trigger. The couples included in the study had normal karyotypes and no severe male factor. Participants/materials, setting, methods Data on the number of oocytes retrieved, ratio of metaphase II (MII) oocytes and fertilization rate, blastocyst development and embryo euploidy were assessed and compared between the GnRHa, hCG and dual trigger treatments. A secondary analysis was performed to evaluate data stratified by patient age. Main results and the role of chance A total of 1085 PGT-A cycles involving 934 patients (38.07 ± 3.01 years) and 2803 biopsied blastocysts were included in the analysis. Dual trigger was used in 726 (77.73%) cycles, GnRHa in 287 (26,4%) cycles and hCG in 72 (7.71%) cycles. The number of oocytes retrieved was significantly higher in the GnRHa group (GnRHa: 18.72 ± 8.59; hCG: 9.88 ± 5.64; dual trigger: 10.58 ± 5.65; p &amp;lt; 0.001) with similar maturation rate (GnRHa: 77.46%; hCG: 74.58%; dual trigger: 76.03%; p = 0.789), fertilization rate (GnRHa: 78.12%; hCG: 76.37%; dual trigger: 79.12%; p = 0.168), blastulation rate (GnRHa: 61.03%; hCG: 58.32%; dual trigger: 58.73%; p = 0,629), high-quality blastocyst rate (GnRHa: 27,88%; hCG: 33,67%; dual trigger: 27,88%; p = 0.543) and embryo euploidy per biopsied embryo (GnRHa: 42.48%; hCG:45.49%; dual trigger:46.98%; p = 0.112). Analysis of the data stratified by age showed similar rates of euploidy rates with the three trigger methods in women younger than 41 years (&amp;lt;35 years: GnRHa 66.50%; hCG 66.67%; dual trigger 56.97% p = 0.290; 35-37 years: GnRHa 49.45% hCG 51.04%; dual trigger 50.23%; p = 0.988; 38-40 years: GnRHa 40.93%; hCG 41.27%; dual trigger 39.13% p = 0.257). However, in patients ≥41 years old the euploidy rate was statistically higher in the GnRHa group (GnRH 31.51%; hCG 18.75%; dual trigger 16.89%; p = 0.032). Limitations, reasons for caution The main limitations of the study are: its retrospective design, the possibility that the trigger method was chosen based on the ovarian response and the limited number of patients in both the hCG trigger and women ≥ 41 years groups. Wider implications of the findings Our results suggest that triggering with GnRHa, hCG or dual trigger leads to comparable euploidy rates. Triggering with GnRHa should be recommended in older patients since it has been associated with a higher rate of embryonic euploidy. Trial registration number Not Applicable

Preimplantation Genetic Testing for Aneuploidy Could Not Improve Cumulative Live Birth Rate Among 705 Couples with Unexplained Recurrent Implantation Failure.

We evaluate whether next-generation sequencing (NGS)-based preimplantation genetic testing for aneuploidy (PGT-A) improves the cumulative pregnancy outcomes of patients with unexplained recurrent implantation failure (uRIF) as compared to conventional in vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI). This was a retrospective cohort study (2015-2022). A total of 705 couples diagnosed with uRIF were included in the study. 229 women transferred blastocysts based on morphological grading (IVF/ICSI) and 476 couples opted for PGT-A to screen blastocysts by NGS. Women were further stratified according to age at retrieval (<38 years and ≥38 years). The primary outcome was the cumulative live-birth rate after all the embryos were transferred in a single oocyte retrieval or until achieving a live birth. Confounders were adjusted using binary logistic regression models. Cumulative live-birth rate was similar between the IVF/ICSI group and the PGT-A group after stratified by age: IVF/ICSI vs PGT-A in the <38 years subgroup (49.7% vs 57.7%, adjusted OR (95% CI) = 1.25 (0.84-1.84), P = 0.270) and in the ≥38 years subgroup (14.0% vs 19.5%, adjusted OR (95% CI) = 1.09 (0.41-2.92), P = 0.866), respectively. Nonetheless, the PGT group had a lower first-time biochemical pregnancy loss rate (17.0% vs 8.7%, P = 0.034) and a higher cumulative good birth outcome rate (35.2% vs 46.4%, P = 0.014) than the IVF/ICSI group in the <38 years subgroup. Other pregnancy outcomes after the initial embryo transfer and multiple transfers following a single oocyte retrieval were all similar between groups. Our results showed no evidence of favorable effects of PGT-A treatment on improving the cumulative live birth rate in uRIF couples regardless of maternal age. Use of PGT-A in the <38 years uRIF patients would help to decrease the first-time biochemical pregnancy loss and increase the cumulative good birth outcome.

Single-cell multi-omics sequencing reveals chromosome copy number inconsistency between trophectoderm and inner cell mass in human reconstituted embryos after spindle transfer.

Is the chromosome copy number of the trophectoderm (TE) of a human reconstituted embryos after spindle transfer (ST) representative of the inner cell mass (ICM)? Single-cell multi-omics sequencing revealed that ST blastocysts have a higher proportion of cell lineages exhibiting intermediate mosaicism than conventional ICSI blastocysts, and that the TE of ST blastocysts does not represent the chromosome copy number of ICM. Preimplantation genetic testing for aneuploidy (PGT-A) assumes that TE biopsies are representative of the ICM, but the TE and ICM originate from different cell lineages, and concordance between TE and ICM is not well-studied, especially in ST embryos. We recruited 30 infertile women who received treatment at our clinic and obtained 45 usable blastocysts (22 from conventional ICSI and 23 reconstituted embryos after ST). We performed single-cell multi-omics sequencing on all blastocysts to predict and verify copy number variations (CNVs) in each cell. We determined the chromosome copy number of each embryo by analysing the proportion of abnormal cells in each blastocyst. We used the Bland-Altman concordance and the Kappa test to evaluate the concordance between TE and ICM in the both groups. The study was conducted at a public tertiary hospital in China, where all the embryo operations, including oocytes retrieval, ST, and ICSI, were performed in the embryo laboratory. We utilized single-cell multi-omics sequencing technology at the Biomedical Pioneering Innovation Center, School of Life Sciences, Peking University, to analyse the blastocysts. Transcriptome sequencing was used to predict the CNV of each cell through bioinformatics analysis, and the results were validated using the DNA methylation library of each cell to confirm chromosomal normalcy. We conducted statistical analysis and graphical plotting using R 4.2.1, SPSS 27, and GraphPad Prism 9.3. Mean age of the volunteers, the blastocyst morphology, and the developmental ratewere similar in ST and ICSI groups. The blastocysts in the ST group had some additional chromosomal types that were prone to variations beyond those enriched in the blastocysts of the ICSI group. Finally, both Bland-Altman concordance test and kappa concordancetest showed good chromosomal concordance between TE and ICM in the ICSI blastocysts (kappa = 0.659, P < 0.05), but not in ST blastocysts (P = 1.000), suggesting that the TE in reconstituted embryos is not representative of ICM. Gene functional annotation (GO and KEGG analyses) suggests that there may be new or additional pathways for CNV generation in ST embryos compared to ICSI embryos. This study was mainly limited by the small sample size and the limitations of single-cell multi-omics sequencing technology. To select eligible single cells, some cells of the embryos were eliminated or not labelled, resulting in a loss of information about them. The findings of this study are innovative and exploratory. A larger sample size of human embryos (especially ST embryos) and more accurate molecular genetics techniques for detecting CNV in single cells are needed to validate our results. Our study justifies the routine clinical use of PGT-A in ICSI blastocysts, as we found that the TE is a good substitute for ICM in predicting chromosomal abnormalities. While PGT-A is not entirely accurate, our data demonstrate good clinical feasibility. This trial was able to provide correct genetic counselling to patients regarding the reliability of PGT-A. Regarding ST blastocysts, the increased mosaicism rate and the inability of the TE to represent the chromosomal copy number of the ICM are both biological characteristics that differentiate them from ICSI blastocysts. Currently, ST is not used clinically on a large scale to produce blastocysts. However, if ST becomes more widely used in the future, our study will be the first to demonstrate that the use of PGT-A in ST blastocysts may not be as accurate as PGT-A for ICSI blastocysts. This study was supported by grants from the National Key R&D Program of China (2018YFA0107601) and the National Key R&D Program of China (2018YFC1003003). The authors declare no conflict of interest. N/A.

P-311 Embryo morphological grade and day of vitrification can impact the outcome of a single euploid frozen embryo transfer (FET)

Abstract Study question How does the embryo morphological grade and day of vitrification affect FET outcomes from in vitro fertilization cycles utilizing preimplantation genetic testing for aneuploidy (PGT-A)? Summary answer The morphological grade of a transferred embryo affects FET outcomes in age groups differently, and Day-5 embryos have significantly better pregnancy outcomes than Day-6 embryos. What is known already The use of PGT-A has shown to increase implantation rates, lower miscarriage rates and promote counseling for a single embryo transfer. Although euploid embryos are the priority for transfer, it is important to examine embryo characteristics that may influence these outcomes within these euploid embryos to maximize the outcome for each FET. Study design, size, duration In this retrospective study, 1,554 FETs transferring one euploid Day-5 or Day-6 embryo between January 2020 and October 2022 were evaluated, with the exclusion of donor oocyte, donor embryo, gestational carrier and Day-7 embryo transfer cycles. The inner cell mass and trophectoderm were each categorized as good (G), fair (F) or poor (P) respectively at the time of embryo cryopreservation. A total of 282 GG, 1,053 FG, and 209 FF embryos were transferred. Participants/materials, setting, methods Embryo grade categories were used to compare Day-5 versus Day-6 transfer outcomes and by age groups. Separating groups as &amp;lt; 34, 34-37, and ≥38 eliminated age as a significant factor. Primary outcomes included rates of positive chemical pregnancy (CP), biochemical loss (BL), clinical uterine gestation (CIG), and fetal cardiac activity (FCA). Independent t-tests compared age, and aggregate data was analyzed by proportional z-tests. A p-value &amp;lt;0.05 was defined as statistically significant with a 95% confidence interval. Main results and the role of chance Age and BL were not significant between transfer days or age groups. Day-5 had higher CP (GG: 84% vs 65%, p &amp;lt; 0.001; FG: 75% vs 59%, p = 0.006; FF: 63% vs 44%, p = 0.019), CIG (GG: 73% vs 57%, p = 0.006; FG: 64% vs 48%, p &amp;lt; 0.001; FF: 55% vs 34%, p = 0.007), and FCA (GG: 66% vs 52%, p = 0.022, FG: 60% vs 40%, p &amp;lt; 0.001, FF: 55% vs 30%, p &amp;lt; 0.001). For patients &amp;lt;34, GG had higher CP compared to FG and FF (74% vs 64%, p = 0.017; 74% vs 46%, p &amp;lt; 0.001), CIG (66% vs 55%s, p = 0.011; 66% vs 38%, p &amp;lt; 0.001) and FCA (61% vs 50%, p = 0.012; 61% vs 38%, p = 0.001). FG had higher CP (64% vs 46%, p = 0.007) and CIG (55% vs 38%, p = 0.008) compared to FF. For patients 34-37, GG and FG had higher CP (78% vs 53, p &amp;lt; 0.001; 70% vs 53%, p = 0.004), CIG (66% vs 42%, p = 0.002; 56% vs 42%, p = 0.029), and FCA (58% vs 35%, p = 0.002; 50% vs 35%, p = 0.011) compared to FF. When compared to FF in patients ≥38, GG and FG had increased CP (79% vs 47%, p &amp;lt; 0.001; 64% vs 47%, p = 0.012) and CIG (65% vs 36%, p = 0.004; 53% vs 36%, p = 0.010). Only GG had higher FCA (56% vs 36%, p = 0.049). Limitations, reasons for caution Even with the exclusion criteria, the retrospective nature of the study may be viewed as a limitation, so the significance of the results should be evaluated further. Wider implications of the findings Based on these results, this study demonstrates the need to compare the characteristics of euploid embryos before selection to maximize the chance of pregnancy. Embryos vitrified on Day-5 or categorized as GG, especially in patients &amp;lt;34, or FG should be prioritized over Day-6 or FF embryos. Trial registration number not applicable

Is it necessary for young patients with recurrent implantation failure to undergo preimplantation genetic testing for aneuploidy?

To determine whether preimplantation genetic testing for aneuploidy (PGT-A) can improve the pregnancy outcomes of patients aged under 38 years who have a history of recurrent implantation failure(RIF). Retrospective cohort study. We retrospectively studied the pregnancy outcomes of RIF patients aged under 38 years from January 2017 to December 2021.178 patients were divided into two groups according to whether they underwent PGT-A: the PGT-A group(n=59)and the control group(n=119).In the PGT-A group, we compared the euploidy rate of the different quality and developmental rate blastocysts. In both groups,the patients were the first frozen-thaw single blastocysts transfer after the diagnosis of RIF. Among the pregnancy outcomes, the clinical pregnancy rate was assessed as the primary outcome. The spontaneous abortion rate and ongoing pregnancy rate were the secondry outcomes. The generalized estimation equation was used to adjust for the blastocysts derived from the same patients. Multivariate logistic analysis models were used to compare the pregnancy outcomes between the two groups. In the PGT-A group, 293 blastocysts obtained from59 patients underwent PGT-A. The proportions of euploidy, aneuploidy and mosaic blastocysts were 56.31%, 25.60% and 18.09%, respectively. A comparison of the euploidy rates of different quality blastocysts showed that the rate of good-quality blastocysts was significantly higher than that of poor-quality blastocysts (67.66% vs 46.88%; odds ratio [OR], 2.203; 95%confidence interval[CI], 0.943-3.612; P=0.002). However, no significant difference was observed in the different developmental rates blastocysts. Compared with Day 5 blastocysts, the euploidy rates of Day 6 and Day 7 blastocysts were not significantly different(61.54%vs51.91%; OR,0.945; 95%CI, 0.445-2.010; P=0.884; and 61.54%vs47.37%; OR, 1.106; 95%CI, 0.774-1.578; P=0.581, respectively).As for the pregnancy outcomes, the clinical pregnancy rate was significantly increase after the use of PGT-A compared with the control group(71.19%vs56.30%; OR, 0.538; 95%CI, 0.262-1.104; P=0.039). However, the spontaneous abortion rates and ongoing pregnancy rates were not significantly different between the control and PGT-A groups (21.43% vs 19.40%; aOR,0.727; 95%CI,0.271-1.945; P=0.525; and55.93% vs 45.38%; aOR, 0.649; 95%CI, 0.329-1.283; P = 0.214,respectively). PGT-A improved the clinical pregnancy rate after blastocyst transfer in RIF patients aged under 38 years.

P-545 The age-related required number of zygotes estimated from prior clinical studies of preimplantation genetic testing for aneuploidy (PGT-A)

Abstract Study question How many eggs will be required to optimize the chances of a live birth with or without PGT-A? Summary answer The number of zygotes required for live birth is higher in women with an advanced age, and the use of PGT-A does not provide improvement. What is known already Women who are undergoing PGT-A often wish to know how many eggs will be required to optimize the chances of a live birth. This important information could be provided as part of prior genetic counseling, but there are no precise data on this at present. If the number of eggs required to give the best chance of a successful live birth was known, treatment plans with or without PGT-A could be better determined. Study design, size, duration We estimated the optimal number of eggs required for IVF treatment with PGT-A to produce at least a single live birth, stratified by maternal age, on the basis of information from prior studies and in current databases. Participants/materials, setting, methods We derived our calculation parameters from three prior large-scale clinical investigations associated with PGT-A. We estimated a live birth rate using the following factors: rate of zygotes that develop a useful blastocyst, euploid rate in PGT-A, and the live birth rate after euploid embryo transfer. All of these factors were assumed to be statistically independent in this study for the purposes of our calculations and the live birth rate per single zygote was calculated. Main results and the role of chance The estimations in our present analyses however indicate a probability of less than 10% that woman over 40 years of age will have a live birth from a single zygote, regardless of whether PGT-A is performed or not. We used a negative binomial distribution approach to calculate how many zygotes are needed to obtain at least one live birth. The plot of these results is provided in Figure 2. To achieve a 50% chance of getting at least one live birth, patients required 8 zygotes at age of 40 and 21 zygotes at the age of 43. Furthermore, to achieve an 80% chance of obtaining a live birth, our calculations estimate that 18 and 47 zygotes would be required at these two ages, respectively, which would be challenging to achieve. On the other hand, by avoiding unnecessary transplants using PGT-A, women may have to wait a shorter period to accomplish a live birth or may be able to avoid wasting their limited remaining reproductive period, particularly if they are older than 42. Limitations, reasons for caution The reference data from PGT-A studies that have estimated of the live birth rate include chromosomal quantitative PCR, microarray analysis, and next generation sequencing (NGS). There is a high possibility that the embryos designated as “euploid” in those studies include mosaic embryos, which represents a limitation of our present meta-analysis. Wider implications of the findings More details on the clinical outcomes of PGT-A will be revealed as clinical studies progress in the future. It is our hope that the results of this present study will assist with future genetic counseling strategies for PGT-A in the meantime. Trial registration number not applicable

Effect of surgical hysteroscopy and PGT-A on pregnancy rates in in vitro fertilization with own eggs

Background: In vitro Fertilization (IVF) is a tool for assisted reproduction used with the aim of increasing pregnancy rates in couples with infertility issues. These procedures may be optimized using techniques for genetical evaluation of the embryo by means of preimplantation genetic testing for aneuploidy (PGT-A) or diagnosis and correction of the uterine cavity such as Hysteroscopy. Objective: The objective of this study was to evaluate the impact of hysteroscopy in contrast with PGT-A analysis with respect to pregnancy rates on IVF cycles. Materials and methods: A study was carried out with Mexican patients during 2018-2021. Patients were divided in two groups: Group 1, patients with guarded prognosis for fertilization; Group 2, patients with guarded prognosis for implantation. The couples evaluated were subjected to different methodologies before IVF. Results: It was found that prior use of PGT-A or Hysteroscopy increase pregnancy rates by 9.4% up to 20.92%. In Group 1 the use of PGT-A/IVF caused a mean pregnancy rate of 77.7%, being favorable the transference of a single embryo. In Group 2, the best combination was Hysteroscopy/IVF with a pregnancy rate of 76.96%. Conclusion: Both of the methodologies prior to the IVF cycle improve pregnancy rates, being recommendable to carry out a PGT-A in patients with a poor genetic prognosis with the transference of a single embryo. Hysteroscopy is recommended when lesions or infectious processes are detected in the uterine cavity, and two-embryo transference is carried out.

Live Birth with or without Preimplantation Genetic Testing for Aneuploidy

BackgroundEmbryo selection with preimplantation genetic testing for aneuploidy (PGT-A) may improve pregnancy outcomes after initial embryo transfer. However, it remains uncertain whether PGT-A improves the cumulative live-birth rate as compared with conventional in vitro fertilization (IVF).MethodsIn this multicenter, randomized, controlled trial, we randomly assigned subfertile women with three or more good-quality blastocysts to undergo either PGT-A or conventional IVF; all the women were between 20 and 37 years of age. Three blastocysts were screened by next-generation sequencing in the PGT-A group or were chosen by morphologic criteria in the conventional-IVF group and then were successively transferred one by one. The primary outcome was the cumulative live-birth rate after up to three embryo-transfer procedures within 1 year after randomization. We hypothesized that the use of PGT-A would result in a cumulative live-birth rate that was no more than 7 percentage points higher than the rate after conventional IVF, which would constitute the noninferiority margin for conventional IVF as compared with PGT-A.ResultsA total of 1212 patients underwent randomization, and 606 were assigned to each trial group. Live births occurred in 468 women (77.2%) in the PGT-A group and in 496 (81.8%) in the conventional-IVF group (absolute difference, −4.6 percentage points; 95% confidence interval [CI], −9.2 to −0.0; P<0.001). The cumulative frequency of clinical pregnancy loss was 8.7% and 12.6%, respectively (absolute difference, −3.9 percentage points; 95% CI, −7.5 to −0.2). The incidences of obstetrical or neonatal complications and other adverse events were similar in the two groups.ConclusionsAmong women with three or more good-quality blastocysts, conventional IVF resulted in a cumulative live-birth rate that was noninferior to the rate with PGT-A. (Funded by the National Natural Science Foundation of China and others; ClinicalTrials.gov number, NCT03118141.)

Preimplantation Genetic Testing for Aneuploidy Improves Live Birth Rates with In Vitro Produced Bovine Embryos: A Blind Retrospective Study.

Approximately one million in vitro produced (IVP) cattle embryos are transferred worldwide each year as a way to improve the rates of genetic gain. The most advanced programmes also apply genomic selection at the embryonic stage by SNP genotyping and the calculation of genomic estimated breeding values (GEBVs). However, a high proportion of cattle embryos fail to establish a pregnancy. Here, we demonstrate that further interrogation of the SNP data collected for GEBVs can effectively remove aneuploid embryos from the pool, improving live births per embryo transfer (ET). Using three preimplantation genetic testing for aneuploidy (PGT-A) approaches, we assessed 1713 cattle blastocysts in a blind, retrospective analysis. Our findings indicate aneuploid embryos have a 5.8% chance of establishing a pregnancy and a 5.0% chance of given rise to a live birth. This compares to 59.6% and 46.7% for euploid embryos (p < 0.0001). PGT-A improved overall pregnancy and live birth rates by 7.5% and 5.8%, respectively (p < 0.0001). More detailed analyses revealed donor, chromosome, stage, grade, and sex-specific rates of error. Notably, we discovered a significantly higher incidence of aneuploidy in XY embryos and, as in humans, detected a preponderance of maternal meiosis I errors. Our data strongly support the use of PGT-A in cattle IVP programmes.

SHOULD PATIENTS WITH ONLY TWO EMBRYOS ELIGBILE FOR BIOPSY AFTER A SINGLE CONTROLLED OVARIAN HYPERSTIMULATION CYCLE UTILIZE PGT-A PRIOR TO TRANSFER SELECTION? cycles without PGT-A.

Preimplantation genetic testing for aneuploidy (PGT-A) is often utilized to enhance implantation rates by enabling the selection of a euploid embryo for transfer. Clinicians may question whether the use of PGT-A is beneficial, especially for patients who produce low numbers of embryos after controlled ovarian hyperstimulation (COH).1 Our study aims to assess the potential benefit and/or risk of utilizing PGT-A prior to embryo transfer in patients with only two available embryos compared to fresh or frozen ET cycles without PGT-A.

Comparison of preimplantation genetic testing for aneuploidy versus intracytoplasmic sperm injection in severe male infertility.

The retrospective cohort study was conducted to evaluate the effectiveness of preimplantation genetic testing for aneuploidy (PGT-A) for severe male factor (SMF) infertility on pregnancy outcomes in comparison with intracytoplasmic sperm injection (ICSI). A total of 206 couples with SMF were included in the study, among which, 102 couples underwent ICSI with next-generation sequencing (NGS)-based PGT-A (the PGT-A group), while 104 underwent ICSI only (the control group). Results showed while no differences were noted in clinical pregnancy rate (CPR) (66.7% versus. 69.9%, p=.64) and ongoing pregnancy rate (OPR) (62.2% versus. 54.7%, p=.29) per transfer between groups, early miscarriage rate (EMR) per transfer was significantly lower (6.7% versus. 21.6%, p=.02) in the PGT-A group. Cumulative OPR per patient remained similar between groups (54.9% versus. 55.8%, p=.90). Results of multivariable logistic regression also demonstrated the use of PGT-A was significantly associated with lower EMR (adjusted OR 0.17, 95%CI 0.05-0.55) in SMF, while it was not related to cumulative OPR. In conclusion, our results showed that NGS-based PGT-A can improve pregnancy outcomes for couples with SMF by significantly decreasing EMR without compromising cumulative OPR, indicating that NGS-based PGT-A could be offered as an appropriate approach for couples with SMF.

Gestational carrier pregnancy outcomes from frozen embryo transfer depending on the number of embryos transferred and preimplantation genetic testing: a retrospective analysis

To compare gestational age, birth weight (BW), and live birth rates in gestational carriers (GC) after the transfer of 1 or 2 frozen embryo(s) with or without preimplantation genetic testing for aneuploidy (PGT-A), with the understanding that several social and economic factors may motivate intended parents to request the transfer of 2 embryos and/or PGT-A when using a GC. Retrospective cohort study SETTING: An assisted reproductive technology practice. All frozen blastocyst transfers with GCs from 2009-2018. One or 2 embryo frozen embryo transfers with and without PGT-A. Live birth, preterm birth, and low BW. A total of 583 frozen embryo transfer cycles with vitrified high-grade blastocysts (grade BB or higher) to GCs were analyzed. Although the live birth rate was significantly greater in frozen embryo transfers with 2 embryos, after single embryo transfer (SET), the mean gestational age and BW of live births were statistically significantly greater than those of double embryo transfer (DET). The rate of multiple births was 1.9% for SET compared to 20.0% for DET per transfer. Only 3.8% of live births from SET experienced low BW and 0.6% had very low or extremely low BW. By comparison, 12.5% of DET live births were low BW and 5% were very low BW. After SET, 13.4% of live births were preterm, compared with 40% in DET. The analysis also included a total of 194 transfers with PGT-A compared to 389 cycles without. Overall, live births per transfer were not significantly different between these latter 2 subgroups. Frozen embryo transfer cycles in GCs with DET were associated with more preterm births and lower birth weights compared with those of SET. Intended parents and GCs should be counseled that DET is associated with greater risks of adverse pregnancy and perinatal outcomes, which mitigates higher live birth rates. The use of PGT-A did not appear to improve the live birth rate.

Sonographic abnormalities in pregnancies conceived following IVF with and without preimplantation genetic testing for aneuploidy (PGT-A)

PurposeTo report the rate of fetal anomalies detected on anatomy ultrasound in pregnant patients who underwent IVF with preimplantation genetic testing for aneuploidy (PGT-A) compared to patients who conceived following IVF with unscreened embryos and age-matched patients with natural conceptions.MethodsRetrospective cohort study at a single maternal-fetal medicine practice. Patients with singleton pregnancies who had a mid-trimester anatomy ultrasound between January 2017 and December 2018 were screened for inclusion. A total of 712 patients who conceived after IVF with or without PGT-A were age-matched with natural conception controls. The primary outcome was the rate of fetal and placental anomalies detected on mid-trimester anatomical survey. Secondary outcomes included the rates of abnormal nuchal translucency (NT), second trimester serum analytes, non-invasive prenatal testing (NIPT), and invasive diagnostic testing.Result(s)There were no differences in the rate of fetal anomalies in patients who underwent IVF with PGT-A compared to patients who conceived following IVF with unscreened embryos and age-matched patients with natural conceptions. Rate of abnormal NT, high-risk NIPT, and abnormal invasive diagnostic testing were also similar. Patients who conceived after IVF with or without PGT-A had higher rates of abnormal placental ultrasound findings and abnormal second trimester serum analytes compared to natural conception controls.ConclusionThe use of PGT-A was not associated with a difference in risk of fetal anomaly detection on a mid-trimester anatomical survey. The results of this study highlight the importance of improved patient counseling regarding the limitations of PGT-A, and of providing standard prenatal care for pregnancies conceived through ART, regardless of whether PGT-A was performed.

EMBRYOLOGIC OUTCOMES IN INTRACYTOPLASMIC SPERM INJECTION (ICSI) CYCLES UTILIZING SPERM SELECTED VIA A MICROFLUIDICS DEVICE COMPARED TO STANDARD SELECTION

Microfluidics sperm sorting allows for improved selection of normal sperm compared with conventional sperm preparation, but blastulation rate after its use has not been studied. We aim to compare the high-quality blastulation rate in cycles in which sperm was chosen via a microfluidics device versus standard sperm selection. Single academic IVF center retrospective cohort study. The IVF/ICSI cycles of 45 patients aged 18-46 years between 2014-20 utilizing a microfluidics chamber (ZyMotTM) with ICSI for sperm selection were compared to the same patients’ previous ICSI cycles in which standard sperm selection was used. Use of microfluidics was based on physician discretion and/or the IVF lab recommendation based on previous low fertilization rate or blastulation rate. For standard selection, after centrifugation and washing by swim-up method sperm was chosen manually based on motility and morphology. In study cycles, motile sperm were manually selected after traversing through a microfluidics chamber. Primary outcome was high-quality blastulation rate (≥3BB by Gardner scoring). Secondary outcomes included fertilization rate, number of high-quality blastocysts frozen, and euploidy rate among IVF cycles using preimplantation genetic testing for aneuploidy (PGT-A). For paired data, McNemar’s test was used for categorical and paired T-test for continuous data. A two-sided p-value of <0.05 was considered statistically significant. Mean patient age, partner age, BMI, and AMH level at the time of standard selection cycle were 37.3 years, 37.7 years, 25.5 kg/m2, and 2.5 ng/mL, respectively. The majority of patients’ primary infertility diagnosis was male factor (42.2%), followed by unexplained (22.2%). 35.6% of patients used the same stimulation protocol for both cycles. The use of PGT-A, mean numbers of oocytes retrieved, and pre- and post-processing sperm parameters were similar. Although the high quality blastulation rate was higher in microfluidics cycles, the difference was not significant. The fertilization rate, euploidy rate and mean number of embryos frozen did not differ significantly (Table 1).Table 1Embryologic OutcomesStandard Selection CyclesMicrofluidics Cyclesp-valueNo. of oocytes retrieved (n ± SEM)10.1 ± 8.811.7 ± 8.60.08Fertilization rate (% ± SEM)54.1 ± 4.657.8 ± 3.50.49High-quality blastulation rate (% ± SEM)25.2 ± 7.338.1 ± 4.70.15Number of high-quality blastocysts frozen (n ± SEM)1.9 ± 0.32.8 ± 0.60.11Euploidy rate (% ± SEM)16.7 ± 8.920.1 ± 6.40.78 Open table in a new tab Microfluidics sperm selection did appear to improve blastulation rate although this was not statistically significant due to the sample size. Further large prospective randomized studies are needed.

A systematic review exploring the patient decision-making factors and attitudes towards pre-implantation genetic testing for aneuploidy and gender selection.

Pre-implantation genetic testing for aneuploidy (PGT-A) is in high demand worldwide, with ongoing debate among medical societies as to which patient groups it should be offered. The psychological aspects for patients regarding its use, lag behind the genomic technological advances, leaving couples with limited decision-making support. The development of this technology also leads to the possibility for its utilization in gender selection. Despite the controversy surrounding these issues, very few studies have investigated the psychological aspects of patients using PGT-A. This systematic review provides an up-to-date analysis of the psychosocial aspects surrounding PGT for aneuploidy and sex selection, as well as decision-making factors. A systematic search of English peer-reviewed journals of three computerized databases were undertaken following PRISMA guidelines. The qualitative data were extracted using thematic analysis. PROSPERO Registration number: CRD42019126439. The main outcome measures were patients' motivations, decision-making factors, attitudes and experiences surrounding the use of PGT for aneuploidy and sex selection. Ten studies were included, four for PGT-A and six for sex selection. Attitudes towards PGT-A were positive, with the main motivating factors being decreasing miscarriage rate, reducing the risk of termination of pregnancy and reducing the time to pregnancy. Consistently raised concerns regarding PGT-A were the financial burden and moral beliefs. The vast majority of patients felt sufficiently knowledgeable to make the decision; however, studies did reveal that a minority mis-interpreted certain potential benefits of PGT-A. Studies investigating PGT for sex selection predominantly reported the main motivation was to achieve gender balance within the family dynamic, with most studies finding no difference between couples using PGT for gender selection to have male or female offspring. Although this systematic review was limited by the small number of studies investigating this topic, a significant minority of patients appeared to misunderstand certain benefits and limitations of PGT-A. Fertility clinics must ensure they provide adequate counseling to all patients using PGT-A. With the use of PGT-A on the rise globally, there is a need to develop decision support tools for couples who have an increasing number of genetic testing options becoming available to them.