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1.
Am J Hum Genet ; 106(4): 525-534, 2020 04 02.
Artigo em Inglês | MEDLINE | ID: mdl-32220293

RESUMO

Despite next-generation sequencing, which now allows for the accurate detection of segmental aneuploidies from in vitro fertilization embryo biopsies, the origin and characteristics of these aneuploidies are still relatively unknown. Using a multifocal biopsy approach (four trophectoderms [TEs] and one inner cell mass [ICM] analyzed per blastocyst; n = 390), we determine the origin of the aneuploidy and the diagnostic predictive value of segmental aneuploidy detection in TE biopsies toward the ICM's chromosomal constitution. Contrary to the prevalent meiotic origin of whole-chromosome aneuploidies, we show that sub-chromosomal abnormalities in human blastocysts arise from mitotic errors in around 70% of cases. As a consequence, the positive-predictive value toward ICM configuration was significantly lower for segmental as compared to whole-chromosome aneuploidies (70.8% versus 97.18%, respectively). In order to enhance the clinical utility of reporting segmental findings in clinical TE biopsies, we have developed and clinically verified a risk stratification model based on a second TE biopsy confirmation and segmental length; this model can significantly improve the prediction of aneuploidy risk in the ICM in over 86% of clinical cases enrolled. In conclusion, we provide evidence of the predominant mitotic origin of segmental aneuploidies in preimplantation embryos and develop a risk stratification model that can help post-test genetic counseling and that facilitates the decision-making process on clinical utilization of these embryos.


Assuntos
Blastocisto/fisiologia , Embrião de Mamíferos/fisiologia , Desenvolvimento Embrionário/genética , Aneuploidia , Aberrações Cromossômicas , Cromossomos/genética , Hibridização Genômica Comparativa/métodos , Feminino , Fertilização In Vitro/métodos , Humanos , Incidência , Gravidez , Diagnóstico Pré-Implantação/métodos
2.
BMC Bioinformatics ; 21(1): 41, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-32007105

RESUMO

BACKGROUND: Haplotyping reveals chromosome blocks inherited from parents to in vitro fertilized (IVF) embryos in preimplantation genetic diagnosis (PGD), enabling the observation of the transmission of disease alleles between generations. However, the methods of haplotyping that are suitable for single cells are limited because a whole genome amplification (WGA) process is performed before sequencing or genotyping in PGD, and true haplotype profiles of embryos need to be constructed based on genotypes that can contain many WGA artifacts. RESULTS: Here, we offer scHaplotyper as a genetic diagnosis tool that reconstructs and visualizes the haplotype profiles of single cells based on the Hidden Markov Model (HMM). scHaplotyper can trace the origin of each haplotype block in the embryo, enabling the detection of carrier status of disease alleles in each embryo. We applied this method in PGD in two families affected with genetic disorders, and the result was the healthy live births of two children in the two families, demonstrating the clinical application of this method. CONCLUSION: Next generation sequencing (NGS) of preimplantation embryos enable genetic screening for families with genetic disorders, avoiding the birth of affected babies. With the validation and successful clinical application, we showed that scHaplotyper is a convenient and accurate method to screen out embryos. More patients with genetic disorder will benefit from the genetic diagnosis of embryos. The source code of scHaplotyper is available at GitHub repository: https://github.com/yzqheart/scHaplotyper.


Assuntos
Testes Genéticos/métodos , Haplótipos , Diagnóstico Pré-Implantação/métodos , Análise de Célula Única/métodos , Blastocisto/citologia , DNA/genética , Feminino , Fertilização In Vitro , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Gravidez
3.
Rev Med Suisse ; 15(668): 1909-1913, 2019 Oct 23.
Artigo em Francês | MEDLINE | ID: mdl-31643150

RESUMO

In Switzerland, since modifications of the law regulating reproductive medicine introduced the 1rst of September 2017, preimplantation genetic testing (PGT) has been legalised. Infertile couples undergoing in vitro fertilization (IVF) can benefit from this technology by detecting which embryos are aneuploid (ie abnormal number of chromosomes, PGT-A). This is performed in order to transfer euploid embryos (normal number of chromosomes) and to optimise success, though data are limited. Couples at risk of transmitting a severe monogenic disease or unbalanced translocation can undergo PGT for monogenic disease or chromosomal structural rearrangements (PGT-M/SR). These tests are subject to strict legal criteria. Their clinical application needs to be approved through a multidisciplinary approach taking into account legal and ethical issues while respecting the autonomy of the couples.


Assuntos
Testes Genéticos/ética , Testes Genéticos/legislação & jurisprudência , Diagnóstico Pré-Implantação/ética , Aneuploidia , Feminino , Fertilização In Vitro , Testes Genéticos/métodos , Humanos , Masculino , Gravidez , Diagnóstico Pré-Implantação/métodos , Suíça
4.
J Assist Reprod Genet ; 36(8): 1561-1569, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31385120

RESUMO

PURPOSE: There is clinical evidence that early cleavage timing parameters predictive of blastocyst development also correlate with embryo implantation potential. The aim of this study is to determine the developmental competency of embryos with delayed blastulation. METHODS: Retrospective study performed from 2015 to 2016 at the Division of Reproductive Endocrinology and Infertility at Northwestern University. RESULTS: A total of 2,292 embryos from 524 patients were included. Day 6 blastocysts had statistically significant longer times for every time point analyzed than day 5 blastocysts (p < 0.001). We found no statistically significant difference in euploidy rates between day 5 (44%) and day 6 (41%) embryos (p = 0.573). t7 and t8 time points were independent predictors of euploidy after controlling for day of biopsy (p < 0.015 and p < 0.014, respectively). Intrauterine pregnancy (IUP) and live birth (LB) were less likely to occur after transferring day 6 embryos (p = 0.0033 and p = 0.0359) without previous genetic testing. However, in embryos that undergo preimplantation genetic testing for aneuploidy (PGT-A), there were no significant differences in IUP or LB rates. CONCLUSION: Early time-lapse points can be used to predict embryo development. Day of blastulation may be an independent predictor IUP, with day 6 blastocysts having lower pregnancy and live birth rates. Our data suggests that day 5 and day 6 PGT-A tested embryos show similar rates of euploidy, suggesting that differences in PR seen in the non-PGT-A tested group may be caused by factors other than aneuploidy. Genetic testing technologies in combination with time-lapse microscopy may provide further information to improve IVF outcomes.


Assuntos
Aneuploidia , Blastocisto/patologia , Implantação do Embrião/fisiologia , Fertilização In Vitro , Testes Genéticos/métodos , Diagnóstico Pré-Implantação/métodos , Imagem com Lapso de Tempo/métodos , Adulto , Transferência Embrionária , Feminino , Humanos , Gravidez , Taxa de Gravidez , Estudos Retrospectivos
5.
J Assist Reprod Genet ; 36(9): 1901-1908, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31352621

RESUMO

PURPOSE: To evaluate the influence of the endometrial receptivity array (ERA) test on the implantation rate (IR) and pregnancy rate (PR) in patients with previous failed euploid embryo transfers (Euploid-ET) or oocyte donation embryo transfers (Donor-ET). METHODS: Single-center retrospective study of patients with ≥ 1 previous failed Euploi-ET (n = 24) or ≥ 2 failed Donor-ET (n = 32) who underwent an ERA test and a post-ERA Euploid-ET/Donor-ET between 2012 and 2018. Controls were patients with ≥ 1 previously failed Euploid-ET (n = 119) or ≥ 2 failed Donor-ET (n = 158) who underwent Euploid-ET/Donor-ET during the same period without performing an ERA test. Only blastocyst stage embryos were included. IR/PR was compared between the post-ERA ET and the last ET in the control group. RESULTS: There was no statistically significant difference regarding IR [55.6% (34.6-76.5%) vs. 65.0% (56.9-73.1%)] nor PR (58.3% vs.70.6%, p = 0.238) in the Euploid-ET ERA vs. Euploid-ET control groups. In the Donor-ET arm, both IR [26.8% (12.3-41.4%) vs. 57.2% (50.1-64.3%)] and PR (34.4% vs. 65.2%, p = 0.001) were significantly lower in the ERA group. Multivariate analysis confirmed that performing an ERA test did not influence the PR in the Euploid-ET arm and was associated with a diminished PR in the Donor-ET arm. In the ERA group, 41.1% patients were non-receptive (NR). No significant difference was found regarding IR/PR in NR vs. receptive patients in both Euploid-ET/Donor-ET arms. CONCLUSIONS: In our sample, the performance of an ERA test did not improve pregnancy outcomes. Future prospective studies in larger samples are needed to confirm the role of the ERA test in Euploid-ET/Donor-ET.


Assuntos
Endométrio/fisiologia , Doação de Oócitos , Diagnóstico Pré-Implantação/métodos , Adulto , Aneuploidia , Blastocisto/fisiologia , Implantação do Embrião , Feminino , Fertilização In Vitro , Humanos , Pessoa de Meia-Idade , Gravidez , Resultado da Gravidez , Taxa de Gravidez , Estudos Retrospectivos , Falha de Tratamento
6.
J Assist Reprod Genet ; 36(8): 1609-1621, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31292818

RESUMO

PURPOSE: This study was to evaluate if spent culture media (SCM) of embryos could be used as a non-invasive tool to achieve aneuploidy screening. Ploidy calls, as well as concordance rates between PGT-A results from trophectoderm (TE) and SCM, were compared. Clinical outcomes of single euploid transfers were also evaluated. METHODS: The study was conducted from March 2017 to June 2018 in a university-based ART center. SCM of day 3 to the day(s) of TE biopsy of all biopsied blastocysts were collected for testing. PGT-A results of SCM were compared with the standard results of TE, with clinical relevance and outcomes examined. RESULTS: NiPGT-A using SCM gave a sensitivity of 81.6%, specificity of 48.3%, positive predictive value of 82.6%, and negative predictive value of 46.7% in ploidy calling. The concordance rates for autosomes and sex determination were 62.1% and 82.4%, respectively. There were 14 single embryo transfer cycles of euploids as determined by TE biopsy. Clinical outcomes not only confirmed 3 false positive results from SCM but also reflected the true ploidy status of the transferred embryo in one case. If ploidy calls were dichotomized without mosaic embryos, the sensitivity and NPV would increase to 91.0% and 66.7% (p = 0.60 and p = 0.25), respectively. CONCLUSIONS: Cell-free DNA found in SCM could provide ploidy information of an embryo as in PGT-A from its TE. Given its potential to reflect the comprehensive chromosomal profile of the whole embryo, more research based on clinical outcomes is required to determine if SCM could be a reliable selection tool in PGT-A.


Assuntos
Aneuploidia , Meios de Cultura/metabolismo , Fertilização In Vitro , Testes Genéticos/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Diagnóstico Pré-Implantação/métodos , Trofoblastos/metabolismo , Técnicas de Cultura Embrionária , Feminino , Humanos , Recuperação de Oócitos , Indução da Ovulação , Gravidez , Estudos Prospectivos , Trofoblastos/citologia
7.
J Assist Reprod Genet ; 36(8): 1591-1597, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31325068

RESUMO

PURPOSE: The aim of our study was to evaluate the influence of different ejaculatory abstinence time frames (several days versus 1 h) on semen parameters, blastocysts ploidy rate, and clinical results in assisted reproduction cycles on sibling oocytes. METHODS: This is a prospective study including 22 preimplantation genetic testing for aneuploidy (PGT-A) cycles performed between November 2015 and December 2018. Male partners with oligoastenoteratozoospermia produced two semen samples on the day of oocyte retrieval: the first one after several days of abstinence and the second, 1 h after the first one. Oocytes from each patient were divided into two groups: those in group 1 were injected with spermatozoa from the first ejaculate (N = 121) and oocytes in group 2 with spermatozoa from the second one (N = 144). Outcomes of aniline blue test, fertilization, blastocyst formation, ploidy rates, and clinical results were compared between the two groups. RESULTS: Semen volume resulted lower in the second sperm retrieval. Sperm concentration, motility, and morphology were similar in the two groups. A total of 106 blasotcysts were biospied. Higher blastocyst euploidy rates resulted in group 2 (43.6%) than in group 1 (27.5%). A higher percentage of mature chromatine was observed in group 2. CONCLUSION: Using spermatozoa from samples with a shorter abstinence could be a simple method to select higher quality spermatozoa, reducing aneuploidy rate in blastocysts. Prospective randomized controlled trials should be performed to confirm the potential advantage of using semen samples with short abstinence period to improve the outcome of assisted reproduction cycles.


Assuntos
Aneuploidia , Blastocisto/fisiologia , Transferência Embrionária/estatística & dados numéricos , Fertilização In Vitro , Oligospermia , Diagnóstico Pré-Implantação/métodos , Espermatozoides/química , Adulto , Blastocisto/citologia , Feminino , Testes Genéticos , Humanos , Masculino , Gravidez , Taxa de Gravidez , Estudos Prospectivos , Análise do Sêmen
8.
J Assist Reprod Genet ; 36(8): 1623-1629, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31165389

RESUMO

PURPOSE: Does blastocyst morphology following euploid elective single embryo transfer (eSET) after preimplantation genetic testing for aneuploidies (PGT-A) via next generation sequencing impact clinical outcome? METHODS: Two hundred ninety-six patients underwent PGT-A. Of 1549 blastocysts, 1410 blastocysts had a conclusive result after PGT-A and were included for analysis. An eSET policy was followed in a frozen embryo replacement cycle. A total of 179 euploid blastocysts were thawed and transferred. Clinical outcomes were categorized in four different embryo quality groups: excellent, good, average and poor. RESULTS: Euploidy rate was 19/36 (52.7%, 95% CI 37-68), 199/470 (42.3%, 95% CI 38-47), 156/676 (23.0%, 95% CI 20-26) and 39/228 (17.1%, 95% CI 13-23) in the excellent, good, average and poor quality blastocyst groups, respectively. Fitted logistic regression analysis taking into account the following covariables: female, age, embryo chromosomal status and day of blastocyst development/biopsy showed that morphology was predictive of the comprehensive chromosome screening result (p < 0.05). A logistic regression analysis was also performed on clinical outcomes taking into account the effect of blastocyst morphology and day of blastocyst development/biopsy. None of the parameters were shown to be significant, suggesting morphology and day of blastocyst development/biopsy do not reduce the competence of euploid embryos (p > 0.05). CONCLUSIONS: After eSET, implantation rate was 80-86%; live birth rate per embryo transfer was 60-73% and clinical miscarriage rate was found to be < 10% and were not significantly affected by the embryo morphology. Results are concordant with those reported when using aCGH and highlights the competence of poor-quality euploid embryos.


Assuntos
Blastocisto/fisiologia , Implantação do Embrião , Fertilização In Vitro/métodos , Testes Genéticos/métodos , Idade Materna , Diagnóstico Pré-Implantação/métodos , Adulto , Aneuploidia , Coeficiente de Natalidade , Blastocisto/citologia , Transferência Embrionária , Desenvolvimento Embrionário , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Ploidias , Gravidez , Taxa de Gravidez
9.
J Assist Reprod Genet ; 36(8): 1599-1607, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31236830

RESUMO

PURPOSE: Preimplantation genetic testing for aneuploidy (PGT-A) has become increasingly controversial since normal euploid births have been reported following transfer of embryos diagnosed as "abnormal." There is an increasing trend in transferring "abnormal" embryos; but it is still unknown how many IVF centers transfer "abnormal" embryos and with what efficiency. METHODS: We performed a worldwide web-survey of IVF centers to elucidate PGT-A related practice patterns including transfer of human embryos found "abnormal" by PGT-A. Participating centers reflected in vitro fertilization (IVF) cycles in the USA, Canada, Europe, Asia, South America, and Africa. RESULTS: One hundred fifty-one IVF centers completed the survey; 125 (83%) reported utilization of PGT-A. Europe had the highest utilization (32.3%), followed by the USA and Canada combined at 29.1%. The leading indications for PGT-A were advanced maternal age (77%), followed by recurrent implantation failure (70%), unexplained pregnancy loss (65%), and sex determination (25%); 14% of respondents used PGT-A for all of their IVF cycles; 20% of IVF units reported transfers of chromosomally "abnormal" embryos, and 56% of these took place in the USA, followed by Asia in 20%. Remarkably, 106 (49.3%) cycles resulted in ongoing pregnancies (n = 50) or live births (n = 56). Miscarriages were rare (n = 20; 9.3%). CONCLUSIONS: The transfers of "abnormal" embryos by PGT-A offered robust pregnancy and live birth chances with low miscarriage rates. These data further strengthen the argument that PGT-A cannot reliably determine which embryos should or should not be transferred and leads to disposal of many normal embryos with excellent pregnancy potential.


Assuntos
Aborto Espontâneo/prevenção & controle , Aneuploidia , Aberrações Cromossômicas , Fertilização In Vitro/métodos , Testes Genéticos/métodos , Nascimento Vivo , Diagnóstico Pré-Implantação/métodos , Aborto Espontâneo/genética , Adulto , Transferência Embrionária , Feminino , Humanos , Internet , Gravidez , Inquéritos e Questionários
10.
Radiología (Madr., Ed. impr.) ; 61(3): 225-233, mayo-jun. 2019. ilus, tab, graf
Artigo em Espanhol | IBECS | ID: ibc-185294

RESUMO

Antecedentes y objetivo: Existe una carencia de métricas cuantitativas de la calidad del hueso trabecular alveolar, factor determinante en implantología. El objetivo de este estudio es desarrollar una metodología con tomografía computarizada multidetector para objetivar la calidad del hueso trabecular y establecer diferencias entre los distintos tipos y el estado de las piezas dentarias mediante procesado de imágenes y análisis estructural. Materiales y métodos: Se analizan 20 pacientes con exploración de tomografía computarizada multidetector dental para la valoración del hueso mandibular y posiciones dentales. El análisis de las imágenes incluyó la segmentación automática de la mandíbula, obtención de secciones perpendiculares a la arcada dentaria y análisis estructural del hueso trabecular de cada sección. Se obtuvieron la ratio entre volumen de hueso y volumen total de la sección, el grosor, la separación y el número trabecular, y la atenuación promedio en unidades Hounsfield. Se analizaron diferencias entre tipos de diente (incisivos, caninos, premolares y molares) y entre estados de las piezas dentarias (ausente o presente). Resultados: Se obtuvieron diferencias estadísticamente significativas entre los tipos y estados de las piezas. Por tipo, los incisivos mostraron mayor ratio de hueso trabecular, con disminución progresiva para caninos, premolares y molares. Por estado, las secciones pertenecientes a dientes ausentes presentaron mayor ratio de hueso que con el diente presente. Conclusiones: La metodología desarrollada permite cuantificar las propiedades estructurales del hueso alveolar a partir de imágenes de tomografía computarizada multidetector. Los resultados obtenidos objetivan el estado del sustrato óseo de cara a la planificación y seguimiento de la colocación de implantes dentales


Background and objective: There is a lack of quantitative measures of the quality of alveolar trabecular bone, an important factor in implantology. This study aimed to develop a method of objectively assessing the quality of trabecular bone by means of image processing and structural analysis of multidetector computed tomography images and to establish differences between tooth types and tooth presence/absence. Materials and methods: We analyzed 20 patients who underwent multidetector computed tomography to evaluate mandibular bone and tooth positioning. Image analysis included automatic segmentation of the mandible, obtainment of sections perpendicular to the dental arch, and structural analysis of the trabecular bone in each section. We calculated the ratio between the volume of bone and the total volume of the section, the thickness, the trabecular number, and the mean attenuation in Hounsfield units. We analyzed the differences among different tooth types (incisors, canines, premolars, and molars) and between present and absent teeth. Results: We found statistically significant differences between different tooth types and between sections in which teeth were present or absent. Incisors had a greater ratio of trabecular bone; the ratio of trabecular bone progressively decreased from the incisors to the canines, premolars, and molars. The ratio of trabecular bone was greater in sections in which teeth were absent than in those in which teeth were present. Conclusions: The method allows to quantify the structural properties of alveolar bone from multidetector computed tomography images. Our results provide an objective picture of the bone substrate that can be useful for planning and following up dental implant procedures


Assuntos
Humanos , Tomografia Computadorizada Multidetectores/métodos , Osso Esponjoso/diagnóstico por imagem , Mandíbula/diagnóstico por imagem , Implantação Dentária Endo-Óssea/métodos , Cuidados Pré-Operatórios/métodos , Diagnóstico Pré-Implantação/métodos , Estudos Retrospectivos
11.
Fertil Steril ; 112(1): 82-88, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31056308

RESUMO

OBJECTIVE: To investigate whether blastocyst biopsy in preimplantation genetic testing (PGT) increases the risk of adverse neonatal outcomes. DESIGN: Retrospective cohort study. SETTING: University-affiliated center. PATIENTS: Live births after blastocyst biopsy combined with frozen ET (PGT group) and frozen blastocyst transfer after in vitro fertilization or intracytoplasmic sperm injection (control group). INTERVENTION(S): Blastocyst biopsy. MAIN OUTCOME MEASURE(S): Gestational age (GA), birth weight (BW), and rates of preterm birth (PB), very preterm birth (VPB), extreme preterm birth (EPB), low birth weight (LBW), very low birth weight (VLBW), and macrosomia. RESULT(S): No significant differences were observed in the sex ratio, GA, PB, VPB, EPB, BW, or rates of LBW, VLBW, and macrosomia between the PGT and control groups for either singletons or twins. However, the cesarean section rate of the PGT group was significantly higher than that of the control group for twins (adjusted odds ratio, 2.383 [1.079, 5.259]). Regarding fluorescence in situ hybridization-PGT neonates, neonatal outcomes, including GA, BW, and rates of PB, VPB, LBW, and VLBW, did not differ between the different groups of biopsied cells (≥10 group and <10 group) for either the grade B or grade C trophectoderm score subgroups; however, in the grade B trophectoderm score subgroup, the rate of boy babies in the ≥10 group was significantly higher than that in the <10 group (83.3% vs. 40.9%). The association between the number of biopsied cells and GA/BW was not statistically significant. CONCLUSION(S): Blastocyst biopsy may not add additional risk to neonatal outcomes when compared with a control group.


Assuntos
Blastocisto/patologia , Transferência Embrionária , Fertilização In Vitro , Testes Genéticos , Diagnóstico Pré-Implantação/métodos , Adulto , Biópsia/efeitos adversos , Peso ao Nascer , Transferência Embrionária/efeitos adversos , Feminino , Fertilização In Vitro/efeitos adversos , Idade Gestacional , Humanos , Hibridização in Situ Fluorescente , Recém-Nascido de Baixo Peso , Recém-Nascido , Recém-Nascido Prematuro , Nascimento Vivo , Valor Preditivo dos Testes , Gravidez , Nascimento Prematuro/etiologia , Estudos Retrospectivos , Medição de Risco , Fatores de Risco , Injeções de Esperma Intracitoplásmicas , Resultado do Tratamento
12.
Fertil Steril ; 112(2): 291-297.e3, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31133385

RESUMO

OBJECTIVE: To investigate the effects of parental mosaicism on their preimplantation embryos. DESIGN: Case series. SETTING: An institute for reproductive and stem cell engineering. PATIENT(S): Sixty-eight mosaic couples. INTERVENTION(S): Assisted reproduction with preimplantation genetic testing (PGT). MAIN OUTCOME MEASURE(S): Karyotypes, embryo-related chromosomal abnormalities, and PGT results. RESULT(S): A total of 209 embryos were obtained from 68 mosaic couples, and 153 (73.21%) of 209 of the total embryos were obtained from 55 mosaic couples with abnormal sex chromosome numbers. Of these 153 embryos, 2 (1.31%) had an abnormal copy number of X chromosome, 1 had mosaicism with 46,XN,+X(mosaic, 40%), 1 (0.65%) had an extra Y chromosome, 3 (1.96%) exhibited both X chromosomal and autosomal abnormalities, and 4 (2.61%) exhibited de novo X chromosome structural abnormalities. A total of 56 (26.79%) of 209 embryos were obtained from mosaic couples (n = 13) with abnormal autosomal structures. Notably, of these 56 embryos, 5 (8.93%) had a 16q21-q24.3 copy number abnormality related to the parental karyotype, with a fragile site at 16q22; 5 (7.14%) exhibited 46,XX,dup(8p23.1-8p11.21) and 46,XY,del(8p22-8p11.21), which were related to the parental karyotype; and 10 (17.86%) were de novo chromosome abnormalities. CONCLUSION(S): Our data demonstrate that the risk of embryo-related chromosome abnormalities in mosaic patients with abnormal sex chromosomes is very low. Therefore, PGT may not need to be recommended for mosaic patients with abnormal copy numbers of sex chromosomes, especially for patients with financial difficulties. By contrast, the mosaic patients with structural abnormalities of autosomes may have a relatively high risk of abnormal embryos with an unbalanced segment of the involved chromosomes. Thus, PGT is highly recommended for mosaic patients with autosomal structure abnormalities, especially those with a fragile site at 16q22.


Assuntos
Fertilização In Vitro , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Mosaicismo , Diagnóstico Pré-Implantação/métodos , Adulto , Blastocisto , Aberrações Cromossômicas/embriologia , Aberrações Cromossômicas/estatística & dados numéricos , Análise Citogenética/métodos , Análise Citogenética/estatística & dados numéricos , Feminino , Fertilização In Vitro/métodos , Fertilização In Vitro/estatística & dados numéricos , Testes Genéticos/métodos , Humanos , Masculino , Mosaicismo/estatística & dados numéricos , Diagnóstico Pré-Implantação/estatística & dados numéricos , Estudos Retrospectivos , Fatores de Risco
13.
Fertil Steril ; 112(2): 283-290.e2, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31103283

RESUMO

OBJECTIVE: To assess whether pregnancies achieved with trophectoderm biopsy for preimplantation genetic testing (PGT) have different risks of adverse obstetric and neonatal outcomes compared with pregnancies achieved with IVF without biopsy. DESIGN: Observational cohort. SETTING: University-affiliated fertility center. PATIENT(S): Pregnancies achieved via IVF with PGT (n = 177) and IVF without PGT (n = 180) that resulted in a live birth. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Maternal outcomes including preeclampsia and placenta previa and neonatal outcomes including birth weight and birth defects. RESULT(S): There was a statistically significant increase in the risk of preeclampsia among IVF+PGT pregnancies compared with IVF without PGT pregnancies, with an incidence of 10.5% versus 4.1% (adjusted odds ratio [aOR] = 3.02; 95% confidence interval [95% CI], 1.10, 8.29). The incidence of placenta previa was 5.8% in IVF+PGT pregnancies versus 1.4% in IVF without PGT pregnancies (aOR = 4.56; 95% CI, 0.93, 22.44). Similar incidences of gestational diabetes, preterm premature rupture of membranes, and postpartum hemorrhage were observed. IVF+PGT and IVF without PGT neonates did not have a significantly different gestational age at delivery or rate of preterm birth, low birth weight, neonatal intensive care unit admission, neonatal morbidities, or birth defects. All trends, including the significantly increased risk of preeclampsia in IVF+PGT pregnancies, persisted upon stratification of analysis to only singleton live births. CONCLUSION(S): To date, this is the largest and most extensively controlled study examining maternal and neonatal outcomes after trophectoderm biopsy. There was a statistically significant three-fold increase in the odds of preeclampsia associated with trophectoderm biopsy. Given the rise in PGT use, further investigation is warranted.


Assuntos
Ectoderma/patologia , Resultado da Gravidez/epidemiologia , Diagnóstico Pré-Implantação , Trofoblastos/patologia , Adulto , Biópsia/efeitos adversos , Estudos de Coortes , Feminino , Fertilização In Vitro/métodos , Fertilização In Vitro/estatística & dados numéricos , Testes Genéticos/métodos , Testes Genéticos/estatística & dados numéricos , Humanos , Recém-Nascido , Pré-Eclâmpsia/epidemiologia , Pré-Eclâmpsia/etiologia , Gravidez , Diagnóstico Pré-Implantação/efeitos adversos , Diagnóstico Pré-Implantação/métodos , Diagnóstico Pré-Implantação/estatística & dados numéricos , Nascimento Prematuro/epidemiologia , Nascimento Prematuro/etiologia , Estudos Retrospectivos
14.
Syst Biol Reprod Med ; 65(3): 258-263, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30977407

RESUMO

The current study describes a successful case of preimplantation genetic diagnosis (PGD) of primary open angle glaucoma (POAG) and verifies the efficiency of next-generation sequencing (NGS)-based haplotyping for PGD of POAG. In this study, we applied NGS as part of PGD to effectively detect POAG prior to embryo implantation and avoid the prospect of pregnancy termination in event of vertical inheritance of POAG. We used the technique of multiple annealing and looping based amplification cycles (MALBAC) to conduct whole genome amplification (WGA) and to reduce the allele dropout (ADO). We also employed Sanger sequencing to directly detect the mutation c.1109 C > T in MYOC and NGS-based single nucleotide polymorphism (SNP) haplotyping to distinguish the chromosomes that carried the mutation. Copy number variation (CNV) analysis was carried out to determine the copy number of embryos' chromosomes. Of the 4 blastocysts obtained in this study, only 2 (sample 5 and 7) could be successfully amplified by WGA. CNV results indicated that chromosomes of both these samples were balanced (46, XN). Sanger sequencing and NGS-based SNP haplotyping confirmed that sample 7 carried the mutation c.1109 C > T in MYOC, while sample 5 did not. Moreover, no ADO was observed. Thus, blastocyst 5 was transferred into the uterus of the patient, and a healthy baby without the MYOC mutation c. 1109C>T was born 39 weeks after transplantation. Our study suggests that NGS-based SNP haplotyping is an effective technique for the PGD of POAG. Abbreviations: PGD: preimplantation genetic diagnosis; POAG: primary open angle glaucoma; NGS: next-generation sequencing; MALBAC: multiple annealing and looping based amplification cycles chemistry; WGA: whole genome amplification; ADO: allele dropout; SNP: single nucleotide polymorphism; CNV: copy number variation; MYOC: Myocilin; OPTN: Optineurin; WDR36: WD repeat domain 36; CYP1B1: Cytochrome P450 1 B Chain; ICSI: intracytoplasmic sperm injection; TFNA: testicular fine-needle aspiration; TE: trophectoderm; PCR: polymerase chain reaction.


Assuntos
Proteínas do Citoesqueleto/genética , Proteínas do Olho/genética , Glaucoma de Ângulo Aberto/diagnóstico , Glicoproteínas/genética , Diagnóstico Pré-Implantação/métodos , Variações do Número de Cópias de DNA , Feminino , Fertilização In Vitro , Glaucoma de Ângulo Aberto/genética , Haplótipos , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Gravidez
15.
Fertil Steril ; 111(6): 1151-1158, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31005312

RESUMO

OBJECTIVE: To evaluate the growth, health, and motor development of children born after preimplantation genetic diagnosis (PGD). DESIGN: Observational cohort study and comparison of 5-year-old children born after PGD to similar aged children born after IVF/intracytoplasmic sperm injection (ICSI) and children from families with a genetic disorder born after natural conception (NC). SETTING: University hospital. PATIENT(S): One hundred three children were included in the PGD group. The two control groups consisted of 90 children born after IVF/ICSI and 58 children born after NC. INTERVENTION(S): PGD. MAIN OUTCOME MEASURE(S): We measured height, weight, body circumferences, body mass index, and blood pressure and performed a dysmorphological and neurological examination. We also collected data about the children's medical history, health care consultations, and motor milestones. RESULT(S): The mean height, weight, and body mass index were comparable for all groups. Six (5.8%) PGD, four (4.4%) IVF/ICSI, and five (8.6%) NC children had a major congenital abnormality. The incidence of acute and chronic illnesses was similar in all groups. Motor milestones were achieved on time, but the IVF/ICSI group had a slightly younger mean sitting age. None of the children had severe neurological problems. CONCLUSION(S): Five-year-old children born after PGD show normal growth, health, and motor development when compared with children born after IVF/ICSI and NC children from families with a genetic disorder. TRIAL REGISTRATION NUMBER: NCT02149485.


Assuntos
Desenvolvimento Infantil , Saúde da Criança , Fertilização In Vitro/efeitos adversos , Doenças Genéticas Inatas/genética , Testes Genéticos , Infertilidade/terapia , Destreza Motora , Diagnóstico Pré-Implantação/métodos , Fatores Etários , Pressão Sanguínea , Estatura , Índice de Massa Corporal , Pré-Escolar , Feminino , Doenças Genéticas Inatas/diagnóstico , Doenças Genéticas Inatas/fisiopatologia , Nível de Saúde , Humanos , Infertilidade/diagnóstico , Infertilidade/fisiopatologia , Masculino , Medição de Risco , Fatores de Risco , Injeções de Esperma Intracitoplásmicas/efeitos adversos , Resultado do Tratamento , Ganho de Peso
16.
J Assist Reprod Genet ; 36(5): 989-994, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30887160

RESUMO

OBJECTIVE: To investigate the usefulness of preimplantation genetic diagnosis (PGD) based on mutated allele revealed by sequencing with aneuploidy and linkage analyses (MARSALA) for a pedigree with X-linked retinitis pigmentosa (XLRP). METHODS: One pathogenic mutation (c.494G > A) of the retinitis pigmentosa GTPase regulator (RPGR) gene was identified in a pedigree affected by XLRP. Then, PGD was carried out for the couple, of which the wife was an XLRP carrier. Three blastocysts were biopsied and then MARSALA was performed by next-generation sequencing (NGS). Prenatal diagnosis was also carried out to confirm the PGD results. RESULTS: Three blastocysts were all unaffected. Then, one of the embryos was chosen randomly to be transferred, and the pregnancy was acquired successfully. The results of prenatal diagnosis were consistent with the PGD results. The fetus did not carry RPGR mutation (c.494G > A) and had normal chromosome karyotype. As a result, a healthy baby free of XLRP condition was born. CONCLUSION: The PGD method based on MARSALA was established and applied to a family with XLRP successfully. MARSALA will be a valid tool, not only for XLRP families but also for families affected with other monogenetic disorders, to prevent transmission of the genetic disease from parents to offspring.


Assuntos
Proteínas do Olho/genética , Doenças Genéticas Ligadas ao Cromossomo X/diagnóstico , Doenças Genéticas Ligadas ao Cromossomo X/genética , Mutação Puntual , Diagnóstico Pré-Implantação/métodos , Retinite Pigmentosa/diagnóstico , Retinite Pigmentosa/genética , Adulto , Aneuploidia , Análise Mutacional de DNA , Feminino , Ligação Genética , Humanos , Masculino , Linhagem , Gravidez , Resultado da Gravidez
17.
J Assist Reprod Genet ; 36(5): 819-826, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30895497

RESUMO

In recent years, a growing body of literature has emerged investigating the clinical utility of spent embryo media (SEM) for preimplantation genetic testing for aneuploidy (PGT-A) (Hammond et al. in Fertil Steril. 107(1):220-8, 2017; Xu et al. in Proc Natl Acad Sci USA. 113(42):11907-12, 2016; Shamonki et al. in Fertil Steril. 106(6):1312-8, 2016; Feichtinger et al. in Reprod BioMed Online. 34(6):583-9, 2017; Vera-Rodriguez et al. in Hum Reprod. 33(4):745-56, 2018; Kuznyetsov et al. in PLoS One. 13(5):e0197262, 2018; Ho et al. in Fertil Steril. 110(3):467-75, 2018; Capalbo et al. in Fertil Steril. 110(5):870-9, 2018). Most of these studies have reported moderate success rates, suggesting the need for improvements in sensitivity and specificity. The concordance between spent media and embryo biopsy or whole embryo was reported to be between 30.4 and 90%, with 50-70% correlation being the most representative (Xu et al. in Proc Natl Acad Sci USA. 113(42):11907-12, 2016; Shamonki et al. in Fertil Steril. 106(6):1312-8, 2016; Feichtinger et al. in Reprod BioMed Online. 34(6):583-9, 2017; Vera-Rodriguez et al. in Hum Reprod. 33(4):745-56, 2018; Kuznyetsov et al. in PLoS One. 13(5):e0197262, 2018; Ho et al. in Fertil Steril. 110(3):467-75, 2018). Here, we will analyze all spent media testing strategies including SEM collection methods, whole genome amplification (WGA) strategies, chromosome copy number detection, and bioinformatics analysis tools. We will propose improvements to further increase the accuracy and sensitivity of the assay before bringing PGT-A with SEM into the clinical sphere.


Assuntos
Aneuploidia , Meios de Cultura/análise , Técnicas de Cultura Embrionária/métodos , Embrião de Mamíferos/metabolismo , Testes Genéticos/métodos , Diagnóstico Pré-Implantação/métodos , Embrião de Mamíferos/citologia , Feminino , Humanos , Gravidez
18.
BMC Med Genomics ; 12(1): 52, 2019 03 18.
Artigo em Inglês | MEDLINE | ID: mdl-30885195

RESUMO

BACKGROUND: Preimplantation genetic testing (PGT) has already been applied in chromosomally balanced translocation carriers to improve the clinical outcome of assisted reproduction. However, traditional methods could not further distinguish embryos carrying a translocation from those with a normal karyotype prior to implantation. METHODS: To solve this problem, we developed a method named "Chromosomal Phasing on Base level" (BasePhasing), which based on Infinium Asian Screening Array-24 v1.0 (ASA) and a specially phasing pipeline. Firstly, by comparing the number of single nucleotide polymorphism (SNP) loci in different minor allele frequencies (MAFs) and in 2Mbp continuous windows of ASA chip and karyomap-12 chip, we verified whether ASA could be adopted for genome-wide haplotype linkage analysis. Besides, the whole gene amplification (WGA) of 3-10 cells of GM16457 cell line was used to verify whether ASA chip could be used for testing of WGA products. Finally, two balanced translocation families were utilized to carry out BasePhasing and to validate the feasibility of its clinical application. RESULTS: The average number of SNP loci in each window of ASA (473.2) was twice of that of Karyomap-12 (201.2). The coincidence rate of SNP loci in genomic DNA and WGA products was about 97%. The 5.3Mbp deletion was detected positively in cell line GM16457 of both genomic DNA and WGA products, and haplotype linkage analysis was performed in genome wide successfully. In the two balanced translocation families, 18 blastocysts were analyzed, in which 8 were unbalanced and the other 10 were balanced or normal chromosomes. Two embryos were transferred back to the patients successfully, and prenatal cytogenetic analysis of amniotic fluid was performed in the second trimester. The results predicted by BasePhasing and prenatal diagnosis were totally consistent. CONCLUSIONS: Infinium ASA bead chip based BasePhasing pipeline shows good performance in balanced translocation carrier testing. With the characteristics of simple operation procedure and accurate results, we demonstrate that BasePhasing is one of the most suitable methods to distinguish between balanced and structurally normal chromosome embryos from translocation carriers in PGT at present.


Assuntos
Testes Genéticos/métodos , Haplótipos , Diagnóstico Pré-Implantação/métodos , Translocação Genética/genética , Linhagem Celular , Feminino , Humanos , Polimorfismo de Nucleotídeo Único , Gravidez
19.
Fertil Steril ; 111(5): 928-935, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30922652

RESUMO

OBJECTIVE: To investigate whether aneuploidy screening in preimplantation genetic testing (PGT) for monogenic diseases improves the ongoing pregnancy/live birth rate of single frozen/thawed embryo transfer (FET) cycles in young women. DESIGN: Retrospective cohort study. SETTING: Single university-based fertility center. PATIENT(S): From January 2016 to December 2017, 569 FET cycles were selected for analysis. The aneuploidy screening (AS) group included 131 FET cycles from 105 oocyte retrieval cycles in 98 patients who underwent PGT for monogenic diseases with aneuploidy screening, and the non-AS group included 438 FET cycles from 280 oocyte retrieval cycles in 266 patients who underwent PGT for monogenic diseases without aneuploidy screening. INTERVENTION(S): The patient population was all under the age of 35 years and underwent PGT for monogenic diseases with and without AS. MAIN OUTCOME MEASURE(S): Ongoing pregnancy/live birth rate, live birth rate, implantation rate, and miscarriage rate. RESULT(S): Aneuploidy screening significantly improved the ongoing pregnancy/live birth rate (61.22% vs. 43.98%), implantation rate (64.29% vs. 50.38%), and live birth rate (53.06% vs. 36.09%) of young women carrying monogenic diseases in the first FET cycles. When adjusted for the parity, number of previous miscarriages, and percentage of infertility, the likelihood of implantation was 1.874 times higher (95% confidence interval 1.126-3.119), and an ongoing pregnancy/live birth was 2.139 times more likely (95% confidence interval 1.295-3.534). In addition, the miscarriage rate was significantly decreased (3.17% vs. 11.94%). In the cumulative pregnancy outcomes, the cumulative ongoing pregnancy/live birth rate both per transfer and per patient were significantly higher in the AS group (62.24% vs. 50.38% and 79.59% vs. 68.80%), but no difference existed after adjusting for the parity, number of previous miscarriage, and percentage of infertility. Nevertheless, aneuploidy screening reduced the time interval from the first ET to the achievement a pregnancy. CONCLUSION(S): Aneuploidy screening in PGT significantly improved the ongoing pregnancy/live birth rate of young women carrying monogenic diseases in the first FET cycles.


Assuntos
Aneuploidia , Testes Genéticos/métodos , Recuperação de Oócitos/métodos , Resultado da Gravidez/genética , Diagnóstico Pré-Implantação/métodos , Adulto , Estudos de Coortes , Feminino , Humanos , Masculino , Gravidez , Resultado da Gravidez/epidemiologia , Estudos Retrospectivos , Adulto Jovem
20.
Taiwan J Obstet Gynecol ; 58(2): 239-243, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30910146

RESUMO

OBJECTIVE: The primary objective of this study was to investigate whether preimplantation genetic testing for aneuploidy (PGT-A) of blastocysts through array comparative genomic hybridization (aCGH) improves live birth rates (LBR) in IVF cycles for patients with high prevalence of aneuploidy. MATERIALS AND METHODS: This study included 1389 blastocysts with aCGH results derived from 296 PGT-A cycles in IVF patients with advanced maternal age (AMA) (n = 87, group A), those with repeated implantation failure (RIF) (n = 82, group B), those with recurrent miscarriage (RM) (n = 82, group C), and oocyte donors (OD) (n = 45, young age, as a control group). Another 61 AMA patients without PGT-A procedures were used as a control group for group A. Vitrification was performed after blastocyst biopsy, and thawed euploid embryos were transferred in a nonstimulated cycle. RESULTS: For the AMA group, a significant increase in LBRs was found in the PGT-A group compared with the non-PGT-A group (54.1% vs. 32.8%, p = 0.018). Consistent LBRs (54.1%, 51.6%, 55.9%, and 57.1%, respectively, in group A, B, C, and young age group) were obtained for all the indications. CONCLUSIONS: LBRs can be improved using PGT-A of blastocysts with aCGH in IVF cycles for patients with a high rate of aneuploidy, especially for patients with AMA.


Assuntos
Aneuploidia , Coeficiente de Natalidade , Fertilização In Vitro/métodos , Testes Genéticos/métodos , Diagnóstico Pré-Implantação/métodos , Aborto Habitual/genética , Adulto , Estudos de Casos e Controles , Hibridização Genômica Comparativa , Transferência Embrionária/efeitos adversos , Transferência Embrionária/métodos , Transferência Embrionária/estatística & dados numéricos , Feminino , Fertilização In Vitro/estatística & dados numéricos , Humanos , Idade Materna , Gravidez , Estudos Retrospectivos , Falha de Tratamento
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