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Character of cell-free genomic DNA in embryo culture medium and the prospect of its clinical application in preimplantation genetic testing

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Reproductive and Developmental Medicine 2022年第1期6卷 51-56页

作者： Cai-xia Lei Xiao-Xi Sun Prenatal Diagnosis Center Obstetrics and Gynecology HospitalFudan UniversityShanghai 200011China Department of Genetics Shanghai Ji Ai Genetics&IVF InstituteShanghai 200011China Department of Endocrinology and Reproductive Medicine Obstetrics and Gynecology HospitalFudan UniversityShanghai 200011China Department of Gynaecology Shanghai Ji Ai Genetics&IVF InstituteShanghai 20001TChina Key Laboratory of Female Reproductive Endocrine Related Diseases Obstetrics and Gynecology Hospital of Fudan UniversityShanghai 200011China

There is increasing evidence that cell-free DNA (cfDNA) in spent culture media (SCM) can be amplified for genetic testing. Therefore, this paper reviews the characteristics of cfDNA, including its fragment size, amount, origin, as well as some factors affecting the success rate of its amplification, together to provide researchers with a more comprehensive perspective on embryonic cfDNA. The origin of cfDNA in SCM is complicated and poses challenges to the interpretation of genetic test results. Advanced molecular techniques should distinguish between embryonic and contaminated DNA to maximize the success rate of amplification and analysis. Recent data showed that the type of culture medium, assisted hatching or not, the type of amplification kit, and fresh or thawed embryos were not related to the success rate of amplification, but the length of culture time might affect the success rate. The longer culture time, the more cfDNA is available in the SCM. Then we focused on the concordance between trophectoderm (TE), inner cell mass, whole embryo, and embryonic cfDNA. Despite successful amplification, the concordance between TE and embryonic cfDNA was low. In summary, non-invasive genetic testing using SCM could represent a major advance in future single embryo selection, however, contamination and timing for media collection are key factors affecting the results, and current non-invasive cfDNA testing should not be directly applied to clinical practice. Further research is needed to improve the methods used for testing techniques and genetic analysis to achieve greater accuracy and trace its origins before it can be used in the clinics.

关键词： Cell-free DNA Embryo culture medium Genomic DNA Non-invasive Pre-implantation genetic testing

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Modified strict sperm morphology threshold aids in the clinical selection of conventional in vitro fertilization(IVF)or intracytoplasmic sperm injection(ICSI)

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Asian Journal of Andrology 2022年第1期24卷 62-66页

作者： Yong Zhu Feng Zhang Hua Cheng Xiao-Xi Sun Feng Jiang Obstetrics and Gynecology Hospital Fudan UniversityShanghai 200011China Shanghai Ji Ai Genetics and IVF Institute Obstetrics and Gynecology HospitalFudan UniversityShanghai 200011China

For infertility treatment,the selection of in vitro fertilization(IVF)or intracytoplasmic sperm injection(ICSI)is decided by multiplying indicators(including fallopian tube factors,semen count,and semen motility),except for sperm morphology.In this study,we conducted a retrospective analysis,from implantation to birth,over a period of 5 years.A total of 1873 couples with primary or secondary fallopian tube factors and an increased defective sperm morphology rate(DSMR)were divided into different groups to receive IVF or ICSI cycles.By comparing the outcomes,we found that the F1 group(DSMR98%,IVF group 3;P98%,ICSI group 3)had higher cleavage rate(P<0.001),biochemical pregnancy rate(P<0.05),clinical pregnancy rate(P<0.05)and live birth rate(P<0.05)than the F3 group.However,the ICSI subgroup had a lower two pronuclei fertilization rate than the IVF subgroup(P<0.05).Our data suggest that the sperm morphology should also be considered when selecting IVF or ICSI combined with other semen parameters before the first assisted reproductive technologies(ART)cycle,especially for males with severe sperm defects.

关键词： clinical outcome in vitro fertilization intracytoplasmic sperm injection sperm morphology

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Predictive Modeling in Reproductive Medicine

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Reproductive and Developmental Medicine 2018年第4期2卷 224-229页

作者： Jing Lin Xiao-Xi Sun Shanghai Ji Ai Genetics and IVF Institute Obstetrics and Gynecology Hospital of Fudan UniversityShanghai 200011China Laboratory of Female Reproductive Endocrine-Related Diseases Shanghai 200011China

The accurate prediction of fertility outcomes is an extremely interesting and challenging task in reproductive medicine. Efforts in this area focus on classic statistical models and newer technologies, including machine learning. The modeling process has three steps, namely, data preparation, model selection and data fitting, and model validation. Here, we present a review of studies on these methods of fertility prediction. Various databases were searched using relevant keywords. Original studies with full-text available on this topic were included for review. Earlier studies explored prediction models for spontaneous pregnancy prognosis, reproductive outcomes after intrauterine insemination and in vitro fertilization, and implantation potential based on embryo morphology and morphokinetic data. Future directions for predictive modeling in reproductive medicine include solving problems presented by big data, identifying novel informative features, balancing predictive power and result interpretability, and validating models with gold-standard study designs.

关键词： Machine Learning Prediction Model Predictive Modeling Regression Model Reproductive Medicine

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Effects of the Interval between Ovulation Induction using Clomiphene Citrate and Frozen Embryo Transfer on Pregnancy Outcome

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Reproductive and Developmental Medicine 2021年第2期5卷 90-96页

作者： Yi-Ning Xu Lu Li Xiao-Xi Sun Department of Reproductive Medicine Shanghai Ji Ai Genetics and IVF InstituteObstetrics and Gynecology Hospital of Fudan UniversityShanghai 200011China

Objective:To explore the best timing for frozen embryo transfer(FET)after ovarian stimulation and egg retrieval using the clomiphene citrate(CC)+human menopausal gonadotropin(hMG)ovulation induction regimen through a retrospective analysis.Methods:Data of patients who underwent CC+hMG ovulation induction and FET from January 2014 to December 2019 were analyzed retrospectively.The patients were divided into three groups according to the interval from egg retrieval to FET:CC1(within 1 menstrual cycle),CC2(2 menstrual cycles),and CC3(≥3 menstrual cycles).Indicators such as hormone levels and pregnancy outcomes were recorded to explore the effect of different intervals on pregnancy outcome.Results:A total of 1,082 transfer cycles were included in this retrospective analysis.The implantation,clinical pregnancy,and live birth rates in the CC1 group were significantly lower than those in the CC2 and CC3 groups(P<0.05).The E2/P4 ratio on progesterone injection day(3 days before thawed embryo transfer)was lower in the CC1 group than in the other groups(P<0.05).After adjusting for all factors using multifactor regression analysis,the interval between egg retrieval and FET was found to be an independent predictor of the implantation,pregnancy,and live birth rates.Conclusion:An interval of more than one menstrual cycle between the day of egg retrieval after ovarian stimulation with the CC+hMG ovulation induction regimen and the day of FET can result in high implantation,clinical pregnancy,and live birth rates,which can lead to an improved pregnancy outcome.

关键词： Assisted Reproduction Clomiphene Citrate Endometrial Receptivity Frozen Embryo Transfer Ovulation Induction Pregnancy Outcome

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Epidemiological Survey and Risk Factor Analysis of Recurrent Spontaneous Miscarriages in Infertile Women at Large Infertility Centers

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Chinese Medical Journal 2017年第17期130卷 2056-2062页

作者： Hai-Yan Wang Jie Qiao Xiao-Xi Sun Shu-Yu Wang xiao-Yan Liang Yun sun Feng-Hua Liu Reproductive Medical Center Department of Obstetrics and Gynecology Peking University Third Hospital Beijing 100191 China Shanghai Ji Ai Diagnosis and Treatment Center for Genetics and Infertility Shanghai 200011 China Reproductive Medical Center Beijing Obstetrics and Gynecology Hospital Beijing 100006 China Reproductive Medical Center The Sixth Affiliated Hospital Sun Yat-sen University Guangzhou Guangdong 510665 China Reproductive Medical Center Shanghai Jiao Tong University School of Medicine Affiliated Renji Hospital Shanghai 200011 China Reproductive Medical Center Maternal and Child Health Hospital of Guangdong Province Guangzhou Guangdong 511400 China

Background：A higher frequency of spontaneous miscarriage has been observed in infertile couples,and there is a higher prevalence of infertility among patients with a history of recurrent spontaneous miscarriages （RSMs;〉2 miscarriages）.This study aimed to determine the proportion of infertile patients with RSM and examine risk factors associated in patients with RSM being treated with assisted reproductive technologies.Methods：This cross-sectional observational study was conducted at six reproductive medicine centers in three cities of China.Data of 751 patients with at least one spontaneous miscarriage were analyzed.Demographic data and etiological factors associated with infertility were compiled and compared between patients with a single spontaneous miscarriage （SSM） and those with RSM.Results：Two hundred （26.6%,95% confidence interval [CI]：23.50-29.95%） patients experienced RSMs and 551 （73.4%） had a single miscarriage.The odds of RSM increased with increasing age （odds ratio [OR] =1.06）,uterine disorders （OR =2.09）,endocrine disorders （OR =2.48）,and immune disorders （OR =2.98）.Higher education level,masters or above,and a pelvic cavity disorder were associated with lower risk of RSM （OR =0.27 and 0.46,respectively）.Late spontaneous miscarriages were more frequent in patients with RSM than in those with a SSM （31.5% vs.14.2%,respectively,P 〈 0.001） and were associated with a history of uterine cavity procedures （OR =2.095） and cervical factors related to infertility （OR =4.136,95% CI：1.012-16.90）.Conclusions：Compared to patients with only a SSM,the conditions of patients with RSM are more complicated.To increase the success rate of assisted reproductive technology,factors including uterus cavity adhesion,cervical relaxation,endocrine disorders,and immune disorders should be treated before assisted reproduction is initiated.These data may provide treatment guidance for infertile patients with a history of RSM.

关键词： Assisted Reproductive Technology Epidemiological Survey Infertility Recurrent Spontaneous Miscarriage Risk Factor

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Current Status and Recent Advances in Preimplantation Genetic Testing for Structural Rearrangements

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Reproductive and Developmental Medicine 2020年第1期4卷 1-6页

作者： Shuo Zhang Cai-xia Lei Xiao-Xi Sun Cong-Jian Xu Department of Genetics Shanghai Ji Ai Genetics and IVF InstituteObstetrics and Gynecology HospitalFudan UniversityShanghai 200011China Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases Obstetrics and Gynecology HospitalFudan UniversityShanghai 200011China

Preimplantation genetic testing(PGT)is an early form of prenatal genetic diagnosis,which can identify the abnormal embryos cultured in vitro,allow only transfer of genetically normal embryos,and improve the pregnancy rate.In recent years,the rapid development of microarrays and next-generation sequencing(NGS)technologies has remarkably accelerated the clinical application of PGT.In particular,a variety of detection methods have emerged and achieved significant progress in PGT for structural rearrangements(PGT-SR).The detection-related abilities of these methods range from the detection of limited chromosome aneuploidy to comprehensive chromosome screening of the whole genome to differentiation of embryos with normal or balanced translocation/inversion karyotypes.In this study,we reviewed PGT-SR-related detection techniques to provide a better reference for clinical application and research.We have also discussed the potential development of novel techniques in the future.

关键词： Chromosome Aneuploidy Inversion Karyotype Preimplantation Genetic Testing Structural Chromosomal Rearrangement Structural Rearrangement Translocation Karyotype

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Integrated bioinformatics analysis to identify key genes and pathways involved in the endometria of patients with recurrent implantation failure undergoing in vitro fertilization and embryo transfer

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Reproductive and Developmental Medicine 2023年第4期7卷 230-237页

作者： Yi-Lun Sui Lu Li Xiao-Xi Sun Shanghai Ji Ai Genetics and IVF Institute Obstetrics and Gynaecology HospitalFudanUniversityShanghai200011China

Objectives: Recurrent implantation failure (RIF) is a challenging clinical problem that occurs in approximately 10% of patients undergoingin vitro fertilization and embryo transfer. Because of the varied criteria for RIF, as well as differences in the time and methods of endometrial biopsy, previous studies have not reached an agreement regarding the key genes and pathways of RIF. In this study, bioinformatic analyses were performed in a homogeneous population with consistent biopsy conditions to explore the molecular mechanisms and identify key RIF-associated genes and pathways in the endometrium.Methods: Seven datasets from the Gene Expression Omnibus database were reviewed in detail, and GSE111974, GSE103465, and GSE26787 were included for further analysis. Common differentially expressed genes (DEGs) were identified using the GEO2R online tool. Gene Ontology and pathway enrichment analyses were performed and a protein–protein interaction (PPI) network was constructed. The RNA and protein expression files of the hub genes were obtained.Results: A total of 402 common DEGs were identified between patients with RIF and controls. The enrichment of these DEGs was mainly associated with tube morphogenesis, vasculature development, and positive regulation of cell migration. Pathway enrichment indicated that pathways involved in cancer, the vascular endothelial growth factor A-vascular endothelial growth factor receptor 2 signaling pathway, and cell cycle pathways were mainly involved. Moreover, a PPI network was constructed, and the three most significant submodules were mainly involved in rRNA processing and the regulation of proteolysis. The following ten DEGs with high degrees of connectivity were identified as hub genes:CDC42,VEGFA,MAP2K1,APP,CYCS,UBE2I,PLEK,LMNA,SMAD2, andATRX.Conclusion: Key genes and pathways identified in this study could improve our understanding of the underlying molecular events in RIF and be used as potential biomarkers and therapeutic targets. Further studies are required to confirm thes

关键词： Bioinformatics Biomarker Gene expression Recurrent implantation failure Pathways

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A homozygous protein-truncating mutation in ACTL7A causes male infertility characterized by fertilization failure

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Reproductive and Developmental Medicine 2022年第3期6卷 169-174页

作者： Yao Chen Biao-Bang Chen Qing Sang Lei Wang Jun-Li Zhao Xiao-Xi Sun Shanghai Ji Ai Genetics and IVF Institute Obstetrics and Gynecology Hospital of Fudan UniversityShanghai 200011China NHC Key Lab of Reproduction Regulation(Shanghai Institute of Planned Parenthood Research) Fudan UniversityShanghai 200032China Institute of Pediatrics Children's Hospital of Fudan Universitythe Institutes of Biomedical Sciencesand the State Key Laboratory of Genetic EngineeringFudan UniversityShanghai 200032China Center for Reproductive Medicine General Hospital of Ningxia Medical UniversityYinchuan 750004China

Objective:This study aimed to screen for novel mutations in ACTL7A and expand the spectrum of known mutations responsible for recurrent fertilization failure.Methods:Whole-exome sequencing was performed on samples from couples who experienced recurrent assisted reproductive technology failure and visited the General Hospital of Ningxia Medical University.Western blotting and quantitative Real-time PCR were used to investigate the effects of the mutation on HEK293T cells.Results:Samples from 12 couples with total fertilization failure or poor fertilization(fertilization rate<20%)were subjected to whole-exome sequencing,and a novel homozygous protein-truncating mutation(c.1101dupC,p.S368Qfs*5)in ACTL7A was identified in a patient with recurrent poor fertilization.The mutant resulted in a truncated protein as well as decreased protein expression level in HEK293T cells.Conclusions:Our findings expand the mutational and phenotypic spectrum of ACTL7A,thus providing a potential diagnostic marker for fertilization failure due to male factors.

关键词： ACTL7A Fertilization failure Male infertility Mutation

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Sperm-specific protein ACTL7A as a biomarker for fertilization outcomes of assisted reproductive technology

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Asian Journal of Andrology 2022年第3期24卷 260-265页

作者： Tian-Ying Yang Ying Chen Guo-Wu Chen Yi-Si sun Zhi-Chao Li xiao-Rong Shen Yi-Ni Zhang Wen He Dan Zhou Hui-Juan Shi Ai-Jie xin Xiao-Xi Sun Shanghai Ji Ai Genetics and IVF Institute Obstetrics and Gynecology HospitalFudan UniversityShanghai 200011China NHC Key Lab of Reproduction Regulation(Shanghai Institute for Biomedical and Pharmaceutical Technologies) School of PharmacyFudan UniversityShanghai 200032China Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases Shanghai 200011China Department of Endocrinology and Reproductive Medicine Obstetrics and Gynecology HospitalFudan UniversityShanghai 200011China

Obtaining high-quality embryos is one of the key factors to improve the clinical pregnancy rate of assisted reproductive technologies(ART).So far,the clinical evaluation of embryo quality depends on embryo morphology.However,the clinical pregnancy rate is still low.Therefore,new indicators are needed to further improve the evaluation of embryo quality.Several studies have shown that the decrease of sperm-specific protein actin-like 7A(ACTL7A)leaded to low fertilization rate,poor embryo development,and even infertility.The aim of this study was to study whether the different expression levels of ACTL7A on sperm can be used as a biomarker for predicting embryo quality.In this study,excluding the factors of severe female infertility,a total of 281 sperm samples were collected to compare the ACTL7A expression levels of sperms with high and low effective embryo rates and analyze the correlation between protein levels and in-vitro fertilization(IVF)laboratory outcomes.Our results indicated that the ACTL7A levels were significantly reduced in sperm samples presenting poor embryo quality.Furthermore,the protein levels showed a significant correlation with fertilization outcomes of ART.ACTL7A has the potential to be a biomarker for predicting success rate of fertilization and effective embryo and the possibility of embryo arrest.In conclusion,sperm-specific protein ACTL7A has a strong correlation with IVF laboratory outcomes and plays important roles in fertilization and embryo development.

关键词： actin-like 7A assisted reproductive technology biomarker embryo quality in‑vitro fertilization outcomes

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Influence of Growth Hormone Supplementation in Patients with Thin Endometrium Undergoing Frozen Embryo Transfer

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Reproductive and Developmental Medicine 2019年第1期3卷 49-53页

作者： Jun-Yi Yang He Li Nan Lu Lu Li Xiao-Xi Sun Shanghai Ji Ai Genetics and IVF Institute Obstetrics and Gynecology Hospital of Fudan UniversityShanghai 200011China Key Laboratory of Female Reproductive Endocrine Related Diseases Obstetrics and Gynecology Hospital of Fudan UniversityShanghai 200011China

Objective:To evaluate the effect of recombinant human growth hormone(rhGH)supplementation during hormone-replacement therapy and frozen-thawed embryo transfer(FET)in patients with thin endometrium.Methods:A retrospective research was conducted on 225 patients,who underwent artificial cycle FET in Shanghai,China,between January 2016 and November 2017.Data from 245 FET cycles were analyzed,of which 184 cycles received rhGH(GH group)and 61 did not(control group).Results:Clinical pregnancy and implantation rates were significantly higher in the GH group than those in the control group(64.7%vs.49.2%,P=0.032;44.8%vs.32.8%,P=0.019,respectively).After logistic regression analysis,rhGH was considered the only significant variable that influenced clinical pregnancy rate,increasing it by 1.89-fold.On the other hand,the presence of rhGH did not seem to affect the early pregnancy loss.Conclusions:Our results indicated that simultaneous addition of rhGH could improve clinical outcomes of FET in patients with thin endometrium,particularly in patients between 30 and 34 years of age.

关键词： Endometrial Receptivity Frozen-Thawed Embryo Transfer Growth Hormone Thin Endometrium

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