Antonio Capalbo

Juno Genetics · reproductive genetics laboratory

The advancement of in vitro culture media has improved the efficacy of ARTs in the treatment of infertility (Chronopoulou and Harper, 2015). Despite this progress, embryos may demonstrate delayed development and reach the blastocyst stage on Day 7. The prevalence of these slow embryos has been reported as 5% and even though they are considered to b...

Purpose Carrier screening (CS) is a term used to describe a genetic test performed on individuals without family history of genetic disorders, to investigate the carrier status for pathogenic variants associated with multiple recessive conditions. The advent of next-generation sequencing enabled simultaneous CS for an increasing number of condition...

STUDY QUESTION How should ART/preimplantation genetic testing (PGT) centres manage the detection of chromosomal mosaicism following PGT? SUMMARY ANSWER Thirty good practice recommendations were formulated that can be used by ART/PGT centres as a basis for their own policy with regards to the management of ‘mosaic’ embryos. WHAT IS KNOWN ALREADY T...

Preimplantation genetic testing for aneuploidy (PGT-A) is used as a frequent add-on for in vitro fertilization (IVF) to improve clinical outcomes. The purpose is to select a euploid embryo following chromosomal testing on embryo biopsies. The current practice includes comprehensive chromosome screening (CCS) technology applied on trophectoderm (TE)...

In IVF cycles, the application of aneuploidy testing at the blastocyst stage is quickly growing, and the latest reports estimate almost half of cycles in the US undergo preimplantation genetic testing for aneuploidies (PGT-A). Following PGT-A cycles, understanding the predictive value of an aneuploidy result is paramount for making informed decisio...

Sir, We read with interest the study by Barad et al. (2022) entitled ‘IVF outcomes of embryos with abnormal PGT-A biopsy previously refused transfer: a prospective cohort study’. Although the definition of positive and negative predictive values across the broad spectrum of aneuploidies detectable in trophectoderm biopsies is certainly of paramount...

The implementation of high-resolution genome-wide methods, usually next-generation sequencing-based, in preimplantation genetic testing (PGT), has led to the frequent detection of embryos with chromosomal mosaicism (whole chromosome and/or segmental aberrations). Despite a growing series of papers showing the birth of healthy babies following the t...

Study question Do different gonadotrophins for controlled-ovarian-stimulation (COS) affect metaphase-II (MII) oocyte competence? Summary answer Euploid blastocyst rate (EBR) per cohort of MII-oocytes, live-birth-rate (LBR) per first vitrified-warmed euploid single-embryo-transfer (SET) and cumulative-LBR are independent from the gonadotrophins use...

Study question Can chromosomal abnormalities beyond aneuploidies (i.e., ploidy and microdeletions, MD) be detected on a single trophectoderm (TE) embryo biopsy using a next-generation sequencing (NGS)-based workflow? Summary answer This NGS-based integrated approach allows accurate detection of ploidy status and the most common microdeletions from...

Study question Would reimbursement of expanded carrier screening (ECS) panels of different gene content be cost-effective in a universal healthcare system? Summary answer Both tested ECS panels would be cost-effective if reimbursed by the Italian national healthcare system for a population without personal/familiar or racial/ethnic preconception g...

RESEARCH QUESTION To develop a methodology for case selection and whole-exome sequencing (WES) analysis in infertile women due to recurrent oocyte maturation defects (OOMD) and/or preimplantation embryo lethality (PREMBL). DESIGN We collected data from IVF patients attending the Istanbul Memorial Hospital (2015-2021).A statistical methodology for...

The December edition of the ESHRE Journal Cub discussed a recent article by Tan et al. (2022) describing the use of metabolic imaging to detect aneuploidy non-invasively in mouse embryos. A strong body of evidence supports that embryonic aneuploidy increases the risk of pregnancy loss (Lee and Kiessling, 2017). The current method to detect aneuploi...

The most important factor associated with oocytes’ developmental competence has been widely identified as the presence of chromosomal abnormalities. However, growing application of genome-wide sequencing (GS) in population diagnostics has enabled the identification of multifactorial genetic predispositions to sub-lethal pathologies, including those...

Epigenetics is the branch of genetics that studies the different mechanisms that influence gene expression without direct modification of the DNA sequence. An ever-increasing amount of evidence suggests that such regulatory processes may play a pivotal role both in the initiation of pregnancy and in the later processes of embryonic and fetal develo...

The December edition of the ESHRE Journal Cub discussed a recent article by Tan et al. (2022) describing the use of metabolic imaging to detect aneuploidy non-invasively in mouse embryos. A strong body of evidence supports that embryonic aneuploidy increases the risk of pregnancy loss (Lee and Kiessling, 2017). The current method to detect aneuploi...

Miscarriage is the spontaneous termination of a pregnancy before 24 weeks of gestation. We studied the genome of euploid miscarried embryos from mothers in the range of healthy adult individuals to understand genetic susceptibility to miscarriage not caused by chromosomal aneuploidies. We developed gp , a pipeline that we used to prioritize 439 uni...

Hereditary cancer syndromes are a heterogeneous group of genetic conditions that are associated with an increased risk of developing cancer during lifespan. In affected women, parenthood may be accompanied by concerns for the offspring, considering the common autosomal dominant inheritance. Moreover, fertility preservation to prevent the detrimenta...

OBJECTIVE To develop a methodology for case selection and whole-exome sequencing (WES) analysis in infertile women due to recurrent oocyte maturation defects(OOMD) and/or preimplantation embryo lethality (PREMBL). DESIGN Retrospective cohort study. SETTING IVF patients attending the Istanbul Memorial Hospital (2015-2021). WES and bioinformatics w...

Chromosome imbalance (aneuploidy) is the major cause of pregnancy loss and congenital disorders in humans. Analyses of small biopsies from human embryos suggest that aneuploidy commonly originates during early divisions, resulting in mosaicism. However, the developmental potential of mosaic embryos remains unclear. We followed the distribution of a...

With an upward trend in delaying parenthood, women across the world face an increasing risk of age-related infertility and involuntary childlessness. Elective oocyte banking strategies offer women the possibility to protect part of their reproductive potential until personal finances, personal relationship, or career have stabilized. Timely collect...

Study question What is the effect of varying diagnostic thresholds on the accuracy of Next Generation Sequencing (NGS)-based preimplantation genetic testing for aneuploidies (PGT-A)? Summary answer When single trophectoderm biopsies are tested, the employment of 80% upper threshold increases mosaic calls and false negative aneuploidy results compa...

Study question Can PGT-A reduce the anxiety generally experienced by infertile women undergoing IVF in the waiting period between embryo transfer and the pregnancy test? Summary answer PGT-A reduces anxiety in infertile women after embryo transfer, probably due to a gain of confidence in their treatment route. What is known already The waiting pe...

Study question How common abnormal cleavage patterns (ACP) are in IVF and what are their consequences on embryo developmental competence? Summary answer ACP might affect up to 25% of the 2PN-zygotes, independently from patients’/cycles’ characteristics, and mostly cause embryo developmental arrest around the 4-to–8-cell transition. What is known...

Study question Do whole-exome sequencing (WES) data from infertile women provide valuable information for the discovery of genes/pathways involved in extreme IVF phenotypes, i.e. oocyte/embryo developmental arrest? Summary answer The development of a specific bioinformatic WES pipeline revealed known and new candidate genes/pathways for isolated o...

Study question Can major de novo genetic and chromosomal abnormalities (i.e., ploidy, microdeletions) be effectively tested on a single embryo biopsy specimen using an integrated NGS approach? Summary answer The integrated NGS workflow provided high accuracy for multilevel chromosome and genetic abnormalities analysis based on single biopsies expa...

Study question Can major de novo genetic and chromosomal abnormalities (i.e., ploidy, microdeletions) be effectively tested on a single embryo biopsy specimen using an integrated NGS approach? Summary answer The integrated NGS workflow provided high accuracy for multilevel chromosome and genetic abnormalities analysis based on single biopsies expa...

Study question: What is the effect of varying diagnostic thresholds on the accuracy of Next Generation Sequencing (NGS)-based preimplantation genetic testing for aneuploidies (PGT-A)? Summary answer: When single trophectoderm biopsies are tested, the employment of 80% upper threshold increases mosaic calls and false negative aneuploidy results comp...

Il nuovo testo di Biologia e Tecniche della Riproduzione edito da Edi.Ermes è diretto agli studenti dei corsi di laurea in Biologia e Biotecnologie, ma anche dei Master e dei Corsi di perfezionamento e ai professionisti operanti nell’ambito della riproduzione umana. Nella prima sezione sono sviluppati argomenti relativi alla biologia della riproduz...

STUDY QUESTION Is it possible to define a set of performance indicators (PIs) for clinical work in ART, which can create competency profiles for clinicians and for specific clinical process steps? SUMMARY ANSWER The current paper recommends six PIs to be used for monitoring clinical work in ovarian stimulation for ART, embryo transfer, and pregnan...

Study question: What is the clinical validity and utility of preconception Expanded Carrier Screening (ECS) application on the management of prospective parents? Summary answer: The high detection rate of at-risk couples (ARCs) and the high proportion opting for IVF/preimplantation genetic testing (PGT) treatment demonstrate the clinical utility...

Purpose of review: Expanded Carrier Screening (ECS) is a genetic test able to detect carriers for a large number of autosomal recessive and X-linked diseases. Its clinical utilization is increasing but some technical aspects for its implementation are still controversial. Recent findings: In the current literature, several aspects of ECS panel i...

Study question: Is there an association between patients' reproductive history and the mean euploidy rates per biopsied blastocysts (m-ER) or the live birth rates (LBRs) per first single vitrified-warmed euploid blastocyst transfers? Summary answer: Patients' reproductive history (as annotated during counselling) showed no association with the m...

Background Next generation sequencing (NGS) has increased detection sensitivity of intermediate chromosome copy number variations (CNV) consistent with chromosomal mosaicism. Recently, this methodology has found application in preimplantation genetic testing (PGT) of trophectoderm (TE) biopsies collected from IVF-generated human embryos. As a conse...

A clear definition of developmentally incompetent preimplantation embryo (DIPE) in literature is still missing, while several scientific societies are discussing this challenging topic. From both a clinical and scientific perspective, the identification of embryos unfit for reproductive purpose is crucial. This aim should be pursued in light of all...

Miscarriage is the spontaneous termination of a pregnancy before 24 weeks of gestation. We studied the genome of euploid miscarried embryos from mothers in the range of healthy adult individuals to understand genetic susceptibility to miscarriage not caused by chromosomal aneuploidies. We developed GP, a pipeline that we used to prioritize 439 uniq...

Background: Our genetic code is now readable, writable and hackable. The recent escalation of genome-wide sequencing (GS) applications in population diagnostics will not only enable the assessment of risks of transmitting well-defined monogenic disorders at preconceptional stages (i.e. carrier screening), but also facilitate identification of multi...

mosaic classification in blastocysts-stage primplantation genetic testing

comparing the two trophectoderm biopsy protocols

A recent study by Munné et al. portrayed a protocol to retrieve in vivo produced blastocysts after IUI and uterine lavage for preimplantation genetic testing (PGT) purposes. The authors claimed this protocol might represent a reasonable future perspective for patients who do not want to undergo IVF, but still want to be informed about their embryos...

Following early studies showing no adverse effects, cleavage stage biopsy by zona drilling using acid Tyrode's solution, and removal of single blastomeres for preimplantation genetic testing (PGT) and identification of sex in couples at risk of X-linked disease, was performed by Handyside and colleagues in late 1989, and pregnancies reported in 199...

Patient-Centered Assisted Reproduction - edited by Alice D. Domar March 2020

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;...

Purpose To report the effects of blastocyst stage aneuploidy testing on clinical, gestational, and neonatal outcomes for patients of advanced maternal age undergoing IVF. Methods This is a single-center observational-cohort study with 2 years follow-up. The study includes a total of 2538 couples undergoing 2905 egg collections (control group), 308...

Objective: To investigate whether the morphodynamic characterization of a euploid blastocyst's development allows a higher prediction of a live birth after single-embryo-transfer (SET). Design: Observational cohort study conducted in two phases: training and validation. Setting: Private in vitro fertilization centers. Patient(s): Euploid bla...

Thyroid hormones are regarded as the major controllers of metabolic rate and oxygen consumption in mammals. Although it has been demonstrated that thyroid hormone supplementation improves bovine embryo development in vitro, the cellular mechanisms underlying these effects are so far unknown. In this study, we investigated the role of thyroid hormon...

Limited translational genomic research data have been reported on the application of exome sequencing and parallel gene testing for preconception carrier screening (PCS). Here, we present individual-level data from a large PCS program in which exome sequencing was routinely performed on either gamete donors (5,845) or infertile patients (8,280) und...

Understanding fertility in young and old Fertility in humans follows a U-curve, with low rates in both teenagers and women of advancing maternal age (mid-30s and above). Gruhn et al. found that this distinct shape originates from chromosomal errors in human eggs, which result in genomic imbalance and pregnancy loss. The error types and chromosomes...

Study question: Can miRNAs be reliably detected in the spent blastocyst media (SBM) after IVF as putative biomarkers of the implantation potential of euploid embryos? Summary answer: Adjustment of the data for blastocyst quality and the day of full-expansion hinders the predictive power of a fast, inexpensive, reproducible and user-friendly prot...

Purpose Surplus cryopreserved affected/aneuploid blastocysts may be obtained after in vitro fertilization (IVF) treatments with preimplantation genetic testing (PGT). These embryos are considered not suitable for transfer and may be discarded. Currently, in Italy, an embryo disposition decision (EDD) is not allowed and the frozen/vitrified blastocy...

Introduction The introduction of highly sensitive Next Generation Sequencing (NGS) platforms for PGT-A applications concurred with an increased detection of segmental aneuploidies. Recent studies focused on understanding whether segmental aneuploidies derive from mitotic or meiotic events, however, definitive data are still lacking. To further cont...

Study question: Which are the clinical benefits and risks of including poor-quality blastocysts (PQBs) in the cohort of biopsied embryos during a cycle with preimplantation genetic testing for aneuploidies (PGT-A)? Summary answer: PQBs show a worse prognosis with respect to sibling non-PQBs, but their clinical use allows an overall 2.6% increase...

Objective: To study whether embryonic cell-free DNA (cfDNA) in spent blastocyst media is representative of the chromosomal constitution of a blastocyst. Design: Pilot prospective blinded study. Setting: In vitro fertilization center and genetics laboratory. Patient(s): A total of 115 trophectoderm (TE) biopsies and spent blastocyst media (SB...

Purpose To investigate the association of cumulus cell (CC)-related expression of a selected cluster of key genes (PTGS2, CAMK1D, HAS2, STC1, and EFNB2) with embryo development to blastocyst. Methods Exploratory study at a private clinic. Eighteen advanced maternal age patients were enrolled (37.3 ± 4.0 years). Seventy-five cumuli were collected,...

Recent advancements in genomic analysis allow testing of an increasing number of genetic features in human preimplantation embryos. Typical single gene mutation and whole chromosomes testing can now be integrated with assessment of mitochondrial DNA and polygenic conditions. Diagnostic expansion into epigenetic and transcriptomic assessment in the...

Objective: To evaluate the economical benefit of preimplantation genetic testing of aneuploidies (PGT-A) when applied in an extended culture and stringent elective single ET framework. Design: Theoretical cost-effectiveness study. Setting: Not applicable. Patients/animal(s): None. Intervention(s): Comparison of the cost-effectiveness betwe...

Objective: To determine whether blastocoel fluid (BF) or spent blastocyst medium (SBM) is a suitable template for genotype and/or karyotype assessment of in vitro fertilization–generated embryos. Design: Prospective blinded study. Setting: Genetic laboratory. Patient(s): From 26 patients undergoing preimplantation genetic testing (PGT) treatments,...

Study question: Are trophectoderm biopsy or other pre-vitrification features or laboratory practices associated with differences in blastocyst post-warming behavior (degeneration, re-expansion and live birth after single embryo transfer (SET))? Summary answer: Blastocyst morphology, day of full development and artificial shrinkage (either laser-...

Study question: Can a second round of biopsy, vitrification and chromosomal testing provide a valid diagnosis where the first attempt fails? Summary answer: The risk of inconclusive chromosomal-assessment after trophectoderm biopsy was 2.5% but a further biopsy and vitrification-warming appeared not to impair the competence of euploid blastocyst...

In the version of this Letter originally published, an author error led to the affiliations for Brendan Payne, Jonathan Coxhead and Gavin Hudson being incorrect. The correct affiliations are: Brendan Payne: 3Wellcome Trust Centre for Mitochondrial Research, Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, UK. 6Institute of...

Preimplantation genetic testing is a methodology aimed at the assessment of the genetic composition of an embryo. This diagnostic approach has been used in assisted reproduction for almost thirty years. During these years, the technologies used for embryo's genetic analysis have been continuously improved allowing the development of more precise, c...

Research question: Biochemical pregnancy loss (BPL), defined as serum beta-human chorionic gonadotropin levels ≥50 IU/l in at least two pregnancy tests, not associated with any ultrasonographical evidence of pregnancy, is often attributed to chromosomal abnormalities; however, no hard evidence exists to support this hypothesis. Are any IVF cycle pa...

Study question: Which is the prevalence of a 47,XXY karyotype in human blastocysts biopsied during preimplantation genetic testing for aneuploidies (PGT-A) cycles? Summary answer: The prevalence of a 47,XXY karyotype amongst male blastocysts without autosomal aneuploides is ~1%. What is known already: The prevalence of Klinefelter syndrome is...

Introduzione: Considerando l’elevato numero di patologie recessive o X-linked è stato stimato che circa il 2-3% delle coppie sono a rischio di concepire un figlio affetto in quanto costituite da portatori sani inconsapevoli di malattie a trasmissione recessiva. In caso di programmazione di una futura gravidanza, può essere utile conoscere lo stato...

Mitochondrial DNA (mtDNA) mutations cause inherited diseases and are implicated in the pathogenesis of common late-onset disorders, but how they arise is not clear1,2. Here we show that mtDNA mutations are present in primordial germ cells (PGCs) within healthy female human embryos. Isolated PGCs have a profound reduction in mtDNA content, with disc...