Management of copy number variants associated with incomplete penetrance and variable expressivity-Results of a French survey.

Increasing interest regarding neurodevelopmental disorders and democratization of chromosomal microarray analysis have led to growing identification of neuro-susceptibility copy number variations (CNVs). These CNVs have incomplete penetrance and variable expressivity (PIEV), which makes phenotypic features hard to predict. The French Consortium "AchroPuce" has provided a list of 17 CNVs that should be considered as PIEV CNVs. This list led to consensual French practices of healthcare professionals in postnatal diagnosis. However, no consensus was established in prenatal diagnosis and fetal pathology. 121 French health professionals were surveyed their opinions and practices regarding reporting of PIEV CNVs to patients, in order to identify key points so as to establish French recommendations. The survey showed that professionals in favor of reporting PIEV CNVs to patients in prenatal diagnosis and fetal pathology (respectively, 76% and 84% of respondents) considered highlighted that multidisciplinary consultation is the main point-of-care management before family survey. This statement is close to recommendations published worldwide. As a consequence, multidisciplinary expertise should be the basis of French recommendations concerning the reporting of PIEV CNVs and genetic counseling in prenatal diagnosis and fetal pathology.

Levels of follicular G-CSF and interleukin-15 appear as noninvasive biomarkers of subsequent successful birth in modified natural in vitro fertilization/intracytoplasmic sperm injection cycles.

OBJECTIVE: To explore oocyte competence for subsequent birth. The modified natural IVF/intracytoplasmic sperm injection (ICSI) cycle was used as an experimental model by measuring levels of cytokines, chemokines, and growth factors in individual follicular fluids (FF). DESIGN: A retrospective blinded study. SETTING: European network of research, Embryo Implantation Control (EMBIC). PATIENT(S): Single FF from 83 women were analyzed during a modified natural IVF/ICSI cycle, and reproducibility of follicular composition was evaluated over two cycles for 15 patients. INTERVENTION(S): Each FF sample was blindly tested to assess levels of 26 factors by bead-based immunoassays. MAIN OUTCOME MEASURE(S): Each mediator was evaluated as a potential biomarker of subsequent birth by multivariate regression analysis. RESULT(S): A combination of both FF G-CSF and IL-15 was the optimal model to predict birth (AUC(ROC), 0.85). Birth rates per cycle were 48.9% (16/33) if two good-prognosis criteria were present (FF G-CSF>12 pg/mL and IL-15<7 pg/mL) and 8% (3/36) and 0% (0/14) if, respectively, one or none were present. FF G-CSF was significantly correlated over two cycles (r=.71), suggesting a possible prognostic value of its documentation. CONCLUSION(S): Combined follicular G-CSF and IL-15 quantification appears as an efficient and noninvasive method to define oocyte competence for subsequent successful conception in modified natural IVF/ICSI cycles.

[Ten years' experience of preimplantation genetic diagnosis in Paris: remaining obstacles and potential solutions]. / Diagnostic pré-implantatoire: dix ans d'expérience en région parisienne: impasse actuelle et solutions à venir.

Preimplantation genetic diagnosis (PGD) has been authorized in France since 1999. Encouraging results have been obtained during the past 10 years in our Paris center, where 832 patients have undergone 1056 IVF-PGD procedures. With the advent of new techniques for the identification of genetic disease markers, our center can now offer PGD procedures for aneuploidy and 75 single-gene diseases. New indications for PGD have also been developed, such as mitochondrial DNA diseases, amyloid neuropathy, pulmonary arterial hypertension, and HLA typing The implantation rate is currently 29,6% and, by 31 December 2009, 151 healthy babies had been born. Unfortunately, demand for PGD procedures far outstrips available technical capacity, and the waiting period is longer than 18 months. Increased funding is urgently needed

Preimplantation genetic diagnosis: state of the art.

Preimplantation genetic diagnosis (PGD) is used to analyze embryos genetically before their transfer into the uterus. It was developed first in England in 1990, as part of progress in reproductive medicine, genetic and molecular biology. PGD offers couples at risk the chance to have an unaffected child, without facing termination of pregnancy. Embryos are obtained by in vitro fertilization with intracytoplasmic sperm injection (ICSI), and are biopsied mostly on day 3; blastocyst biopsy is mentioned as a possible alternative. The genetic analysis is performed on one or two blastomeres, by fluorescent in situ hybridization (FISH) for cytogenetic diagnosis, or polymerase chain reaction (PCR) for molecular diagnosis. Genetic analysis of the first or second polar body can be used to study maternal genetic contribution. Only unaffected embryos are transferred into the uterus. To improve the accuracy of the diagnosis, new technologies are emerging, with comparative genomic hybridization (CGH) and microarrays. In Europe, depending on national regulations, PGD is either prohibited, or allowed, or practiced in the absence of recommendations. The indications are chromosomal abnormalities, X-linked diseases or single gene disorders. The number of disorders being tested increases. In Europe, data collection from the year 2004 reports that globally 69.6% of cycles lead to embryo transfer and implantation rate is 17%. European results from the year 2004 show a clinical pregnancy rate of 18% per oocyte retrieval and 25% per embryo transfer, leading to 528 babies born. The cohort studies concerning the paediatric follow-up of PGD babies show developmental outcomes similar to children conceived after IVF-ICSI. Recent advances include human leucocyte antigen (HLA) typing for PGD embryos, when an elder sibling is affected with a genetic disorder and needs stem cell transplantation. The HLA-matched offspring resulting can give cord blood at birth. Preimplantation genetic screening (PGS) consists in euploid embryo selection; it could be used for advanced maternal age, repeated implantation failure, single embryo transfer or idiopathic recurrent pregnancy loss. These applications are controversial. PGD for inherited cancer predispositions is discussed and social sexing remains prohibited in Europe. PGD requires a close collaboration between obstetricians, fertility specialists, IVF laboratory and human geneticists. It needs intensive effort, expensive techniques and is demanding for the patients, but it offers tremendous opportunity for couples whose previous child has exhibited genetic abnormalities. The debate on certain indications is ongoing.

In vitro fertilization and embryo transfer in seminatural cycles for patients with ovarian aging.

OBJECTIVE: To investigate whether seminatural cycle is a reasonable management for ovarian aging patients. DESIGN: Prospective study. SETTING: ART Unit, Clamart, France. PATIENT(S): Seventy-five women, 158 cycles. INTERVENTION(S): Infertile women who presented with ovarian aging (defined as low ovarian reserve and characterized by cycle day 3 high FSH, high E2, and/or low inhibin B and/or previous cycle cancellations due to poor ovarian response to COH) were studied. Patients were offered up to three cycles. Treatment was scheduled as follows. From cycle day 8 onward the selection of the dominant follicle was monitored by ultrasound and hormonal measurements. When the dominant follicle appeared, patients received GnRH antagonist and, thereafter, hMG to support further follicular development. MAIN OUTCOME MEASURE(S): Implantation rate and clinical pregnancy. RESULT(S): Twenty-eight of 158 cycles were cancelled (17.7%). Oocyte pickups were performed in 119 (75.3%) cycles, 91 (57.6%) mature oocytes were retrieved, and 67 (42.4%) embryos transferred. Nineteen clinical pregnancies were obtained; the cumulative pregnancy rate per patient, after 3 cycles, was 35.2%. CONCLUSION(S): Use of a seminatural cycle is a reasonable management for patients with ovarian aging who have ovulatory menstrual cycles. It achieves a high implantation rate (28.3%).

Dynamics of serum anti-Müllerian hormone levels during the luteal phase of controlled ovarian hyperstimulation.

BACKGROUND: To investigate the dynamics of serum anti-Müllerian hormone (AMH) levels during the luteal phase of controlled ovarian hyperstimulation (COH) and its possible association with follicle development. METHODS: We prospectively studied 34 women undergoing COH with GnRH agonist and FSH. On the day of hCG (dhCG), serum AMH, estradiol (E2), progesterone and hCG levels were measured, and ovarian follicles were sorted into three size classes: <12, 12-15 and 16-22 mm. Hormonal measurements were repeated 4 days (hCG + 4) and 7 days (hCG + 7) after hCG. RESULTS: From dhCG to hCG + 4, we observed a decline in serum AMH levels (-64 +/- 3%; P < 0.0001), which paralleled that of E2 levels. From hCG + 4 to hCG + 7, an increase in AMH levels occurred (82 +/- 28%; P < 0.02), whose magnitude was correlated with the number of < 12 mm follicles (r = 0.68; P < 0.0001) but not with other follicle size classes nor with the remaining hormone levels. CONCLUSIONS: After hCG, AMH levels initially decline, presumably as an effect of follicle luteinization, then increase during the mid-luteal phase. Although the mechanisms implicated in the mid-luteal AMH increase are unclear, its positive association with small follicle count, but not with luteal progesterone and E2 levels, supports the hypothesis that AMH levels might reflect luteal follicle development.

[The French experience with preimplantation genetic diagnosis (PGD)]. / L'expérience française du diagnostic génétique pré-implantatoire (DPI). Bilan des premières naissances.

CONTEXT: Context Preimplantation genetic diagnosis (PGD) consists in the genetic analysis of one or two embryo cells obtained following embryo biopsy on the third day of culture. This diagnostic technique is reserved for couples with an identified risk of transmitting a serious and incurable disease and hence avoids the distress of having to revert to an abortion. METHOD: Since the publication ten years ago of the first preimplantation genetic diagnosis by the team in the UK, the number of PGD centres has progressed, but slowly. This is due to the need to associate the efforts of an in vitro fertility laboratory and a genetic laboratory capable of providing a diagnosis only on one or two embryo cells. This technique, controlled by the 1994 bio-ethical laws, first appeared in France in the year 2000. RESULTS: The indications, the techniques used and the first 12 births obtained in the first year of activity in the first two official centres in France are reported. In particular, no error in diagnosis following the PGD was noted with the systematic blood sample taken from the umbilical cord at birth. CONCLUSION: This diagnosis is of interest for couples who present risk of genetic transmission. The advantages and inconveniences of this method must be weighed against the prenatal diagnosis.

[Preimplantation genetic diagnosis: update of the Parisian group]. / Le diagnostic génétique préimplantatoire: premier bilan du groupe parisien.

To report the birth of the first fourteen infants conceived after preimplantation genetic diagnosis (PGD) in our unit. Fifty-nine couples were enrolled between January 2000 and July 2001. They had a total of 71 oocyte pick-up cycles. The collected oocytes were inseminated by intracytoplasmic sperm injection. The resulting embryos were biopsied on the third day of development and the genetic analysis was performed on the same day. Most of the embryo transfers were carried out on the fourth day. The 71 oocyte pick-up cycles yielded 872 oocytes of which 731 were suitable for intracytoplasmic sperm injection. Among the 505 embryos obtained, 421 embryos were biopsied and genetic diagnosis was performed for 312 (74%) of these 127 embryos were transferred during the course of 58 transfer procedures. There were 18 biochemical and 12 ongoing (7 singles, 4 twins and 1 triple) pregnancies. Sixteen infants have been born and 2 are expected. PGD has gained a place among the choices offered to couples at risk of transmission of a serious and incurable genetic disease.