Adeno-associated virus-mediated gene therapy in a patient with Canavan disease using dual routes of administration and immune modulation.

Gene replacement therapy is a rational therapeutic strategy and clinical intervention for neurodegenerative disorders like Canavan disease, a leukodystrophy caused by biallelic mutations in the aspartoacylase (ASPA) gene. We aimed to investigate whether simultaneous intravenous (i.v.) and intracerebroventricular (i.c.v.) administration of rAAV9-CB6-ASPA provides a safe and effective therapeutic strategy in an open-label, individual-patient, expanded-access trial for Canavan disease. Immunomodulation was given prophylactically prior to adeno-associated virus (AAV) treatment to prevent an immune response to ASPA or the vector capsid. The patient served as his own control, and change from baseline was assessed by clinical pathology tests, vector genomes in the blood, antibodies against ASPA and AAV capsids, levels of cerebrospinal fluid (CSF) N-acetylaspartate (NAA), brain water content and morphology, clinical status, and motor function tests. Two years post treatment, the patient's white matter myelination had increased, motor function was improved, and he remained free of typical severe epilepsy. NAA level was reduced at 3 months and remained stable up to 4 years post treatment. Immunomodulation prior to AAV exposure enables repeat dosing and has prevented an anti-transgene immune response. Dual-route administration of gene therapy may improve treatment outcomes.

Modification of late human embryo development after blastomere removal on day 3 for preimplantation genetic testing.

The purpose of our study was to use a time-lapse monitoring (TLM) system to determine if day 3 blastomere biopsy for preimplantation genetic testing (PGT) had an impact on subsequent morphokinetic parameters at the morula and blastocyst stages. In this retrospective monocentric study conducted between May 2013 and August 2017, we compared late morphokinetic parameters in embryos undergoing day 3 blastomere biopsy for PGT and in control non-biopsied embryos obtained in intracytoplasmic sperm injection (ICSI) cycles for male infertility. All embryos in both groups were cultured in a TLM system. The biopsy group was composed of 1691 embryos (386 PGT cycles). The control group was composed of 2578 embryos (786 ICSI cycles). Early morphokinetic parameters up to day 3 were similar in both groups. Concerning late morphokinetic parameters, the onset of compaction (tSC), fully-compacted morula stage (tM), onset of cavitation/early blastulation (tSB), and blastocyst stages (tB and tEB) appeared significantly earlier in the biopsy group than in the control group. We found that late morphokinetic events at the morula and the blastocyst stages occurred significantly earlier in biopsied embryos than in control non-biopsied-embryos. The mechanisms underlying these modifications of embryo development after biopsy should be investigated in order to determine precisely, and this phenomenon could be associated with embryo, fetal, and offspring development.Abbreviations: TLM: time-lapse monitoring; PGT: preimplantation genetic testing; ICSI: intracytoplasmic sperm injection; tSC: the onset of compaction; tM: fully-compacted morula stage; tSB: onset of cavitation/early blastulation; tB and tEB: blastocyst stages; OHSS: ovarian hyperstimulation syndrome.

Can time-lapse parameters predict embryo ploidy? A systematic review.

Embryo morphology assessment performs relatively poorly in predicting implantation. Embryo aneuploidy screening (PGS) has recently improved, but its clinical value is still debated, and the development of a cheap non-invasive method for the assessment of embryo ploidy status is a highly desirable goal. The growing implementation of time-lapse devices led some teams to test the effectiveness of morphokinetic parameters as predictors of embryo ploidy, with conflicting results. The aim of this study was to conduct a comprehensive review of the literature on the predictive value of morphokinetic parameters for embryo ploidy status. A systematic search on PubMed was conducted using the following key words: time-lapse, morphokinetic, aneuploidy, IVF, preimplantation genetic screening, PGS, chromosomal status. A total of 13 studies were included in the analysis. They were heterogeneous in design, patients, day of embryo biopsy, statistical approach and outcome measures. No single or combined morphokinetic parameter was consistently identified as predictive of embryo ploidy status. In conclusion, the available studies are too heterogeneous for firm conclusions to be drawn on the predictive value of time-lapse analysis for embryo aneuploidy screening. Hence, morphokinetic parameters should not be used yet as a surrogate for PGS to determine embryo ploidy in vitro.

Comparison of embryo morphokinetics after in vitro fertilization-intracytoplasmic sperm injection in smoking and nonsmoking women.

OBJECTIVE: To evaluate the impact of female smoking on early embryo development in vitro using a time-lapse system. DESIGN: Retrospective analysis of prospectively collected database. SETTING: Assisted reproductive technology (ART) program. PATIENT(S): A total of 135 infertile couples undergoing IVF-intracytoplasmic sperm injection (ICSI) cycle. INTERVENTION(S): Embryo culture in a time-lapse incubator (Embryoscope, Unisense Fertilitech). MAIN OUTCOME MEASURE(S): Embryo morphokinetic parameters. RESULT(S): Of 135 women, 139 and 729 oocytes were cultured in 23 smokers and 112 nonsmokers, respectively. Basal characteristics and ovarian stimulation parameters were comparable in both groups. Serum antimüllerian hormone level and antral follicle count were significantly lower in smokers than in nonsmokers. Conventional embryologic parameters were comparable in both groups, but time-lapse analysis showed that most cleavage events occurred significantly later in smokers than in nonsmokers, leading to poor cycle outcome in smokers. CONCLUSION(S): Female smoking impairs early embryo development, as reflected by time-lapse analysis. This might contribute to the dramatic decrease in pregnancy rates (PRs) in smokers undergoing IVF. Time lapse is a useful tool for the identification of deleterious components impairing early embryo development.