The aim of our study was to discover whether genomic analysis of maternal exomes is capable of identifying new target genes to improve infertility diagnosis in cases with recurrent preimplantation developmental embryo arrest. Genetic analysis. The study was conducted at Istanbul Memorial Hospital ART and Reproductive Genetics Unit between December 2018-November 2019 in cooperation with Igenomix, Italy. Ten women, five with consanguinity history, four being the offspring of first-cousin marriage with a history of recurrent embryo developmental failure in multiple IVF cycles were recruited. WES was performed (Agilent SureSelect whole-exome capture and Illumina sequencing technology). Variant calling against the reference genome GRCh38 was done using Freebayes and identified on average 436k high quality variants per samples. According to Ensembl classification 2.8% are expected to have high (0.25%) or moderate (2.56%) disruptive impact in the gene product. Variants were filtered on a per-individual base using a number of criteria (frequency <0.05% in the 1000 Genomes and gnomeAD; severity as estimated by Ensembl; the functional effect using the CADD score above the 90 percentile and variants location in genes highly intolerant to loss of function, pLI>0.9. Finally variants retained had to be in genes relevant to the early embryonic development (3600 gene list). To control for false positives, we run the same filtering on 100 replicates of 10 random samples from the publicly available Human Genome Diversity Project data set, and we filtered out variants falling in genes showing up in 50% of the hundred replicates, controlling for random occurrence of hits. Overall, 1700 unique variants in 1281 unique genes were retained after filtering, most involved in lethal embryonic pathways. Thirty-one unique retained variants have high impact and among them sixteen are splice variants and nine are stop-gains. Each sample carries on average 185.9 (10.0 s.d.) potentially detrimental variants. Of particular relevance two individuals had pathogenic variants in SPAG5, an essential component of the mitotic spindle required for normal chromosome segregation and progression into anaphase. Furthermore, two individual showed pathogenetic variants in the zinc finger protein 91 (ZFP91). The knockdown of ZFP91 reduces FOXA1 polyubiquitination and cellular progression in embryonic and cancer cells. Finally, three samples share the G allele of the rs1217009744 variants in homozygosis in the SHANK3 gene. Exome analysis of women with recurrent embryo arrest successfully identifies genomic variants lethal at the embryonic stage, thus providing a diagnostic tool. However, functional genomics studies and validation in an independent cohort of patients with preimplantation embryo arrest phenotype and of different ethnicity is required to corroborate these findings. The generation of polygenic models will also further contribute to increasing discovery rate and to the development of more general and powerful predictive models for this phenotype.

OBJECTIVE:Tricuspid annular movement and velocities before and after thrombolytic therapy were investigated for the detection of right ventricular (RV) involvement in RCA (right coronary artery)-related acute inferior myocardial infarction (IMI).METHODS:Patients with RCA-related acute IMI were evaluated for this pilot prospective cohort study. Annular movement was measured by TAPSE (tricuspid annular plane systolic excursion), and annular velocities were measured by tissue Doppler echocardiography. Data collected before and after thrombolysis and angiography. Diagnosis of RV myocardial infarction (RVMI) was defined by co-presence of electrocardiographic and angiographic criteria. Chi-square and Student's t-tests were used in statistical analysis.RESULTS:Thirty-one patients were included. Before thrombolysis, annular velocities and TAPSE were found significantly higher in patients without RVMI than in patients with RVMI. Comparison of tricuspid systolic velocity (Sa) and movement before and after thrombolytic therapy in patients without RVMI revealed no significant difference (21.6±2.1 mm vs. 21.8±2.0 mm p>0.05 and 136.1±8.8 mm/s vs. 137.5±9.0 mm/s p>0.05, for TAPSE and Sa respectively). Contrarily, in patients with RVMI, TAPSE and systolic velocity increased significantly after thrombolysis compared with pre-thrombolysis (16.2±2.0 mm vs. 17.6±1.8 mm p=0.001 and 110.0±12.6 mm/s vs. 113.08±12.7 mm/s p=0.027 for TAPSE and Sa respectively). Diastolic velocities did not change significantly after thrombolysis in patients with RVMI.CONCLUSION:Tricuspid annular movement and velocity measurement by echocardiography may contribute to echocardiographic diagnosis of RV involvement in RCA-related IMI. Patients without RVMI have significantly higher annular velocities and TAPSE than in patients with RVMI before thrombolysis. Only in IMI patients with RVMI, significant increases in TAPSE and Sa were observed after thrombolysis.