National Rapid Genome Sequencing in Neonatal Intensive Care.

Importance: National implementation of rapid trio genome sequencing (rtGS) in a clinical acute setting is essential to ensure advanced and equitable care for ill neonates. Objective: To evaluate the feasibility, diagnostic efficacy, and clinical utility of rtGS in neonatal intensive care units (NICUs) throughout Israel. Design, Setting, and Participants: This prospective, public health care-based, multicenter cohort study was conducted from October 2021 to December 2022 with the Community Genetics Department of the Israeli Ministry of Health and all Israeli medical genetics institutes (n = 18) and NICUs (n = 25). Critically ill neonates suspected of having a genetic etiology were offered rtGS. All sequencing, analysis, and interpretation of data were performed in a central genomics center at Tel-Aviv Sourasky Medical Center. Rapid results were expected within 10 days. A secondary analysis report, issued within 60 days, focused mainly on cases with negative rapid results and actionable secondary findings. Pathogenic, likely pathogenic, and highly suspected variants of unknown significance (VUS) were reported. Main Outcomes and Measures: Diagnostic rate, including highly suspected disease-causing VUS, and turnaround time for rapid results. Clinical utility was assessed via questionnaires circulated to treating neonatologists. Results: A total of 130 neonates across Israel (70 [54%] male; 60 [46%] female) met inclusion criteria and were recruited. Mean (SD) age at enrollment was 12 (13) days. Mean (SD) turnaround time for rapid report was 7 (3) days. Diagnostic efficacy was 50% (65 of 130) for disease-causing variants, 11% (14 of 130) for VUS suspected to be causative, and 1 novel gene candidate (1%). Disease-causing variants included 12 chromosomal and 52 monogenic disorders as well as 1 neonate with uniparental disomy. Overall, the response rate for clinical utility questionnaires was 82% (107 of 130). Among respondents, genomic testing led to a change in medical management for 24 neonates (22%). Results led to immediate precision medicine for 6 of 65 diagnosed infants (9%), an additional 2 (3%) received palliative care, and 2 (3%) were transferred to nursing homes. Conclusions and Relevance: In this national cohort study, rtGS in critically ill neonates was feasible and diagnostically beneficial in a public health care setting. This study is a prerequisite for implementation of rtGS for ill neonates into routine care and may aid in design of similar studies in other public health care systems.

[GENOMIC COUNSELING - CHALLENGES AND OPPORTUNITIES].

INTRODUCTION: Medical genetics is a field that is continually evolving. In addition to enabling the discovery and understanding of the mechanisms underlying many diseases, the decoding of the human genome and swiftly advancing technology have also made it possible to integrate medical genetics into new clinical fields and acknowledge it as a significant profession across all clinical specialties. For genetic teams, these changes present numerous difficulties, particularly in the specific context of genetic counseling. The development of models of genetic counseling by teams that are not geneticists, adaptation to the reality where genetics is available to the general public, and models of genetic counseling by teams that are not geneticists are some of the challenges and suggested solutions discussed in detail in this article.

Limb girdle muscular disease caused by HMGCR mutation and statin myopathy treatable with mevalonolactone.

Myopathy is the main adverse effect of the widely prescribed statin drug class. Statins exert their beneficial effect by inhibiting HMG CoA-reductase, the rate-controlling enzyme of the mevalonate pathway. The mechanism of statin myopathy is yet to be resolved, and its treatment is insufficient. Through homozygosity mapping and whole exome sequencing, followed by functional analysis using confocal microscopy and biochemical and biophysical methods, we demonstrate that a distinct form of human limb girdle muscular disease is caused by a pathogenic homozygous loss-of-function missense mutation in HMG CoA reductase (HMGCR), encoding HMG CoA-reductase. We biochemically synthesized and purified mevalonolactone, never administered to human patients before, and establish the safety of its oral administration in mice. We then show that its oral administration is effective in treating a human patient with no significant adverse effects. Furthermore, we demonstrate that oral mevalonolactone resolved statin-induced myopathy in mice. We conclude that HMGCR mutation causes a late-onset severe progressive muscular disease, which shows similar features to statin-induced myopathy. Our findings indicate that mevalonolactone is effective both in the treatment of hereditary HMGCR myopathy and in a murine model of statin myopathy. Further large clinical trials are in place to enable the clinical use of mevalonolactone both in the rare orphan disease and in the more common statin myopathy.

Teaching clinicians practical genomic medicine: 7 years' experience in a tertiary care center.

PURPOSE: Increased implementation of complex genetic technologies in clinical practice emphasizes the urgency of genomic literacy and proficiency for medical professionals. We evaluated our genomic education model. METHODS: We assessed the 5-day, extended format program, encompassing lectures, videos, interactive tests, practice cases, and clinical exercises. Pre- and post questionnaires assessed knowledge change, using t-tests to compare groups. Satisfaction on program completion and after 3 years were evaluated. Implementation in other centers determined acceptability. RESULTS: During 2012-2018, 774 clinicians from multiple disciplines and career stages attended 35 programs; 334 (43%) attended the 5-day extended format. Evaluations showed significant improvement of genomic literacy (mean 15.05/100 points, p < 0.001). Residents initially had higher scores than specialists (pre: 66.3 ± 17.3 vs. 58.7 ± 16.6, respectively, p = 0.002); both significantly improved, with specialists "catching up" (post: 79.1 ± 17.2 vs. 75.7 ± 15.9, nonsignificant (NS)); there was a similar trend between fellows and subspecialists (pre: 70 ± 18 vs. 59.4 ± 16.4, respectively, p = 0.007; post: 78.6 ± 16.4 vs. 73.2 ± 17.7, respectively, NS). Younger specialists (≤10 years residency) had significantly higher pre- and post scores. Absolute improvement in scores did not depend on medical specialties. CONCLUSION: Our program is effective in improving genomics literacy for clinicians, irrespective of career length or expertise, and could be a model for improving skills in practical genomics for all medical professionals.

B4GALT1-congenital disorders of glycosylation: Expansion of the phenotypic and molecular spectrum and review of the literature.

A congenital disorder of glycosylation due to biallelic mutations in B4GALT1 has been previously reported in only three patients with two different mutations. Through homozygosity mapping followed by segregation analysis in an extended pedigree, we identified three additional patients homozygous for a novel mutation in B4GALT1, expanding the phenotypic spectrum of the disease. The patients showed a uniform clinical presentation with intellectual disability, marked pancytopenia requiring chronic management, and novel features including pulmonary hypertension and nephrotic syndrome. Notably, affected individuals exhibited a moderate elevation of Man3GlcNAc4Fuc1 on serum N-glycan analysis, yet two of the patients had a normal pattern of transferrin glycosylation in repeated analysis. The novel mutation is the third disease-causing variant described in B4GALT1, and the first one within its transmembrane domain.

Chromosomal microarray findings in pregnancies with an isolated pelvic kidney.

Objective To examine the risk for abnormal chromosomal microarray analysis (CMA) results among fetuses with an apparently isolated pelvic kidney. Methods Data from all CMA analyses performed due to an isolated pelvic kidney reported to the Israeli Ministry of Health between January 2013 and September 2016 were retrospectively obtained. Risk estimation was performed comparing the rate of abnormal observed CMA findings to the general population risk, based on a systematic review encompassing 9272 cases and on local data of 5541 cases. Results Of 120 pregnancies with an isolated pelvic kidney, two gain-of-copy number variants suggesting microduplication syndromes were demonstrated (1.67%). In addition, three variants of unknown significance were detected (2.5%). Conclusion The risk for clinically significant CMA findings among pregnancies with an isolated single pelvic kidney was not significantly different compared to both control populations. The results of our study question the practice of routine CMA analysis in fetuses with an isolated pelvic kidney.

Microarray analysis in pregnancies with isolated echogenic bowel.

INTRODUCTION: Fetal echogenic bowel is a frequent sonographic finding, demonstrated in about 1% of pregnancies. The advised evaluation of fetal echogenic bowel includes maternal serology, genetic testing for cystic fibrosis, detailed sonographic anatomic survey, and invasive prenatal testing for fetal chromosomal aberrations. The objective of our study was to evaluate the risk for clinically significant chromosomal microarray analysis (CMA) findings in pregnancies with isolated echogenic bowel. METHODS: Data from all CMA analyses performed due to isolated echogenic bowel reported to the Israeli Ministry of Health between January 2013 and September 2016 were retrospectively obtained. Risk estimation was performed comparing the rate of abnormal microarray findings to the control population, based on a systematic review of 9272 pregnancies and a large local cohort of 5541 fetuses with normal ultrasound, undergoing CMA testing due to maternal request. RESULTS: Of 103 CMA analyses performed due to isolated echogenic bowel, two (1.94%) pathogenic findings were detected (47,XYY and 16p11.2 duplication). This risk was not significantly elevated compared to the control groups. In addition, three variants of unknown significance were demonstrated. CONCLUSIONS: To our best knowledge, our study is the first report describing the rate of clinically significant copy number variants in pregnancies with isolated echogenic bowel. According to our results, it seems that pregnancies with isolated echogenic bowel do not have an increased risk for abnormal CMA compared to fetuses with no evidence of sonographic anomalies. Our findings suggest that the consideration to perform CMA analysis in such pregnancies should not differ from any pregnancy with normal ultrasound.

Progressive hereditary spastic paraplegia caused by a homozygous KY mutation.

Twelve individuals of consanguineous Bedouin kindred presented with autosomal recessive progressive spastic paraplegia evident as of age 0-24 months, with spasticity of lower limbs, hyperreflexia, toe walking and equinus deformity. Kyphoscolisois was evident in older patients. Most had atrophy of the lateral aspects of the tongue and few had intellectual disability. Nerve conduction velocity, electromyography and head and spinal cord magnetic resonance imaging were normal in tested subjects. Muscle biopsy showed occasional central nuclei and fiber size variability with small angular fibers. Genome-wide linkage analysis identified a 6.7Mbp disease-associated locus on chromosome 3q21.3-3q22.2 (LOD score 9.02; D3S1290). Whole-exome sequencing identified a single homozygous variant within this locus, c.51_52ins(28); p.(V18fs56*) in KY, segregating in the family as expected and not found in 190 Bedouin controls. High KY transcript levels were demonstrated in muscular organs with lower expression in the CNS. The phenotype is reminiscent of kyphoscoliosis seen in Ky null mice. Two recent studies done independently and parallel to ours describe somewhat similar phenotypes in one and two patients with KY mutations. KY encodes a tranglutaminase-like peptidase, which interacts with muscle cytoskeletal proteins and is part of a Z-band protein complex, suggesting the disease mechanism may resemble myofibrillar myopathy. However, the mixed myopathic-neurologic features caused by human and mouse Ky mutations are difficult to explain by loss of KY sarcomere stabilizing function alone. KY transcription in CNS tissues may imply that it also has a role in neuromotor function, in line with the irregularity of neuromuscular junction in Ky null mutant mice.

Clinical aspects of prenatally detected congenital heart malformations and the yield of chromosomal microarray analysis.

OBJECTIVE: The yield of chromosomal microarray analysis (CMA) for prenatally detected congenital heart defects (CHD) is 6.6% to 19.2%. We evaluated the yield of CMA in cases of prenatally detected CHD in regard to specific clinical characteristics. METHODS: Data from 192 cases of CHD including type, clinical and familial background, workup performed during the pregnancy, and pregnancy outcomes were collected. RESULTS: Fetal echocardiography was performed in all cases; 61.4% of CHD were suspected by ultrasound. There was a positive family history (FH) in 15.7%. Abnormal nuchal translucency or umbilical cord anomalies were detected in 1.7% and 5.9%, respectively, and 55.1% were isolated cases. In 11 of 96 cases in which genetic testing was performed, karyotype and CMA were abnormal (11.5%). The detection rate of CMA (performed in 72 cases) was 9.7%. The yield of CMA was similar in simple cases, isolated cases, and cases with a positive FH. CMA was abnormal in 7.3% of ventricular septal defect cases. CONCLUSION: Most cases of prenatally detected CHD had no additional extra-cardiac, sonographic findings suggesting increased risk for CHD. The yield of CMA testing was significant in all clinical scenarios including simple heart malformations, isolated cases, and cases with a positive FH. © 2016 John Wiley & Sons, Ltd.

The physiologic and therapeutic role of heparin in implantation and placentation.

Implantation, trophoblast development and placentation are crucial processes in the establishment and development of normal pregnancy. Abnormalities of these processes can lead to pregnancy complications known as the great obstetrical syndromes: preeclampsia, intrauterine growth restriction, fetal demise, premature prelabor rupture of membranes, preterm labor, and recurrent pregnancy loss. There is mounting evidence regarding the physiological and therapeutic role of heparins in the establishment of normal gestation and as a modality for treatment and prevention of pregnancy complications. In this review, we will summarize the properties and the physiological contributions of heparins to the success of implantation, placentation and normal pregnancy.

Meconium-stained amniotic fluid--risk factors and immediate perinatal outcomes among SGA infants.

OBJECTIVE: To detect factors that are associated with meconium-stained amniotic fluid (MSAF) among deliveries of small for gestational age (SGA) neonates and to identify perinatal outcomes of deliveries of SGA infants complicated with MSAF. METHODS: A population-based study comparing deliveries of SGA neonates with and without MSAF was conducted. Deliveries occurred during the years 1988-2007 at the Soroka University Medical Center. Risk factors for MSAF among SGA infants were evaluated. Incidence of adverse pregnancy outcomes were compared between deliveries of SGA neonates with and without MSAF. RESULTS: During the study period 9583 deliveries were of SGA neonates. Of these, 16.6% (n = 1597) were complicated with MSAF. Among SGA neonates, older maternal age, multiparty, lack of prenatal care and weight were significantly associated with MSAF. Having delivered an SGA infant with MSAF was associated with decreased rates of induction of labor and increased rates of labor dystocia, delivery by cesarean section and fetal distress. Using multivariable regression models, having delivered an SGA infant with MSAF was independently associated with fetal distress. CONCLUSION: Among SGA neonates, deliveries complicated with MSAF are associated with additional adverse pregnancy outcomes.

Antenatal diagnosis of idiopathic arterial calcification: a systematic review with a report of two cases.

INTRODUCTION: Idiopathic antenatal calcification is a rare, generally lethal, condition with unclear etiology. MATERIALS AND METHODS: Around 200 cases, most of them undergoing postnatal diagnosis, are reported in literature. The majority of the affected infants die before the age of 6 months, and very few have survived for more than 1 year. Five cases of spontaneous resolution of the disease are described. DISCUSSION: An autosomal recessive pattern of inheritance has been suggested for this condition, with some gene mutations which have been recently discovered. Therapy with bisphosphonates has been suggested, with conflicting evidence regarding the utility in the regression of the disease. CONCLUSION: The main purpose of the present report is to provide the available knowledge on this subject through a systematic review of the literature. In addition, we describe two cases of antenatal idiopathic arterial calcification in which antenatal diagnosis was achieved.

A homozygous deletion of 8q24.3 including the NIBP gene associated with severe developmental delay, dysgenesis of the corpus callosum, and dysmorphic facial features.

We have identified by microarray-based comparative genomic hybridization analysis (aCGH), a homozygous deletion of 8q24.3 [arr cgh 8q24.3(140,879,937 --> 141,021,392)x0 mat pat] in a patient with dysmorphic facial features, dysgenesis of the corpus callosum, and severe mental retardation. The deletion was inherited from asymptomatic, consanguineous parents, each of them being heterozygous for the same deletion. The only gene known to map to this segment is the NIBP gene, and so far no clinical manifestations have been found in association with this gene mutation in homozygous or heterozygous state in humans. Our findings suggest that a homozygous deletion in the NIBP gene results in an autosomal recessive condition with multiple abnormalities and severe delay. In addition, the child inherited a 781-kb deletion on 4q32.2 from the mother that contains the SPOCK3 gene. We suggest that this heterozygous deletion is likely to be non-contributory to the phenotype.

Routine cervical dilatation during elective cesarean delivery - Is it really necessary?

OBJECTIVE: To examine the necessity of routine cervical dilatation during elective cesarean delivery (ECD). MATERIAL AND METHODS: A retrospective cohort study including all ECD during 2005 was performed, comparing post operative complications between patients with and without cervical dilatation. RESULTS: Out of 666 ECD, 348 underwent routine cervical dilatation. No significant differences were found between the cervical dilatation and the comparison group regarding postpartum febrile morbidity (5.1 and 3.1%, respectively; p = 0.071), hospitalisation duration (4.1 +/- 1.4 and 4.1 +/- 2.0 days; p = 0.95), wound infection (0.9% and 1.25%, p = 0.451) or anemia rate (9.50 +/- 0.73 and 9.54 +/- 0.65, p = 0.91). Nevertheless, among patients following a previous vaginal delivery, cervical dilatation was significantly associated with post-operative fever (OR = 5.8; 95%CI 1.2-38.0; p = 0.021). CONCLUSION: Routine cervical dilatation during ECD does not reduce post operative morbidity. Moreover, among patients with a previous vaginal delivery cervical dilatation is a risk factor for febrile morbidity.

The clinical significance of bleeding during the second trimester of pregnancy.

OBJECTIVE: This study aimed at investigating the clinical importance and pregnancy outcome in women suffering from bleeding during the second half of their pregnancies. METHODS: A population-based study including all deliveries between the years 1988 and 2005 was conducted. Comparison was performed between patients with and without vaginal bleeding during the second half of pregnancy. Pregnancies, which terminated before 22 weeks, multiple gestations and women lacking prenatal care were excluded from the analysis. Stratified analyses, using the Mantel-Haenszel technique, and a multiple logistic regression model were performed to control for confounders. RESULTS: During the study period, 175,093 singleton deliveries occurred in our institute. Of these, 2,010 (1.1%) were complicated with bleeding upon admission during the second half of pregnancy. The cases were mostly attributed to placental abruption (63.5%; n = 1,276) and placenta previa (36.5%; n = 734). Independent risk factors associated with bleeding, using a backward, stepwise multivariate analysis were oligohydramnios, polyhydramnions, [odds ratio (OR) = 1.6; 95% confidence interval (CI) 1.2-2.0; P = 001 and 1.5; 1.2-1.8; P < 0.01, respectively], suspected intra uterine growth restriction (IUGR, 3.2; 2.6-4.0; P < .001), gestational age, previous abortions and maternal age. These patients subsequently were more likely to deliver by cesarean section (CS, 72.9 vs. 12.1%, OR = 19.5; 95% CI 17.6-19.9; 14.9 vs. 1.1%; P < 0.001). Perinatal mortality among patients admitted due to second half bleeding was significantly higher as compared to patients without bleeding (P < .001). CONCLUSION: Bleeding upon admission during the second half of pregnancy is an independent risk factor for perinatal mortality. Careful surveillance, including fetal monitoring, is suggested in these cases in order to reduce the adverse perinatal outcome.

Reduced third-trimester levels of soluble human leukocyte antigen G protein in severe preeclampsia.

OBJECTIVE: Recently, lower maternal plasma human leukocyte antigen (HLA)-G protein levels in preeclampsia (PE) in the first and second trimester was reported. Thus, we sought to evaluate the levels of HLA-G protein in patients with severe PE during the third trimester. STUDY DESIGN: In this prospective case control study, amniotic fluid and maternal and cord blood samples were aspirated from 50 pregnant women during the third trimester. The study group included 26 pregnant women diagnosed with severe PE and 24 women without PE serving as controls. A soluble HLA-G-specific enzyme-linked immunosorbent assay was used to measure protein levels. Statistical analysis included the Student t test and simple regression analysis. RESULTS: Maternal serum HLA-G levels in PE pregnancies were found to be significantly lower as compared with normal pregnancies (10.97 +/- 6.55 vs 36.05 +/- 34.53 microg/mL; P = .003). CONCLUSION: A reduced level of maternal HLA-G protein was associated with severe PE during the third trimester. This finding may suggest an essential role for HLA-G in normal and preeclamptic pregnancies.

Trial of labor and vaginal birth after cesarean section in patients with uterine Müllerian anomalies: a population-based study.

OBJECTIVE: The aim of our study was to determine the success rate of vaginal birth after cesarean section among patients with Müllerian anomalies in comparison to the success rate of vaginal birth after cesarean section in patients with normal uterus with emphasis on the rate of uterine rupture. STUDY DESIGN: A retrospective population-based study was designed, including all patients with a previous cesarean section that attempted vaginal birth after cesarean section during the study period. Women with known Müllerian anomalies were included in the study group. The control group consisted of women with normal uterus. The rates of vaginal birth after cesarean section, uterine rupture, maternal morbidity, and perinatal outcome were compared between the groups. RESULTS: Of 5571 eligible patients, 165 (2.96%) had Müllerian anomalies. The rate of vaginal birth after cesarean section was significantly lower among patients with Müllerian anomalies than in patients with normal uterus, 37.6% (62/165) vs 50.7% (2740/5406), respectively (P = .0009). During the study period, there were 10 cases of uterine rupture, all in patients with normal uterus. The major indication for repeated cesarean delivery among Müllerian anomalies patients was malpresentation, 58.3% (60/103) vs 14.4% (385/2666) in patients with normal uterus (P < .001). CONCLUSION: A trial of vaginal birth after cesarean section in patients with uterine Müllerian malformations and cephalic presentation is not associated with a higher rate of maternal morbidity and uterine rupture.