Lissencephaly (LIS) is a spectrum of cortical malformations including agyria, pachygyria and subcortical band heterotopia, which arises from aberrant neuronal migration and is associated with severe neurodevelopmental impairments. Despite advancements in prenatal imaging, diagnosing LIS remains challenging. Genetic factors play a crucial role in LIS, involving multiple genes and signalling pathways, yet research on prenatal diagnosis and the genetic basis is still limited. This study aimed to assess the diagnostic yield of whole exome sequencing (WES) in LIS and to examine genotype-phenotype correlations, addressing the challenge of 'phenotype lag' in prenatal LIS diagnosis. This study included 20 fetuses with LIS suggested by prenatal imaging and 20 children with LIS diagnosed after birth; all cases were diagnosed by magnetic resonance imaging and underwent genetic testing. In addition, a literature review was conducted and 80 studies were included, of which 1 was used to compare detection efficacy and 79 studies totalling 210 cases were used to assess genotype-phenotype correlation. In the prenatal cohort, 85.0% (17/20) of cases exhibited concurrent anomalies, predominantly ventriculomegaly (50.0%) and microcephaly (25.0%). In the postnatal cohort, the most common phenotypes were epilepsy (80.0%, 16/20) and global developmental delay (65.0%, 13/20), with half of the cases (10/20) showing no abnormalities in the prenatal period. The diagnostic yields were 55.0% (11/20) and 65.0% (13/20), respectively, with PAFAH1B1 point mutations or 17p13.3 microdeletions being the predominant genetic variant in both cohorts, accounting for 31.3% (prenatal) and 25.5% (postnatal) of cases, respectively. DARS2 and NPRL3 were reported to be associated with LIS for the first time in this study. Literature synthesis revealed an overall diagnostic yield of 79.04%, dominated by PAFAH1B1 (26.3%), DYNC1H1 (11.9%), and DCX (10.2%). By reviewing the prenatal images, up to 48.05% (74/154) of the cases had no specific findings in the prenatal period, and the most common presentations were ventriculomegaly/hydrocephalus (52.63%) and head circumference anomalies (29.82%). This study highlights the significant genetic heterogeneity, phenotypic complexity and diagnostic challenges of LIS by integrating data from our cohort and the published literature. We developed a comprehensive genetic aetiology classification framework for LIS and identified novel associations with non-canonical genes such as NPRL3 and DARS2. With a high molecular diagnostic yield of 79.04%, we recommend WES as the first-line genetic test. Furthermore, the establishment of an integrated prenatal imaging-molecular diagnostic system, along with a postnatal multidisciplinary model, is crucial for improving prognosis assessment, clinical decision-making and genetic counselling.

Fetal microcephaly (FMIC) is a neurodevelopmental disorder with heterogeneous etiologies and uncertain prenatal prognosis. Discrepancies between prenatal and postnatal head circumference (HC) measurements may confound ultrasound-based diagnosis, underscoring the need for genetic stratification to improve risk assessment. This prospective cohort study analyzed data from 301 fetuses with suspected FMIC collected between 2014 and 2022. Trio-prenatal exome sequencing (pES) was performed in 301 fetuses with suspected FMIC and normal results on karyotyping and chromosomal microarray analysis. Diagnostic yield, molecular spectrum, and pathway enrichment were analyzed. Clinical follow-up was conducted to correlate genetic findings with postnatal neurodevelopmental outcomes. Molecular diagnoses were achieved in 41 cases (13.6

Accurate parental haplotype information is crucial for noninvasive prenatal diagnosis (NIPD) of recessive single-gene disorders (SGD) (NIPD-SGD). However, conventional approaches rely on complex experimental techniques or trio-based SNP linkage analysis requiring family members, which limit clinical application. Here, we present a novel direct haplotyping based NIPD approach termed DiHNIPD, utilizing single-tube long fragment read (stLFR) sequencing. First, parental genome-wide haplotypes were reconstructed by stLFR-based whole genome sequencing (WGS). Second, SNPs within and surrounding the target gene in maternal plasma were identified by WGS. Finally, fetal haplotypes were determined by implementing a parental haplotype-assisted hidden Markov model combined with the Viterbi algorithm. The DiHNIPD results were further confirmed by invasive prenatal diagnosis. This study recruited 23 couples at risk of having a fetus with SGD. DiHNIPD directly phased all parental haplotypes of the target gene and accurately deduced 23 fetal genotypes, achieving 100% concordance with diagnostic results. These results demonstrate that DiHNIPD is a sensitive, user-friendly and inexpensive strategy for NIPD-SGD. It eliminates the need for expensive devices and complex procedures and does not rely on family members. This method shows significant promise for clinical use in high-risk pregnancies without prior offspring.

Objectives: To characterize the prenatal phenotypic spectrum, genetic findings, and pregnancy outcomes of fetal renal agenesis (RA), and to clarify the complementary roles of chromosomal microarray analysis (CMA) and whole exome sequencing (WES) in phenotype-stratified prenatal evaluation. Methods: This retrospective study included 203 RA fetuses between March 2017 and November 2025. All cases underwent genome-wide copy number variant (CNV) analysis, and selected cases underwent WES. Detection rates were compared across subgroups by laterality, isolated vs. non-isolated phenotype, fetal sex, and presence of extrarenal anomalies. Pregnancy outcomes and postnatal imaging follow-up were collected when available. A systematic literature review of prenatal genetic testing in RA fetuses was performed. Results: Among 203 fetuses, unilateral RA accounted for 92.6% of cases, and 65.0% were isolated. Chromosomal abnormalities were identified in 15 fetuses (7.4%), including aneuploidies and pathogenic or likely pathogenic (P/LP) CNVs. WES identified P/LP single nucleotide variants in 8 of 127 cases (6.3%), increasing to 8.7% when variants with potential clinical relevance were included. Diagnostic yield of WES was significantly higher in bilateral RA, non-isolated cases, and fetuses with extrarenal anomalies. Postnatal follow-up confirmed RA in most liveborn cases, although additional phenotypes emerged in some children. Literature synthesis identified recurrent CNVs at 16p11.2 and 22q11.21 and frequent involvement of FRAS1, FREM2, GFRA1, and GREB1L. Conclusions: RA shows marked phenotypic and genetic heterogeneity. CMA remains a first-tier prenatal test, while WES provides substantial incremental yield in bilateral, non-isolated, or extrarenal-associated RA. Integrated, phenotype-driven testing with longitudinal follow-up supports improved prognostication and genetic counseling.

BackgroundDandy-Walker malformation (DWM) is a rare congenital brain defect whose mechanism is still not fully understood. Genetic studies have suggested that NADH Dehydrogenase 1 alpha Subcomplex 4 (NDUFA4) may be associated with DWM; however, the functional consequences of NDUFA4 dysregulation in experimental neural models remain unclear. In this study, we investigated the cellular effects of NDUFA4 deficiency, rather than aiming to establish a causal mechanism for DWM.MethodsNDUFA4-related differentially expressed proteins were analyzed using iTRAQ tandem mass spectrometry, followed by functional, pathway, and protein interaction network analyses. Then, voltage-dependent anion Channel 1 (VDAC1), apoptosis-, endoplasmic reticulum (ER) stress-related proteins, and ETC Complex IV activity were assessed in NDUFA4 knockout mice. After NDUFA4 and VDAC1 knockdown, cell proliferation, apoptosis, mitochondrial status, and ER function were evaluated by CCK-8, Edu staining, flow cytometry, and Western blot in C8-D1A cells. In addition, the interaction between NDUFA4 and VDAC1 was evaluated using immunofluorescence and coimmunoprecipitation.ResultsNDUFA4 knockout upregulated VDAC1, ER stress- and apoptosis-related proteins, and inhibited ETC complex IV activity in mice. NDUFA4 knockdown inhibited proliferation and promoted apoptosis, ER stress, and mitochondrial damage in C8-D1A cells, and these changes were partially reversed by VDAC1 knockdown. Moreover, NDUFA4 and VDAC1 colocalized in C8-D1A cells and mouse cerebellar tissue, and NDUFA4 was found to interact with VDAC1.ConclusionsNDUFA4 deletion was associated with VDAC1 upregulation, mitochondrial damage, ER stress, and apoptosis-related changes in our experimental models. These findings may provide insight into cellular changes potentially relevant to DWM; however, they do not establish a direct causal relationship.

RNA sequencing (RNA-seq) provides transcript-level evidence for interpreting variants of uncertain significance in monogenic disorders, but its prenatal application is limited by sample availability and processing time. This study evaluated uncultured amniocytes as a practical substrate for prenatal RNA-seq. We performed RNA-seq on 15 matched paired cultured and uncultured amniocyte samples and compared their transcriptomic profiles using principal component analysis, differential expression analysis, and pathway enrichment. We then analyzed 77 uncultured amniocyte samples collected between 16+6 and 29+6 weeks of gestation to disease-related gene expression coverage and gestational-age-associated variation. Five selected cases were further analyzed to evaluate transcript-level findings relevant to variant interpretation. Cultured and uncultured amniocytes showed distinct transcriptomic profiles, with 11,234 differentially expressed genes and culture-associated transcriptional and pathway changes. Uncultured amniocytes expressed an average of 63.4% of genes across disease-associated panels, and most disease-related genes were consistently expressed across gestational stages, although stage-associated differences in gene detectability were observed. In selected cases, uncultured amniocyte RNA-seq detected aberrant expression, aberrant splicing, allelic imbalance, and expressed sequence variants. These findings support the feasibility of prenatal RNA-seq using uncultured amniocytes and further evaluation of this approach as a complementary functional assay for prenatal variant interpretation.

BACKGROUND:Congenital heart disease (CHD) is the leading cause of birth defect-related mortality. Genetic mutations, including those in the MAPK1 gene, play a critical role in the development of CHD. However, the exact mechanisms by which MAPK1 mutations contribute to CHD remain unclear. METHODS:The MAPK1 mutation (c.1061 T > G, p.F354C) was introduced into P19 murine embryonal carcinoma cells using CRISPR/Cas9 technology. Differentiation was induced with 1 % dimethyl sulfoxide (DMSO), and cell proliferation and apoptosis were assessed using the Cell Counting Kit-8, flow cytometry, and Hoechst staining assays, respectively. Transcriptome sequencing was conducted to explore downstream targets associated with the MAPK1 mutation, and the effects of NOD-like receptor (NLR) pathway inhibition on the phenotype of MAPK1-mutant cells were examined using NOD-IN-1. RESULTS:MAPK1 mutation notably reduced cell viability, promoted apoptosis, and impaired cardiomyocyte differentiation in P19 cells. Western blot analyses revealed decreased protein levels of Bcl-2 and cilium formation markers (p-GSK3βand DZIP1), alongside elevated levels of Bax in MAPK1-mutant cells. Additionally, the mRNA expression of cardiomyocyte differentiation markers, including cTnT, GATA4, MEF2C, and αMHC, was reduced in P19 cells with the MAPK1 mutation. Cells harboring the MAPK1 mutation exhibited clear chromatin condensation and formed fewer and smaller embryoid bodies. Mechanistically, MAPK1 mutation upregulated the protein expression of NOD1 and NOD2 and increased the phosphorylation of RIP2. Treatment with the NLR pathway inhibitor NOD-IN-1 significantly alleviated the detrimental effects of the MAPK1 mutation. CONCLUSION:MAPK1 gene mutation promotes the NLR signaling pathway and affects CHD development, providing new insights into the treatment of CHD.

OBJECTIVE:To assess the diagnostic utility of exome sequencing (ES) in macrocephalic fetuses. METHODS:Fetuses with macrocephaly (head circumference (HC) ≥ +2 SD) and negative chromosomal microarray results were included, who had available trio-ES data. Molecular diagnoses were systematically analyzed. Subgroup analyses were performed on the ES diagnostic yield based on gestational age, HC Z-scores, associated anomalies, and growth parameters. RESULTS:Molecular diagnoses were established in 34 out of 87 macrocephalic fetuses (39.1%) through trio-ES. These diagnoses revealed that the variants predominantly affect key signaling pathways, including mTOR, RASopathies and Sotos syndrome. The detection rate was significantly higher in non-isolated compared to isolated macrocephaly cases (65.0%, 26/40 vs. 17.0%, 8/47; p < 0.001). The most frequent anomalies associated with genetic diagnoses included micromelia (100.0%, 14/14), megalencephaly (100.0%, 2/2), and ventriculomegaly (60.0%, 6/10). Subgroup analysis identified higher diagnostic yields in fetuses diagnosed before 32 gestational weeks, with HC Z-scores ≥ +3 SD, micromelia, and absence of large-for-gestational-age (LGA). CONCLUSIONS:Exome sequencing significantly enhances the detection of monogenic disorders in macrocephalic fetuses compared with CMA, irrespective of isolated or non-isolated cases. These clinical features and phenotypes are essential for assessing monogenic disorders and for prenatal counseling and evaluations of macrocephalic fetuses.

Chromatin topology can impact gene regulation, but how evolutionary divergence in chromatin topology has shaped gene regulatory landscapes for distinctive human traits remains poorly understood. CTCF sites determine chromatin topology by forming domains and loops. Here, we show evolutionary divergence in CTCF-mediated chromatin topology at the domain and loop scales during primate evolution, elucidating distinct mechanisms for shaping regulatory landscapes. Human-specific divergent domains lead to a broad rewiring of transcriptional landscapes. Divergent CTCF loops concord with species-specific enhancer activity, influencing enhancer connectivity to target genes in a concordant yet constrained manner. Under this concordant mechanism, we establish the role of human-specific CTCF loops in shaping transcriptional isoform diversity, with functional implications for disease susceptibility. Furthermore, we validate the function of these human-specific CTCF loops using human forebrain organoids. This study advances our understanding of genetic evolution from the perspective of genome architecture.

This study aims to assess the genetic burden of fetal congenital diaphragmatic hernia (CDH) and identify prenatal, perinatal, and postnatal predictors to improve early diagnosis, monitoring, and intervention. This study included 130 CDH fetuses who underwent invasive prenatal diagnosis, with fetal prognosis evaluated using imaging parameters such as observed-to‐expected lung-to-head ratio (o/e LHR), observed‐to‐expected total lung volume (o/e TLV), and percent predicted lung volume (PPLV). Clinical outcomes included neonatal outcomes, extracorporeal membrane oxygenation (ECMO) requirement, and post-neonatal prognosis. Logistic regression and receiver operating characteristic (ROC) curve analyses were used to evaluate prognostic indicators and construct predictive models. Chromosomal microarray analysis (CMA) and exome sequencing (ES) yielded diagnostic rates of 7.7

INTRODUCTION:Congenital duodenal obstruction (CDO) is one of the most common fetal gastrointestinal anomalies, but previous prenatal studies tend to investigate CDO as a portion of gastrointestinal obstruction. Few studies describe the genetic findings of CDO, especially copy number variants, in fetal cohorts with a relatively large sample size. The study aims to investigate the detection rate of genetic causes at different levels and to explore the potential influencing factors of perinatal outcomes of fetuses with CDO. MATERIAL AND METHODS:This retrospective study analyzed karyotype, chromosomal microarray analysis, and trio-whole exome sequencing (trio-WES) results of singleton fetuses suspected of CDO in a tertiary center between January 2014 and September 2023. In addition, perinatal outcomes and postnatal medical records of enrolled cases were followed up and analyzed. RESULTS:A total of 98 fetuses were included in the study, of which 69 (70.4%) were classified as isolated CDO and 29 (29.6%) were classified as non-isolated. The overall rate of genetic anomalies was 20.4% (20/98). Trisomy 21 (9/98 [9.2%]) and microduplication of the 17q12 region (3/98 [3.2%]) were the most common chromosomal numerical abnormalities and pathogenic copy number variants found in this cohort, respectively. Compared to chromosomal microarray analysis, no additional pathogenic or likely pathogenic variants were found in seven cases undergoing trio-WES. The rate of chromosomal numerical and structural abnormalities was significantly higher in the non-isolated group (8/29 [27.6%] vs. 6/69 [8.7%], p < 0.05). In terms of perinatal outcomes, the live birth rate was significantly higher in the isolated group (52/69 [75.4%] vs. 15/29 [51.7%], p < 0.05) largely due to fewer terminations of pregnancy. All cases with positive genetic results elected to terminate the pregnancy, and 82% of those with negative results opted to continue the pregnancy. Neonatal mortality was significantly higher in the non-isolated group (2/15 [13.3%] vs. 0/52 [0.0%], p < 0.05). The overall neonatal survival rate was 97.0% (65/67). CONCLUSIONS:The present study highlights the value of prenatal diagnostic testing for fetuses suspected of CDO, both in isolated and non-isolated cases. Genetic diagnostic outcomes exert substantial influence on pregnancy decision-making. Perinatal outcome and short-term prognosis of affected fetuses are reasonably favorable when known genetic causes are excluded.

Objective: Duplex kidney is a relatively frequent form of urinary system abnormality. This study aimed to elucidate the value of chromosomal microarray analysis (CMA) and whole exome sequencing (WES) for duplex kidney and the perinatal outcomes of duplex kidney fetuses.Methods: This retrospective cohort study included 63 patients with duplex kidney diagnosed using antenatal ultrasound between August 2013 and January 2023. We reviewed the clinical characteristics, genetic test results, and pregnancy outcomes of the patients.Results: Among the 63 cases based on the inclusion criteria, the CMA detected seven (11.1%) clinically significant variants and nine variants of uncertain significance (VUS), and the pathogenic/likely pathogenic (P/LP) copy number variations (CNVs) in the recurrent region that were associated with prenatal duplex kidney included 17q12, 17p13.3, and 22q11.2. No significant disparity was observed in the CMA detection rate between the unilateral and bilateral groups, or between the isolated and non-isolated groups. WES identified three (50%) P/LP single-gene variants in six fetuses with duplex kidney. We detected the following pathogenic genes in the duplex kidney fetuses: KMT2D, SMPD4, and FANCI. Pregnancy termination in cases where clinically significant variants were detected by genetic testing was different in statistical significance from that in cases with negative results (9/10, 90.0% vs 8/48, 16.7%, P < 0.001).Conclusion: This study elucidated the value of CMA and WES for fetal duplex kidney, proving that CMA and WES may be useful tools in prenatal diagnosis and genetic counseling.

Objective This single-center retrospective study aimed to investigate the genetic factors contributing to neonatal and pediatric pulmonary hypertension in a Chinese population using trio whole-exome sequencing (trio-WES). Method This retrospective analysis reviewed the clinical and genetic profiles of children under 18 years of age diagnosed with pulmonary hypertension between March 2017 and March 2022. The diagnosis of pediatric pulmonary hypertension was confirmed through echocardiography and catheterization. Trio-WES was performed on the patients and their parents after obtaining informed consent. Results A total of 51 children with neonatal and pediatric pulmonary hypertension were included, comprising 20 with pediatric pulmonary arterial hypertension and 31 with persistent pulmonary hypertension of the newborn. Trio-WES detected 16 pathogenic or likely pathogenic variants in 14 patients across ten genes, including: BMPR2 (n = 2), CHD7 (n = 2), FOXF1 (n = 2), MED13L (n = 1), TNNI3 (n = 2), ALMS1 (n = 1), KMT2D (n = 2), NKX2-1 (n = 1), NONO (n = 1), and CACNA1E (n = 1). In addition, two patients exhibited de novo pathogenic copy number variations. Conclusion Our findings demonstrate the significant diagnostic value of trio-WES in pediatric pulmonary hypertension, supporting its recommendation for these patients.

BACKGROUND:Right aortic arch (RAA) is a common congenital aortic arch abnormality. Fetuses with RAA frequently have good outcomes after birth. However, chromosomal abnormalities and genetic syndromes suggest poor prognosis for these patients. So far the underlying genetic etiology is still not identified in most RAA patients based on traditional genetic techniques and a problem is still debated whether fetuses with isolated RAA should be referred for CMA. Our study aims to investigate the genetic etiology of fetuses with right aortic arch (RAA) by chromosomal microarray analysis (CMA) and whole exome sequencing (WES) and evaluate the efficacy of CMA in fetal isolated RAA. RESULTS:Among these 153 fetuses, 99 (64.7%) with isolated RAA and 54 (35.3%) with non-isolated RAA; 25.5% (39/153) with additional intracardiac anomalies (ICA), and 19.0% (29/153) with extracardiac anomalies (ECA). Tetralogy of Fallot (n = 10) and persistent left superior vena cava (n = 11) are the most common ICA and ECA, respectively. CMA detected 15 clinically significant copy number variations (CNVs) in 14 cases (9.2%); microdeletion of 22q11.21 was the most common pathogenic CNVs (7.8%). The chromosomal abnormalities rate was higher in non-isolated RAA and RAA with ICA groups than in isolated RAA group (16.7% vs. 5.1%; 20% vs. 5.1%, both p < 0.05). From five cases further undergoing WES, a diagnostic variant in MTOR gene (c.7255G > A, de novo) was first reported in prenatal, extending the prenatal manifestation of Smith-Kingsmore syndrome (OMIM: 616638); a clinically relevant variant c.3407A > T in STAG2 was identified, being inherited from the healthy mother. Moreover, the premature birth and termination rates were higher in non-isolated RAA group than in isolated RAA group (11.1% vs. 1.0%; 37.0% vs. 2.0%, both p < 0.01). CONCLUSIONS:We demonstrate that CMA and WES are useful diagnostic tools for fetal RAA, particularly non-isolated RAA, and all fetuses with RAA should be referred for CMA. The data probably aids in prenatal diagnosis and prenatal counseling of fetal RAA.

BACKGROUND:Polycystic renal disease is a frequent congenital anomaly of the kidneys, but research using chromosomal microarray analysis and exome sequencing in fetuses with polycystic renal disease remains sparse, with most studies focusing on the multisystem or genitourinary system. OBJECTIVE:This study aimed to assess the detection rate of detectable genetic causes of fetal polycystic renal disease at different levels, novel disease-causing variants, and genotype-phenotype correlations. STUDY DESIGN:This study included 220 fetal polycystic renal disease cases from January 2014 to June 2022. Cases were divided into the following 3 groups: isolated multicystic dysplastic kidneys, nonisolated multicystic dysplastic kidneys, and suspected polycystic kidney disease group. We reviewed data on maternal demographics, ultrasonographic results, chromosomal microarray analysis/exome sequencing results, and pregnancy outcomes. RESULTS:In our cohort, chromosomal microarray analysis identified 19 (8.6%) fetuses carrying chromosomal abnormalities, and the most common copy number variation was 17q12 microdeletion (7/220; 3.2%). Furthermore, 94 families chose to perform trio-exome sequencing testing, and 21 fetuses (22.3%) were found to harbor pathogenic/likely pathogenic variants. There was a significant difference in the live birth rate among the 3 groups (91/130 vs 46/80 vs 1/10; P<.001). Among 138 live birth cases, 106 (78.5%) underwent postnatal ultrasound review, of which 95 (89.6%) had a consistent prenatal-postnatal ultrasound diagnosis. CONCLUSION:For both isolated and nonisolated polycystic renal disease, our data showed high detection efficiency with both testing tools. The detection of novel pathogenic variants expands the known disease spectrum of polycystic renal disease-associated genes while enriching our understanding of the genotype-phenotype correlation. Therefore, we consider it feasible to perform chromosomal microarray analysis+exome sequencing testing in fetal polycystic renal disease. Moreover, prenatal-postnatal ultrasound concordance was greater, the live birth rate was higher, and prognosis was better when known genetic disorders were excluded, indicating that genetic testing results significantly influenced pregnancy decisions.

Backgroud A systematic analysis was conducted to investigate the molecular etiology of fetal cleft lip and/or palate (CL/P) and the association between various types of CL/P and copy number variations (CNVs), as well as their impact on birth outcomes. Methods In this retrospective study conducted between January 2016 and July 2022, a cohort of pregnancies diagnosed with fetal CL/P was enrolled and comprehensive clinical data for all cases were extracted from our medical record database, including demographic data about the pregnancies, ultrasound findings, results of Chromosomal microarray (CMA), as well as relevant pregnant and perinatal outcomes. Results Among the 358 cases, 32 clinically significant variants in 29 (8.1%) fetuses with CL/P were detected by CMA. In 338 singleton pregnancies, the diagnostic yield of CMA in the context of CL/P fetuses was determined to be 7.7% (26/338). CP cases exhibited a relatively higher prevalence of pathogenic/likely pathogenic CNVs at a rate of 25% (3/12), followed by CLP cases at 8.0% (23/288). Notably, the CL group did not demonstrate any pathogenic/likely pathogenic CNV findings among the examined cases (0/38). The diagnostic rate of clinically significant variants was notably higher in the non-isolated CL/P group than in the isolated CL/P group (11/33, 33.3% vs. 15/305, 4.9%, p < 0.001). Within the remaining 20 twin pregnancies, three clinically significant variants (15%) were observed. Conclusions This study provides powerful evidence supporting the efficacy of CMA as a valuable tool for facilitating the prenatal genetic diagnosis of fetal CL/P. The presence of CP and CLP in fetal cases demonstrated a relatively higher incidence of pathogenic/likely pathogenic CNVs. Moreover, when these cases were accompanied by additional ultrasound abnormalities, the likelihood of identifying diagnostic CNVs significantly increased. Conversely, cases of CL alone might not be associated with positive CNVs. The present data may significantly enhance prenatal diagnosis accuracy and facilitate informed genetic counseling for cases of fetal CL/P.

The vast neuronal diversity in the human neocortex is vital for high-order brain functions, necessitating elucidation of the regulatory mechanisms underlying such unparalleled diversity. However, recent studies have yet to comprehensively reveal the diversity of neurons and the molecular logic of neocortical origin in humans at single-cell resolution through profiling transcriptomic or epigenomic landscapes, owing to the application of unimodal data alone to depict exceedingly heterogeneous populations of neurons. In this study, we generated a comprehensive compendium of the developing human neocortex by simultaneously profiling gene expression and open chromatin from the same cell. We computationally reconstructed the differentiation trajectories of excitatory projection neurons of cortical origin and inferred the regulatory logic governing lineage bifurcation decisions for neuronal diversification. We demonstrated that neuronal diversity arises from progenitor cell lineage specificity and postmitotic differentiation at distinct stages. Our data paves the way for understanding the primarily coordinated regulatory logic for neuronal diversification in the neocortex.

Purpose 17q12 copy number variants (CNVs) have variable presentations and incomplete penetrance, challenging prenatal counseling and management. This study aims to investigate the intrauterine phenotype. Methods We included 48 fetuses diagnosed with 17q12 microdeletion or microduplication by chromosomal microarray analysis. Results For 17q12 deletion, renal anomalies were found in 35 fetuses (35/37, 94.6%), with hyperechogenic kidneys (HEK, 28/37, 75.7%) and multicystic dysplastic kidneys (17/37, 45.9%) being the most common findings. Duodenal obstruction (DO) was most frequently combined in 17q12 duplication fetuses. In addition, cardiac abnormalities were the first reported prenatal phenotype in 17q12 duplication fetuses. Conclusion Our study shows that HEK and DO are the most predominant presentations of 17q12 deletion and duplication, respectively, and cardiac structural abnormalities may be associated with the latter. Although 17q12 CNVs have incomplete penetrance and variable expressivity and may be mainly involved in neurodevelopmental disorders, their short-term prognosis appears positive.

BACKGROUND:Acid ceramidase (ACDase) deficiency is an ultrarare autosomal recessive lysosomal disorder caused by pathogenic N-acylsphingosine amidohydrolase (ASAH1) variants. It presents with either Farber disease (FD) or spinal muscular atrophy with progressive myoclonic epilepsy (SMA-PME).OBJECTIVE:The study aims to identify a novel splice site variant in a hydrops fetus that causes ASAH1-related disorder, aid genetic counseling, and accurate prenatal diagnosis.METHODS:We report a case of hydrops fetalis with a novel homozygous mutation in ASAH1 inherited from non-consanguineous parents. We performed copy number variation sequencing (CNV-Seq) and whole exome sequencing (WES) on the fetus and family, respectively. Minigene splicing analyses were conducted to confirm the pathogenic variants.RESULTS:WES data revealed a splice site variant of the ASAH1 (c.458-2A>T), which was predicted to affect RNA splicing. Minigene splicing analyses found that the c.458-2A>T variant abolished the canonical splicing of intron 6, thereby activating two cryptic splicing products (c.456_458ins56bp and c.458_503del).CONCLUSIONS:Overall, we identified a novel splice site variant in the mutational spectrum of ASAH1 and its aberrant effect on splicing. These findings highlight the importance of ultrasonic manifestation and family history of fetal hydrops during ASAH1-related disorders and could also aid genetic counseling and accurate prenatal diagnosis. To the best of our knowledge, this is the shortest-lived account of ASAH1-related disorders in utero with severe hydrops fetalis.

Background The molecular mechanism of fetal cystic hygroma (CH) is still unclear, and no study has previously reported the transcriptome changes of single cells in CH. In this study, single-cell transcriptome sequencing (scRNA-seq) was used to investigate the characteristics of cell subsets in the lesion tissues of CH patients.Methods Lymphoid tissue collected from CH patients and control donors for scRNA-seq analysis. Differentially expressed gene enrichment in major cell subpopulations as well as cell-cell communication were analyzed. At the same time, the expression and interactions of important VEGF signaling pathway molecules were analyzed, and potential transcription factors that could bind to KDR (VEGFR2) were predicted.Results The results of scRNA-seq showed that fibroblasts accounted for the largest proportion in the lymphatic lesions of CH patients. There was a significant increase in the proportion of lymphatic endothelial cell subsets between the cases and controls. The VEGF signaling pathway is enriched in lymphatic endothelial cells and participates in the regulation of cell-cell communication between lymphatic endothelial cells and other cells. The key regulatory gene KDR in the VEGF signaling pathway is highly expressed in CH patients and interacts with other differentially expressed EDN1, TAGLN, and CLDN5 Finally, we found that STAT1 could bind to the KDR promoter region, which may play an important role in promoting KDR up-regulation.Conclusion Our comprehensive delineation of the cellular composition in tumor tissues of CH patients using single-cell RNA-sequencing identified the enrichment of lymphatic endothelial cells in CH and highlighted the activation of the VEGF signaling pathway in lymphoid endothelial cells as a potential modulator.Simple summary The molecular and cellular pathogenesis of fetal cystic hygroma (CH) remains largely unknown. This study examined the distribution and gene expression signature of each cell subpopulation and the possible role of VEGF signaling in lymphatic endothelial cells in regulating the progression of CH by single-cell transcriptome sequencing. The enrichment of lymphatic endothelial cells in CH and the activation of the VEGF signaling pathway in lymphatic endothelial cells provide some clues to the pathogenesis of CH from the perspective of cell subpopulations.