Mutations in ZNHIT3 are strongly associated with progressive encephalopathy with edema, hypsarrhythmia and optic atrophy (PEHO syndrome), characterized by severe cerebellar atrophy and profound intellectual disability; however, their role in cerebellar development remains unknown. By developing spatiotemporally-regulated conditional Znhit3 knockout mice, we discovered that Znhit3 is essential for granule cell progenitor survival, proliferation, differentiation, and migration. Knockout of Znhit3 caused loss of granule cell progenitors due to apoptosis, premature cell-cycle exit, and migration arrest and resulted in progressive anterior-lobe atrophy and motor deficits. The granule cell progenitor-autonomous defects secondarily impaired Purkinje cell alignment, dendritic maturation, and synaptic organization. Transcriptomic analyses revealed activation of the p53/p21 pathway, rRNA processing defects, and nucleolar stress. Genetic or pharmacologic inhibition of p53/p21 signaling rescued granule cell progenitor development and restored cerebellar architecture in the Znhit3-knockout mice. Thus, ZNHIT3 is a critical regulator of ribosome biogenesis and cerebellar growth, suggesting nucleolar stress-p53/p21 signaling as a potential therapeutic target in ZNHIT3-related disorders.

Mental disorders are a major global health challenge, exacerbated by a narrow range of effective treatments and inadequate responses. The discovery of novel drug targets is necessary. We leveraged genome-wide association study summary statistics for 6 mental disorders (anorexia nervosa (AN), post-traumatic stress disorder (PTSD), attention deficit hyperactivity disorder (ADHD), major depressive disorder (MDD), schizophrenia (SCZ), and bipolar disorder (BIP), sample sizes ranging from 72,517 to 500,199) and quantitative trait loci summary statistics of 713 proteins in cerebrospinal fluid (CSF) to investigate genetic relationships between CSF protein and the mental disorders. Two-sample Mendelian randomization (MR) analysis with the Wald ratio method revealed that the expression of 17, 13, 16, 24, 36, and 27 proteins had potential causal effects on the 6 mental disorders AN, PTSD, ADHD, MDD, SCZ, and BIP, respectively (P < 0.05). Among these, Bayesian colocalization suggested a shared genetic etiology among several proteins (posterior probability > 0.70), including PGD with AN, LGALS9 with PTSD, SERPING1 with MDD, AGRP with SCZ, and FOLH1 and HGFAC with BIP, whereas no protein showed colocalization with ADHD. Protein-protein interaction was obtained to identify potential molecular interactions among these proteins. A total of 18 drug compunds that target these proteins were obtained in DrugBank. This study provides genetics-informed target prioritization for mental disorders.

Ozone (O3) pollution is increasingly recognized as a serious environmental threat associated with elevated dementia risks. However, evidence regarding the impact of prenatal exposure to O3 on the morphological development of the brain remains limited. This nationwide population-based study investigated associations between prenatal O3 exposure and brain volume in 3294 infants aged 0-1 year without major anomalies, who underwent magnetic resonance imaging for non-neurological indications at hospitals within the China Neonatal Neuro-Critical Care Network group. Prenatal O3 exposure was estimated using geocoded residential addresses linked to 1 km × 1 km air pollution datasets. Linear mixed-effects models revealed that each 10 μg/m3 increase in prenatal O3 exposure was associated with a decrease of 1.14% (95% CI: 0.38-1.88) in total brain volume, 1.15% (95% CI: 0.40-1.89) in gray matter volume, 1.13% (95% CI: 0.37-1.88) in white matter volume, 1.11% (95% CI: 0.35-1.87) in cerebral blood volume, and 1.12% (95% CI: 0.36-1.88) in cerebrospinal fluid. No significant association was observed for the apparent diffusion coefficient [-0.08% (95% CI: -0.43-0.27)]. The associations remained significant in dual-pollutant models that additionally adjusted for prenatal PM2.5 exposure. Mediation analysis confirmed a direct effect of prenatal O3 exposure on brain morphology, as birth weight accounted for only 3.47% to 3.54% of the total effect. Our findings underscore the imperative for stricter air quality regulations to protect pregnant individuals and mitigate potential neurodevelopmental impairment originating as early as the fetal stage.

Next-generation sequencing technologies have been widely applied in diagnosing genetic disorders in pediatric patients. However, the cancer predisposition and tumor characteristics in individuals who have germline pathogenic variants remain unclear. We analyzed exome sequencing data from 75,602 pediatric patients referred for genetic testing between January 2016 and January 2025, tracking cancer as a secondary finding. The most common reasons for genetic testing were symptoms related to the nervous system, metabolic disorders and immune dysfunction. Among 110,692 variants of 139 tumor susceptibility genes, we identified 501 (456 single-nucleotide variants, 45 copy number variations, 0.45%) pathogenic or likely pathogenic (P/LP) and 3,848 (3,650 single-nucleotide variants, 198 copy number variations, 3.5%) variants of uncertain significance leaning toward likely pathogenic variants. Of 411 patients with tumors (203 with preexisting tumors and 208 with new tumors diagnosed during follow-up), 134 (32.6%) harbored causative germline P/LP variants in genes such as NF1 (13.1%), TSC2 (5.8%), RB1 (4.6%) and WT1 (3.2%). Critically, prospective follow-up of 64,187 patients without initial tumors revealed a significantly higher incidence of malignant tumors in those carrying P/LP variants (3.23 per 1,000 person-years) compared with those with variants of uncertain significance leaning toward likely pathogenic or other variants (0.236 and 0.272 per 1,000 person-years, respectively). These findings underscore the importance of proactive genetic counseling and surveillance for pediatric patients with pathogenic germline variants.

Necrotizing enterocolitis (NEC) is a severe gastrointestinal disease affecting premature infants, yet its precise pathogenic mechanisms remain unclear. While immunothrombosis, an interplay between inflammation and coagulation, is well recognized in systemic inflammatory diseases, its role in NEC pathogenesis and local intestinal regulation has been incompletely characterized. Here, we combine multi-omics profiling, histopathological analyses, and functional studies using clinical NEC samples and neonatal mouse models. We demonstrate prominent intestinal thromboinflammation in NEC, histologically characterized by immunothrombosis with infiltrating CD177⁺ neutrophils, activated platelets, and fibrin deposition. We show that extensive formation of CD177⁺ neutrophil-platelet aggregates (NPAs) amplifies intestinal injury through neutrophil extracellular trap (NET)-mediated thromboinflammation. Blocking CD177⁺ NPAs or inhibiting NET formation significantly alleviates NEC severity in mouse models. Clinically, elevated CD177⁺ NPA levels correlate with NEC severity and increased thrombotic biomarkers, highlighting their potential as diagnostic and prognostic biomarkers. Importantly, prophylactic administration of low-molecular-weight heparin (LMWH) disrupts NPA formation, reduces NET burden, and improves survival in experimental NEC. Collectively, these findings establish CD177⁺ NPAs as central drivers of NEC pathogenesis, support a thromboinflammatory framework for NEC, and highlight LMWH therapy and CD177⁺ NPA monitoring as possible clinically translatable strategies for neonatal care.

Preterm birth influences early functional brain maturation at term-equivalent age. Using resting-state functional magnetic resonance imaging from a large Chinese neonatal cohort (62 term-born and 107 preterm neonates), we examined static and dynamic network organization using multi-level graph-theoretical analyses. Preterm neonates exhibited reduced global integration and segregation, reflected by lower global efficiency, clustering coefficient, and local efficiency, together with increased characteristic path length. Widespread nodal and modular alterations were observed across multiple networks. Dynamic analyses revealed selective edge-level disturbances involving the right parahippocampal gyrus and its connections with default mode, visual, and limbic networks, despite limited group differences in global or nodal dynamic metrics. Developmental analyses further showed associations between specific global metrics and postmenstrual age at scan. Network measures were also associated with prenatal factors, including multiple pregnancy and cesarean delivery. These findings characterize early alterations in static and dynamic functional network organization after preterm birth and highlight prenatal factors potentially related to neonatal brain development.

DYRK1A syndrome is a neurodevelopmental disorder caused by DYRK1A haploinsufficiency. We generated a human induced pluripotent stem cell (iPSC) line, FDIBSi002-A, from a 4-year-old female patient carrying a de novo heterozygous c.1042G>A (p.G348R) mutation in DYRK1A. Peripheral blood mononuclear cells (PBMCs) were reprogrammed using non-integrating episomal vectors. The established iPSC line exhibited a normal karyotype (46, XX), expressed pluripotency markers, and demonstrated trilineage differentiation potential. This patient-specific cell line provides a valuable model for investigating the pathogenic mechanisms of DYRK1A-related intellectual disability and for drug screening.

Bronchopulmonary dysplasia (BPD) is the most common chronic lung disease in preterm infants. Although inflammation is broadly recognized as a key contributor to BPD, the precise roles and underlying mechanisms of immune cells in BPD pathogenesis remain incompletely understood. Moreover, the absence of reliable early diagnostic tools impedes the prediction of disease progression and the implementation of timely interventions. We first analyzed public blood transcriptomic data from preterm infants (<32 weeks gestational age) to explore immune alterations in BPD. Subsequently, we isolated neutrophils from infants at postnatal day 14 for transcriptomic and flow cytometric analyses. Longitudinal complete blood count (CBC) data were retrospectively collected at birth, and on days 2, 7, 14, 21, and 28, to validate these findings; machine learning models were subsequently developed to predict disease severity, with performance evaluated using the area under the receiver operating characteristic curve (AUC). Finally, single-cell transcriptome analysis was performed on public lung data from hyperoxia-exposed neonatal mice, followed by experimental validation. Blood transcriptomic profiling revealed increased neutrophils and downregulated T cell response pathways in BPD patients. Neutrophils from BPD infants exhibited enhanced inflammatory signaling and downregulated T-cell activation pathways compared to preterm controls. Flow cytometry confirmed an increased presence of polymorphonuclear myeloid-derived suppressor cell-like (PMN-MDSC-like) neutrophils in BPD, further supporting the presence of immunosuppression-associated features. Longitudinal CBC data (n = 381 infants) confirmed increased neutrophils and decreased lymphocyte levels in BPD, which correlated with disease severity. Based on this, we developed machine learning models, which exhibited promising internal-test performance in predicting moderate-to-severe BPD (AUC = 0.9328). In BPD mouse lungs, single-cell analysis showed increased neutrophils with elevated immunosuppressive signatures, including Cd274 and Lgals3, and decreased CD4+ and CD8+ T cells. Flow cytometry validated these findings; neutrophil depletion in vivo attenuated alveolar simplification and led to an increase in CD4+ and CD8+ T cells in hyperoxia-exposed neonatal mice. Our study identifies neutrophil-associated immune signatures in human BPD and supports a contributory role of neutrophils in hyperoxia-induced alveolar simplification in mice. Furthermore, we demonstrate the potential of longitudinal CBC-based biomarkers for disease severity stratification.

BACKGROUND & AIMS:Hirschsprung disease (HSCR) is a congenital enteric neuropathy with distal aganglionosis and dysmotility. Germline mutations explain many familial cases, but most sporadic cases lack a molecular explanation. We sought noncanonical mechanisms and blood-accessible biomarkers. METHODS:We performed RNA-sequencing (RNA-seq) on paired aganglionic and ganglionic colon biopsy specimens from 103 sporadic HSCR patients, with control colon samples (n = 22). Blood whole-genome sequencing (WGS) was available for 41 patients with RNA-seq profiles. We assessed PR/SET domain 9 (PRDM9) in patient tissues, perturbed prdm9 in zebrafish and mice, and performed mechanistic profiling in PRDM9-knockout human-induced pluripotent stem cell-derived enteric neural crest cells. Mosaic promoter deletions (MPDs) were called from colon WGS (n = 30; matched blood, n = 27) and summarized as a blood MPD score in blood WGS cohorts (discovery: 89 HSCR and 43 controls; validation: 165 HSCR and 42 controls), alone and combined with the polygenic risk score. RESULTS:A predominant subgroup (79.6% [82 of 103]) showed coordinated repression of neurogenesis programs. Promoter motif enrichment implicated PRDM9, which we localized to normal enteric nervous system but found down-regulated in aganglionic tissue with promoter hypermethylation. Prdm9 perturbation reduced HuC/D-positive differentiated enteric neurons and impaired motility in zebrafish and mice. In enteric neural crest cells, PRDM9 loss redistributed DNA double-strand breaks toward promoters and enhancers, generated MPDs at neurogenesis loci, and correlated with transcriptional repression and impaired neuronal differentiation. Blood MPD score discriminated HSCR from controls in a discovery cohort (area under the receiver operating characteristic curve [AUROC], 0.78) and an independent validation cohort (AUROC, 0.82), and improved with polygenic risk score (AUROC, 0.89 and 0.91, respectively). CONCLUSIONS:PRDM9 deficiency links ectopic DNA breaks to MPDs and impaired enteric neuronal differentiation in a predominant molecular subgroup within sporadic HSCR and enables complementary, noninvasive molecular stratification.

Neonatal jaundice (NJ) might increase the risk of autism spectrum disorder (ASD) in children. This study examined whether alterations in the gut microbiota could explain the link between NJ and ASD. We analyzed three cohorts: NJ cohort 1 comprised 68 neonates with NJ and 68 healthy controls (HCs); NJ cohort 2 included 56 infants with NJ and 14 HCs; and the ASD cohort consisted of 43 children with ASD and 31 typically developing children. Fecal samples were collected aseptically. We performed 16S rRNA sequencing (NJ cohort 1), liquid chromatography with tandem mass spectrometry metabolomics (NJ cohort 1 and ASD cohort), and shotgun metagenomics (NJ cohort 2 and ASD cohort). We characterized the gut DNA virome, quantified bile acid metabolism genes, and integrated multi-omics data using causal mediation and machine learning causal inference. Both NJ and ASD were associated with increased diversity of bile acid metabolism genes, suggesting biomarker potential. The gut DNA virome was also identified as a potential biomarker. Causal mediation analysis showed that the gut DNA virome influences bile acid metabolism genes in both conditions. Using machine learning-based causal modeling, we further found that gut human betaherpesviruses and human mastadenoviruses contribute to NJ and ASD, respectively, mediated by gut bile acid-metabolizing bacteria. These findings suggest that perturbations in the virome and bile acid-metabolizing bacteria may explain the link between NJ and ASD. Our results indicate that NJ and ASD are associated with bile acid metabolism alterations, which are also influenced by the gut DNA virome. Dysbiosis of the gut DNA virome and bile acid-metabolizing bacteria may mechanistically link NJ and ASD.IMPORTANCEHuman epidemiological studies have established an association between perinatal pathogenic infections and autism spectrum disorder (ASD), and the gut microbiota plays an extremely important role in this relationship. Neonatal jaundice (NJ) may increase the risk of ASD in children. However, it remains unclear whether alterations in the gut microbiota affect the association between NJ and ASD. Both NJ and ASD are linked to altered gut bile acid metabolism and significantly elevated gene diversity among bile acid metabolism enzymes, and these relationships are influenced by the gut virome. Gut human betaherpesviruses and human mastadenoviruses influence the development of NJ and ASD, respectively, by influencing the abundance of gut bile acid-metabolizing microbes. Alterations of the gut virome and bile acid-metabolizing bacteria appear to explain the link between NJ and ASD. There is a lack of effective treatment options for ASD. We found that both NJ and ASD are linked to altered bile acid metabolism. Gaining a comprehensive understanding of the role of the bile acid-gut microbiota axis in the pathogenesis of NJ and ASD, as well as regulating this axis, may be crucial for developing novel preventive and therapeutic strategies for ASD.

Background: Duplications at the 19q13.42 region are rare, and the relationship between 19q13.42 duplications and neurodevelopmental phenotypes remains poorly characterized due to the paucity of cases with well-documented clinical features. Methods: Cases with 19q13.42 duplications were reviewed from a cohort of over seventy-five thousand pediatric patients. These patients, suspected of having genetic diseases, underwent next-generation sequencing (NGS) at two collaborating medical centers between 2016 and December 2024. Cases were identified and subjected to further phenotypic and genomic analysis. The reported cases of 19q13. 42 duplications were also searched in public resources. Results: Twenty patients, including 10 cases from our study and 10 cases from public resources with 19q13.42 duplication were enrolled. The duplication breakpoints detected by NGS in internal cases were nonrecurrent and the duplication sizes ranged from 76.864 to 724.194 Kb, which were verified by quantitative real-time polymerase chain reaction (qPCR). Neurodevelopmental disorders (NDDs) were the primary clinical phenotype, present in 94.1% (16/17) of patients. All 16 cases harbored duplications involving PRKCG, of which 12 contained complete gene duplications and 4 involved partial duplications. Genotypephenotype correlation analysis and the smallest region of overlap (SRO) mapping indicates that PRKCG is a candidate gene underlying the associated NDD phenotypes. Conclusions: We observed recurrent neurodevelopmental features in a subset of reported 19q13.42 duplications and propose PRKCG as a candidate gene warranting follow-up functional studies. These findings provide a preliminary genotype-phenotype framework for this region, though additional cases with comprehensive clinical phenotyping are required to validate and refine these observations.

Neonatal jaundice (NJ) affects 60-80% of neonates, yet the underlying microbial mechanisms remain elucidated, despite known links between gut dysbiosis and bilirubin and bile acid (BA) metabolism. Through two-stage shotgun metagenomic-metabolomic analysis of 150 fecal samples from 120 neonates, we identified key taxa linked to bile acid (BA) metabolism in moderate-to-severe NJ. Furthermore, multi-omics integration revealed significant interkingdom correlations among gut phages, bacteria, and BAs. Dysbiosis featured enriched Streptococcus and Escherichia, depleted Bifidobacterium animalis, and group-specific phage signatures. In the independent clinical validation cohort, jaundice intervention normalized the dysbiotic profile, demonstrating significant suppression of pathogenic taxa concomitant with restoration of B. animalis abundance. In vitro, B. animalis subsp. lactis Y103-OTU5 remodeled BA via deconjugation. In a phenylhydrazine hydrochloride (PHZ)-induced murine model of hemolytic jaundice, oral administration of isolated B. animalis subsp. lactis Y103-OTU5 significantly attenuated hyperbilirubinemia and hepatic inflammation, likely via Cyp7a1/Cyp7b1-dependent modulation of BA synthesis and detoxification pathways. Structural equation modeling revealed a tripartite regulatory network: phages indirectly modulated BA through bacterial remodeling, while B. animalis directly regulated BA pathways, positioning it as a potential therapeutic candidate for hemolysis-associated neonatal jaundice. Collectively, these findings reveal a gut phage-bacteria-BA network in NJ, highlighting B. animalis as a therapeutic candidate with dual modulation of BA metabolism and phage-bacteria interactions.

ABSTRACT A substantial proportion of patients with X‑‐linked inhibitor of apoptosis (XIAP) deficiency develop severe and treatment‑‐refractory Crohn's disease (CD). Although hematopoietic stem cell transplantation (HSCT) remains the only curative option for these patients, its outcomes are suboptimal, with a long‑‐term survival rate of only 50%. Therefore, identifying novel therapeutic targets is crucial to bridge this unmet clinical need. Here, we demonstrate that the abundance of tuft cells is reduced in both XIAP‐deficient CD patients and Xiap knockout (Xiap−/−) mice. Mechanistically, XIAP deficiency reduces TLE4 ubiquitination, resulting in elevated TLE4 protein levels and consequent suppression of Wnt/β‑‐catenin–ASCL2 signaling, which is critical for secretory lineage differentiation. Tuft cell deficiency may increase susceptibility to microbial dysregulation, thereby promoting intestinal inflammation. Furthermore, we demonstrate that JAK inhibition promotes tuft cell regeneration and ameliorates mucosal inflammation in Xiap−/− mice. Consistently, in an XIAP‑‐deficient CD patient, treatment with a selective JAK1 inhibitor effectively increased tuft cell proportion and alleviated colonic symptoms. In conclusion, our study identifies tuft cell deficiency as a trigger of intestinal pathology in XIAP‑‐deficient Crohn's disease and suggests JAK inhibition as a promising therapeutic strategy.

Background:The optimal management of patent ductus arteriosus (PDA), particularly hemodynamically significant PDA (hsPDA), in extremely preterm infants remains highly controversial. This study aimed to evaluate whether early ibuprofen for hsPDA was associated with clinically meaningful outcomes in extremely preterm infants and whether this association was modified by clinical tolerance to PDA-related hemodynamic changes. Methods:This prospective cohort study used data from the Chinese Multicenter Collaboration Platform for PDA in Extremely Preterm Infants and included infants with a gestational age ≤29+6 weeks who were admitted between November 2024 and September 2025 and diagnosed with hsPDA by echocardiography on postnatal days 3-5. A target trial emulation framework was applied, with the day of echocardiography as time zero and a 2-day grace period. Ibuprofen initiation within the grace period defined treatment. Inverse probability of treatment weighting was used to balance baseline characteristics and emulate random treatment assignment. Infants were stratified as clinically tolerant and intolerant based on respiratory and hemodynamic support at the time of echocardiography. Outcomes included mortality and major prematurity-related morbidities. Results:A total of 239 infants were included, of whom 65 received ibuprofen within the grace period and 174 did not. In the overall cohort, ibuprofen was associated with a lower, but not statistically significant, risk of mortality [risk ratio (RR), 0.51; 95% confidence interval (CI): 0.25-1.02]. In analyses stratified by clinical tolerance, ibuprofen was associated with a reduced risk of mortality only in the intolerant subgroup (RR, 0.18; 95% CI: 0.06-0.51), but not in the tolerant subgroup (RR, 0.96; 95% CI: 0.23-4.00). Conclusions:The effect of ibuprofen for hsPDA in extremely preterm infants appears heterogeneous, with differences in mortality associations observed across clinical tolerance subgroups. These findings are exploratory and warrant confirmation in future adequately powered randomized trials.

Preterm infants are susceptible to hyperglycemia within the neonatal period, leading to increased mortality and morbidity. Nevertheless, the risk factors contributing to hyperglycemia in this population remain incompletely elucidated. To address this issue, we employed machine learning algorithms to identify the risk factors for hyperglycemia detected by continuous glucose monitoring (CGM) in preterm neonates. In this retrospective observational study, we included 66 preterm infants (< 37 weeks’ gestation) who were hospitalized at the Children’s Hospital of Fudan University from June 2015 to May 2023 and underwent CGM initiated within 7 days after birth during the neonatal period. We evaluated maternal, neonatal, and postnatal factors associated with hyperglycemia frequency, defined as CGM glucose > 10 mmol/L and quantified by the proportion of CGM time above this threshold. Pairwise correlation, structural equation modeling (SEM), and a histogram-based gradient boosting regression tree (HGBRT) with SHAP-based feature ranking were used for analysis. Hyperglycemia (> 10 mmol/L) was detected in 17 of 66 CGM-monitored preterm neonates (25.76

Adverse exposures during early life, including environmental and genetic factors, can have profound and long-lasting effects on health in later life. Birth cohorts provide invaluable platforms for studying these effects, as they enable the collection of longitudinal data across multiple life stages. This paper explores the integration of genomic research into large birth cohorts. By utilizing genome-wide association studies, multi-omics approaches, and advanced epidemiological techniques, birth cohorts can integrate genetic, environmental, and molecular data to enhance our understanding of the biological underpinnings of maternal and child health. However, several challenges persist. These include ethical concerns regarding data privacy and informed consent, the analytical complexity in multi-modal datasets, the slow translation of research findings into clinical practice, the inevitable attrition in long-term follow-up, and the scarcity of multi-generational enrollment. Despite these challenges, integrating genomic research into birth cohorts is a critical step toward personalized healthcare and effective disease prevention strategies, providing essential insights into the origins of maternal and child health outcomes.