Zhongzhong Chen

While the natural light–dark cycle is crucial for health, modern lifestyles are associated with increased artificial light exposure, including during reproductive windows. Although maternal light exposure has been linked to adverse offspring outcomes, comprehensive multisystem evaluations remain scarce. In this study, we established a rat model of continuous light exposure to simulate maternal preconception chronodisruption. We found that preconception chronodisruption was associated with multisystem abnormalities in offspring, including increased body weight, diminished cardiac function, thickened vessel walls, elevated blood pressure, and insulin resistance. Behavioral tests revealed stress-coping and behavioral despair-related responses along with impaired cognitive function. The m6A methylation patterns in maternal oocytes were altered and enriched in developmental pathways related to the brain, muscle, and cardiovascular systems. Collectively, preconception exposure to continuous light was associated with physiological and behavioral deficits in offspring, and these deficits were potentially related to m6A RNA modification changes in maternal oocytes. These findings underscore the importance of preconception environmental factors and suggest a potential link between oocyte epigenetics and developmental programming.

Hypospadias is one of the most common birth defects in China and a key feature of differences in sex development (DSD), yet its genetic etiology remains largely unresolved. Current diagnostic approaches using DSD-targeted gene panels have a low rate of definitive diagnoses (5.5%), highlighting the need for more comprehensive genetic investigation. In this study, we performed next-generation sequencing (NGS) on the largest trio-based cohort of hypospadias to date, comprising 106 pediatric cases and their parents (92 trios). We achieved a definitive genetic diagnosis in 6.6% of patients, identifying pathogenic variants in canonical DSD genes such as AR, NR5A1 and WT1 . By incorporating a broader spectrum of potentially clinically significant variants, we increased the overall genetic identification rate to 33.0% (35/106). Strikingly, trio analysis uncovered a significant burden of de novo loss-of-function (LoF) variants (2.2-fold enrichment, P =0.001), primarily driven by variants in genes associated with ciliopathies (10.35-fold, P =0.016), a previously underappreciated gene class in hypospadias. Furthermore, we identified and functionally validated two high-confidence risk genes, PRKCZ and HRNR , based on recurrent de novo variants. Functional assays confirmed that these variants disrupt key biological mechanisms, including cell proliferation, migration, and androgen signaling. Our large-scale trio approach substantially expands the genetic landscape of hypospadias, demonstrates the critical value of trio-based sequencing for improving diagnostic yield, and decisively implicates ciliary genes in its pathogenesis.

Objective: Steroid‐induced osteonecrosis of the femoral head (SONFH) is a refractory skeletal disorder influenced by genetic and environmental factors. However, conclusive pathogenic genetic evidence remains elusive due to the limited exploration of rare damaging variants. In this study, we aimed to identify rare variants associated with SONFH. Methods: We conducted whole‐exome sequencing in an SONFH case‐control study comprising 174 patients with systemic lupus erythematosus (SLE) taking steroids, followed by comparing the cases with an ethnically matched healthy population and validation in an additional SONFH cohort of 246 patients without SLE. Rare damaging variants were identified via genetic burden analysis and confirmed by Sanger sequencing and pedigree studies. The functional assessments of the erythrocytes on target variants were performed. Results: We identified 10 heterozygous ANK1 and EPB41 rare variants in 9 (10.8%) of 83 patients with SONFH compared with 0 of the 91 non‐SONFH controls (0%). The burden effect of them remained robust after adjusted analysis (Firth's logistic regression‐adjusted odds ratio [aOR] _ANK1 11.3; aOR _EPB41 32.3; both P < 0.05). The variants were detected in 10 of the validated SONFH cohort (4.1%) with 246 non‐SLE conditions. Functional analyses revealed that the variants result in membrane dysfunctions in the patients’ erythrocytes, characterized by reduced ANK1 expression, abnormal morphology, and increased hypotonic hemolysis. Conclusion: These findings suggest that the rare variants in ANK1 and EPB41 are novel SONFH disease risk factors that compromise erythrocyte membrane integrity, exacerbating microcirculatory damage in the presence of glucocorticoid as a second hit.

Cryptorchidism, characterized by undescended testes, is associated with infertility and increased cancer risk through complex, multifactorial pathophysiological mechanisms involving interconnected alterations in testicular microenvironment, including but not limited to elevated temperature, hormonal dysregulation, altered vascular perfusion, and immune responses. These factors interact synergistically to drive testicular pathology. Using a surgically induced bilateral cryptorchid mouse model established at postnatal day 21 (PND21), we investigated phase-specific pathological mechanisms through analyses at prepubertal (PND35) and sexually mature (PND70) phases. Our transcriptome analysis revealed distinct molecular signatures at different developmental phases, with prepubertal cryptorchid testes showing 2570 differentially expressed genes predominantly enriched in immunoproteasome components and inflammatory pathways, while sexually mature testes exhibited 883 differentially expressed genes primarily related to extracellular matrix (ECM) remodeling and oncogenic pathways. Prepubertal molecular changes indicated immunoproteasome activation and inflammatory responses, whereas mature-phase alterations were characterized by ECM reorganization and fibrotic remodeling. Functional analysis demonstrated prepubertal enrichment in spermatogenesis regulation and interferon responses, while mature-phase signatures were associated with apoptosis, epithelial–mesenchymal transition, and inflammatory signaling cascades. Phase-specific oncogenic pathway correlations revealed distinct mechanisms: metabolic reprogramming and epigenetic regulation in prepubertal testes versus structural remodeling and invasion-related pathways in mature testes. Molecular validation confirmed elevated PI3K-Akt and NF-κB signaling at both developmental phases, identifying these as potential therapeutic targets. This first phase-resolved characterization of cryptorchidism pathology provides insights into developmental phase-specific mechanisms and suggests timing-dependent therapeutic strategies. Although differing from human congenital cryptorchidism in developmental timing and etiology, our surgically induced model recapitulates anatomical testicular malposition with multiple inseparable pathophysiological alterations, and the identified molecular signatures reflect integrated responses to the complex cryptorchid microenvironment.

Previous studies have demonstrated that peri-conceptional inositol supplementation could effectively ameliorate the recurrence of neural tube defects (NTDs); though, the mechanism remains unclear. In the current investigation, we detected the myo-inositol (MI) levels in maternal plasma and embryonic tissues in a Chinese population with high prevalence of NTDs and found maternal MI deficiency increased NTD susceptibility in this area. Pregnant mice were randomly divided into 2 groups. The control group was treated with 0.9% saline, while the experimental group was treated with IMPase inhibitor by intraperitoneal injection on E7.5 to generate the maternal MI-deficient mouse model, followed by comprehensive phenotypic and molecular analysis at E13.5. Corresponding in vitro experiments treated neural stem cells (NSCs) for 16 h with IMPase inhibitor, MI supplementation, or specific PI3K and mTOR inhibitors to systematically investigate the downstream signaling pathways. We observed aberrant activation PI3K/Akt/mTOR signaling and reduced autophagy in a maternal MI deficiency mouse model after ruling out the genetic perturbations. Further in vitro kinase assay showed that MI negatively regulated PI3K activity, in a dose-dependent manner, which possibly due to the hydrogen bond interactions between amino acid residues in the ATP-binding site of PI3K and inositol. MI deficiency NSCs also presented PI3K/AKT/mTOR pathway activation concurrent with reduced autophagy level; MI supplementation or inhibition of the PI3K/Akt/mTOR pathway could rescue MI deficiency-induced autophagy impairment. Finally, we validated decreased autophagy levels as well as hyperactivation of the PI3K/Akt/mTOR pathway by MI deficient human NTD embryonic neural tissues. Our results suggested that maternal MI deficiency might increase susceptibility to NTDs. Low levels of MI might impair autophagy in the developing neural tube involving upregulation of PI3K/Akt/mTOR pathway.

With improved survival rates among congenital heart disease (CHD) patients, pulmonary arterial hypertension (PAH) linked to CHD becomes more prevalent in both children and adults. PAH remains a significant contributor to morbidity and mortality in this population. Although genome-wide association studies (GWAS) have identified potential genetic variants with PAH risk and prognosis, the identification of circulating biomarkers with causal roles in CHD-PAH remains unclear. We employed the summary data-based Mendelian randomization (SMR) method, integrating expression profile data from the Gene Expression Omnibus (GEO) database related to CHD-PAH. This approach aimed to pinpoint genes causally associated with risk of CHD-PAH. We used a two-sample Mendelian randomization (MR) approach to efficiently screen for circulating proteins affecting CHD-PAH, leveraging publicly available genetic data from the UK biobank Pharma Proteomics Project (UKB-PPP) (54,219 UKB participants). Genetic determinants (cis-SNPs) of circulating proteins were used as instruments, and MR analyses assessed the influence of these proteins on CHD-PAH susceptibility in the largest PAH GWAS (2085 cases and 9659 controls). We conducted colocalization analyses to ensure shared genetic signals between circulating proteins and PAH and performed immune cell infiltration analysis to understand immune regulatory mechanisms in CHD-PAH. We found that a 1 SD increase in circulating S100 calcium binding protein A3 (S100A3) levels correlated with a reduced PAH risk (OR: 0.073, 95% CI: 0.020-0.267; p = 0.00799). Sensitivity analyses including various cis-SNPs, provided consistent estimates for S100A3 (inverse variance weighted (IVW) OR: 0.085, 95% CI: 0.032-0.225; p = 7.5 × 10-7 and MR-Egger OR: 0.212, 95% CI: 0.013-3.376; p = 0.387). Colocalization analyses confirmed a shared genetic signal for S100A3 and PAH, with a posterior probability of 99.9%. Transcriptomic investigations further highlighted S100A3's protective role in CHD-PAH. Our study using SMR and GEO data identified S100A3 as a gene associated with a reduced risk of PAH in CHD patients. Elevated circulating levels of S100A3 were linked to a reduced PAH risk, and transcriptomic evidence further supported its protective function in CHD-PAH.

This study aims to evaluate the relationship between mandibular developmental abnormalities and swallowing function in children with Pierre Robin Sequence (PRS). Swallowing function was assessed by a Modified Kubota Drinking Test (MKDT). Pre- and postoperative CT scans of PRS patients who underwent Mandibular Distraction Osteogenesis (MDO) were analyzed through three-dimensional (3D) digital reconstruction technology. Mandibular and airway evaluation parameters were measured, including the distance between bilateral mandibular angular, the length of bilateral mandibular ramus, mandibular notch angle (α), mandibular angle (β), mandibular body angle (γ), and the lateral and longitudinal dimensions of the posterior lingual airway. Results showed that the length of the bilateral mandibular rami and posterior lingual airway dimensions were significantly reduced postoperatively compared to controls (p < 0.01). After MDO, the length of mandibular rami and lateral retroglossal airway dimensions increased, α and β angles increased, while γ angle decreased (p < 0.05). Notably, the distance between bilateral mandibular angles, mandibular rami length, and lateral retroglossal airway dimensions had the strongest impact on swallowing score. In conclusion, mandibular width, length, and airway dimensions were closely linked to swallowing function in PRS patients. MDO effectively improved mandibular hypoplasia, improved swallowing dysfunction, and significantly enhanced quality of life for the patients.

Right ventricular volume overload (RVVO) is a common hemodynamic abnormality in patients with congenital heart disease (CHD) and frequently leads to pathological cardiac remodeling. Our previous research demonstrated that RVVO disrupts the metabolic maturation of cardiomyocytes. Mitochondrial metabolic maturation, a crucial process in postnatal cardiomyocyte development, remains poorly understood under RVVO conditions. In this study, an mouse RVVO model was established on postnatal day 7 by creating a fistula between the abdominal aorta and inferior vena cava, confirmed by abdominal ultrasound and echocardiography. Transcriptomic analyses revealed significant downregulation of genes linked to mitochondrial metabolic maturation. Transmission electron microscopy showed impaired mitochondrial structure and maturation markers, while Seahorse assays demonstrated a marked reduction in oxidative phosphorylation rates in RVVO cardiomyocytes. These findings collectively indicated that RVVO restricted mitochondrial metabolic maturation in the postnatal RV. Targeting mitochondrial metabolic maturation could offer a promising therapeutic strategy to mitigate RVVO-induced pathological remodeling.

Previous studies have indicated an elevated risk of infertility in certain birth defects, including congenital heart disease (CHD), hypospadias, cryptorchidism, and disorders of sexual development (DSD). Although the identification of chromosomal abnormalities or chromosomal aberrations (CAs) is crucial for the diagnosis of these conditions, the assessment of CAs in these disorders remains unclear, and few large-scale studies have been conducted at multiple centers. The aim of the current study was to systematically evaluate the prevalence of CAs in CHD, hypospadias, cryptorchidism, and DSD. Studies reporting CAs in these birth defects were retrospectively analyzed from 1991-2023, using online databases such as PubMed and Google scholar as well as preprints and references from related literature. Comprehensive screening, data acquisition, and systematic assessments of the identified literature were performed. Ultimately, searches yielded a total of 7,356 samples from 14 published articles on CHD, 298 hypospadias cases from 4 published articles, 1,681 cryptorchidism cases from 4 published articles, and 2,876 DSD cases from 7 published articles. Carrier rates of CAs varied widely among these studies and conditions. A retrospective analysis revealed that CHD was associated with the highest carrier rate (26%) for CAs, followed by DSD (21%), hypospadias (9%), and cryptorchidism (5%). A subtype analysis of CAs indicated a higher prevalence of numerical abnormalities among the reported cases. Therefore, considering CAs in birth defects associated with infertility is imperative. This provides a foundation for the further clinical implementation of chromosomal screening and enhancing high-risk screening for individuals in the real world.