Abstract As pervasive environmental contaminants, nanoplastics (NPs) have garnered increasing concern due to their potential health hazards. However, the impact of gestational and lactational exposure to NPs on offspring vascular structural injury remains unexplored. In this study, we used BALB/c mice to model gestational and lactational exposure to 100 nm polystyrene nanoplastics (PS-NPs) at doses of 0.1, 1, and 10 mg/l. Our findings revealed that PS-NPs exposure significantly reduced offspring body weight in both sexes. Histopathological analysis demonstrated that exposure induced multiorgan damage in offspring at postnatal day 21 (PND 21), with partial recovery by PND 90. Critically, PS-NPs exposure induced cardiovascular toxicity in offspring, which included acute vascular injuries (disordered elastic fibers and evident calcium salt deposition) at PND 21 that were largely resolved by PND 90, alongside more persistent issues of abnormal lipid deposition in the aortic root and dyslipidemia observed at both time points. Integrated biomarker response indices, principal component analysis, and hierarchical clustering confirmed dose-dependent toxicity in PS-NPs-exposed offspring. In vitro studies further showed that PS-NPs induced lipid accumulation in 3 major cardiovascular cell types: endothelial cells, smooth muscle cells, and macrophages. Our study firstly reveals that maternal PS-NPs exposure throughout gestation and lactation induces vascular injury and lipid deposition in mouse offspring, indicating that PS-NP pollution represents a critical environmental risk factor for offspring cardiovascular adverse outcomes later in life.
Abstract Attention-deficit hyperactivity disorder (ADHD) is a neurodevelopmental disorder characterized by inattention, hyperactivity, and impulsivity. Methylphenidate (MPH), a first-line medication, is often used long-term and not routinely discontinued in individuals planning to have children. Guided by the Paternal Origins of Health and Disease framework, in this study, we investigated intergenerational effects of paternal MPH exposure and reversibility of sperm epigenetic changes. Male BALB/c mice received MPH (15 mg/kg) or vehicle for 28 days and were mated with untreated females immediately after treatment (F0-T1) or after a 40-day washout period (F0-T2). Behavioral tests and brain gene expression analyses were conducted in the offspring (F1), and sperm small noncoding RNA (sncRNA) profiles were examined in F0 males. Male offspring in the F0-T1-MPH group showed increased open-arm entry into the elevated plus maze and reduced expression of Maoa, Maob, and Bdnf in the medial prefrontal cortex. These changes were absent in the offspring derived from the F0-T2-MPH group. Sperm sncRNA sequencing identified 5,684 differentially expressed fragments in F0-T1-MPH mice, including marked alterations in miRNA-derived fragments, such as miR-34c-5p and miR-30e-5p. The target genes of these altered miRNAs were enriched in ADHD-related pathways, including axon guidance and Rap1 signaling. After washout, the number of differentially expressed sncRNA fragments decreased to 665, whereas the number of altered miRNA-derived fragments decreased from 383 to 7. These findings suggest that paternal MPH exposure may influence offspring behavior and brain gene expression through sperm miRNA alterations and that these effects are largely reversible following drug withdrawal.
Abstract Perfluorooctanesulfonic acid (PFOS) is epidemiologically linked to gestational diabetes mellitus (GDM), but the mechanisms remain undefined. We hypothesized that PFOS exposure impairs pancreatic β-cell function through mitochondrial dysfunction and deficits in ATP/cAMP signaling. Pregnant and non-pregnant rats received PFOS (50 μg/mL in drinking water) for 17 days (gestation day [GD] 4–20 in pregnant dams). PFOS-exposed pregnant dams exhibited fasting hypoinsulinemia (65% decrease) with normal fasting glucose. Glucose tolerance was impaired (31% increase in glucose AUC), with blunted insulin secretion (52% decrease in insulin AUC). In contrast, non-pregnant PFOS-exposed rats showed no changes in fasting glucose or insulin but developed glucose intolerance (25% increase in glucose AUC) with only modest early insulin suppression, revealing greater vulnerability in pregnancy. Pancreatic islet morphology and insulin content remained unchanged in PFOS-exposed dams. Isolated islets from PFOS-exposed dams and INS-1 β-cells treated with PFOS (10–100 μM) showed reduced glucose-stimulated insulin secretion despite normal Ins1 and Ins2 expression and insulin content. In INS-1 cells, PFOS selectively reduced Complex II subunits, depleted ATP (32–39%) and cAMP (57–74%), and impaired insulin secretion under low and high glucose. Co-treatment with the mitochondria-targeted antioxidant MitoQ (0.1μM) fully restored ATP, cAMP, and insulin secretion to control levels. These findings demonstrate that PFOS exposure causes gestational glucose intolerance and β-cell secretory failure, accompanied by reduced mitochondrial Complex II subunit abundance and ATP/cAMP depletion. The complete rescue by MitoQ highlights oxidative mitochondrial dysfunction as a mechanistic contributor linking PFOS exposure and GDM risk.
Abstract Synthetic messenger RNA (mRNA) formulated in lipid nanoparticles (LNPs) is increasingly used for vaccines and therapeutics, yet the consequences of high-dose systemic exposure for bone marrow hematopoiesis remain poorly defined. We previously reported that intramuscular mRNA/LNP administration transiently alters circulating reticulocytes and bone marrow erythroid cellularity in mice; here, we investigated the mechanistic basis and cross-species generalizability of this hematopoietic bias in murine and non-human primate models. Animals received intramuscular LNP-formulated mRNA encoding SARS-CoV-2 spike protein or firefly luciferase at deliberately supra-clinical, upper-bound doses, and type I interferon signaling was interrogated by interferon-α/β receptor blockade. Bone marrow was assessed by histopathology, bulk and single-cell RNA sequencing, and trajectory analysis. LNP-formulated mRNA induced reversible erythroid-lineage suppression with concomitant granulocytic expansion, irrespective of the encoded antigen. These effects were absent after naked mRNA and only modestly induced by LNP alone, indicating that formulation of synthetic mRNA within the LNP was required to elicit the full phenotype. Single-cell analysis showed biased hematopoietic stem and progenitor cell differentiation toward granulopoiesis with broad induction of interferon-stimulated genes, and the bias resolved within 2 wk. Interferon-α/β receptor blockade mitigated both changes, demonstrating type I interferon dependence. High-dose LNP-formulated synthetic mRNA can therefore drive reversible, type I interferon-dependent hematopoietic lineage bias in preclinical animal models. This study identifies type I interferon signaling as the driver of a reversible, dose-dependent shift in bone marrow hematopoiesis under upper-bound exposure to LNP-formulated synthetic mRNA, informing preclinical safety evaluation of the rapidly expanding class of nucleic acid-based therapeutics.
Abstract Humans are exposed to complex mixtures of endocrine-disrupting chemicals (EDCs) in the real world. Most prior studies, however, focus on individual EDCs. Here, we investigated the ovarian impact of organic pollutant mixtures extracted from the Raritan River, a heavily industrialized watershed in central New Jersey, using a three-dimensional (3D) in vitro ovarian follicle culture system. Ultra-high-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS) identified organic pollutant mixtures dominated by alkylphenols, pharmaceuticals, per- and polyfluoroalkyl substances (PFAS), and phthalates, together with previous reports demonstrating that the Raritan River also contains hydrophobic persistent organic pollutants (POPs), including polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and dioxins. In vitro exposure to the Raritan River-derived EDC mixture did not affect follicle growth and ovulation but dose-dependently increased preovulatory testosterone (T) secretion while suppressing postovulatory progesterone (P4) production. Transcriptomic analysis, gene set enrichment analysis (GSEA), reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and in situ RNA hybridization revealed widespread reprogramming of follicular somatic cell gene expression, including altered steroidogenic pathways and activation of aryl hydrocarbon receptor (AhR) signaling. An established AhR agonist, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), reproduced the phenotype of EDC mixture-induced hyperandrogenism, which was reversed by a selective AhR inhibitor. Together, this study demonstrates that real-world EDC mixtures, particularly AhR-activating chemicals, disrupt ovarian steroidogenesis and induce hyperandrogenism, highlighting EDC mixtures as an underrecognized mechanism potentially linking aquatic pollution to impaired women’s reproductive health.
