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Raw data were imported for baseline filtering, peak detection, integration, retention time correction, and peak alignment to generate a data matrix containing retention time, m/z, and peak intensity. Subsequently, feature peaks were searched against the metabolite database, matching both MS and MS/MS spectral information. The MS mass error was set to less than 10 ppm, and metabolites were identified based on the MS/MS matching score.MetaboLightsPublicLiquid Chromatography MS - negative - reverse-phaseLiquid Chromatography MS - positive - reverse-phaseChromatographic separation was performed using a Thermo Scientific Vanquish Horizon UHPLC system equipped with an Accucore C30 column (100 mm × 2.1 mm i.d., 2.6 µm; Thermo). The mobile phase consisted of (A) 50% acetonitrile in water (containing 0.1% formic acid and 10 mmol/L ammonium acetate) and (B) acetonitrile/isopropanol/water (10/88/2, v/v/v) (containing 0.02% formic acid and 2 mmol/L ammonium acetate). The flow rate was 0.4 mL/min, the column temperature was maintained at 40°C, and the injection volume was 3 µL.Bisphenol mixture-induced DGKB downregulation promotes premature human neurogenesis through the activation of MAPK pathway.HAO LIsichuan universityOrganoidmass spectrometry assayLipid extraction was performed using a methanol/water and methyl tert-butyl ether (MTBE)-based protocol. Briefly, samples were homogenized with 280 µL methanol/water (2:5, v/v) and 400 µL MTBE. After ultrasonication and centrifugation, the upper organic phase was collected, dried under nitrogen, and reconstituted in 100 µL isopropanol/acetonitrile (1:1, v/v). Pooled quality control (QC) samples were prepared by mixing equal volumes (20 µL) of supernatant from each sample to assess system stability.Homo sapienshttps://www.ebi.ac.uk/metabolights/MTBLS15671Yan Huang.Hao Li. 17882586286@163.com.Pinli Zeng.Qian Bu. Sichuan University. buqian7978@scu.edu.cn.Raw mass spectrometry data were processed using LipidSearch software (Thermo, CA) for peak detection, alignment, and lipid identification based on MS/MS fragmentation analysis (MS mass error < 10 ppm). Subsequent statistical analyses and visualization were performed using the Majorbio Cloud Platform. Differentially expressed lipids (DELs) were identified based on Student’s t-test (P < 0.05), variable importance in projection (VIP) > 1, and FC > 1.5 or FC < 0.67.Group17882586286@163.comHuman cortical organoids from the control and 1000 nM treatment groups were randomly harvested on day 36. For each biological replicate (n = 5 per group, with 5-6 organoids per sample), the organoids were collected and immediately processed for lipid extraction. Metabolomicspooled quality control sampleuntargeted analysisLipidSearchLipidomicsHomo sapiensOrganoidbisphenol Sexperimental sampleThermo Scientific Vanquish UHPLC SystemOrganoidsbisphenol AFThermo Scientific Q Exactive HF-Xbisphenol Epooled quality control sampleuntargeted analysisLipidSearchOrganoidLipidomicsHomo sapiensbisphenol Sexperimental sampleThermo Scientific Vanquish UHPLC SystemOrganoidsbisphenol AFThermo Scientific Q Exactive HF-Xbisphenol EMass spectrometric detection was conducted on a Q Exactive HF-X Orbitrap mass spectrometer (Thermo Fisher Scientific, USA) equipped with a heated electrospray ionization (HESI) source. Data were acquired in both positive and negative electrospray ionization modes using a data-dependent acquisition (DDA) mode. The mass scan range was m/z 200-2000. The spray voltage was set at 3000 V (positive) and -3000 V (negative), with sheath gas at 60 arb, auxiliary gas at 20 arb, and ion source heater temperature at 370°C. Normalized collision energy was set at 20, 40, and 60 V.falseBisphenol mixture-induced DGKB downregulation promotes premature human neurogenesis through the activation of MAPK pathway The widespread industrial transition from bisphenol A (BPA) to structural analogs such as BPS, BPE, and BPAF has raised significant concerns regarding their potential health risks. However, their neurodevelopmental risks and underlying molecular mechanisms, particularly when present as complex mixtures, remain poorly defined in human-specific contexts. In this study, we employed human induced pluripotent stem cell (iPSC)-derived cortical organoids to investigate the developmental neurotoxicity of a bisphenol mixture (BPS, BPE, and BPAF at 1:1:1) at environmentally relevant concentrations (10–1000 nM). The mixture induced premature neuronal differentiation, characterized by a significant expansion of the CTIP2+ deep-layer neuronal population. Integrated analyses of network toxicology, transcriptomics, and lipidomics identified a novel diacylglycerol (DG)-mediated signaling hub. Specifically, we identified the transcriptional downregulation of diacylglycerol kinase beta (DGKB) as a critical molecular initiating event (MIE), that drives DG accumulation. This lipidomic disruption selectively activated the PKCα/JNK signaling axis, leading to the induction of the transcription factor FOS, which promoted transcription of neural differentiation genes (ASCL1, NEUROD1, TBR2, and CTIP2). Collectively, our findings reveal a DG-mediated PKCα/JNK/FOS axis underlying bisphenol mixture-induced premature neuronal differentiation, providing mechanistic insights into the developmental neurotoxicity of emerging bisphenol alternatives.2026-09-292026-09-15MTBLS15671