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ObjectiveThis study investigated how cumulative environmental exposures influence offspring behaviour and inflammation-related molecular signatures in the brain and peripheral immune system.MethodsA novel "triple-hit" mouse model was developed using C57Bl/6JAusB mice (N = 70), combining preconceptual social stress, antenatal high-fat diet, and a postnatal immune challenge (poly(I:C), 10 mg/kg). At 12 weeks, offspring underwent behavioural tests relevant to neurodevelopmental disorders (NDDs), including the Elevated Plus Maze, 3-Chamber Social Preference, Self-Grooming, and Marble Burying. A composite NDD-risk index was calculated. Single-cell RNA sequencing (scRNA-seq) and bulk proteomics were performed on male triple-hit offspring to identify differentially expressed genes and proteins associated with inflammatory pathways.ResultsMale triple-hit offspring showed elevated NDD-related behavioural risk and social deficits, not observed in females. scRNA-seq revealed altered inflammatory and ribosomal pathways in brain glia and peripheral immune cells. Proteomic analysis showed decreased abundance of proteins involved in inflammation, translation, chromatin remodelling, and synaptic function in both brain and blood.ConclusionCombined environmental stressors may drive male-specific behavioural and inflammatory changes relevant to NDDs. The identification of overlapping inflammatory signatures in brain and peripheral immune cells supports a role for shared immune mechanisms in brain–immune axis dysfunction. However, these pathway-level findings should be interpreted as preliminary hypotheses and warrant independent validation to confirm their mechanistic significance.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00011-025-02152-y.

BackgroundOvarian cancer (OC) remains the deadliest gynecological cancer, primarily due to late-stage diagnosis and high rates of chemotherapy resistance and recurrence. Lack of representative preclinical models complicate the challenges of discovering effective therapies, especially for platinum-resistant OC. Patient-derived xenograft (PDX) models maintain the genetic characteristics of the original tumor and are ideal for testing candidate therapies in vivo, but their high cost limits their feasibility for high-throughput drug screening. Organoid models mimic the tumor’s 3D structure and preserve intra-tumoral heterogeneity. While organoids established directly from primary patient tumors are the optimal model for personalized drug response studies, the supply of primary tissue is often limited. Patient-derived xenograft tumors can be passaged in mice and provide a renewable source of cancer cells for organoids. This study aimed to determine if PDX-derived organoids (PDXOs) can reflect patient responses to chemotherapy similarly to primary patient-derived organoids (PDOs).Methods3D Organoid models were established from the malignant ascites of five high grade serous ovarian cancer patients: two platinum-sensitive, two platinum-resistant, and one platinum-refractory, along with their matched PDX samples from ascites and solid tumor. Organoid viability after 72-hour treatment with paclitaxel (PTX), carboplatin (CBDCA), or their combination was compared between organoids derived directly from the patient or from the PDX models. The in vitro drug responses of PDXOs and PDOs were then compared to defined patient clinical responses: platinum-sensitive (initial response to standard platinum/paclitaxel therapy lasting > 6 months post-treatment), platinum-resistant (initial response to standard chemotherapy lasting < 6 months), or platinum-refractory (no initial response to standard chemotherapy).ResultsIn drug response assays, PDXOs and PDOs demonstrated similar sensitivity to standard chemotherapy and also reliably reflected patient responses based on the clinical designation of platinum sensitivity. While organoids derived from the ascites were smaller with a denser morphology, their drug response mirrored that of the organoids derived from solid tumor. Platinum-sensitive cases exhibited significant reductions (around 50% reduction) in organoid viability when treated with carboplatin, paclitaxel, or their combination. Platinum-resistant or refractory organoids showed little to no reduction in viability with carboplatin or paclitaxel monotherapy or the combination. Organoids derived from one platinum-resistant case did show a small but significant reduction in viability with single-agent paclitaxel, suggesting that organoid models might predict response to second-line paclitaxel therapy.ConclusionThis study demonstrates that PDXOs respond to drugs similarly to PDOs and confirms that both models effectively mirror patient response to standard chemotherapy. This highlights the potential of PDXOs as renewable models for screening novel therapies and developing personalized strategies in OC.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12885-025-14811-8.

AbstractThe NOTCH pathway regulates cell proliferation, differentiation, and stem cell maintenance. Thus, aberrant NOTCH activation plays a key role in cancer initiation, progression, and chemoresistance. Mutations and amplification of NOTCH pathway genes have been identified in high-grade serous ovarian cancers and are associated with poor clinical outcomes. Among the four NOTCH receptors, NOTCH3 alterations were strongly correlated with poor overall survival. Previously, we identified auranofin, an oral gold salt therapeutic compound, as a novel NOTCH pathway inhibitor that disrupts the DNA binding of RBPJ, the major downstream transcriptional effector of the NOTCH pathway. In this study, we surveyed the response of eight ovarian cancer cell lines to auranofin and found IC50 values ranging from 1.7 to 12 μmol/L, with NOTCH3-negative SKOV3 cells having the highest IC50 value. In NOTCH-dependent OVCAR3 cells, auranofin synergized with cisplatin to enhance cell death. Importantly, auranofin treatment led to a dose-dependent decrease in RBPJ occupancy at the NOTCH-dependent promoters, HES1 and HES4. Furthermore, knocking down NOTCH3 in OVCAR3 cells significantly decreased sensitivity to auranofin, further supporting the notion that NOTCH3 signaling is a major target of auranofin. Moreover, auranofin increased cisplatin efficacy in an OVCAR3-derived xenograft mouse model. Using eight patient-derived cancer organoid models, we found that auranofin increased cisplatin efficacy in killing cancer organoids generated from clinically platinum-sensitive patients but also restored platinum response in a subset of organoid models developed from platinum-resistant patients. These studies underscore the potential of auranofin to improve platinum-based cancer therapy, particularly in NOTCH3-expressing cancers.Significance:NOTCH signaling underlies cancer initiation, progression, and chemoresistance. Our study revealed the potential of auranofin as a NOTCH pathway inhibitor to enhance the efficacy of platinum-based ovarian cancer therapy.