The functional implication of ATF6α in castration-resistant prostate cancer cells.

Stress in the endoplasmic reticulum (ER) may perturb proteostasis and activates the unfolded protein response (UPR). UPR activation is frequently observed in cancer cells and is believed to fuel cancer progression. Here, we report that one of the three UPR sensors, ATF6α, was associated with prostate cancer (PCa) development, while both genetic and pharmacological inhibition of ATF6α impaired the survival of castration-resistance PCa (CRPC) cells. Transcriptomic analyses identified the molecular pathways deregulated upon ATF6α depletion, and also discovered considerable disparity in global gene expression between ATF6α knockdown and Ceapin-A7 treatment. In addition, combined analyses of human CRPC bulk RNA-seq and single-cell RNA-seq (scRNA-seq) public datasets confirmed that CRPC tumors with higher ATF6α activity displayed higher androgen receptor (AR) activity, proliferative and neuroendocrine (NE) like phenotypes, as well as immunosuppressive features. Lastly, we identified a 14-gene set as ATF6α NE gene signature with encouraging prognostic power. In conclusion, our results indicate that ATF6α is correlated with PCa progression and is functionally relevant to CRPC cell survival. Both specificity and efficacy of ATF6α inhibitors require further refinement and evaluation.

M2 Microglia Extracellular Vesicle miR-124 Regulates Neural Stem Cell Differentiation in Ischemic Stroke via AAK1/NOTCH.

BACKGROUND: Small extracellular vesicles (sEVs) derived from M2 microglia (M2-microglia-derived small extracellular vesicles [M2-sEVs]) contribute to central nervous system repair, although the underlying mechanism remains unknown. In this study, we aimed to identify the mechanism through which microRNA-124 (miR-124) carried in sEVs promotes neural stem cell (NSC) proliferation and neuronal differentiation in the ischemic mouse brain. METHODS: M2-sEVs with or without miR-124 knockdown were injected intravenously for 7 consecutive days after transient middle cerebral artery occlusion surgery. The atrophy volume, neurological score, and degree of neurogenesis were examined at different time points after ischemic attack. NSCs treated with different sEVs were subjected to proteomic analysis. Target protein concentrations were quantified, and subsequent bioinformatic analysis was conducted to explore the key signaling pathways. RESULTS: M2-sEV transplantation promoted functional neurological recovery following transient middle cerebral artery occlusion injury. M2-sEV treatment decreased the brain atrophy volume, neurological score, and mortality rate. The effect was reserved by knockdown of miR-124 in M2-sEVs. M2-sEVs promoted proliferation and differentiation of mature neuronal NSCs in vivo. Proteomic analysis of NSC samples treated with M2-sEVs with and without miR-124 knockdown revealed that AAK1 (adaptor-associated protein kinase 1) was the key responding protein in NSCs. The binding of AAK1 to Notch promoted the differentiation of NSCs into neurons rather than astrocytes. CONCLUSIONS: Our data suggest that AAK1/Notch is the key pathway in NSCs that responds to the miR-124 carried within M2-sEVs in the ischemic brain. M2-sEVs carrying ample quantities of miR-124 promote functional recovery after ischemic stroke by enhancing NSC proliferation and differentiation. Targeting of M2-sEVs could represent a potential therapeutic strategy for brain recovery.

Association of exposure to per- and polyfluoroalkyl substances with hemoglobin and hematocrit during pregnancy.

BACKGROUND: Per- and polyfluoroalkyl substances (PFASs) are common environmental contaminants and are widely detected in humans. Previous studies have linked PFASs exposure to adverse birth outcomes. However, the association between maternal exposure to PFASs and hemoglobin (Hb) and hematocrit (HCT) remains unclear. OBJECTIVES: We aimed to explore the relationship between PFASs exposure with Hb and HCT during pregnancy. METHODS: The present birth cohort study included 1044 pregnant women from Wuhan, China. Maternal HCT and Hb were measured in the first, second and third trimesters, and 13 PFASs were detected in the cord sera. Mixed linear models and general linear regression were applied to analyze the association between each single PFASs and Hb and HCT. Weighted quantile sum (WQS) regressions were used to investigate the association between PFASs mixture and Hb and HCT during pregnancy. RESULTS: In single-PFAS models, 10 PFASs were positively associated with HCT and Hb across pregnancy (a 10-fold increase in PFASs was associated with 1.47-3.54 % change in HCT and 1.46-3.20 % change in Hb (All P-FDR < 0.05). In addition, Hb and HCT were more positively related to PFASs in the second and third trimesters rather than the first trimester. The association between PFASs exposure and maternal HCT and Hb was not significant in the iron supplementation group, whereas significant in the non-iron supplementation group. A significant interaction between iron supplementation and non-iron supplementation was also detected. WQS regressions showed that perfluorononanoic acid (PFNA) and perfluorohexane sulfonate (PFHxS) contributed most to the association between PFASs and HCT and Hb in the second and third trimesters, respectively. CONCLUSION: Maternal PFASs exposure was positive with serum Hb and HCT. Moreover, maternal iron supplementation may play a modifying effect in influencing the relationship between PFASs and HCT and Hb.

Signal characteristics of normal adult bone marrow in whole-body diffusion-weighted imaging.

BACKGROUND: Knowledge of the signal characteristics of normal adult bone marrow in whole-body diffusion-weighted (DW) images (WB-DWI) is essential for correctly interpreting DW images in clinical practice; however, these factors have not yet been clearly determined. PURPOSE: To evaluate the signal characteristics of normal adult bone marrow in WB-DWI, to correlate these characteristics with age and gender, and to determine the causes of these phenomena. MATERIAL AND METHODS: Ninety-eight healthy volunteers underwent WB-DWI (b = 0 and 800 s/mm(2)). Two radiologists visually evaluated the signal characteristics of bone marrow in DW images separately. One radiologist measured the apparent diffusion coefficient (ADC) of the thoracic and lumbar vertebrae, bilateral femur (including head, neck, and proximal and distal femoral shaft), bilateral humeral head, ilium, and scapula. The signal characteristics of normal bone marrow were analyzed. RESULTS: The visual evaluation results of DW images indicated that hyperintensity of bone marrow was more frequently seen in women aged 21-50 years (68.4%) than in men aged 21-50 years (3.3%) (P < 0.001), men aged 51-81 years (5.9%) (P < 0.001), and women aged 51-81 years (15.4%) (P = 0.001). However, no statistically significant difference was found between men and women aged 51-81 years (P = 0.565). The ADC of bone marrow was significantly higher in women than in men aged 21-50 years. Bone marrow ADC showed significant negative correlation with age in women but not in men. CONCLUSION: The signal intensity of bone marrow varies with age and gender in DW images. ADC and the T2 shine-through effect contributed to the bone marrow signal intensity in DW images, and the latter effect may predominate.

Normal bone marrow signal characteristics in whole-body diffusion-weighted images of amateur marathon runners.

Background: Marathon training can reverse bone marrow conversion; however, little is known about the normal bone marrow whole-body diffusion-weighted imaging (WB-DWI) signal characteristics of amateur marathon runners. If marathon training can cause diffuse hyperintensity of bone marrow on WB-DWI is essential for correctly interpreting the diffusion-weighted (DW) images. This study sought to evaluate the WB-DWI signal characteristics of normal bone marrow in amateur marathon runners. Methods: In this prospective cross-sectional study, 30 amateur marathon runners who had trained for over 3 years for regular or half-marathon races and had a running frequency of more than 20 days a month at a distance of more than 100 km per month from the Chengde Marathon Outdoor Sports Association in Hebei, China, and 30 age- and gender-matched, healthy volunteers (the control group) who had no long-term heavy-load sports history were recruited between April 2021 to September 2021. All the subjects underwent WB-DWI (b-value: 0, 800 s/mm2) and lumbar vertebral transverse relaxation time (T2) mapping. The bone marrow WB-DWI signal characteristics were analyzed visually and statistically by chi-square (χ2) tests. The apparent diffusion coefficient (ADC), DWI signal intensity, and T2 values of the bone marrow were quantitatively and statistically analyzed by the independent sample t-test and Mann-Whitney U test. Results: No subjects were excluded from the study. The bone marrow of 30 of the 60 subjects (aged 30-50 years) showed diffuse hyperintensity in the DW images. However, in all 60 subjects, the humeral heads, femoral heads, and great trochanters had low signals. The frequency of diffuse bone marrow DWI hyperintensity was significantly higher in the male amateur marathon runners (50%) than the male controls (5%, P=0.003), but no such significant difference was found between the female amateur marathon runners (100%) and female controls (90%, P>0.99). The DW signal intensity ratios of bone marrow to muscle (SIRBM-muscle) were significantly higher in the male amateur marathon runners than the male controls in the thoracic vertebrae (4.68 vs. 3.57, P=0.021), lumbar vertebrae (4.49 vs. 3.01, P<0.001), sacrum (3.67 vs. 2.62, P=0.002), and hip (3.45 vs. 2.50, P=0.002), but were only significantly higher in the female amateur marathon runners than the female controls in the thoracic vertebrae (7.69 vs. 5.87, P=0.029) and hip (4.76 vs. 3.92, P=0.004). The mean T2 values of the lumbar vertebrae were significantly higher in the male amateur marathon runners than the male controls (116.76 vs. 97.63 ms, P=0.001), but no such significant difference was observed between the female amateur marathon runners and the corresponding controls (118.58 vs. 124.10 ms, P=0.386). Conclusions: Marathon training resulted in diffuse hyperintensity in the bone marrow based on WB-DWI in 50% of the male amateur marathon runners aged 30-50 years. Thus, when WB-DWI is used for bone marrow disease screening, marathon training history should be considered to avoid false-positive diagnoses.

Environmental monobutyl phthalate exposure promotes liver cancer via reprogrammed cholesterol metabolism and activation of the IRE1α-XBP1s pathway.

Humans are widely exposed to phthalates, a major chemical plasticizer that accumulates in the liver. However, little is known about the impact of chronic phthalate exposure on liver cancer development. In this study, we applied a long-term cell culture model by treating the liver cancer cell HepG2 and normal hepatocyte L02 to environmental dosage of monobutyl phthalate (MBP), the main metabolite of phthalates. Interestingly, we found that long-term MBP exposure significantly accelerated the growth of HepG2 cells in vitro and in vivo, but barely altered the function of L02 cells. MBP exposure triggered reprogramming of lipid metabolism in HepG2 cells, where cholesterol accumulation subsequently activated the IRE1α-XBP1s axis of the unfolded protein response. As a result, the XBP1s-regulated gene sets and pathways contributed to the increased aggressiveness of HepG2 cells. In addition, we also showed that MBP-induced cholesterol accumulation fostered an immunosuppressive microenvironment by promoting tumor-associated macrophage polarization toward the M2 type. Together, these results suggest that environmental phthalates exposure may facilitate liver cancer progression, and alerts phthalates exposure to patients who already harbor liver tumors.

Integrated analysis of single-cell and bulk transcriptomics develops a robust neuroendocrine cell-intrinsic signature to predict prostate cancer progression.

Neuroendocrine prostate cancer (NEPC) typically implies severe lethality and limited treatment options. The precise identification of NEPC cells holds paramount significance for both research and clinical applications, yet valid NEPC biomarker remains to be defined. Methods: Leveraging 11 published NE-related gene sets, 11 single-cell RNA-sequencing (scRNA-seq) cohorts, 15 bulk transcriptomic cohorts, and 13 experimental models of prostate cancer (PCa), we employed multiple advanced algorithms to construct and validate a robust NEPC risk prediction model. Results: Through the compilation of a comprehensive scRNA-seq reference atlas (comprising a total of 210,879 single cells, including 66 tumor samples) from 9 multicenter datasets of PCa, we observed inconsistent and inefficient performance among the 11 published NE gene sets. Therefore, we developed an integrative analysis pipeline, identifying 762 high-quality NE markers. Subsequently, we derived the NE cell-intrinsic gene signature, and developed an R package named NEPAL, to predict NEPC risk scores. By applying to multiple independent validation datasets, NEPAL consistently and accurately assigned NE feature and delineated PCa progression. Intriguingly, NEPAL demonstrated predictive capabilities for prognosis and therapy responsiveness, as well as the identification of potential epigenetic drivers of NEPC. Conclusion: The present study furnishes a valuable tool for the identification of NEPC and the monitoring of PCa progression through transcriptomic profiles obtained from both bulk and single-cell sources.

Targeting the IRE1α-XBP1s axis confers selective vulnerability in hepatocellular carcinoma with activated Wnt signaling.

Liver-specific Ern1 knockout impairs tumor progression in mouse models of hepatocellular carcinoma (HCC). However, the mechanistic role of IRE1α in human HCC remains unclear. In this study, we show that XBP1s, the major downstream effector of IRE1α, is required for HCC cell survival both in vitro and in vivo. Mechanistically, XBP1s transactivates LEF1, a key co-factor of ß-catenin, by binding to its promoter. Moreover, XBP1s physically interacts with LEF1, forming a transcriptional complex that enhances classical Wnt signaling. Consistently, the activities of XBP1s and LEF1 are strongly correlated in human HCC and with disease prognosis. Notably, selective inhibition of XBP1 splicing using an IRE1α inhibitor significantly repressed the viability of tumor explants as well as the growth of tumor xenografts derived from patients with distinct Wnt/LEF1 activities. Finally, machine learning algorithms developed a powerful prognostic signature based on the activities of XBP1s/LEF1. In summary, our study uncovers a key mechanistic role for the IRE1α-XBP1s pathway in human HCC. Targeting this axis could provide a promising therapeutic strategy for HCC with hyperactivated Wnt/LEF1 signaling.

4-hydroxybenzophenone exposure inhibits mouse hippocampal neural stem cell proliferation in vitro by upregulating Cxcl1.

Benzophenones are widely used in industry and commonly added in many personal care products. However, the neurotoxicity, in particular neurodevelopmental toxicity, of benzophenone family chemicals and metabolites has not been fully elucidated. Our recent mechanistic study in mice showed that early life exposure to a major benzophenone metabolite, 4-hydroxybenzophenone (4HBP), disrupted endoplasmic reticulum (ER) proteostasis and evoked inflammatory response in hippocampal neural stem cells (NSCs), leading to cognitive dysfunction. Despite so, detailed inflammatory cytokine(s) that possibly mediate this toxicity remains to be defined and validated. In this study, we confirmed that 4HBP treatment inhibited the viability and sphere growth of mouse NSCs in vitro. Importantly, re-interrogation of the transcriptomic data of NSCs treated with 4HBP identified the top upregulated genes, wherein the chemokine Cxcl1 ranked first. Immunofluorescent staining and qRT-PCR validated the robust induction of Cxcl1 on the protein and mRNA levels upon 4HBP treatment. Furthermore, siRNA-mediated knockdown of Cxcl1 transiently blocked its expression and led to enhanced NSCs viability in the presence of 4HBP. Together, these in vitro results indicated that the adverse effect of benzophenones on NSCs is mediated, at least in part, by induction of the chemokine Cxcl1.

Characterization of Molecular Subtypes in Head and Neck Squamous Cell Carcinoma With Distinct Prognosis and Treatment Responsiveness.

Head and neck squamous cell carcinoma (HNSCC) is one of the most aggressive malignancies with complex phenotypic, etiological, biological, and clinical heterogeneities. Previous studies have proposed different clinically relevant subtypes of HNSCC, but little is known about its corresponding prognosis or suitable treatment strategy. Here, we identified 101 core genes from three prognostic pathways, including mTORC1 signaling, unfold protein response, and UV response UP, in 124 pairs of tumor and matched normal tissues of HNSCC. Moreover, we identified three robust subtypes associated with distinct molecular characteristics and clinical outcomes using consensus clustering based on the gene expression profiles of 944 HNSCC patients from four independent datasets. We then integrated the genomic information of The Cancer Genome Atlas (TCGA) HNSCC cohort to comprehensively evaluate the molecular features of different subtypes and screen for potentially effective therapeutic agents. Cluster 1 had more arrested oncogenic signaling, the highest immune cell infiltration, the highest immunotherapy and chemotherapeutic responsiveness, and the best prognosis. By contrast, Cluster 3 showed more activated oncogenic signaling, the lowest immune cell infiltration, the lowest immunotherapy and chemotherapy responsiveness, and the worst prognosis. Our findings corroborate the molecular diversity of HNSCC tumors and provide a novel classification strategy that may guide for prognosis and treatment allocation.

The signal intensity characteristics of normal bone marrow in diffusion weighted imaging at various menstrual status women.

PURPOSE: Diffuse hyperintensities of the bone marrow in whole-body diffusion-weighted (DW) imaging (DWI) have been encountered more frequently in females aged 21-50 compared to elder females or men. Therefore, we aimed to visually evaluate DWI among pre-, peri- and postmenopausal women and to verify whether it correlates also quantitatively with hormonal status. METHOD: The prospective study was approved by our institutional review board and informed consent was obtained in a total of 70 healthy premenopausal, perimenopausal, and postmenopausal women aged 40-58 years from February 2017 to October 2017. The bone marrow DW imaging signal characteristics were visually evaluated in comparison to the erector spinae muscle. Imaging data were acquired using a 1.5 T MRI yielding signal intensity values from a DWI-pulse sequence (b-value of 800 s/mm2; apparent diffusion coefficient (ADC) maps from b-values of 0-800 s/mm2), and a T2 mapping sequence covering the L2-L4 lumbar vertebrae. Serous estradiol (E2), follicle stimulating hormone (FSH), and luteinizing hormone (LH) were measured through venous blood assay. The relationship of the mean DW signal intensity (SIDWI) with T2 values, female hormone level, and mean ADC were analyzed using Spearman's rho test. RESULTS: The proportion of diffuse DWI hyperintensities of the bone marrow was significantly higher in premenopausal (91% (21/23)) women compared to peri- (75% (18/24)) and postmenopausal (8% (2/23)) women. A positive correlation was observed for the mean SIDWI (median [interquartile range], 47.33 [30.14]) and mean T2 (mean ± SD, 121.01 ± 13.54) (r = 0.438, p < 0.001) as well as for the mean SIDWI and E2 (median [interquartile range], 52.45 [92.78]) (r = 0.407, p < 0.001). A negative correlation was observed for the mean SIDWI and serous FSH (median [interquartile range], 15.55 [42.08]) as well as for the mean SIDWI and serous LH (median [interquartile range], 6.96 [31.06]) (r = -0.557, p < 0.001; r = -0.535, p < 0.001; respectively), but no significant correlation was found for mean SIDWI and mean ADC (mean ± SD, 599.36 ± 82.70) (r = 0.099, p = 0.415). A negative correlation was also encountered for the mean T2 values and serous FSH (r = -0.339, p = 0.004) as well as for the mean T2 values and serous LH (r = -0.281, p = 0.018). CONCLUSIONS: The mean SIDWI correlates positively with mean T2 and serous E2 values, while there's no significant correlation with mean ADC, indicating that T2 shine-through effects might interfere with bone marrow signaling on DW images. Knowledge of the bone marrow signal characteristics changing in DW images in close relationship with menstrual status is essential to correctly interpret DWI in clinical practice.

Maternal Benzophenone Exposure Impairs Hippocampus Development and Cognitive Function in Mouse Offspring.

Benzophenones are widely supplemented in personal care products, but little is known about its neurodevelopmental toxicity. The previous epidemiological study discovered a negative correlation between maternal exposure to a benzophenone metabolite 4-hydroxybenzophenone (4HBP) and child's neurodevelopment, yet the causal relationship and detailed mechanism remain to be defined. Here, it is reported that prenatal, but not postnatal, exposure to environmentally relevant level of 4HBP impairs hippocampus development and causes cognitive dysfunction in offspring mice. Transcriptomic analyses reveal that 4HBP induces the endoplasmic reticulum stress-induced apoptotic signaling and inflammatory response in hippocampal neural stem cells. Mechanistically, 4HBP exposure activates protein kinase R-like ER kinase (PERK) signaling, which induces CHOP, inhibits IκB translation, and transactivates p65, thereby promoting inflammation and apoptosis on multiple levels. Importantly, genetic or pharmacological inhibition of PERK pathway significantly attenuates 4HBP-induced NFκB signaling and neurodevelopmental abnormalities in mice and in a human brain organoid model. The study uncovers the neurodevelopmental toxicity of BP and cautions its exposure during pregnancy.

Post-Traumatic Stress and School Adaptation in Adolescent Survivors Five Years after the 2010 Yushu Earthquake in China.

(1) Background: The devastating Ms 7.1 earthquake struck Yushu city, China, in 2010, leading to serious consequences and damage in the central Tibetan Plateau. This study aimed to assess school adaptation and post-traumatic stress disorder (PTSD) symptoms of adolescent survivors five years after the Yushu earthquake. (2) Methods: A large-scale, school-based mental health survey was conducted 5 years after the earthquake among Tibetan students in the city of Yushu using the Adolescent's School Adaptation Scale (ASAS) and the PTSD Checklist. (3) Results: A total of 1976 questionnaires were collected. A total of 30.7% of Tibetan adolescents had poor school adaptation and 19.5% were estimated as having probable PTSD. Logistic regression showed that females (OR = 0.73, 95% CI: 0.60-0.89), senior students (OR = 0.48, 95% CI: 0.39-0.59), and those who participated in post-disaster reconstruction (OR = 0.68, 95% CI: 0.54-0.85) were less likely to have poor school adaptation, while a positive association was observed among those buried under a collapsed building (OR = 1.47, 95% CI: 1.04-2.09) and those who experienced bereavement (OR = 1.77, 95% CI: 1.27-2.45). Students who had experienced bereavement were also more likely to have PTSD (OR = 1.60, 95% CI: 1.12-2.28). (4) Conclusions: The post-traumatic effects of the Yushu earthquake on Tibetan adolescents were severe and long-lasting. Sustainable long-term mental health services to help adolescents to restructure their mental health are necessary.

Differentiating benign from malignant bone tumors using fluid-fluid level features on magnetic resonance imaging.

OBJECTIVE: To analyze different fluid-fluid level features between benign and malignant bone tumors on magnetic resonance imaging (MRI). MATERIALS AND METHODS: This study was approved by the hospital ethics committee. We retrospectively analyzed 47 patients diagnosed with benign (n = 29) or malignant (n = 18) bone tumors demonstrated by biopsy/surgical resection and who showed the intratumoral fluid-fluid level on pre-surgical MRI. The maximum length of the largest fluid-fluid level and the ratio of the maximum length of the largest fluid-fluid level to the maximum length of a bone tumor in the sagittal plane were investigated for use in distinguishing benign from malignant tumors using the Mann-Whitney U-test and a receiver operating characteristic (ROC) analysis. Fluid-fluid level was categorized by quantity (multiple vs. single fluid-fluid level) and by T1-weighted image signal pattern (high/low, low/high, and undifferentiated), and the findings were compared between the benign and malignant groups using the χ(2) test. RESULTS: The ratio of the maximum length of the largest fluid-fluid level to the maximum length of bone tumors in the sagittal plane that allowed statistically significant differentiation between benign and malignant bone tumors had an area under the ROC curve of 0.758 (95% confidence interval, 0.616-0.899). A cutoff value of 41.5% (higher value suggests a benign tumor) had sensitivity of 73% and specificity of 83%. CONCLUSION: The ratio of the maximum length of the largest fluid-fluid level to the maximum length of a bone tumor in the sagittal plane may be useful to differentiate benign from malignant bone tumors.