The therapeutic effect of a novel GAPDH inhibitor in mouse model of breast cancer and efficacy monitoring by molecular imaging.

BACKGROUND: Breast cancer is a serious threat to women's health with high morbidity and mortality. The development of more effective therapies for the treatment of breast cancer is strongly warranted. Growing evidence suggests that targeting glucose metabolism may be a promising cancer treatment strategy. We previously identified a new glyceraldehyde-3-phosphate dehydrogenase (GAPDH) inhibitor, DC-5163, which shows great potential in inhibiting tumor growth. Here, we evaluated the anticancer potential of DC-5163 in breast cancer cells. METHODS: The effects of DC-5163 on breast cancer cells were investigated in vitro and in vivo. Seahorse, glucose uptake, lactate production, and cellular ATP content assays were performed to examine the impact of DC-5163 on cellular glycolysis. Cell viability, colony-forming ability, cell cycle, and apoptosis were assessed by CCK8 assay, colony formation assay, flow cytometry, and immunoblotting respectively. The anticancer activity of DC-5163 in vivo was evaluated in a mouse breast cancer xenograft model. RESULTS: DC-5163 suppressed aerobic glycolysis and reduced energy supply of breast cancer cells, thereby inhibiting breast cancer cell growth, inducing cell cycle arrest in the G0/G1 phase, and increasing apoptosis. The therapeutic efficacy was assessed using a breast cancer xenograft mouse model. DC-5163 treatment markedly suppressed tumor growth in vivo without inducing evident systemic toxicity. Micro-PET/CT scans revealed a notable reduction in tumor 18F-FDG and 18F-FLT uptake in the DC-5163 treatment group compared to the DMSO control group. CONCLUSIONS: Our results suggest that DC-5163 is a promising GAPDH inhibitor for suppressing breast cancer growth without obvious side effects. 18F-FDG and 18F-FLT PET/CT can noninvasively assess the levels of glycolysis and proliferation in tumors following treatment with DC-5163.

Characterization of an imported HIV-1 A1/A7/G recombinant in China.

BACKGROUND: International migration has accelerated the HIV-1 spread across national borders, gradually reducing the restrictions on the geographical distribution of HIV-1 subtypes. Subtypes A and G are globally recognized as the third and sixth most dominant HIV-1 genotypes, mainly prevalent in Africa, but rarely detected in China. Here we reported an imported HIV-1 recombinant which was composed of sub-subtypes A1 and A7 of subtype A and subtype G genes in a Chinese female. This virus was the first HIV-1 recombinant including A7 genes reported in the world. CASE PRESENTATION: The near full-length genome (NFLG) was obtained from the plasma sample of the female in an HIV-1 molecular epidemiological survey with 853 participants in China. Phylogenetic analyses showed that this NFLG sequence contains three A7 segments, four G segments and one A1 segment with seven breakpoints, and all these segments were closely related to HIV-1 references circulating in Africa. The evidence from epidemiological investigation indicated that this female participant had a more-than-two-years heterosexual contact history with a fixed partner from Nigeria, a country in west Africa, which further supported the results of phylogenetic analyses. By the Bayesian phylogenetic analyses, the times of most recent common ancestors (tMRCA) of the partial pol gene (nt2308-3284, A7 region) and full-length vpr-vpu plus partial env gene (nt5534-6858, G region) were estimated around 1989 and 1984, respectively. CONCLUSIONS: In this study, by using the NFLG sequencing, we identified an imported HIV-1 A1/A7/G recombinant which was estimated to originate around 1980s in Africa and introduced into China with international migration. This study highlighted the complexity of the global HIV-1 epidemic, the necessity of using genome sequences to determine HIV-1 genotypes and the importance of real-time monitoring of HIV-1 infection among international migrants and travelers.

Methodological and reporting quality of systematic reviews on health effects of air pollutants were higher than extreme temperatures: a comparative study.

BACKGROUND: An increasing number of systematic reviews (SRs) in the environmental field have been published in recent years as a result of the global concern about the health impacts of air pollution and temperature. However, no study has assessed and compared the methodological and reporting quality of SRs on the health effects of air pollutants and extreme temperatures. This study aims to assess and compare the methodological and reporting quality of SRs on the health effects of ambient air pollutants and extreme temperatures. METHODS: PubMed, Embase, the Cumulative Index to Nursing and Allied Health Literature (CINAHL), Cochrane Library, Web of Science, and Epistemonikos databases were searched. Two researchers screened the literature and extracted information independently. The methodological quality of the SRs was assessed through A Measurement Tool to Assess Systematic Reviews 2 (AMSTAR 2). The reporting quality was assessed through Preferred Reporting Items of Systematic reviews and Meta-Analyses (PRISMA). RESULTS: We identified 405 SRs (286 for air pollution, 108 for temperature, and 11 for the synergistic effects). The methodological and reporting quality of the included SRs were suboptimal, with major deficiencies in protocol registration. The methodological quality of SRs of air pollutants was better than that of temperature, especially in terms of satisfactory explanations for any heterogeneity (69.6% v. 45.4%). The reporting quality of SRs of air pollution was better than temperature, however, adherence to the reporting of the assessment results of risk of bias in all SRs (53.5% v. 34.3%) was inadequate. CONCLUSIONS: Methodological and reporting quality of SRs on the health effect of air pollutants were higher than those of temperatures. However, deficiencies in protocol registration and the assessment of risk of bias remain an issue for both pollutants and temperatures. In addition, developing a risk-of-bias assessment tool applicable to the temperature field may improve the quality of SRs.

Suitable coverage and slope guided by soil and water conservation can prevent non-point source pollution diffusion: A case study of grassland.

Non-point source pollution caused by surface runoff has been a popular hydrological and environmental safety issue and has attracted extensive attention from global scholars. To identify the optimal vegetation coverage of Festuca arundinacea grassland for controlling soil erosion and purifying surface runoff, bare land was chosen as the control in this experiment. Simulated rainfall experiments were carried out with three levels of coverage (low coverage, moderate coverage and full coverage) under four slope conditions (flat slope, gentle slope, medium slope, and steep slope) and at four rainfall intensities (moderate rainfall, heavy rainfall, rainstorm and heavy rainstorm). The comprehensive evaluation results suggested that the capacity of Festuca arundinacea grassland for reducing the surface runoff, sediment yield, suspended solids (SS), total nitrogen (TN), total phosphorus (TP) and chemical oxygen demand (COD) decreased with increasing rainfall intensity and slope but increased with increasing vegetation coverage. Structural equation model (SEM) results suggested that there were positive relationships between the vegetation coverage and purification capacity index and negative correlations between the rainfall intensity and slope and the purification capacity index. The response surface analysis results suggested that the optimal vegetation coverage should be higher than 84% and that the slope should be smaller than 10° for controlling soil erosion and avoiding pollution via diffusion with surface runoff in Festuca arundinacea grassland. This study proposes recommendations for the vegetation configuration pattern in the development and management of runoff purification systems.

Mechanisms of allicin exposure for the sludge fermentation enhancement: Focusing on the fermentation processes and microbial metabolic traits.

As a frequently used product with antimicrobial activity, consumed allicin might be discharged and concentrated in waste-activated sludge (WAS). However, the influence of allicin (as an exogenous pollutant) on WAS fermentation has not been clearly revealed. This study aimed to disclose the impacts of allicin on volatile fatty acid (VFA) generation during WAS fermentation. The results showed that the appropriate presence of allicin (10 mg/g TSS) significantly enhanced the VFA yield (1894 versus 575 mg COD/L in the control) with increased acetate proportion (24.3%). Further exploration found that allicin promoted WAS solubilization, hydrolysis and acidification simultaneously. Metagenomic analysis revealed that the key genes involved in extracellular hydrolysis metabolism (i.e., CAZymes), membrane transport (i.e., gtsA and ytfT), substrate metabolism (i.e., yhdR and pfkC) and fatty acid synthesis (i.e., accA and accD) were all highly expressed. Allicin also induced the bacteria to produce more signalling molecules and regulate cellular functions, thereby enhancing the microbial adaptive and regulatory capacity to the unfavourable environment. Moreover, the variations in fermentative microbes and their contributions to the upregulation of functional genes (i.e., ytfR, gltL, INV, iolD and pflD) for VFA generation were disclosed. Overall, the simultaneous stimulation of functional microbial abundances and metabolic activities contributed to VFA production in allicin-conditioned reactors.

Use of family disability service by families with young children with disabilities.

AIM: To investigate which families with young children with disabilities used disability services and when they used services to inform policy on service delivery. METHOD: We used linked administrative data from different ministries in Alberta to describe families' use of disability services when their children were between the ages of 3 and 8 years old. Disability was investigated on the basis of the presence of a severe special education code for children, and level of special education code. The outcome was the use of family disability services. RESULTS: Of 31 346 children, 24 761 (79.0%) had no special education code, 3982 (12.7%) had a mild special education code, and 2603 (8.3%) had a severe special education code. Level of special education code was associated with child characteristics and service use. Children with severe special education codes generally were more likely to report service use and have poor outcomes than those with less severe codes. Of note, 26% of children with severe special education codes used family disability services. In addition, among children with severe special education codes, many years of severe coding (compared with fewer years) had the strongest association with family disability service use (prevalence ratio 5.50; 95% confidence interval 4.10-7.37). Associations with family disability service use were seen with mental health, health care, and educational achievement. Interactions between child characteristics and service use were observed. INTERPRETATION: This study provides evidence that families were more likely to use disability services when they were involved with other services, and that use interacts with various factors. The findings highlight the importance of considering service eligibility, referral, and integration.

Ampelopsin Inhibits Breast Cancer Cell Growth through Mitochondrial Apoptosis Pathway.

Ampelopsin, a flavonoid with a wide variety of biological activities, has been proposed to be a potent antitumor agent. However, the mechanism by which Ampelopsin shows anti-breast cancer activity remains unclear. Therefore, this study will explore the mechanism of Ampelopsin's anti-breast cancer activity by culturing MDA-MB-231 and MCF-7 breast cancer cells. Cell Counting Kit-8 (CCK-8) method and plate cloning method were used to detect the proliferation inhibition of breast cancer cells. Fluorescence microscopy was used to detect mitochondrial membrane potential (MMP). 2',7'-Dichlorodihydrofluorescein diacetate (DCFH-DA) method was used to determine the content of intracellular reactive oxygen species (ROS). Hoechst 33258 staining was used to detect the apoptotic morphological changes. Transmission electron microscope was used to observe the mitochondrial structure. Western blot was used to detect the protein expression of Bax and Bcl-2. The results showed that Ampelopsin could significantly inhibit the proliferation of breast cancer cells, and promote cells apoptosis. In addition, the occurrence of apoptosis in breast cancer cells was associated with mitochondrial dysfunction, including the loss of mitochondrial membrane potential, the production of large amounts of reactive oxygen species, and the up-regulation of Bax/Bcl-2 expression. In conclusion, Ampelopsin-induced mitochondria damage leads to loss of mitochondria membrane potential, overproduction of ROS and activation of Bax, increasing mitochondria membrane permeability and ultimately inducing breast cell apoptosis. These findings provided a new perspective on the role of Ampelopsin in breast cancer prevention and treatment.

Accurate and nondestructive detection of apple brix and acidity based on visible and near-infrared spectroscopy.

Rapid, nondestructive and accurate detection of internal qualities of the apple is an important research interest. In this study, the brix, acidity and brix/acidity ratio of the apple were rapidly detected by visible and near-infrared spectroscopy (VIS-NIRS). By scanning spectra and measuring the reference values of brix and acidity of apple samples, the relationship models between the spectra and brix, acidity, brix/acidity ratio were, respectively, established. Sample division, characteristic wavelength optimization, and modeling methods were compared systematically, and the optimal prediction model of each quality index was determined. The experimental results show that the competitive adaptive reweighted sampling method can effectively select characteristic wavelengths, which not only improves the prediction speed, but also greatly enhances the prediction accuracy. The established partial least squares models based on these selected characteristic wavelengths all have high accuracy and robustness for the three quality indices. The determination coefficients of the models are 0.9899, 0.9615, 0.9535, and the relative percent deviation are 9.9269, 5.0987, 4.6374, respectively. All this work proves that VIS-NIRS can be used for rapid and nondestructive detection of the internal qualities of an apple.

Comparative transcriptome analyses provide novel insights into etiolated shoot development of walnut (Juglans regia L.).

MAIN CONCLUSION: In general, genes promoting IAA, CTK GA and ethylene biosynthesis were upregulated, while genes participating in ABA, chlorophyll and starch biosynthesis pathways performed opposite tendency during etiolation. Etiolation as a method for rejuvenation plays an important role in the vegetative propagation of woody plants. However, the molecular mechanism of etiolated shoot development remains unclear. In this study, we investigated changes at different etiolation stages of Juglans regia. The histology and transcriptome of J. regia were analysed using etiolated stems, which were treated in darkness for 30, 60, 90 days. The results showed that the ratios of pith (Pi) diameter/stem diameter (D), cortex (Co) width/D, and phloem (Ph) width/D increased, while the ratio of xylem (Xy) width/D decreased after etiolation, and the difference in these ratios between etiolated stems and the control was more significant at 60 days than 90 days. Differentially expressed genes (DEGs) were significantly enriched in pathways such as plant hormone biosynthesis and signal transduction, chlorophyll biosynthesis and degradation, and starch and sucrose metabolism. The difference in the contents of indole-3-acetic acid (IAA), abscisic acid (ABA), sugar and chlorophyll between etiolated stems and the control increased with increasing treatment duration; in contrast, the concentrations of gibberellin (GA), zeatin (ZT), and starch, as well as the difference between the etiolated stems and control were lowest at 60 days among the three stages. On the whole, the positive effect of etiolation on the rejuvenation of walnut stems changed as the treatment period increased. The present investigation lays a foundation for future studies on the effect of etiolation on rejuvenation and for promoting the efficiency of vegetative propagation.

Confidence intervals of the difference between areas under two ROC curves in matched-pair experiments.

In a matched-pair study, when outcomes of two diagnostic tests are ordinal/continuous, the difference between two correlated areas under ROC curves (AUCs) is usually used to compare the overall discriminatory ability of two diagnostic tests. This article considers confidence interval (CI) construction problems of difference between two correlated AUCs in a matched-pair experiment, and proposes 13 hybrid CIs based on variance estimates recovery with the maximum likelihood estimation, Delong's statistic, Wilson score statistic (WS) and WS with continuity correction, the modified Wald statistic (MW) and MW with continuity correction and Agresti-Coull statistic, and three Bootstrap-resampling-based CIs. For comparison, we present traditional parametric and nonparametric CIs. Simulation studies are conducted to assess the performance of the proposed CIs in terms of empirical coverage probabilities, empirical interval widths, and ratios of the mesial noncoverage probabilities to the noncoverage probabilities. Two examples from clinical studies are illustrated by the proposed methodologies. Empirical results evidence that the hybrid Agresti-Coull CI with the empirical estimation (EAC) behaved most satisfactorily because its coverage probability was quite close to the prespecified confidence level with short interval width. Hence, we recommend the usage of the EAC CI in applications.

Membrane-Free Hybrid Capacitive Deionization System Based on Redox Reaction for High-Efficiency NaCl Removal.

Capacitive deionization (CDI) is a promising technology for desalination due to its advantages of low driven energy and environmental friendliness. However, the ion removal capacity (IRC) of CDI is insufficient for practical application because such a capacity is limited by the available surface area of the carbon electrode for ion absorption. Thus, the development of a novel desalination technology with high IRC and low cost is vital. Here, a membrane-free hybrid capacitive deionization system (HCDI) with hollow carbon@MnO2 (HC@MnO2) to capture sodium via redox reaction and hollow carbon sphere with net positive surface charges (PHC) for chloride adsorption is introduced. The as-obtained HC@MnO2 with unique structure and high conductivity can improve the utilization of MnO2 pseudocapacitive electrodes. Meanwhile, the PHC can selectively adsorb Cl- and prevent the adsorption of Na+ due to electrostatic repulsion. As expected, the membrane-free HCDI system demonstrates excellent desalination performance. The system's IRC and maximum removal rate are 30.7 mg g-1 and 7.8 mg g-1 min-1, respectively. Moreover, the proposed system has a low cost because of the absence of expensive ion exchange membranes (IEM), which is suitable for practical application. The excellent performance of this HCDI makes it a promising desalination technology for future use.

The sex expression and sex determining mechanism in Pistacia species.

Generally, Pistacia species are dioecious, but monoecious strains in several populations have been found, providing excellent models for studying sex differentiation and sex determination mechanisms. Although the mechanisms of sex determination and sex evolution have been extensively studied, related research on heterozygous woody plants is limited. Here, we discuss the expressions of various sex types, which showed broad diversity and complex instability. We have also reviewed the sex determination systems in the plant kingdom and the morphological, cytological, physiological, and molecular aspects of the sex-linked markers in Pistacia trees. Moreover, hypotheses to explain the origin of monoecy are discussed, which is more likely to be the interaction between sex-related genes and environment factors in female plants. Besides, further prospects for the utilization of monoecious resources and the research directions of sex determination mechanism are proposed. This study provides important information on sex expression and provides more insights into sex differentiation and determination.

Iron Plaque: A Barrier Layer to the Uptake and Translocation of Copper Oxide Nanoparticles by Rice Plants.

The waterlogging environment generally results in the deposition of iron plaque on plant roots, which may impact the fate of metal-based nanoparticles. Here, we investigated the influence of iron plaque on the uptake, translocation, and transformation of copper oxide nanoparticles (CuO NPs) in rice plants. The results show that the presence of iron plaque dramatically reduced the Cu contents in roots and shoots by 89% and 78% of those without iron plaque under 100 mg/L CuO NP treatment. Meanwhile, the Cu accumulation in plants was negatively related to the amount of iron plaque. X-ray absorption near edge structure (XANES) analysis demonstrated lower percentage of CuO but higher proportion of Cu(I) in shoots exposed to CuO NPs with the formation of iron plaque. Furthermore, micro X-ray fluorescence (µ-XRF) combined with µ-XANES revealed that the iron plaque in the root epidermis and exodermis consisted of goethite and ferrihydrite, which hindered the uptake of CuO NPs by roots. However, a few CuO NPs were still absorbed by roots via root hairs or lateral roots, and further translocated to shoots. But eventually, more than 90% of total Cu(II) was reduced to Cu(I)-cysteine and Cu2O in leaf veins of rice plants with iron plaque.

Retraction: Yang, C., et al. miR-126 Functions as a Tumor Suppressor in Osteosarcoma by Targeting Sox2. Int. J. Mol. Sci. 2014, 15, 423-437, doi:10.3390/ijms15010423.

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Steroid receptor co-activator-3 promotes osteosarcoma progression through up-regulation of FoxM1.

Increasing evidence suggests that the three homologous members of steroid receptor co-activator (SRC) family (SRC-1, SRC-2, and SRC-3) play key roles in enhancing cell proliferation in various human cancers, such as breast, prostate, and hepatocellular carcinoma. However, the function of SRC-3 in osteosarcoma remains largely unexplored. In the current study, we found that SRC-3, but not SRC-1 and SRC-2, was dramatically up-regulated in human osteosarcoma tissues, compared with adjacent normal tissues. To explore the functions of SRC-3 in osteosarcoma, in vitro studies were performed in MG63 and U2OS cells. SRC-3 overexpression promoted osteosarcoma cell proliferation, whereas knockdown of SRC-3 inhibits its proliferation. In support of these findings, we further demonstrated that SRC-3 up-regulated FoxM1 expression through co-activation of C/EBPγ. Together our results show that SRC-3 drives osteosarcoma progression and imply it as a therapeutic target to abrogate osteosarcoma.

Electroencephalogram-based objective assessment of cognitive function level associated with age-related hearing loss.

Age-Related Hearing Loss (ARHL) is a common problem in aging. Numerous longitudinal cohort studies have revealed that ARHL is closely related to cognitive function, leading to a significant risk of cognitive decline and dementia. This risk gradually increases with the severity of hearing loss. We designed dual auditory Oddball and cognitive task paradigms for the ARHL subjects, then obtained the Montreal Cognitive Assessment (MoCA) scale evaluation results for all the subjects. Multi-dimensional EEG characteristics helped explore potential biomarkers to evaluate the cognitive level of the ARHL group, having a significantly lower P300 peak amplitude coupled with a prolonged latency. Moreover, visual memory, auditory memory, and logical calculation were investigated during the cognitive task paradigm. In the ARHL groups, the alpha-to-beta rhythm energy ratio in the visual and auditory memory retention period and the wavelet packet entropy value within the logical calculation period were significantly reduced. Correlation analysis between the above specificity indicators and the subjective scale results of the ARHL group revealed that the auditory P300 component characteristics could assess attention resources and information processing speed. The alpha and beta rhythm energy ratio and wavelet packet entropy can become potential indicators to determine working memory and logical cognitive computation-related cognitive ability.

Pseudotargeted lipidomics analysis of scoparone on glycerophospholipid metabolism in non-alcoholic steatohepatitis mice by LC-MRM-MS.

As the inflammatory subtype of nonalcoholic fatty liver disease (NAFLD), the progression of nonalcoholic steatohepatitis (NASH) is associated with disorders of glycerophospholipid metabolism. Scoparone is the major bioactive component in Artemisia capillaris which has been widely used to treat NASH in traditional Chinese medicine. However, the underlying mechanisms of scoparone against NASH are not yet fully understood, which hinders the development of effective therapeutic agents for NASH. Given the crucial role of glycerophospholipid metabolism in NASH progression, this study aimed to characterize the differential expression of glycerophospholipids that is responsible for scoparone's pharmacological effects and assess its efficacy against NASH. Liquid chromatography-multiple reaction monitoring-mass spectrometry (LC-MRM-MS) was performed to get the concentrations of glycerophospholipids, clarify mechanisms of disease, and highlight insights into drug discovery. Additionally, pathologic findings also presented consistent changes in high-fat diet-induced NASH model, and after scoparone treatment, both the levels of glycerophospholipids and histopathology were similar to normal levels, indicating a beneficial effect during the observation time. Altogether, these results refined the insights on the mechanisms of scoparone against NASH and suggested a route to relieve NASH with glycerophospholipid metabolism. In addition, the current work demonstrated that a pseudotargeted lipidomic platform provided a novel insight into the potential mechanism of scoparone action.

Changes in seminal plasma microecological dynamics and the mechanistic impact of core metabolite hexadecanamide in asthenozoospermia patients.

Asthenozoospermia (AZS) is a prevalent contributor to male infertility, characterized by a substantial decline in sperm motility. In recent years, large-scale studies have explored the interplay between the male reproductive system's microecology and its implications for reproductive health. Nevertheless, the direct association between seminal microecology and male infertility pathogenesis remains inconclusive. This study used 16S rDNA sequencing and multi-omics analysis to conduct a comprehensive investigation of the seminal microbial community and metabolites in AZS patients. Patients were categorized into four distinct groups: Normal, mild AZS (AZS-I), moderate AZS (AZS-II), and severe AZS (AZS-III). Microbiome differential abundance analysis revealed significant differences in microbial composition and metabolite profiles within the seminal plasma of these groups. Subsequently, patients were classified into a control group (Normal and AZS-I) and an AZS group (AZS-II and AZS-III). Correlation and cross-reference analyses identified distinct microbial genera and metabolites. Notably, the AZS group exhibited a reduced abundance of bacterial genera such as Pseudomonas, Serratia, and Methylobacterium-Methylorubrum in seminal plasma, positively correlating with core differential metabolite (hexadecanamide). Conversely, the AZS group displayed an increased abundance of bacterial genera such as Uruburuella, Vibrio, and Pseudoalteromonas, with a negative correlation with core differential metabolite (hexadecanamide). In vitro and in vivo experiments validated that hexadecanamide significantly enhanced sperm motility. Using predictive metabolite-targeting gene analysis and single-cell transcriptome sequencing, we profiled the gene expression of candidate target genes PAOX and CA2. Protein immunoblotting techniques validated the upregulation protein levels of PAOX and CA2 in sperm samples after hexadecanamide treatment, enhancing sperm motility. In conclusion, this study uncovered a significant correlation between six microbial genera in seminal plasma and the content of the metabolite hexadecanamide, which is related to AZS. Hexadecanamide notably enhances sperm motility, suggesting its potential integration into clinical strategies for managing AZS, providing a foundational framework for diagnostic and therapeutic advancements.

Deoxyuridine-rich cytoplasmic DNA antagonizes STING-dependent innate immune responses and sensitizes resistant tumors to anti-PD-L1 therapy.

DNA damage and cytoplasmic DNA induce type-1 interferon (IFN-1) and potentiate responses to immune checkpoint inhibitors. Our prior work found that inhibitors of the DNA damage response kinase ATR (ATRi) induce IFN-1 and deoxyuridine (dU) incorporation by DNA polymerases, akin to antimetabolites. Whether and how dU incorporation is required for ATRi-induced IFN-1 signaling is not known. Here, we show that ATRi-dependent IFN-1 responses require uracil DNA glycosylase (UNG)-initiated base excision repair and STING. Quantitative analyses of nine distinct nucleosides reveals that ATRi induce dU incorporation more rapidly in UNG wild-type than knockout cells, and that induction of IFN-1 is associated with futile cycles of repair. While ATRi induce similar numbers of micronuclei in UNG wild-type and knockout cells, dU containing micronuclei and cytoplasmic DNA are increased in knockout cells. Surprisingly, DNA fragments containing dU block STING-dependent induction of IFN-1, MHC-1, and PD-L1. Furthermore, UNG knockout sensitizes cells to IFN-γ in vitro , and potentiates responses to anti-PD-L1 in resistant tumors in vivo . These data demonstrate an unexpected and specific role for dU-rich DNA in suppressing STING-dependent IFN-1 responses, and show that UNG-deficient tumors have a heightened response to immune checkpoint inhibitors. STATEMENT OF SIGNIFICANCE: Antimetabolites disrupt nucleotide pools and increase dU incorporation by DNA polymerases. We show that unrepaired dU potentiates responses to checkpoint inhibitors in mouse models of cancer. Patients with low tumor UNG may respond to antimetabolites combined with checkpoint inhibitors, and patients with high tumor UNG may respond to UNG inhibitors combined with checkpoint inhibitors.

miR-126 functions as a tumor suppressor in osteosarcoma by targeting Sox2.

Osteosarcoma (OS) is the most common malignant bone tumor in children and young adults, the early symptoms and signs of which are non-specific. The discovery of microRNAs (miRNAs) provides a new avenue for the early diagnosis and treatment of OS. miR-126 has been reported to be highly expressed in vascularized tissues, and is recently widely studied in cancers. Herein, we explored the expression and significance of miR-126 in OS. Using TaqMan RT-PCR analysis, we analyzed the expression of miR-126 in 32 paired OS tumor tissues and 4 OS cell lines and found that miR-126 was consistently under-expressed in OS tissues and cell lines compared with normal bone tissues and normal osteoblast cells (NHOst), respectively. As miR-126 is significantly decreased in OS tissues and cell lines, we sought to compensate for its loss through exogenous transfection into MG-63 cells with a miR-126 mimic. Ectopic expression of miR-126 inhibited cell proliferation, migration and invasion, and induced apoptosis of MG-63 cells. Moreover, bioinformatic prediction suggested that the sex-determining region Y-box 2 (Sox2) is a target gene of miR-126. Using mRNA and protein expression analysis, luciferase assays and rescue assays, we demonstrate that restored expression of Sox2 dampened miR-126-mediated suppression of tumor progression, which suggests the important role of miR-126/Sox2 interaction in tumor progression. Taken together, our data indicate that miR-126 functions as a tumor suppressor in OS, which exerts its activity by suppressing the expression of Sox2.