ABSTRACT
T cells rewire their metabolic activities to meet the demand of immune responses, but how to coordinate the immune response by metabolic regulators in activated T cells is unknown. Here, we identified autocrine VEGF-B as a metabolic regulator to control lipid synthesis and maintain the integrity of the mitochondrial inner membrane for the survival of activated T cells. Disruption of autocrine VEGF-B signaling in T cells reduced cardiolipin mass, resulting in mitochondrial damage, with increased apoptosis and reduced memory development. The addition of cardiolipin or modulating VEGF-B signaling improved T cell mitochondrial fitness and survival. Autocrine VEGF-B signaling through GA-binding protein α (GABPα) induced sentrin/SUMO-specific protease 2 (SENP2) expression, which further de-SUMOylated PPARγ and enhanced phospholipid synthesis, leading to a cardiolipin increase in activated T cells. These data suggest that autocrine VEGF-B mediates a signal to coordinate lipid synthesis and mitochondrial fitness with T cell activation for survival and immune response. Moreover, autocrine VEGF-B signaling in T cells provides a therapeutic target against infection and tumors as well as an avenue for the treatment of autoimmune diseases.
Subject(s)
Autocrine Communication , Cardiolipins , Mitochondria , Signal Transduction , T-Lymphocytes , Vascular Endothelial Growth Factor B , Mitochondria/metabolism , Mitochondria/immunology , Animals , Mice , Autocrine Communication/immunology , T-Lymphocytes/immunology , T-Lymphocytes/metabolism , Signal Transduction/immunology , Cardiolipins/immunology , Cardiolipins/metabolism , Vascular Endothelial Growth Factor B/genetics , Vascular Endothelial Growth Factor B/metabolism , Vascular Endothelial Growth Factor B/immunology , Lymphocyte Activation , PPAR gamma/metabolism , PPAR gamma/immunology , PPAR gamma/genetics , HumansABSTRACT
Glioblastoma multiforme (GBM) is a highly malignant brain tumor, and glioblastoma stem cells (GSCs) are the primary cause of GBM heterogeneity, invasiveness, and resistance to therapy. Sirtuin 3 (SIRT3) is mainly localized in the mitochondrial matrix and plays an important role in maintaining GSC stemness through cooperative interaction with the chaperone protein tumor necrosis factor receptor-associated protein 1 (TRAP1) to modulate mitochondrial respiration and oxidative stress. The present study aimed to further elucidate the specific mechanisms by which SIRT3 influences GSC stemness, including whether SIRT3 serves as an autophagy substrate and the mechanism of SIRT3 degradation. We first found that SIRT3 is enriched in CD133+ GSCs. Further experiments revealed that in addition to promoting mitochondrial respiration and reducing oxidative stress, SIRT3 maintains GSC stemness by epigenetically regulating CD133 expression via succinate. More importantly, we found that SIRT3 is degraded through the autophagy-lysosome pathway during GSC differentiation into GBM bulk tumor cells. GSCs are highly dependent on glutamine for survival, and in these cells, we found that glutamine deprivation triggers autophagic SIRT3 degradation to restrict CD133 expression, thereby disrupting the stemness of GSCs. Together our results reveal a novel mechanism by which SIRT3 regulates GSC stemness. We propose that glutamine restriction to trigger autophagic SIRT3 degradation offers a strategy to eliminate GSCs, which combined with other treatment methods may overcome GBM resistance to therapy as well as relapse.
ABSTRACT
RATIONALE AND OBJECTIVE: To investigate the influence of the deep learning image reconstruction (DLIR) on the image quality and quantitative analysis of pulmonary nodules under ultra-low dose lung CT conditions. MATERIALS AND METHODS: This was a prospective study with patient consent and included 56 patients with suspected pulmonary nodules. Patients were examined by both standard-dose CT (SDCT) and ultra-low-dose CT (ULDCT). SDCT images were reconstructed with adaptive statistical iterative reconstruction-V 40% (ASIR-V40%) (group A), while ULDCT images were reconstructed using ASIR-V40% (group B) and high-strength DLIR (DLIR-H) (group C). The three image sets were analyzed using a commercial computer aided diagnosis (CAD) software. Parameters such as nodule length, width, density, volume, risk, and classification were measured. The CAD quantitative data of different nodule types (solid, calcified, and subsolid nodules) and nodule image quality scores evaluated by two physicians on a 5-point scale were compared. RESULT: The radiation dose in ULDCT was 0.25 ± 0.08mSv, 7.2% that of the 3.48 ± 1.08mSv in SDCT (P < 0.001). 104 pulmonary nodules were detected (51/53 solid, 26/24 calcified and 27/27 subsolid in Groups A and (B&C), respectively). Group B had lower density for solid, calcified nodules, and lower volume and risk for subsolid nodules than Group A, while Group C had lower density for calcified nodules (P < 0.05), There were no significant differences in other parameters among the three groups (P > 0.05). Group A and C had similar image quality for nodules and were higher than Group B (P < 0.05). CONCLUSION: DLIR-H significantly improves image quality than ASIR-V40% and maintains similar nodule detection and characterization with CAD in ULDCT compared to SDCT.
Subject(s)
Deep Learning , Lung Neoplasms , Radiation Dosage , Radiographic Image Interpretation, Computer-Assisted , Solitary Pulmonary Nodule , Tomography, X-Ray Computed , Humans , Female , Male , Prospective Studies , Middle Aged , Tomography, X-Ray Computed/methods , Lung Neoplasms/diagnostic imaging , Radiographic Image Interpretation, Computer-Assisted/methods , Aged , Solitary Pulmonary Nodule/diagnostic imaging , Multiple Pulmonary Nodules/diagnostic imaging , AdultABSTRACT
Glioblastoma (GBM) cells require large amounts of iron for tumor growth and progression, which makes these cells vulnerable to destruction via ferroptosis induction. Mitochondria are critical for iron metabolism and ferroptosis. Sirtuin-3 (SIRT3) is a deacetylase found in mitochondria that regulates mitochondrial quality and function. This study aimed to characterize SIRT3 expression and activity in GBM and investigate the potential therapeutic effects of targeting SIRT3 while also inducing ferroptosis in these cells. We first found that SIRT3 expression was higher in GBM tissues than in normal brain tissues and that SIRT3 protein expression was upregulated during RAS-selective lethal 3 (RSL3)-induced GBM cell ferroptosis. We then observed that inhibition of SIRT3 expression and activity in GBM cells sensitized GBM cells to RSL3-induced ferroptosis both in vitro and in vivo. Mechanistically, SIRT3 inhibition led to ferrous iron and ROS accumulation in the mitochondria, which triggered mitophagy. RNA-Sequencing analysis revealed that upon SIRT3 knockdown in GBM cells, the mitophagy pathway was upregulated and SLC7A11, a critical antagonist of ferroptosis via cellular import of cystine for glutathione (GSH) synthesis, was downregulated. Forced expression of SLC7A11 in GBM cells with SIRT3 knockdown restored cellular cystine uptake and consequently the cellular GSH level, thereby partially rescuing cell viability upon RSL3 treatment. Furthermore, in GBM cells, SIRT3 regulated SLC7A11 transcription through ATF4. Overall, our study results elucidated novel mechanisms underlying the ability of SIRT3 to protect GBM from ferroptosis and provided insight into a potential combinatorial approach of targeting SIRT3 and inducing ferroptosis for GBM treatment.
Subject(s)
Ferroptosis , Glioblastoma , Sirtuin 3 , Humans , Amino Acid Transport System y+/genetics , Cystine , Ferroptosis/genetics , Glioblastoma/genetics , Glutathione , Indans , Iron , Mitophagy , Sirtuin 3/geneticsABSTRACT
Atractylodis rhizoma (AR) is an herb and food source with great economic, medicinal, and ecological value. Atractylodes chinensis (DC.) Koidz. (AC) and Atractylodes lancea (Thunb.) DC. (AL) are its two botanical sources. The commercial fraud of AR adulterated with Atractylodes japonica Koidz. ex Kitam (AJ) frequently occurs in pursuit of higher profit. To quickly determine the content of adulteration in AC and AL powder, two spectroscopic techniques, near-infrared spectroscopy (NIRS) and hyperspectral imaging (HSI), were introduced. The partial least squares regression (PLSR) algorithm was selected for predictive modeling of AR adulteration levels. Preprocessing and feature variable extraction were used to optimize the prediction model. Then data and image feature fusions were developed to obtain the best predictive model. The results showed that if only single-spectral techniques were considered, NIRS was more suitable for both tasks than HSI techniques. In addition, by comparing the models built after the data fusion of NIRS and HSI with those built by the single spectrum, we found that the mid-level fusion strategy obtained the best models in both tasks. On this basis, combined with the color-texture features, the prediction ability of the model was further optimized. Among them, for the adulteration level prediction task of AC, the best strategy was combining MLF data (at CARS level) and color-texture features (C-TF), at which time the R2T, RMSET, R2P, and RMSEP were 99.85%, 1.25%, 98.61%, and 5.06%, respectively. For AL, the best approach was combining MLF data (at SPA level) and C-TF, with the highest R2T (99.92%) and R2P (99.00%), as well as the lowest RMSET (1.16%) and RMSEP (2.16%). Therefore, combining data and image features from NIRS and HSI is a potential strategy to predict the adulteration content quickly, non-destructively, and accurately.
ABSTRACT
BACKGROUND: To evaluate the value of a deep learning-based computer-aided diagnostic system (DL-CAD) in improving the diagnostic performance of acute rib fractures in patients with chest trauma. MATERIALS AND METHODS: CT images of 214 patients with acute blunt chest trauma were retrospectively analyzed by two interns and two attending radiologists independently firstly and then with the assistance of a DL-CAD one month later, in a blinded and randomized manner. The consensusdiagnosis of fib fracture by another two senior thoracic radiologists was regarded as reference standard. The rib fracture diagnostic sensitivity, specificity, positive predictive value, diagnostic confidence and mean reading time with and without DL-CAD were calculated and compared. RESULTS: There were 680 rib fracture lesions confirmed as reference standard among all patients. The diagnostic sensitivity and positive predictive value of interns weresignificantly improved from (68.82%, 84.50%) to (91.76%, 93.17%) with the assistance of DL-CAD, respectively. Diagnostic sensitivity and positive predictive value of attendings aided by DL-CAD (94.56%, 95.67%) or not aided (86.47%, 93.83%), respectively. In addition, when radiologists were assisted by DL-CAD, the mean reading time was significantly reduced, and diagnostic confidence was significantly enhanced. CONCLUSIONS: DL-CAD improves the diagnostic performance of acute rib fracture in chest trauma patients, which increases the diagnostic confidence, sensitivity, and positive predictive value for radiologists. DL-CAD can advance the diagnostic consistency of radiologists with different experiences.
Subject(s)
Deep Learning , Rib Fractures , Thoracic Injuries , Wounds, Nonpenetrating , Humans , Rib Fractures/diagnostic imaging , Tomography, X-Ray Computed/methods , Retrospective Studies , Sensitivity and Specificity , Wounds, Nonpenetrating/diagnostic imaging , ComputersABSTRACT
RATIONALE AND OBJECTIVES: Infrapatellar fat pad (IPFP) proton density-weighted images (PdWI) hyperintense regions on MRI are an important imaging feature of knee osteoarthritis (KOA) and are thought to represent inflammation which may induce knee pain. The aim of the study was to compare the intravoxel incoherent motion diffusion-weighted imaging (IVIM-DWI) findings of PdWI hyperintense regions of IPFP between symptomatic and asymptomatic KOA and to determine whether IVIM-DWI parameters can be used as an objective biomarker for symptomatic KOA. MATERIALS AND METHODS: In total, 84 patients with symptomatic KOA, 43 asymptomatic KOA persons, and 30 healthy controls with MRI were retrospectively reviewed. Demographic, IPFP-synovitis, Western Ontario and McMaster Osteoarthritis Index (WOMAC) pain sub-score, IPFP volume and depth and quantitative parameters of IVIM-DWI were collected. The chi-square test, Binary logistic regression and receiver operating characteristic curve (ROC) analysis were used for diagnostic performance comparison. RESULTS: The IPFP volume and depth were statistically significant differences between the non-KOA and sKOA groups (p<0.05). The IPFP PdWI hyperintense regions demonstrated significantly higher values of D and D* in the symptomatic KOA compared to those in the asymptomatic KOA (1.51±0.47 vs. 1.73±0.40 for D and 19.24±6.44 vs. 27.09±9.75 for D*) (both p<0.05). Multivariate logistic regression analyses showed that Higher D and D* values of IPFP hyperintense region were significantly associated with higher risks of knee pain (OR: 1.97; 95% CI: 1.21-3.19; p=0.006 for D and OR: 1.24; 95% CI: 1.09-1.41; p=0.001 for D*). Sensitivity and specificity of D value for symptomatic KOA were 80.28% and 83.33%, with an AUC of 0.78 (0.68-0.86). D* value had the sensitivity with 92.96% and a specificity of 58.33%, with an AUC of 0.82 (0.73-0.89) for symptomatic KOA. CONCLUSION: IVIM-DWI can be used as an additional functional imaging technique to study IPFP with signal abnormalities on PdWI, and the D and D* values may have potential value to predict the symptom in mild-to-moderate KOA patients.
Subject(s)
Osteoarthritis, Knee , Humans , Osteoarthritis, Knee/diagnostic imaging , Retrospective Studies , Diffusion Magnetic Resonance Imaging , Magnetic Resonance Imaging/methods , Pain , Adipose Tissue/diagnostic imaging , MotionABSTRACT
Antioxidant transcription factor NRF2 plays a pivotal role in cell ferroptosis. KLK lung adenocarcinoma (LUAD) is a specific molecular subtype of Kras-mutant LUAD. The activation of mutant Kras in combination with the inactivation of Lkb1 and Keap1 abnormally increases NRF2 expression, while high NRF2 confers KLK LUAD cell resistance to ferroptosis. This study assessed the inhibition of NRF2-GSH axis to sensitize a small molecule RSL3 to induce KLK LUAD cell ferroptosis and then explored the underlying molecular mechanisms. The data showed that the NRF2-GSH inhibition sensitized RSL3 induction of KLK LUAD cell ferroptosis in vitro, while RSL3 treatment reduced level of NRF2 protein in KLK LUAD during ferroptosis. Moreover, RSL3 treatment inhibited activity of the NRF2-GSH signaling during in KLK LUAD cell ferroptosis in vitro and in vivo. Mechanistically, the RSL3 reduction of NRF2 expression was through the promotion of NRF2 ubiquitination in KLK LUAD cells. In addition, RSL3 was able to directly bind to USP11, a recently identified de-ubiquitinase of NRF2, and inactivate USP11 protein to induce NRF2 protein ubiquitination and degradation in KLK LUAD cells. These data revealed a novel mechanism of RSL3 induction in KLK LUAD cell ferroptosis by suppression of the USP11-NRF2-GSH signaling. Future study will confirm RSL3 as a novel therapeutic approach in control of KLK lung adenocarcinoma.
ABSTRACT
Sirtuin 3 (SIRT3) deacetylase is a key regulator for chemoresistance in acute myeloid leukemia (AML) cells due to its capability of modulating mitochondrial metabolism and reactive oxygen species (ROS). SIRT3 is de-SUMOylated by SUMO-specific peptidase 1 (SENP1), which enhances its deacetylase activity. Therefore, dysregulation of SIRT3 SUMOylation may lead to fortified chemoresistance in AML. Indeed, SIRT3 de-SUMOylation was induced by chemotherapeutic agents, which in turn, exacerbated resistance against chemotherapies in AML by activating SIRT3 via preventing its proteasome degradation. Furthermore, RNA-seq revealed that expression of a collection of genes was altered by SIRT3 de-SUMOylation including inhibition of transcription factor Hes Family BHLH Transcription Factor 1 (HES1), a downstream substrate of Notch1 signaling pathway, leading to increased fatty acids oxidation (FAO). Moreover, the SENP1 inhibitor momordin-Ic or HES1 overexpression synergized with cytarabine to eradicate AML cells in vitro and in xenograft mouse models. In summary, the current study revealed a novel role of SIRT3 SUMOylation in the regulation of chemoresistance in AML via HES1-dependent FAO and provided a rationale for SIRT3 SUMOylation and FAO targeted interventions to improve chemotherapies in AML.
Subject(s)
Leukemia, Myeloid, Acute , Sirtuin 3 , Animals , Drug Resistance, Neoplasm/genetics , Fatty Acids/metabolism , Humans , Leukemia, Myeloid, Acute/drug therapy , Leukemia, Myeloid, Acute/genetics , Leukemia, Myeloid, Acute/metabolism , Mice , Sirtuin 3/genetics , Sirtuin 3/metabolism , Sumoylation , Transcription Factor HES-1/genetics , Transcription Factor HES-1/metabolismABSTRACT
Our previous studies show that the mitotic phosphorylation of SUMO-specific protease 3 (SENP3) can inhibit its de-SUMOylation activity in G2/M phase of the cell cycle. Inhibition of SENP3 plays a critical role in the correct separation of sister chromatids in mitosis. The mutation of mitotic SENP3 phosphorylation causes chromosome instability and promotes tumorigenesis. In this study, we find that the mutation of mitotic SENP3 phosphorylation in tumor cells can suppress tumor growth in immune-competent mouse model. We further detect an increase of CD8+ T cell infiltration in the tumors, which is essential for the anti-tumor effect in immune-competent mouse model. Moreover, we find that mitotic SENP3 activation increases micronuclei formation, which can activate cGAS signaling-dependent innate immune response. We confirmed that cGAS signaling mediates the mitotic SENP3 activation-induced anti-tumor immunity. We further show that p53 responding to DNA damage activates mitotic SENP3 by inhibiting phosphorylation, and further increases cellular senescence as well as the related innate immune response in tumor cells. Furthermore, TCGA database demonstrates that the SENP3 expression positively correlates with the induction of innate immune response as well as the survival of the p53 mutant pancreatic cancer patients. Together, these data reveal that mitotic SENP3 activation in tumor cells can promote host anti-tumor immune response by coupling with cGAS signaling.
Subject(s)
Neoplasms , Peptide Hydrolases , Animals , Cysteine Endopeptidases/genetics , Cysteine Endopeptidases/metabolism , Mice , Neoplasms/genetics , Nucleotidyltransferases/genetics , Nucleotidyltransferases/metabolism , Peptide Hydrolases/metabolism , Sumoylation , Tumor Suppressor Protein p53/genetics , Tumor Suppressor Protein p53/metabolismABSTRACT
The metabolic program is altered during macrophage activation and influences macrophage polarization. Glutaminolysis promotes accumulation of α-ketoglutarate (αKG), leading to Jumonji domain-containing protein D3 (Jmjd3)-dependent demethylation at H3K27me3 during M2 polarization of macrophages. However, it remains unclear how αKG accumulation is regulated during M2 polarization of macrophages. This study shows that SENP1-Sirt3 signaling controls glutaminolysis, leading to αKG accumulation during IL-4-stimulated M2 polarization. Activation of the SENP1-Sirt3 axis augments M2 macrophage polarization through the accumulation of αKG via glutaminolysis. We also identify glutamate dehydrogenase 1 (GLUD1) as an acetylated protein in mitochondria. The SENP1-Sirt3 axis deacetylates GLUD1 and increases its activity in glutaminolysis to promote αKG production, leading to M2 polarization of macrophages. Therefore, SENP1-Sirt3 signaling plays a critical role in αKG accumulation via glutaminolysis to promote M2 polarization.
Subject(s)
Macrophage Activation , Sirtuin 3 , Ketoglutaric Acids/metabolism , Macrophages/metabolism , Signal Transduction , Sirtuin 3/metabolismABSTRACT
Although oncogenicity of the stem cell regulator SOX9 has been implicated in many solid tumors, its role in lymphomagenesis remains largely unknown. In this study, SOX9 was overexpressed preferentially in a subset of diffuse large B-cell lymphomas (DLBCLs) that harbor IGH-BCL2 translocations. SOX9 positivity in DLBCL correlated with an advanced stage of disease. Silencing of SOX9 decreased cell proliferation, induced G1/S arrest, and increased apoptosis of DLBCL cells, both in vitro and in vivo. Whole-transcriptome analysis and chromatin immunoprecipitation-sequencing assays identified DHCR24, a terminal enzyme in cholesterol biosynthesis, as a direct target of SOX9, which promotes cholesterol synthesis by increasing DHCR24 expression. Enforced expression of DHCR24 was capable of rescuing the phenotypes associated with SOX9 knockdown in DLBCL cells. In models of DLBCL cell line xenografts, SOX9 knockdown resulted in a lower DHCR24 level, reduced cholesterol content, and decreased tumor load. Pharmacological inhibition of cholesterol synthesis also inhibited DLBCL xenograft tumorigenesis, the reduction of which is more pronounced in DLBCL cell lines with higher SOX9 expression, suggesting that it may be addicted to cholesterol. In summary, our study demonstrated that SOX9 can drive lymphomagenesis through DHCR24 and the cholesterol biosynthesis pathway. This SOX9-DHCR24-cholesterol biosynthesis axis may serve as a novel treatment target for DLBCLs.
Subject(s)
Cholesterol/genetics , Immunoglobulin Heavy Chains/genetics , Lymphoma, Large B-Cell, Diffuse/genetics , Nerve Tissue Proteins/genetics , Oxidoreductases Acting on CH-CH Group Donors/genetics , Proto-Oncogene Proteins c-bcl-2/genetics , SOX9 Transcription Factor/genetics , Biosynthetic Pathways , Gene Expression Regulation, Neoplastic , Humans , Mutation , Oncogene Proteins, Fusion/genetics , Oncogenes , TranscriptomeABSTRACT
Metabolic programming and mitochondrial dynamics along with T cell differentiation affect T cell fate and memory development; however, how to control metabolic reprogramming and mitochondrial dynamics in T cell memory development is unclear. Here, we provide evidence that the SUMO protease SENP1 promotes T cell memory development via Sirt3 deSUMOylation. SENP1-Sirt3 signalling augments the deacetylase activity of Sirt3, promoting both OXPHOS and mitochondrial fusion. Mechanistically, SENP1 activates Sirt3 deacetylase activity in T cell mitochondria, leading to reduction of the acetylation of mitochondrial metalloprotease YME1L1. Consequently, deacetylation of YME1L1 suppresses its activity on OPA1 cleavage to facilitate mitochondrial fusion, which results in T cell survival and promotes T cell memory development. We also show that the glycolytic intermediate fructose-1,6-bisphosphate (FBP) as a negative regulator suppresses AMPK-mediated activation of the SENP1-Sirt3 axis and reduces memory development. Moreover, glucose limitation reduces FBP production and activates AMPK during T cell memory development. These data show that glucose limitation activates AMPK and the subsequent SENP1-Sirt3 signalling for T cell memory development.
Subject(s)
AMP-Activated Protein Kinases/metabolism , CD8-Positive T-Lymphocytes/immunology , Cysteine Endopeptidases/metabolism , Immunologic Memory , Mitochondria/metabolism , Sirtuin 3/metabolism , T-Lymphocytes/metabolism , ATPases Associated with Diverse Cellular Activities/metabolism , Acetylation , Allografts , Animals , Cell Line, Tumor , Cell Survival/genetics , Colonic Neoplasms/immunology , Fructosediphosphates/metabolism , GTP Phosphohydrolases/metabolism , Glucose/deficiency , Immunologic Memory/genetics , Metabolomics , Metalloendopeptidases/metabolism , Mice , Mice, Inbred C57BL , Microscopy, Electron, Transmission , Mitochondria/genetics , Mitochondria/ultrastructure , Mitochondrial Dynamics/genetics , Mitochondrial Proteins/metabolism , Oxidative Phosphorylation , Sirtuin 3/antagonists & inhibitors , Sirtuin 3/genetics , Sumoylation , T-Lymphocytes/immunologyABSTRACT
In response to DNA damage, p53-mediated signaling is regulated by protein phosphorylation and ubiquitination to precisely control G2 checkpoint. Here we demonstrated that protein SUMOylation also engaged in regulation of p53-mediated G2 checkpoint. We found that G2 DNA damage suppressed SENP3 phosphorylation at G2/M phases in p53-dependent manner. We further found that the suppression of SENP3 phosphorylation was crucial for efficient DNA damage/p53-induced G2 checkpoint and G2 arrest. Mechanistically, we identified Cdh1, a subunit of APC/C complex, was a SUMOylated protein at G2/M phase. SENP3 could de-SUMOylate Cdh1. DNA damage/p53-induced suppression of SENP3 phosphorylation activated SENP3 de-SUMOylation of Cdh. De-SUMOylation promoted Cdh1 de-phosphorylation by phosphatase Cdc14B, and then activated APC/CCdh1 E3 ligase activity to ubiquitate and degrade Polo-like kinase 1 (Plk1) in process of G2 checkpoint. These data reveal that p53-mediated inhibition of SENP3 phosphorylation regulates the activation of Cdc14b-APC/CCdh1-Plk1 axis to control DNA damage-induced G2 checkpoint.
ABSTRACT
Nuclear factor erythroid-2 related factor 2 (NRF2) is a pivotal transcription factor that maintains cellular redox homeostasis and facilitates the development of malignant tumor phenotypes. At the molecular level, NRF2 promotes de novo serine synthesis and SUMOylation affects its function. Our results indicated that the SUMO1 acceptor site of NRF2 is the conserved lysine residue 110 (K110), and that NRF2 SUMOylation deficiency inhibited tumorigenesis in hepatocellular carcinoma (HCC). Mechanistically, NRF2 SUMOylation promoted de novo serine synthesis in HCC by enhancing the clearance of intracellular reactive oxygen species (ROS) and up-regulating phosphoglycerate dehydrogenase (PHGDH). More importantly, serine starvation increased the level of NRF2 SUMOylation, leading to sustained HCC growth. Collectively, our results indicate the presence of a novel NRF2 SUMOylation-mediated signaling process that maintains HCC tumorigenesis in normal conditions and in response to metabolic stress.
Subject(s)
Carcinoma, Hepatocellular/pathology , Liver Neoplasms/pathology , NF-E2-Related Factor 2/metabolism , Serine/metabolism , Animals , Carcinoma, Hepatocellular/metabolism , Cell Line, Tumor , Female , Hep G2 Cells , Humans , Liver Neoplasms/metabolism , Mice , NF-E2-Related Factor 2/chemistry , Neoplasm Transplantation , Oxidative Stress , Phosphoglycerate Dehydrogenase/metabolism , Reactive Oxygen Species/metabolism , Stress, Physiological , SumoylationABSTRACT
PURPOSE: Our purpose was to combine intravoxel incoherent motion diffusion-weighted MR imaging (IVIM-DWI) and magnetic resonance spectroscopy (MRS) to differentiate osteoporotic fractures from osteolytic metastatic vertebral compression fractures (VCFs). METHODS: A total of 70 patients with VCFs were included and divided into two groups, according to their causes of fractures based on pathological findings or clinical follow-up. All patients underwent conventional sagittal T1WI, T2WI, STIR, IVIM-DWI, and single-voxel MRS. The diffusion coefficient (D), pseudo diffusion (D*), and perfusion fraction (f) parameters from IVIM-DWI and the lipid water ratio (LWR) and fat fraction (FF) parameters from MRS were obtained and compared among groups. Furthermore, the diagnostic performance of MRS, IVIM-DWI, and IVIM-DWI combined with MRS for differentiation between osteoporotic and osteolytic metastatic VCFs was assessed by using receiver operating characteristic (ROC) curve analysis. RESULTS: Compared with the osteoporotic group, the metastatic group had significantly lower values for f, D, and FF, but higher D* (all P < 0.05). The area under the receiver operating characteristic (ROC) curve of MRS, IVIM-DWI, and IVIM-DWI combined with MRS were 0.73, 0.88, and 0.94, respectively. Among these, the IVIM-DWI combined with MRS showed the highest sensitivity, specificity, and accuracy, which are 90.63% (29/32), 97.37 % (37/38), and 94.29% (66/70), respectively. CONCLUSIONS: IVIM-DWI combined with MRS can be more accurate and efficient for differentiation between osteoporotic and osteolytic metastatic VCFs than single MRS or IVIM-DWI.
Subject(s)
Diffusion Magnetic Resonance Imaging/methods , Fractures, Compression/diagnostic imaging , Magnetic Resonance Spectroscopy/methods , Osteoporotic Fractures/diagnostic imaging , Spinal Fractures/diagnostic imaging , Spinal Neoplasms/diagnostic imaging , Aged , Diagnosis, Differential , Diffusion Magnetic Resonance Imaging/standards , Female , Fractures, Compression/metabolism , Humans , Lumbar Vertebrae/diagnostic imaging , Lumbar Vertebrae/injuries , Lumbar Vertebrae/metabolism , Magnetic Resonance Spectroscopy/standards , Male , Middle Aged , Osteoporotic Fractures/metabolism , Spinal Fractures/metabolism , Spinal Neoplasms/metabolism , Thoracic Vertebrae/diagnostic imaging , Thoracic Vertebrae/injuries , Thoracic Vertebrae/metabolismABSTRACT
Sirt3, as a major mitochondrial nicotinamide adenine dinucleotide (NAD)-dependent deacetylase, is required for mitochondrial metabolic adaption to various stresses. However, how to regulate Sirt3 activity responding to metabolic stress remains largely unknown. Here, we report Sirt3 as a SUMOylated protein in mitochondria. SUMOylation suppresses Sirt3 catalytic activity. SUMOylation-deficient Sirt3 shows elevated deacetylation on mitochondrial proteins and increased fatty acid oxidation. During fasting, SUMO-specific protease SENP1 is accumulated in mitochondria and quickly de-SUMOylates and activates Sirt3. SENP1 deficiency results in hyper-SUMOylation of Sirt3 and hyper-acetylation of mitochondrial proteins, which reduces mitochondrial metabolic adaption responding to fasting. Furthermore, we find that fasting induces SENP1 translocation into mitochondria to activate Sirt3. The studies on mice show that Sirt3 SUMOylation mutation reduces fat mass and antagonizes high-fat diet (HFD)-induced obesity via increasing oxidative phosphorylation and energy expenditure. Our results reveal that SENP1-Sirt3 signaling modulates Sirt3 activation and mitochondrial metabolism during metabolic stress.
Subject(s)
Cysteine Endopeptidases/metabolism , Mitochondria/metabolism , Mutation , Obesity/metabolism , Signal Transduction , Sirtuin 3/metabolism , Sumoylation , Acetylation , Animals , Cysteine Endopeptidases/genetics , Dietary Fats/adverse effects , Dietary Fats/pharmacology , HEK293 Cells , Humans , Male , Mice , Mice, Mutant Strains , Mitochondria/genetics , Mitochondria/pathology , Obesity/chemically induced , Obesity/genetics , Obesity/pathology , Sirtuin 3/geneticsABSTRACT
Myeloid-derived suppressor cells (MDSC) can suppress immunity and promote tumorigenesis, and their abundance is associated with poor prognosis. In this study, we show that SUMO1/sentrin-specific peptidase 1 (SENP1) regulates the development and function of MDSC. SENP1 deficiency in myeloid cells promoted MDSC expansion in bone marrow, spleen, and other organs. Senp1-/- MDSC showed stronger immunosuppressive activity than Senp1+/+ MDSC; we observed no defects in the differentiation of myeloid precursor cell in Senp1-/- mice. Mechanistically, SENP1-mediated regulation of MDSC was dependent on STAT3 signaling. We identified CD45 as a specific STAT3 phosphatase in MDSC. CD45 was SUMOylated in MDSC and SENP1 could deconjugate SUMOylated CD45. In Senp1-/- MDSC, CD45 was highly SUMOylated, which reduced its phosphatase activity toward STAT3, leading to STAT3-mediated MDSC development and function. These results reveal a suppressive function of SENP1 in modulating MDSC expansion and function via CD45-STAT3 signaling axis. SIGNIFICANCE: These findings show that increased SUMOylation of CD45 via loss of SENP1 suppresses CD45-mediated dephosphorylation of STAT3, which promotes MDSC development and function, leading to tumorigenesis.
Subject(s)
Cysteine Endopeptidases/metabolism , Myeloid-Derived Suppressor Cells/metabolism , Animals , Carcinogenesis , Carcinoma, Lewis Lung/metabolism , Carcinoma, Lewis Lung/pathology , Cysteine Endopeptidases/biosynthesis , Cysteine Endopeptidases/deficiency , Female , HEK293 Cells , Humans , Leukocyte Common Antigens/metabolism , Melanoma, Experimental/metabolism , Melanoma, Experimental/pathology , Mice , Mice, Knockout , Myeloid-Derived Suppressor Cells/cytology , Myeloid-Derived Suppressor Cells/pathology , STAT3 Transcription Factor/metabolism , SumoylationABSTRACT
BACKGROUND: Natural products (NPs) are evolutionarily chosen "privileged structures" that have a profound impact upon the anticancer drug discovery and development progress. However, the search for new drugs based on structure modification of NPs has often been hindered due to the tedious and complicated synthetic pathways. Fortunately, Mizoroki-Heck reaction and copper-catalyzed alkyne-azide cycloaddition (CuAAC) could provide perfect strategies for selective modification on NPs even in the presence of liable functionalities. OBJECTIVE: Here, we used oridonin, an ent-kaurane diterpenoid that showed a wide range of biological activities, as a parent molecule for the generation of analogues with anticancer activity. METHODS: Derivatives of oridonin were generated based on the structure-activity relationship study of oridonin and synthesized via Mizoroki-Heck reaction and CuAAC. The cytotoxicity of new oridonin derivatives were evaluated on both cancer cells and normal cells. Furthermore, the apoptotic effect and cell cycle arrest effect of the selected potent analogue were evaluated by flow cytometry and western blotting analysis. RESULTS: Two series of novel C-14 and C-17 modified derivatives of oridonin were obtained via Heck reaction and copper-catalyzed alkyne-azide cycloaddition (CuAAC), respectively. In vitro antiproliferative activities showed that the introduction of C-14 (2-triazole)acetoxyl- moiety could retain or enhance cytotoxicity, whereas the introduction of C-17 phenyl ring might exert negative effect. Further studies demonstrated that derivative 23 exhibited broad-spectrum antiproliferative activity, effectively overcame drug-resistance and showed weak cytotoxicity on non-cancer cells. Preliminary mechanistic studies indicated that 23 might cause G2/M phase arrest and induce apoptosis in PC-3 cells. CONCLUSION: Mizoroki-Heck reaction and CuAAC are perfect strategies for structure modification of complex natural products. The introduction of C-14 (2-triazole)acetoxyl- moiety could retain or enhance the cytotoxicity of oridonin, the introduction of C-17 phenyl group might exert negative effect on its cytotoxicity.
Subject(s)
Antineoplastic Agents/pharmacology , Diterpenes, Kaurane/pharmacology , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/chemistry , Apoptosis/drug effects , Cell Cycle/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Click Chemistry , Diterpenes, Kaurane/chemical synthesis , Diterpenes, Kaurane/chemistry , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , Humans , Molecular Conformation , Structure-Activity RelationshipABSTRACT
OBJECTIVE: To perform quantitative measurement based on the standardized uptake value (SUV) of Tc-99m methylene diphosphonate (MDP) in the normal pelvis using a single-photon emission tomography (SPECT)/computed tomography (CT) scanner. MATERIAL AND METHODS: This retrospective study was performed on 31 patients with cancer undergoing bone SPECT/CT scans with 99mTc-MDP. SUVmax and SUVmean of the normal pelvis were calculated based on the body weight. SUVmax and SUVmean of the bilateral anterior superior iliac spine, posterior superior iliac spine, facies auricularis ossis ilii, ischial tuberosity, and sacrum were also calculated. Furthermore, the correlation of SUVmax and SUVmean of all parts of pelvis with weight, height, and CT was assessed. RESULTS: The data for 31 patients (20 women and 11 men; mean age 58.97±9.12 years; age range 37-87 years) were collected. SUVmax and SUVmean changed from 1.65±0.40 to 3.8±1.0 and from 1.15±0.25 to 2.07±0.58, respectively. The coefficient of variation of SUVmax and SUVmean ranged from 0.22 to 0.31. SUVmax and SUVmean had no statistically significant difference between men and women. SUVmax and SUVmean also showed no significant correlation with weight and height. However, part of SUVmax and SUVmean showed a significant correlation with CT. In addition, SUVmax and SUVmean of the bilateral ischial tuberosity showed a significant correlation with CT values. CONCLUSIONS: Determination of the SUV value of the normal pelvis with 99m Tc-MDP SPECT/CT is feasible and highly reproducible. SUVs of the normal pelvis showed a relatively large variability. As a quantitative imaging biomarker, SUVs might require standardization with adequate reference data for the participant to minimize variability.