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BACKGROUND: In developmental and pathological tissues, nascent vessel networks generated by angiogenesis require further pruning/regression to delete nonfunctional endothelial cells (ECs) by apoptosis and migration. Mechanisms underlying EC apoptosis during vessel pruning remain elusive. TMEM215 (transmembrane protein 215) is an endoplasmic reticulum-located, 2-pass transmembrane protein. We have previously demonstrated that TMEM215 knockdown in ECs leads to cell death, but its physiological function and mechanism are unclear. METHODS: We characterized the role and mechanism of TMEM215 in EC apoptosis using human umbilical vein endothelial cells by identifying its interacting proteins with immunoprecipitation-mass spectrometry. The physiological function of TMEM215 in ECs was assessed by establishing a conditional knockout mouse strain. The role of TMEM215 in pathological angiogenesis was evaluated by tumor and choroidal neovascularization models. We also tried to evaluate its translational value by delivering a Tmem215 small interfering RNA (siRNA) using nanoparticles in vivo. RESULTS: TMEM215 knockdown in ECs induced apoptotic cell death. We identified the chaperone BiP as a binding partner of TMEM215, and TMEM215 forms a complex with and facilitates the interaction of BiP (binding immunoglobin protein) with the BH (BCL-2 [B-cell lymphoma 2] homology) 3-only proapoptotic protein BIK (BCL-2 interacting killer). TMEM215 knockdown triggered apoptosis in a BIK-dependent way and was abrogated by BCL-2. Notably, TMEM215 knockdown increased the number and diminished the distance of mitochondria-associated endoplasmic reticulum membranes and increased mitochondrial calcium influx. Inhibiting mitochondrial calcium influx by blocking the IP3R (inositol 1,4,5-trisphosphate receptor) or MCU (mitochondrial calcium uniporter) abrogated TMEM215 knockdown-induced apoptosis. TMEM215 expression in ECs was induced by physiological laminar shear stress via EZH2 downregulation. In EC-specific Tmem215 knockout mice, induced Tmem215 depletion impaired the regression of retinal vasculature characterized by reduced vessel density, increased empty basement membrane sleeves, and increased EC apoptosis. Moreover, EC-specific Tmem215 ablation inhibited tumor growth with disrupted vasculature. However, Tmem215 ablation in adult mice attenuated lung metastasis, consistent with reduced Vcam1 expression. Administration of nanoparticles carrying Tmem215 siRNA also inhibited tumor growth and choroidal neovascularization injury. CONCLUSIONS: TMEM215, which is induced by blood flow-derived shear stress via downregulating EZH2, protects ECs from BIK-triggered mitochondrial apoptosis mediated by calcium influx through mitochondria-associated ER membranes during vessel pruning, thus providing a novel target for antiangiogenic therapy.
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Enterovirus 71 (EV71) is a neurotropic enterovirus associated with hand, foot, and mouth disease (HFMD) fatalities. In this study, we investigated the impact of EV71 on plasmacytoid dendritic cells (pDCs) and CD4+ T cells. The results showed that pDCs were promptly activated, secreting interferon (IFN)-α and inducing CD4+ T cell proliferation and differentiation during early EV71 infection. This initiated adaptive immune responses and promoted proinflammatory cytokine production by CD4+ T cells. Over time, viral nucleic acids and proteins were synthesized in pDCs and CD4+ T cells. Concurrently, the cholinergic anti-inflammatory pathway (CAP) was activated, exhibiting an anti-inflammatory role. With constant viral stimulation, pDCs and CD4+ T cells showed reduced differentiation and cytokine secretion. Defects in pDCs were identified as a key factor in CD4+ T cell tolerance. CAP had a more significant regulatory effect on CD4+ T cells than on pDCs and was capable of inhibiting inflammation in these cells.
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Enterovirus Humano A , Infecções por Enterovirus , Humanos , Neuroimunomodulação , Regulação para Cima , Interferon-alfa/metabolismo , Diferenciação Celular , Infecções por Enterovirus/metabolismo , Linfócitos T CD4-Positivos , Células DendríticasRESUMO
BACKGROUND: Conventional chemotherapy has poor efficacy in triple-negative breast cancer (TNBC) which is highly heterogeneous and aggressive. Imaging-guided therapy is usually combined with diverse treatment modalities, could realize the integration of diagnosis and treatments. Therefore, the primary challenge for combinational therapy is designing proper delivery systems to accomplish multiple synergistic effects. RESULTS: Herein, a facile nanoplatform was manufactured to fulfill the all-in-one approaches for TNBC combinational therapy. Fe3+-based metal-phenolic networks (MPNs) with bovine serum albumin (BSA) modification served as drug delivery carriers to encapsulate bleomycin (BLM), forming BFE@BSA NPs. The self-assembly mechanism, pH-responsive drug release behavior, and other physicochemical properties of this system were characterized. The potential of BFE@BSA NPs as photothermal transduction agents and magnetic resonance imaging (MRI) contrast agents was explored. The synergistic anti-tumor effects consisting of BLM-induced chemotherapy, Fenton reactions-mediated chemodynamic therapy, and photothermal therapy-induced apoptosis were studied both in vitro and in vivo. Once internalized into tumor cells, released BLM could cause DNA damage, while Fenton reactions were initiated to produce highly toxic â¢OH. Upon laser irradiation, BFE@BSA NPs could convert light into heat to achieve synergistic effects. After intravenous administration, BFE@BSA NPs exhibited great therapeutic effects in 4T1 tumor xenograft model. Moreover, as T1-weighted MRI contrast agents, BFE@BSA NPs could provide diagnosis and treatment monitoring for individualized precise therapy. CONCLUSIONS: A nano-system that integrated imaging and combinational therapy (chemotherapy, chemodynamic therapy and photothermal therapy) were developed to kill the tumor and monitor therapeutic efficacy. This strategy provided an all-in-one theranostic nanoplatform for MRI-guided combinational therapy against TNBC.
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Nanopartículas , Neoplasias , Neoplasias de Mama Triplo Negativas , Linhagem Celular Tumoral , Meios de Contraste , Portadores de Fármacos/uso terapêutico , Humanos , Imageamento por Ressonância Magnética , Nanopartículas/química , Neoplasias/tratamento farmacológico , Fototerapia/métodos , Terapia Fototérmica , Soroalbumina Bovina/uso terapêutico , Neoplasias de Mama Triplo Negativas/diagnóstico por imagem , Neoplasias de Mama Triplo Negativas/tratamento farmacológicoRESUMO
Current perception and monitoring systems, such as human recognition, are affected by several environmental factors, such as limited light intensity, weather changes, occlusion of targets, and public privacy. Human recognition using radar signals is a promising direction to overcome these defects; however, the low signal-to-noise ratio of radar signals still makes this task challenging. Therefore, it is necessary to use suitable tools that can efficiently deal with radar signals to identify targets. Reservoir computing (RC) is an efficient machine learning scheme that is easy to train and demonstrates excellent performance in processing complex time-series signals. The RC hardware implementation structure based on nonlinear nodes and delay feedback loops endows it with the potential for real-time fast signal processing. In this paper, we numerically study the performance of the optoelectronic RC composed of optical and electrical components in the task of human recognition with noisy micro-Doppler radar signals. A single-loop optoelectronic RC is employed to verify the application of RC in this field, and a parallel dual-loop optoelectronic RC scheme with a dual-polarization Mach-Zehnder modulator (DPol-MZM) is also used for performance comparison. The result is verified to be comparable with other machine learning tools, which demonstrates the ability of the optoelectronic RC in capturing gait information and dealing with noisy radar signals; it also indicates that optoelectronic RC is a powerful tool in the field of human target recognition based on micro-Doppler radar signals.
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Algoritmos , Radar , Humanos , Processamento de Sinais Assistido por Computador , Monitorização Fisiológica , Aprendizado de MáquinaRESUMO
KEY MESSAGE: Map-based cloning revealed that two novel soybean distorted trichome mutants were due to loss function of GmNAP1 gene, which affected the trichome morphology and pavement cell ploidy by regulating actin filament assembly. Trichomes increase both biotic and abiotic stress resistance in soybean. In this study, Gmdtm1-1 and Gmdtm1-2 mutants with shorter trichomes and bigger epidermal pavement cells were isolated from an ethyl methylsulfonate mutagenized population. Both of them had reduced plant height and smaller seeds. Map-based cloning and bulked segregant analysis identified that a G-A transition at the 3' boundary of the sixth intron of Glyma.20G019300 in the Gmdtm1-1 mutant and another G-A transition mutation at the 5' boundary of the fourteenth intron of Glyma.20G019300 in Gmdtm1-2; these mutations disrupted spliceosome recognition sites creating truncated proteins. Glyma.20G019300 encodes a Glycine max NCK-associated protein 1 homolog (GmNAP1) in soybean. Further analysis revealed that the GmNAP1 involved in actin filament assembling and genetic information processing pathways during trichome and pavement cell development. This study shows that GmNAP1 plays an important role in soybean growth and development and agronomic traits.
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Glycine max/genética , Tricomas/genética , Regulação da Expressão Gênica de Plantas , Íntrons/genética , Mutação/genética , Proteínas de Plantas/genéticaRESUMO
We experimentally and numerically demonstrated the generation of a (3, 1) vector signal by a single Mach-Zehnder modulator (MZM) without pre-coding. The MZM is driven by the (3, 1) modulated signal after photoelectric conversion by the "square law" of a photodetector. Although the phase changes, the corresponding constellation distribution is consistent with that of the regular signal. Our proposed scheme effectively avoids the pre-coding process with a simple architecture. The bit-error-ratio (BER) results indicate that the (3, 1) signal has a better BER performance than the pre-coded quadrature phase shift keying vector signal, and both are below ${3.8}\times {{10}^{ - 3}}$3.8×10-3 after 25 km optical fiber transmission.
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Leucemia Mieloide Aguda , Criança , Humanos , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Decitabina/uso terapêutico , Histona-Lisina N-Metiltransferase/genética , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/genética , Complexo de Proteínas Formadoras de Poros Nucleares/genética , Proteínas de Fusão Oncogênica/genética , Proteínas de Fusão Oncogênica/metabolismoRESUMO
An approach was presented for electrical sensing of deoxyribonucleic acid (DNA)-hybridization in solution using a two-port network based on single-walled suspended carbon nanotube (SWCNT) membrane. A single stranded probe DNA (5'-NH2-(CH2)6-CGC CGA TTG GAC AAA ACT TAA A-3') was immobilized on the SWCNT membrane. A solution with the complementary single stranded DNA (D'FITC: 5'-FITC-T TTA AGT TTT GTC CAA TCG GCG-3') in various concentrations was then dropped on the membrane. The two-port network composed of the suspended SWCNT membrane and its underneath gate were characterized by coupling with a vector network analyzer. The resonance frequency of transmission coefficient S21 was observed to be around 10 MHz. The resonance frequency shifts with DNA-hybridization, and the sensing limit was approximately 50 nM. The advantages of this approach include low-noise frequency output, solution based real time detection and capable of on-chip integration, etc.
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Técnicas Biossensoriais/métodos , Nanotubos de Carbono/química , Hibridização de Ácido Nucleico/métodos , Eletroquímica , EletrodosRESUMO
Strawberries rapidly deteriorate postharvest, necessitating effective measures to extend their shelf life. This study focused on developing an eco-friendly chitosan-based protective film for strawberry preservation. Strawberries were treated with a coating solution containing varying concentrations of hawthorn leaf extract (HLE) (0.4%, 0.7%, and 1.0%), 1.5% chitosan (CH), and 1% acetic acid. The results demonstrated that coating strawberry fruit with 1% CH-HLE notably delayed fruit spoilage. In-depth analysis revealed that, compared with the uncoated strawberry fruits, the 1% CH-HLE coating effectively reduced weight loss, the respiration intensity, malondialdehyde (MDA) levels, and superoxide anion (O2·-) production. Additionally, the coated strawberries exhibited improved firmness, total soluble solids (TSS), vitamin C (Vc) content, titratable acidity (TA), and total phenolic compound (TPC) content. The enzyme activities of superoxide dismutase (SOD) and catalase (CAT) in the CH-HLE-coated strawberries were greater than those in their uncoated counterparts. The application of a 1% CH-HLE coating successfully delayed spoilage and extend the shelf life of the strawberries by approximately 4-5 days. These findings suggest that CH-HLE has significant potential as a resource for protecting fruits and vegetables, offering an environmentally sustainable solution for postharvest preservation.
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Quitosana , Crataegus , Fragaria , Conservação de Alimentos/métodos , Quitosana/farmacologia , Frutas , Extratos Vegetais/farmacologiaRESUMO
Background: There are few reports of infantile mitochondrial DNA depletion syndrome (MDDS) caused by variants in RRM2B and the correlation between genotype and phenotype has rarely been analyzed in detail. This study investigated an infantile patient with MDDS, from clinical characteristics to genetic causes. Methods: Routine physical examinations, laboratory assays, which included gas chromatography-mass spectrometry of blood and urine, and MRI scans were performed to obtain an exact diagnosis. Whole-exome sequencing was used to pinpoint the abnormal gene and bioinformatic analyses were performed on the identified variant. Results: The case presented with progressive neurologic deterioration, failure to thrive, respiratory distress and lactic acidosis. Sequencing revealed that the patient had a homozygous novel missense variant, c.155T>C (p.Ile52Thr), in exon 2 of the RRM2B gene. Multiple lines of bioinformatic evidence suggested that this was a likely detrimental variant. In addition, reported RRM2B variants were compiled from the relevant literature to analyze disease etiology. We found a distinctive distribution of genotypes across disease manifestations of different severity. Pathogenic alleles of RRM2B were significantly enriched in MDDS cases. Conclusion: The novel variant is a likely genetic cause of MDDS. It expands our understanding of the pathogenic variant spectrum and the contribution of the RRM2B gene to the disease spectrum of MDDS.
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Immunogenic dying tumor cells hold promising prospects as cancer vaccines to activate systemic immunity against both primary and metastatic tumors. Especially, X-ray- induced dying tumor cells are rich in highly immunogenic tumor-associated antigens and self-generated dsDNA as potent adjuvants. However, we found that the X-ray induction process can result in the excessive exposure of phosphatidylserine in cancer vaccines, which can specifically bind with the MerTK receptor on macrophages, acting as a "checkpoint" to facilitate immune silence in the tumor microenvironment. Therefore, we developed a novel strategy combining X-ray-induced cancer vaccines with UNC2250, a macrophage MerTK "checkpoint inhibitor," for treating peritoneal carcinomatosis in colon cancer. By incorporating UNC2250 into the treatment regimen, immunosuppressive efferocytosis of macrophages, which relies on MerTK-directed recognition of phosphatidylserine on vaccines, was effectively blocked. Consequently, the immune analysis revealed that this combination strategy promoted the maturation of dendritic cells and M1-like repolarization of macrophages, thereby simultaneously eliciting robust adaptive and innate immunity. This innovative approach utilizing X-ray-induced vaccines combined with a checkpoint inhibitor may provide valuable insights for developing effective cancer vaccines and immunotherapies targeting colon cancer.
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Background: Combined Oxidative Phosphorylation Deficiency 23 (COXPD23) is a rare mitochondrial disease caused by mutations in the GTPBP3 gene. The rare incidence of the disease and the high clinical heterogeneity pose challenges in making a precise diagnosis. Investigations into the rare COXPD23 patients are of pathophysiological and etiological value. In this study, we investigated the genotype-phenotype relationship in a COXPD23 patient from a Manchu family, with GTPBP3 mutations. Methods: Routine physical examinations, laboratory assays and imaging analyses were performed. The metabolic profiles of amino acids in blood, acylcarnitine in blood and organic acids in urine were used to determine the presence of inherited metabolic diseases. Genetic variations in the family were investigated using whole-exome sequencing and Sanger sequencing. Splicing disruption by a mutation was predicted and verified using a minigene assay. Results: The patient presented with severe lactic acidosis, neurological symptoms, multiple symmetrical lesions in the brain and serious mitochondrial energy metabolism disturbances. The c.689A > C (p.Q230P) and c.809-1_809delinsA compound heterozygous mutations were detected in GTPBP3. The novel c.809-1_809delinsA mutation was located at the splicing site of exon 7 and intron 6 and multiple tools predicted that it would disrupt the normal splicing. The minigene assay proved that the novel mutation resulted in two aberrant transcripts that created premature termination codons. Conclusions: The clinical manifestations, brain imaging change, mitochondrial metabolism disturbances and the detection and validation of the GTPBP3 mutations expand the profile of COXPD23 and the pathogenic mutation spectrum. Our study improves the understanding of the pathophysiology and etiology of COXPD23.
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The origin and functionality of long noncoding RNA (lncRNA) remain poorly understood. Here, we show that multiple quantitative trait loci modulating distinct domestication traits in soybeans are pleiotropic effects of a locus composed of two tandem lncRNA genes. These lncRNA genes, each containing two inverted repeats, originating from coding sequences of the MYB genes, function in wild soybeans by generating clusters of small RNA (sRNA) species that inhibit the expression of their MYB gene relatives through post-transcriptional regulation. By contrast, the expression of lncRNA genes in cultivated soybeans is severely repressed, and, consequently, the corresponding MYB genes are highly expressed, shaping multiple distinct domestication traits as well as leafhopper resistance. The inverted repeats were formed before the divergence of the Glycine genus from the Phaseolus-Vigna lineage and exhibit strong structure-function constraints. This study exemplifies a type of target for selection during plant domestication and identifies mechanisms of lncRNA formation and action.
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Domesticação , Regulação da Expressão Gênica de Plantas , Glycine max , Hemípteros , Locos de Características Quantitativas , RNA Longo não Codificante , Glycine max/genética , RNA Longo não Codificante/genética , Animais , Hemípteros/genética , Doenças das Plantas/genética , RNA de Plantas/genéticaRESUMO
It is challenging to perform path planning tasks in complex marine environments as the unmanned surface vessel approaches the goal while avoiding obstacles. However, the conflict between the two subtarget tasks of obstacle avoidance and goal approaching makes the path planning difficult. Thus, a path planning method for unmanned surface vessel based on multiobjective reinforcement learning is proposed under the complex environment with high randomness and multiple dynamic obstacles. Firstly, the path planning scene is set as the main scene, and the two subtarget scenes including obstacle avoidance and goal approaching are divided from it. The action selection strategy in each subtarget scene is trained through the double deep Q-network with prioritized experience replay. A multiobjective reinforcement learning framework based on ensemble learning is further designed for policy integration in the main scene. Finally, by selecting the strategy from subtarget scenes in the designed framework, an optimized action selection strategy is trained and used for the action decision of the agent in the main scene. Compared with traditional value-based reinforcement learning methods, the proposed method achieves a 93% success rate in path planning in simulation scenes. Furthermore, the average length of the paths planned by the proposed method is 3.28% and 1.97% shorter than that of PER-DDQN and dueling DQN, respectively.
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Aprendizagem , Reforço Psicológico , Algoritmos , Simulação por Computador , PolíticasRESUMO
Inflammatory bowel disease (IBD) is a type of recurrent intestinal diseases. Natural product molecules have been gradually developed into an important source of anti-inflammatory drugs for treating IBD owing to their high anti-inflammatory activity, well known safety, structural specificity and therapeutic mechanism diversity. However, most of the natural products are restricted by poor solubility in actual application. How to achieve satisfactory bioavailability during the treatment of IBD is one of the urgent problems to be solved in the current research. Micro/nano drug delivery systems could improve the solubility of drugs with targeted delivery of anti-inflammatory drugs to the colon with responsive release property. Therefore, using micro/nano drug delivery systems, the problems mentioned above involving natural product molecules in the treatment of IBD could be solved. According to the compositions of the intestinal tract and inflammatory characteristics of IBD, the strategies of using micro/nano drug delivery systems for natural products could be summarized in two steps: targeted delivery and responsive release. In this review, the targeted and responsive release strategies of the micro/nano drug delivery systems combined with their anti-inflammatory effects in IBD animal models to illustrate that the proposed strategies could be potential treatments for symptomatic IBD are described.
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Produtos Biológicos , Doenças Inflamatórias Intestinais , Animais , Sistemas de Liberação de Fármacos por Nanopartículas , Produtos Biológicos/farmacologia , Produtos Biológicos/uso terapêutico , Doenças Inflamatórias Intestinais/tratamento farmacológico , Preparações Farmacêuticas , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêuticoRESUMO
AIMS: Diabetes mellitus is a common chronic disease of glucose metabolism. Endothelial dysfunction is an early event in diabetes complicated by cardiovascular disease. This study aimed to reveal the expression of BASP1 and its biological roles in endothelial cell dysfunction in diabetes complicated by cardiovascular disease. MATERIALS AND METHODS: By analyzing the databases related to diabetes complicated with coronary heart disease, BASP1 was screened out as an upregulated gene. Human umbilical vein endothelial cells (HUVECs) and primary mouse aortic endothelial cells were treated with high glucose to establish cell models of diabetes-related endothelial dysfunction, and the expression changes of BASP1 were verified by RT-qPCR, western blot, and immunofluorescence. BASP1 was silenced or overexpressed by siRNA or overexpression plasmid, and its effects on cell migration, apoptosis, tube formation, inflammatory response, and ROS were detected. The possible signaling pathway of BASP1 was found and the mechanism of BASP1 on promoting the progression of endothelial dysfunction was explored using the EGFR inhibitor, gefitinib. RESULTS: Bioinformatics analysis indicated that the expression of BASP1 in patients with diabetes mellitus and concomitant coronary heart disease was increased. High glucose induced the upregulation of BASP1 expression in endothelial cells, and showed a time-dependent relationship. Silencing of BASP1 alleviated the damage of high glucose to endothelial cells. BASP1 regulated EGFR positively. The promoting effect of BASP1 on endothelial cell apoptosis may be achieved by regulating the EGFR pathway. CONCLUSION: BASP1 promotes endothelial cell injury induced by high glucose in patients with diabetes, which may be activated by activating the EGFR pathway.
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Doenças Cardiovasculares , Diabetes Mellitus , Animais , Humanos , Camundongos , Apoptose , Doenças Cardiovasculares/metabolismo , Diabetes Mellitus/metabolismo , Receptores ErbB/metabolismo , Receptores ErbB/farmacologia , Glucose/farmacologia , Glucose/metabolismo , Células Endoteliais da Veia Umbilical Humana/metabolismo , Proteínas de Membrana/metabolismo , Proteínas do Tecido Nervoso , Proteínas Repressoras/metabolismo , Proteínas Repressoras/farmacologia , Transdução de SinaisRESUMO
The present study aimed to determine the capsular serotype distribution and antimicrobial drug resistance patterns of Haemophilus influenzae from children in the Kunming region of China. This information could guide policymakers in clinical treatment. In the present study, H. influenzae isolates were tested for their serotypes, antimicrobial susceptibility pattern, and presence of ß-lactamases. One-hundred forty-eight H. influenzae strains isolated from children 0-2 years old were investigated for capsular types by glass slide agglutination and molecular methods, and biotyped by the biochemical reactions. The drug resistance-encoding genes TEM-1, ROB-1, and the ftsI gene mutations PBP3-3, and PBP3-BLN were detected with real-time quantitative polymerase chain reaction (qPCR). The prevalence of ß-lactamase-producing strains (60.3%) was significantly higher (p < 0.05) than non-enzyme-producing strains. ß-Lactamase-producing strains were multidrug resistant to various antibiotics such as ampicillin, tetracycline, sulfamethoxazole/trimethoprim, chloramphenicol, cefuroxime, and cefaclor. Among ß-lactamase-producing strains, the detection rates of the TEM-1, PBP3-BLN, PBP3-s, and ROB-1 were 54.1%, 18.9%, 11.8%, and 6.9%, respectively. The biotyping results show that most H. influenzae strains were of type II and III. Non-typeable H. influenzae (NTHi) accounted for 89.3% of the strains. NTHi strains were the most prevalent in this region; most belonged to biological types II and III. ß-Lactamase-positive ampi-cillin-resistant (BLPAR) strains were prevalent among H. influenzae isolates in this region.
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Infecções por Haemophilus , Haemophilus influenzae , Criança , Humanos , Recém-Nascido , Lactente , Pré-Escolar , Sorogrupo , Haemophilus influenzae/genética , Infecções por Haemophilus/tratamento farmacológico , Infecções por Haemophilus/epidemiologia , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , beta-Lactamases/genética , Testes de Sensibilidade Microbiana , Resistência a MedicamentosRESUMO
OBJECTIVES: Kawasaki disease (KD) is a systemic vasculitis that is caused by immunological dysregulation in children exposed to pathogens like Epstein-Barr virus (EBV). Myocardial ischemia or infarction due to coronary artery lesions (CALs) might be lethal. However, it is unclear how pathogens, immunomodulation, and CALs interact, particularly in KD patients co-infected with the most widespread virus, EBV. METHODS: We investigated pathogen carriage and fundamental clinical data in 281 KD patients. Immunological differences between CALs and non-CALs in KD patients under different conditions were analyzed. Then, the effect of infection by different pathogens on the immune response was excluded, and most EBV co-infected KD patients were included to assess the incidence of CALs, the level of immune modulation, and regulatory mechanisms in different EBV infection states. RESULTS: Our results showed multiple pathogenic infections occur in KD patients, with EBV being the most prevalent. The incidence of CALs in the EBV-DNA (+) acute infection group, EBV-DNA (-) acute infection group, and EBV latent infection group was 0 (0/6), 27.27% (3/11) and 41.67% (10/24), respectively. The two groups were younger and had increased IL-6 levels and B cells, decreasing CD8+ T cells than the EBV-DNA (+) acute infection group. Interestingly, the increased B cells were not associated with immunoglobulin release. Additionally, these patients down-regulated α7 nicotinic acetylcholine receptor (α7nAChR) and downstream molecule PI3K/AKT/mTOR while activating the NF-κB. CONCLUSION: Patients with different EBV infection statuses exhibit different incidences of CALs. In acute EBV-DNA (-) infected and latent EBV-infected patients, the number of CD8+ T cells decreased and downregulated CD8+ T cells' α7nAChR and PI3K/AKT/mTOR, which may associate with CALs, while the expression of NF-κB and the pro-inflammatory factor IL-6 was upregulated by inhibiting the anti-inflammatory molecule α7nAChR.
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Infecções por Vírus Epstein-Barr , Síndrome de Linfonodos Mucocutâneos , Criança , Humanos , Receptor Nicotínico de Acetilcolina alfa7 , Linfócitos T CD8-Positivos , Vasos Coronários , DNA , Infecções por Vírus Epstein-Barr/complicações , Herpesvirus Humano 4 , Interleucina-6 , Síndrome de Linfonodos Mucocutâneos/complicações , NF-kappa B , Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-akt , Serina-Treonina Quinases TORRESUMO
During vascular development, endothelial cells (ECs) undergo arterialization in response to genetic programs and shear stress-triggered mechanotransduction, forming a stable vasculature. Although the Notch receptor is known to sense shear stress and promote EC arterialization, its downstream mechanisms remain unclear. In this study, the Notch downstream miR-342-5p was found to respond to shear stress and promote EC arterialization. Shear stress upregulated miR-342-5p in a Notch-dependent manner in human umbilical vein endothelial cells (HUVECs). miR-342-5p overexpression upregulated the shear stress-associated transcriptomic signature. Moreover, miR-342-5p upregulated arterial markers and promoted EC arterialization in a Matrigel plug assay and retinal angiogenesis model. In contrast, miR-342-5p knockdown downregulated arterial markers, compromised retinal arterialization, and partially abrogated shear stress and Notch activation-induced arterial marker upregulation. Mechanistically, miR-342-5p overexpression suppressed MYC to repress EC proliferation and promote arterialization, achieved by promoting MYC protein degradation by targeting the EYA3. Consistently, EYA3 overexpression rescued miR-342-5p-mediated MYC downregulation and EC arterialization. In vivo, miR-342-5p expression was notably decreased in the ligated artery in a hindlimb ischemia model, and an intramuscular injection of miR-342-5p promoted EC arterialization and improved perfusion. In summary, miR-342-5p, a mechano-miR, mediates the effects of shear stress-activated Notch on EC arterialization and is a potential therapeutic target for ischemic diseases.
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Human cancers induce a chaotic, dysfunctional vasculature that promotes tumor growth and blunts most current therapies; however, the mechanisms underlying the induction of a dysfunctional vasculature have been unclear. Here, we show that split end (SPEN), a transcription repressor, coordinates rRNA synthesis in endothelial cells (ECs) and is required for physiological and tumor angiogenesis. SPEN deficiency attenuated EC proliferation and blunted retinal angiogenesis, which was attributed to p53 activation. Furthermore, SPEN knockdown activated p53 by upregulating noncoding promoter RNA (pRNA), which represses rRNA transcription and triggers p53-mediated nucleolar stress. In human cancer biopsies, a low endothelial SPEN level correlated with extended overall survival. In mice, endothelial SPEN deficiency compromised rRNA expression and repressed tumor growth and metastasis by normalizing tumor vessels, and this was abrogated by p53 haploinsufficiency. rRNA gene transcription is driven by RNA polymerase I (RNPI). We found that CX-5461, an RNPI inhibitor, recapitulated the effect of Spen ablation on tumor vessel normalization and combining CX-5461 with cisplatin substantially improved the efficacy of treating tumors in mice. Together, these results demonstrate that SPEN is required for angiogenesis by repressing pRNA to enable rRNA gene transcription and ribosomal biogenesis and that RNPI represents a target for tumor vessel normalization therapy of cancer.