ABSTRACT
Ferroptosis, a form of regulated cell death that is driven by iron-dependent phospholipid peroxidation, has been implicated in multiple diseases, including cancer1-3, degenerative disorders4 and organ ischaemia-reperfusion injury (IRI)5,6. Here, using genome-wide CRISPR-Cas9 screening, we identified that the enzymes involved in distal cholesterol biosynthesis have pivotal yet opposing roles in regulating ferroptosis through dictating the level of 7-dehydrocholesterol (7-DHC)-an intermediate metabolite of distal cholesterol biosynthesis that is synthesized by sterol C5-desaturase (SC5D) and metabolized by 7-DHC reductase (DHCR7) for cholesterol synthesis. We found that the pathway components, including MSMO1, CYP51A1, EBP and SC5D, function as potential suppressors of ferroptosis, whereas DHCR7 functions as a pro-ferroptotic gene. Mechanistically, 7-DHC dictates ferroptosis surveillance by using the conjugated diene to exert its anti-phospholipid autoxidation function and shields plasma and mitochondria membranes from phospholipid autoxidation. Importantly, blocking the biosynthesis of endogenous 7-DHC by pharmacological targeting of EBP induces ferroptosis and inhibits tumour growth, whereas increasing the 7-DHC level by inhibiting DHCR7 effectively promotes cancer metastasis and attenuates the progression of kidney IRI, supporting a critical function of this axis in vivo. In conclusion, our data reveal a role of 7-DHC as a natural anti-ferroptotic metabolite and suggest that pharmacological manipulation of 7-DHC levels is a promising therapeutic strategy for cancer and IRI.
Subject(s)
Dehydrocholesterols , Ferroptosis , Humans , Cell Membrane/metabolism , Cholesterol/biosynthesis , Cholesterol/metabolism , CRISPR-Cas Systems/genetics , Dehydrocholesterols/metabolism , Genome, Human , Kidney Diseases/metabolism , Mitochondrial Membranes/metabolism , Neoplasm Metastasis , Neoplasms/metabolism , Neoplasms/pathology , Phospholipids/metabolism , Reperfusion Injury/metabolismABSTRACT
The African swine fever virus (ASFV) type II topoisomerase (Topo II), pP1192R, is the only known Topo II expressed by mammalian viruses and is essential for ASFV replication in the host cytoplasm. Herein, we report the structures of pP1192R in various enzymatic stages using both X-ray crystallography and single-particle cryo-electron microscopy. Our data structurally define the pP1192R-modulated DNA topology changes. By presenting the A2+-like metal ion at the pre-cleavage site, the pP1192R-DNA-m-AMSA complex structure provides support for the classical two-metal mechanism in Topo II-mediated DNA cleavage and a better explanation for nucleophile formation. The unique inhibitor selectivity of pP1192R and the difunctional mechanism of pP1192R inhibition by m-AMSA highlight the specificity of viral Topo II in the poison binding site. Altogether, this study provides the information applicable to the development of a pP1192R-targeting anti-ASFV strategy.
ABSTRACT
We have recently demonstrated that Jumonji domain-containing protein D3 (JMJD3), a histone demethylase of histone H3 on lysine 27 (H3K27me3), is protective against renal fibrosis, but its role in acute kidney injury (AKI) remains unexplored. Here, we report that JMJD3 activity is required for renal protection and regeneration in murine models of AKI induced by ischemia/reperfusion (I/R) and folic acid (FA). Injury to the kidney upregulated JMJD3 expression and induced expression of H3K27me3, which was coincident with renal dysfunction, renal tubular cell injury/apoptosis, and proliferation. Blocking JMJD3 activity by GSKJ4 led to worsening renal dysfunction and pathological changes by aggravating tubular epithelial cell injury and apoptosis in both murine models of AKI. JMJD3 inhibition by GSKJ4 also reduced renal tubular cell proliferation and suppressed expression of cyclin E and phosphorylation of CDK2, but increased p21 expression in the injured kidney. Furthermore, inactivation of JMJD3 enhanced I/R- or FA-induced expression of TGF-ß1, vimentin, and Snail, phosphorylation of Smad3, STAT3, and NF-κB, and increased renal infiltration by F4/80 (+) macrophages. Finally, GSKJ4 treatment caused further downregulation of Klotho, BMP-7, Smad7, and E-cadherin, all of which are associated with renal protection and have anti-fibrotic effects. Therefore, these data provide strong evidence that JMJD3 activation contributes to renal tubular epithelial cell survival and regeneration after AKI.
Subject(s)
Acute Kidney Injury , Histones , Animals , Mice , Acute Kidney Injury/genetics , Acute Kidney Injury/pathology , Cell Proliferation , Histones/metabolism , Kidney/metabolism , PhosphorylationABSTRACT
IMPORTANCE: African swine fever (ASF) is an acute, hemorrhagic, and severe porcine infectious disease caused by African swine fever virus (ASFV). ASF outbreaks severely threaten the global pig industries and result in serious economic losses. No safe and efficacious commercial vaccine is currently available except in Vietnam. To date, large gaps in the knowledge concerning viral biological characteristics and immunoevasion strategies have hindered the ASF vaccine design. In this study, we demonstrate that pD129L negatively regulates the type I interferon (IFN) signaling pathway by interfering with the interaction of the transcriptional coactivator p300 and IRF3, thereby inhibiting the induction of type I IFNs. This study reveals a novel immunoevasion strategy employed by ASFV, shedding new light on the intricate mechanisms for ASFV to evade the host immune responses.
Subject(s)
African Swine Fever Virus , African Swine Fever , E1A-Associated p300 Protein , Interferon Regulatory Factor-3 , Interferon Type I , Animals , African Swine Fever/virology , Interferon Type I/metabolism , Interferon-beta/metabolism , Swine , Transcription Factors/metabolism , Vaccines/metabolism , E1A-Associated p300 Protein/metabolism , Interferon Regulatory Factor-3/metabolism , Immune EvasionABSTRACT
Lysyl oxidase-like 2 (LOXL2) is an extracellular copper-dependent enzyme that plays a central role in fibrosis by catalyzing the crosslinking and deposition of collagen. Therapeutic LOXL2 inhibition has been shown to suppress liver fibrosis progression and promote its reversal. This study investigates the efficacy and underlying mechanisms of human umbilical cord-derived exosomes (MSC-ex) in LOXL2 inhibition of liver fibrosis. MSC-ex, nonselective LOX inhibitor ß-aminopropionitrile (BAPN), or PBS were administered into carbon tetrachloride (CCl4)-induced fibrotic livers. Serum LOXL2 and collagen crosslinking were assessed histologically and biochemically. MSC-ex's mechanisms on LOXL2 regulation were investigated in human hepatic stellate cell line LX-2. We found that systemic administration of MSC-ex significantly reduced LOXL2 expression and collagen crosslinking, delaying the progression of CCl4-induced liver fibrosis. Mechanically, RNA-sequencing and fluorescence in situ hybridization (FISH) indicated that miR-27b-3p was enriched in MSC-ex and exosomal miR-27b-3p repressed Yes-associated protein (YAP) expression by targeting its 3' untranslated region in LX-2. LOXL2 was identified as a novel downstream target gene of YAP, and YAP bound to the LOXL2 promoter to positively regulate transcription. Additionally, the miR-27b-3p inhibitor abrogated the anti-LOXL2 abilities of MSC-ex and diminished the antifibrotic efficacy. miR-27b-3p overexpression promoted MSC-ex mediated YAP/LOXL2 inhibition. Thus, MSC-ex may suppress LOXL2 expression through exosomal miR-27b-3p mediated YAP down-regulation. The findings here may improve our understanding of MSC-ex in liver fibrosis alleviation and provide new opportunities for clinical treatment.
Subject(s)
Mesenchymal Stem Cells , MicroRNAs , Humans , Collagen/metabolism , In Situ Hybridization, Fluorescence , Liver Cirrhosis/chemically induced , Liver Cirrhosis/drug therapy , Liver Cirrhosis/metabolism , Mesenchymal Stem Cells/metabolism , MicroRNAs/metabolismABSTRACT
African swine fever (ASF) is a highly contagious disease of domestic pigs and wild boar with high morbidity and mortality caused by African swine fever virus (ASFV). Due to the lack of commercial vaccines and treatments for ASF, cleaning and disinfection remain one of the most effective biosecurity measures to control ASF. Our previous studies have shown that ASFV can be inactivated by 0.25 to 5% highly complexed iodine (HPCI) in 5 to 30 min. This study evaluated the synergistic inactivation effects of HPCI combined with compound organic acids (COAs) against ASFV. The results showed that the inactivation rates of HPCI, COAs, and HPCI+COAs on the reporter ASFV expressing the green fluorescent protein increased in dose- and time-dependent manners. The best inactivation effects were obtained when the compatibility ratio of HPCI and COAs was 5:1, and the ideal temperature was 25°C. Furthermore, there were no significant differences when comparing the efficacy of HPCI combined with COAs (HPCI+COAs) in inactivating wild-type ASFV and the reporter ASFV (P > 0.05). ASFV of 104.0 50% tissue culture infective dose (TCID50)/mL was completely inactivated by 0.13% HPCI (0.0065% effective iodine), 0.06% COAs, or 0.13% HPCI+COAs (approximately 0.0054% effective iodine), respectively, while 106.0 TCID50/mL ASFV was completely inactivated by 1.00% HPCI (0.05% effective iodine), 0.50% COAs, or 1.00% HPCI+COAs (0.042% effective iodine), respectively. It was found that the combination index (CI) of HPCI and COAs was less than 1 under different conditions. This study demonstrated that HPCI+COAs could synergistically inactivate ASFV and represent an effective compound disinfectant for the control of ASF. IMPORTANCE African swine fever (ASF) is a highly contagious disease of swine with high morbidity and mortality caused by African swine fever virus (ASFV). Due to the lack of commercial vaccines and treatment available for ASF, effective disinfectants and the proper use of them are essential to inactivate ASFV. The significance of this research is in searching for an ideal disinfectant that has the advantages of low toxicity and nonpollution and can inactivate ASFV efficiently. In this study, we demonstrated that HPCI+COAs had synergistic effects on inactivating ASFV. Thus, HPCI+COAs could be used as an effective disinfectant for the control of ASF.
Subject(s)
African Swine Fever Virus , African Swine Fever , Disinfectants , Iodine , African Swine Fever/prevention & control , Animals , Disinfectants/pharmacology , Iodine/pharmacology , Sus scrofa , SwineABSTRACT
BACKGROUND: Orthokeratinized odontogenic cyst (OOC), a newly designated entity of odontogenic cysts, is an intraosseous jaw cyst that is entirely or predominantly lined by orthokeratinized squamous epithelium. The aim of this study was to report a large series of OOC to substantiate its clinicopathologic profiles and to investigate PTCH1 mutations in OOCs. METHOD: The clinicopathologic features of 167 OOCs from 159 patients were analyzed and the immunohistochemical expression of markers related to cell differentiation and proliferation was evaluated. Furthermore, PTCH1 mutations were analyzed in 14 fresh samples of OOC. RESULTS: OOCs occurred mostly in the third and fourth decades (60.4%) with a male predilection (66.7%). The lesions developed more often in the mandible than maxilla, primarily in the posterior mandible and ramus. Eight patients (5.0%) showed multiple locations of either bilateral posterior mandible (n = 6) or both the maxilla and mandible. Radiographically, the majority of OOCs (91.2%) showed a well-demarcated, unilocular radiolucency with 14 multilocular cases (8.8%). A follow-up of 131 patients (123 treated by enucleation with or without marsupialization and eight by peripheral ostectomy) revealed no recurrence during an average period of 4.56 years after surgery. Immunohistochemistry indicated lower proliferative activity and a varying epithelial differentiation pattern in OOC compared with odontogenic keratocysts (OKC). No PTCH1 mutation was detected, except for three known single nucleotide polymorphisms. CONCLUSION: The clinicopathological and molecular differences between OOC and OKC justified their separation, and unlike OKCs, OOCs did not harbor PTCH1 mutations, suggesting different pathogenesis underlying these two jaw cysts.
Subject(s)
Odontogenic Cysts , Odontogenic Tumors , Patched-1 Receptor/genetics , Epithelium/pathology , Female , Humans , Immunohistochemistry , Male , Mutation , Odontogenic Cysts/genetics , Odontogenic Cysts/pathology , Odontogenic Tumors/genetics , Odontogenic Tumors/pathologyABSTRACT
BACKGROUND: Cup-shaped ear is a common congenital auricle deformity. There are many specific surgical methods, such as V-Y method, Barsky method, Musgrave method, etc., but there is no unified treatment method. The authors used the outer helix reconstruction method for cup-shaped ear and achieved remarkable therapeutic effects. MATERIALS AND METHODS: The authors performed the outer helix reconstruction on 30 patients with cup-shaped ear. The authors followed up the patients after the stage II operation. The authors used the SPSAU data science analysis platform (https://spssau.com/) for statistical analysis of the data. RESULT: The mean follow-up time was 14.43±4.5 months. The mean preoperative perimeter of auricle was 8.19±0.56 cm. The mean postoperative perimeter of auricle was 10.82±0.49 cm. The mean perimeter of the healthy auricle was 10.89±0.44 cm. Through data analysis, the authors found that the comparison between preoperative and postoperative auricle perimeters was statistically significant (P<0.05). The comparison between postoperative auricle perimeter and the perimeter of the healthy auricle was not statistically significant (P>0.05). In terms of postoperative satisfaction of patients and their families, the satisfaction rate was 100%. In terms of postoperative complications, there were 1 case of incision dehiscence, 0 cases of incision infection, 2 cases of incision hematoma, and 0 cases of postoperative skin flap ischemia and necrosis. CONCLUSION: The outer helix reconstruction method is suitable for most cup-shaped ears, and the operation is simple and the effect is remarkable. It is worthy of promotion and application in plastic surgery clinical.
Subject(s)
Ear Auricle , Research Design , Humans , Ear, External/surgery , Ear Auricle/surgery , Postoperative Period , Surgical Wound DehiscenceABSTRACT
Coronaviruses are common human viruses and include the severe acute respiratory syndrome coronavirus (SARS-CoV), the middle east respiratory syndrome coronavirus and the SARS-CoV-2. Coronaviruses mainly bind to transmembrane receptor proteins on the human cell membrane through spike proteins (S-proteins), thus releasing the RNA of the virus into the interior of the host cell to cause an infection. In this article, we discuss the mechanism and production of cyclodextrin-soluble angiotensin-converting enzyme 2 (CD-sACE2) inclusion compounds in the treatment of SARS-CoV-2 infections by blocking S-proteins. On the basis of the current research evidence, we believe that CD-sACE2 inclusion compounds have the potential to treat COVID-19. We hope that our article can provide a theoretical basis for later experiments.
Subject(s)
Angiotensin-Converting Enzyme 2/metabolism , COVID-19 Drug Treatment , COVID-19/metabolism , Cyclodextrins/therapeutic use , Humans , Pandemics , Pneumonia, Viral/drug therapy , Pneumonia, Viral/metabolism , Pneumonia, Viral/virology , SARS-CoV-2/drug effects , Spike Glycoprotein, Coronavirus/metabolismABSTRACT
BACKGROUND Axillary osmidrosis (AO) is common in plastic surgery. But there is no perfect way to treat AO. We systematically compared the efficacy of 10 AO treatments with network meta-analysis in order to provide reference for the clinical treatment of axillary odor. MATERIAL AND METHODS Chinese and English databases were searched by computer. Some relevant studies were collected for network meta-analysis. RESULTS We identified 56 studies, including a total of 8618 patients for meta-analysis. The network meta-analysis showed that 21 out of 45 pairs of 10 AO treatments had no statistical significance. In statistical comparison, subcutaneous curettage and swelling suction subcutaneous pruning were better than a single treatment. In addition, the effects of both laser and electric ion therapy were inferior to those of other treatments. The order of therapeutic effects predicted by surface under the cumulative ranking (SUCRA), curve was swelling aspiration+subcutaneous pruning >subcutaneous pruning >subcutaneous curettage+subcutaneous pruning >spindle excision >botulinum toxin A injection >swelling aspiration >subcutaneous curettage >YAG laser therapy >CO2 laser therapy >electric ion therapy. CONCLUSIONS In operative treatment of AO, swelling aspiration+subcutaneous pruning is the best operative treatment, and botulinum toxin A injection is the best in non-operative treatment. Overall, the effect of surgical treatment was more significant than that of non-surgical treatment.
Subject(s)
Apocrine Glands/drug effects , Apocrine Glands/surgery , Odorants/prevention & control , Sweat Gland Diseases/therapy , Apocrine Glands/physiopathology , Axilla , Botulinum Toxins, Type A/therapeutic use , Curettage , Humans , Network Meta-Analysis , Patient Satisfaction , Quality of Life , Randomized Controlled Trials as Topic , Sweat Gland Diseases/physiopathology , Sweat Gland Diseases/surgery , Sweating/physiology , Treatment OutcomeABSTRACT
OBJECTIVE: Odontogenic keratocysts (OKCs) are benign jaw lesions with high growth potential and propensity for recurrence. Our previous study revealed that PTCH1 mutations, which were frequently detected in sporadic OKCs, might be underestimated due to the masking effect of the stromal components within the tested tissues. We aimed to confirm these results in larger scale and further present the unbiased view of the genomic basis of sporadic OKCs except PTCH1. MATERIALS AND METHODS: We analyzed PTCH1 mutations in additional 19 samples. Using whole-exome sequencing (WES), we further characterized the mutational landscape of five sporadic OKC samples lacking PTCH1 mutation and loss of heterozygosity (LOH). RESULTS: Combined with our previously reported 19 cases, thirty of 38 (79%) cases harbored PTCH1 mutations. Through whole-exome sequencing and integrative analysis, 22 novel mutations were confirmed among five PTCH1-negative samples. No recurrent mutations were identified in the WES samples and validation cohort of 10 OKCs. CONCLUSIONS: Our data further confirmed the frequent PTCH1 mutation and other rare genetic alterations in sporadic OKCs, highlighting the central role of SHH signaling pathway. In PTCH1-negative cases, other rare mutations scattered in a subset of OKCs were independent of the SHH pathway. These results suggested that an SHH inhibitor may be effective to treat the majority of OKCs.
Subject(s)
Basal Cell Nevus Syndrome , Odontogenic Cysts/genetics , Patched-1 Receptor/genetics , Adult , Female , Humans , Male , Middle Aged , Mutation , Neoplasm Recurrence, Local , Receptors, Cell Surface , Exome SequencingABSTRACT
Lung cancer remains to be the leading cause of cancer-related mortality worldwide. Finding new noninvasive biomarkers for lung cancer is still a significant clinical challenge. Exosomes are membrane-bound, nano-sized vesicles that are released by various living cells. Studies on exosomal proteomics may provide clues for developing clinical assays. In this study, we performed semi-quantitative proteomic analysis of proteins that were purified from exosomes of NCI-H838 non-small cell lung cancer cell line, with total cellular membrane proteins as control. In the exosomes, LC-MS/MS by data-independent analysis mode identified 3235 proteins. THBS1, ANXA6, HIST1H4A, COL18A1, MDK, SRGN, ENO1, TUBA4A, SLC3A2, GPI, MIF, MUC1, TALDO1, SLC7A5, ICAM1, HSP90AA1, G6PD, and LRP1 were found to be expressed in exosomes at more than 5-fold higher level as compared to total cellular membrane proteins. A well-known cancer biomarker, MUC1, is expressed at 8.98-fold higher in exosomes than total cellular membrane proteins. Subsequent analysis of plasma exosomes from non-small cell lung cancer (NSCLC) patients by a commercial electrochemiluminescence immunoassay showed that exosomal MUC1 level is 1.5-fold higher than healthy individuals (mean value 1.55 ± 0.16 versus mean value 1.05 ± 0.06, p = 0.0213). In contrast, no significant difference of MUC1 level was found between NSCLC patients and healthy individuals' plasma (mean value 5.48 ± 0.65 versus mean value 4.16 ± 0.49). These results suggest that certain proteins, such as MUC1, are selectively enriched in the exosome compartment. The mechanisms for their preferential localization and their biological roles remain to be studied.
Subject(s)
Biomarkers, Tumor/genetics , Carcinoma, Non-Small-Cell Lung/genetics , Exosomes/genetics , Mucin-1/genetics , Carcinoma, Non-Small-Cell Lung/pathology , Chromatography, Liquid , Humans , Mucin-1/ultrastructure , Neoplasm Proteins/genetics , Proteomics , Tandem Mass SpectrometryABSTRACT
BACKGROUND: The Epstein-Barr virus (EBV) is closely associated with several types of malignancies. EBV is normally present in the latent state in the peripheral blood B cell compartment. The EBV latent-to-lytic switch is required for virus spread and virus-induced carinogenesis. Immunosuppression or DNA damage can induce the reactivation of EBV replication. EBV alone is rarely sufficient to cause cancer. In this study, we investigated the roles of host microRNAs and environmental factors, such as DNA-damage agents, in EBV reactivation and its association with lymphomagenesis. METHODS: We first analyzed the publicly available microRNA array data containing 45 diffuse large B-cell lymphoma patients and 10 control lymph nodes or B cells with or without EBV infection. In situ hybridization for miR-18a and immunohistochemitry were performed to evaluate the correlation between the expression of miR-18a and nuclear EBV protein EBNA1 in lymphoid neoplasm. The proliferative effects of miR-18a were investigated in EBV-positive or -negative lymphoid neoplasm cell lines. EBV viral load was measured by a quantitative real-time EBV PCR and FISH assay. The genomic instability was evaluated by CGH-array. RESULTS: In this study, we analyzed the publicly available microRNA array data and observed that the expression of the miR-17-92 cluster was associated with EBV status. In situ hybridization for miR-18a, which is a member of the miR-17-92 cluster, showed a significant upregulation in lymphoma samples. miR-18a, which shares the homolog sequence with EBV-encoded BART-5, promoted the proliferation of lymphoma cells in an EBV status-dependent manner. The DNA-damaging agent UV or hypoxia stress induced EBV activation, and miR-18a contributed to DNA damaging-induced EBV reactivation. In contrast to the promoting effect of ATM on the lytic EBV reactivation in normoxia, ATM inhibited lytic EBV gene expression and decreased the EBV viral load in the prescence of hypoxia-induced DNA damage. miR-18a reactivated EBV through inhibiting the ATM-mediated DNA damage response (DDR) and caused genomic instability. CONCLUSIONS: Taken together, these results indicate that DNA-damaging agents and host microRNAs play roles in EBV reactivation. Our study supported the interplay between host cell DDR, environmental genotoxic stress and EBV.
Subject(s)
Epstein-Barr Virus Infections/genetics , Herpesvirus 4, Human/physiology , Host Microbial Interactions/genetics , Lymphoma, Large B-Cell, Diffuse/genetics , MicroRNAs/metabolism , Ataxia Telangiectasia Mutated Proteins/genetics , Ataxia Telangiectasia Mutated Proteins/metabolism , Carcinogenesis/genetics , Cell Hypoxia/genetics , Cell Line, Tumor , DNA Damage/radiation effects , DNA Replication/genetics , DNA, Viral/genetics , Datasets as Topic , Epstein-Barr Virus Infections/drug therapy , Epstein-Barr Virus Infections/virology , Female , Gene Expression Regulation, Neoplastic , Genomic Instability , Herpesvirus 4, Human/isolation & purification , Humans , Lymphoma, Large B-Cell, Diffuse/pathology , Lymphoma, Large B-Cell, Diffuse/virology , Male , Middle Aged , Ultraviolet Rays/adverse effects , Up-Regulation , Viral Load , Virus Activation/genetics , Virus Replication/geneticsABSTRACT
IFN regulatory factor (IRF) 3 as an important member of IRF family, is required for the host antiviral response. In mammals, IRF3 is known to be a critical player in regulating the transcription of IFN and IFN-stimulated genes (ISGs). However, only a few studies investigated the characteristics of IRF3 genes in fish. In this study, IRF3 from miiuy croaker was identified and characterized in bioinformatics and functions. Miiuy croaker IRF3 had conserved DBD, IAD and SRD domains with other vertebrates IRF3 genes, also miiuy croaker IRF3 had relatively conserved gene synteny and gene structures with other fish IRF3 genes. Evolutionary analysis showed IRF3 genes in mammals underwent positive selection, while IRF3 in fish underwent purifying selection. Expression analysis showed miiuy croaker IRF3 was expressed in all tested tissues and up-regulated expressed in infected liver and kidney; and up-regulated expression of miiuy croaker IRF3 was observed in head kidney macrophages which stimulated with poly(I:C) indicating that miiuy croaker IRF3 participated in the immune response to defense against poly(I:C) infection. Furthermore, luciferase reporter assay showed that overexpression of miiuy croaker IRF3 can activate the production of ISRE and IFNα, suggesting that miiuy croaker IRF3 acted as transcription activators in immune responses and maybe activate IFN signaling pathway. Immunofluorescence assay showed miiuy craoker IRF3 was localized in the cytoplasm in Hela cells. Overall, we systematically and comprehensively analyzed the bioinformatics and functions of miiuy croaker IRF3, which provided further insights into the transcriptional regulation of IRF3 gene in fish and valuable information for the study of evolution of IRF3 genes.
Subject(s)
Fish Proteins/metabolism , Genome , Interferon Regulatory Factor-3/metabolism , Perciformes/immunology , Amino Acid Sequence , Animals , Computational Biology , Evolution, Molecular , Fish Proteins/chemistry , Fish Proteins/genetics , Gene Expression Regulation/drug effects , Interferon Regulatory Factor-3/chemistry , Interferon Regulatory Factor-3/genetics , Organ Specificity , Perciformes/genetics , Perciformes/microbiology , Phylogeny , Poly I-C/pharmacology , Sequence Alignment/veterinary , SyntenyABSTRACT
The laboratory of genetics and physiology 2 (LGP2) is a member of retinoic acid-inducible gene I (RIG-I)-like receptors (RLR receptors), which may participate in the immune regulation process. The role of LGP2 on modulating signaling was ambiguous, some researchers suggested that the regulation mechanism of LGP2 to melanoma differentiation-associated gene 5 (MDA5) and retinoic acid inducible gene-I (RIG-I) were different. In this study, the bioinformatics and functions of LGP2 from miiuy croaker (mmLGP2) were characterized. Comparative genomic analysis showed that the evolution of LGP2 in mammals was more conserved than it in fish. LGP2 contains three structural domains: ResIII, HelicaseC and RD, and ResIII structural domain of LGP2 was extremely conservative. The mmLGP2 was ubiquitously expressed in the tested miiuy croaker tissues and the expressions were significantly upregulated after stimulation with poly(I:C), indicating that LGP2 might participate in the immune response, especially antiviral immunity. Furthermore, immunofluorescence of miiuy croaker LGP2 presents in the cytoplasm in Hela cells. The overexpression of mmLGP2 can activate ISRE, but cannot activate NF-κB luciferase reporter, implying that mmLGP2 might act as a positive regulator in immune responses through activating ISRE to induce the expression of IFN. The research of mmLGP2 will enrich the information of fish LGP2, and the functional experiments will be helpful for the future research about fish immune systems.
Subject(s)
Evolution, Molecular , Fish Diseases/immunology , Fish Proteins/genetics , Immunity, Innate , Perciformes/immunology , RNA Helicases/genetics , Amino Acid Sequence , Animals , DNA, Complementary/genetics , DNA, Complementary/metabolism , Fish Diseases/genetics , Fish Diseases/microbiology , Fish Proteins/chemistry , Fish Proteins/metabolism , Gene Expression Regulation/immunology , HeLa Cells , Humans , Immunity, Innate/drug effects , Lipopolysaccharides/pharmacology , Perciformes/classification , Perciformes/genetics , Phylogeny , Poly I-C/pharmacology , RNA Helicases/chemistry , RNA Helicases/metabolism , Staphylococcal Infections/genetics , Staphylococcal Infections/immunology , Staphylococcal Infections/veterinary , Staphylococcus aureus/physiology , Vibrio/physiology , Vibrio Infections/genetics , Vibrio Infections/immunology , Vibrio Infections/veterinaryABSTRACT
Being indispensable pattern recognition receptors in innate immune responses in host protection, Toll-like receptors (TLRs) play an important role in pathogen recognition. Fish TLRs exhibit high variety and distinct features, although little is known about their function on ligand recognition and signaling pathway in fish. This paper reports the evolutionary spectrum of the TLR1 subfamily (referred to as TLR1, TLR6, and TLR10) as determined using the comparative genomic approach. We hypothesized that the TLR1 subfamily underwent two rounds of gene duplication events; the first duplication occurred prior to the divergence of amphibians, and the second one occurred prior to the divergence of eutherians. To further study the function of fish TLR1, we identified miiuy croaker (Miichthys miiuy) TLR1 (mmiTLR1) and determined its potential ability to perceive Vibrio anguillarum and lipopolysaccharide stimulation. Data further suggested that mmiTLR1 is dependent on TIRAP and MyD88 for signal transmission. In addition, immunocytochemistry showed the speculative interaction between MyD88 and mmiTLR1 TIR domain. Overall, we systematically and comprehensively analyzed evolution of TLR1 subfamily and the function of mmiTLR1, which will provide the basis for future scientific research on fish TLRs.
Subject(s)
Adaptor Proteins, Signal Transducing/genetics , Fish Diseases/genetics , Fish Proteins/genetics , Immunity, Innate , Perciformes , Toll-Like Receptor 1/genetics , Vibrio Infections/veterinary , Adaptor Proteins, Signal Transducing/metabolism , Animals , Fish Diseases/immunology , Fish Diseases/microbiology , Fish Proteins/chemistry , Fish Proteins/metabolism , Lipopolysaccharides/pharmacology , Myeloid Differentiation Factor 88/genetics , Myeloid Differentiation Factor 88/metabolism , Phylogeny , Protein Conformation , Sequence Analysis, DNA/veterinary , Signal Transduction , Toll-Like Receptor 1/chemistry , Toll-Like Receptor 1/metabolism , Vibrio/physiology , Vibrio Infections/genetics , Vibrio Infections/immunology , Vibrio Infections/microbiologyABSTRACT
Schizencephaly is a rare malformation of cortical development resulting from cell migration defects that occur unilaterally or bilaterally. The type of the schizencephalic cleft can be open lip or closed lip. Patients suffering from refractory seizures secondary to schizencephaly should be considered for surgical treatment. In this paper, we retrospectively analyzed two patients with confirmed schizencephaly and intractable seizures. The evaluation methods included a medical history assessment, a neurological examination and magnetic resonance imaging (MRI). Continuous intracranial video-electroencephalogram (vEEG) monitoring with surface electrodes and deep electrodes was evaluated to confirm the epileptogenic zones associated with the schizencephalic lesions. Cortical electrical stimulation was performed to evaluate the neurophysiology of the relevant brain regions. Epileptic focus resection was performed close to the schizencephalic cleft according to the results of intracranial EEG and stimulation while preserving neurological functions. MRI revealed bilateral open lip schizencephaly in one patient and closed lip schizencephaly in the other patient. The epileptogenic zones were localized close to the schizencephalic clefts. The seizure outcome was Engel's class Ia in both patients at 1-year follow-up. No significant neurological deficits were found, and their activities of daily life were significantly improved. We conclude that abnormal cortex near the schizencephalic clefts may display an extrinsic epileptogenicity. Accurate localization of the epileptogenic zones using intracranial EEG and electrical stimulation can lead to a seizure-free outcome in patients with refractory epilepsy associated with schizencephaly.