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1.
Eur J Immunol ; 2020 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-31954378

RESUMO

The B-cell CLL/lymphoma 6 (Bcl6) oncogenic repressor is a master regulator of humoral immunity and B-cell lymphomagenesis. Whereas much research has focused on its regulation and function of germinal center B cells and T cells, the role of Bcl6 in regulating the functions of innate immunity is not well defined. Here, we demonstrated that experimental autoimmune encephalomyelitis (EAE) is exacerbated in LysM Cre+/ - Bcl6fl/fl mice. Although other cells such as neutrophils might be involved in this conditional mutant mouse model, we found that the disease pathology is mainly associated with a biased M1 macrophage activity and an enhanced encephalitogenic CD4+ Th17 cell response. In addition, LPS-induced sepsis mice exhibited an enhanced M1 and inhibited M2 response, further confirming that Bcl6 has an important role in regulating macrophage polarization. Mechanistically, Bcl6 interacts with IκBζ and interferes its binding to the Il-6 (interleukin-6) promotor in macrophages, leading to a suppressed transcription of Il-6. These findings have demonstrated that Bcl6 exerts its regulatory function mainly by repressing Il-6 expression in macrophages. Thus, our study presents a novel role for Bcl6 in regulating immune response and inflammation. Interaction between Bcl6 and IκBζ in macrophages may provide a potential therapeutic target for autoimmune inflammatory disease. This article is protected by copyright. All rights reserved.

3.
Autophagy ; 16(3): 531-547, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31204559

RESUMO

Mutations in the macroautophagy/autophagy gene WDR45 cause ß-propeller protein-associated neurodegeneration (BPAN); however the molecular and cellular mechanism of the disease process is largely unknown. Here we generated constitutive wdr45 knockout (KO) mice that displayed cognitive impairments, abnormal synaptic transmission and lesions in several brain regions. Immunohistochemistry analysis showed loss of neurons in prefrontal cortex and basal ganglion in aged mice, and increased apoptosis in prefrontal cortex, recapitulating a hallmark of neurodegeneration. Quantitative proteomic analysis showed accumulation of endoplasmic reticulum (ER) proteins in KO mouse. At the cellular level, accumulation of ER proteins due to WDR45 deficiency resulted in increased ER stress and impaired ER quality control. The unfolded protein response (UPR) was elevated through ERN1/IRE1 or EIF2AK3/PERK pathway, and eventually led to neuronal apoptosis. Suppression of ER stress or activation of autophagy through MTOR inhibition alleviated cell death. Thus, the loss of WDR45 cripples macroautophagy machinery in neurons and leads to impairment in organelle autophagy, which provides a mechanistic understanding of cause of BPAN and a potential therapeutic strategy to treat this genetic disorder.Abbreviations: 7-ADD: 7-aminoactinomycin D; ASD: autistic spectrum disorder; ATF6: activating transcription factor 6; ATG: autophagy-related; BafA1: bafilomycin A1; BCAP31: B cell receptor associated protein 31; BPAN: ß-propeller protein-associated neurodegeneration; CCCP: carbonyl cyanide m-chlorophenylhydrazone; CDIPT: CDP-diacylglycerol-inositol 3-phosphatidyltransferase (phosphatidylinositol synthase); DDIT3/CHOP: DNA-damage inducible transcript 3; EIF2A: eukaryotic translation initiation factor 2A; EIF2AK3/PERK: eukaryotic translation initiation factor 2 alpha kinase 3; ER: endoplasmic reticulum; ERN1/IRE1: endoplasmic reticulum to nucleus signaling 1; GFP: green fluorescent protein; HIP: hippocampus; HSPA5/GRP78: heat shock protein family A (HSP70) member 5; KO: knockout; LAMP1: lysosomal-associated membrane 1; mEPSCs: miniature excitatory postsynaptic currents; MG132: N-benzyloxycarbonyl-L-leucyl-L-leucyl-L-leucinal; MIB: mid-brain; MTOR: mechanistic target of rapamycin kinase; PCR: polymerase chain reaction; PFA: paraformaldehyde; PFC: prefrontal cortex; PRM: parallel reaction monitoring; RBFOX3/NEUN: RNA binding protein, fox-1 homolog [C. elegans] 3; RTN3: reticulon 3; SEC22B: SEC22 homolog B, vesicle trafficking protein; SEC61B: SEC61 translocon beta subunit; SEM: standard error of the mean; SNR: substantia nigra; SQSTM1/p62: sequestosome 1; TH: tyrosine hydroxylase; Tm: tunicamycin; TMT: tandem mass tag; TUDCA: tauroursodeoxycholic acid; TUNEL: terminal deoxynucleotidyl transferase dUTP nick-end labeling; UPR: unfolded protein response; WDR45: WD repeat domain 45; WT: wild type; XBP1: X-box binding protein 1.

4.
Sheng Wu Gong Cheng Xue Bao ; 35(12): 2295-2307, 2019 Dec 25.
Artigo em Chinês | MEDLINE | ID: mdl-31880137

RESUMO

In the 1960s, scientists first raised the idea of curing genetic diseases using gene therapy. This new conceptual strategy aimed to achieve a much longer therapeutic effect by introducing exogenous genetic materials into the patients. After more than five decades of ups and downs, gene therapy has been brought into a new era by those milestone breakthroughs in the 21st century. Here we reviewed and summarized the history and breakthroughs of gene therapy, including some critical clinical trials, approved drugs, and emerging gene editing techniques. We believe that with their unique advantages over traditional therapies, more gene therapies will become practical approaches to genetic diseases and benefit the entire human race.


Assuntos
Sistemas CRISPR-Cas , Terapia Genética , Edição de Genes , Humanos
5.
Int J Biol Macromol ; 2019 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-31733256

RESUMO

Nanoparticles have been successfully used for immobilization of different enzymes. The enzyme immobilized by nanomaterials has promising application in the biotechnological industry. The ß-glucosidase is very important in industrial field. The immobilization of ß-glucosidase by nanomaterials increases its activity and reusability. In this work, the ß-glucosidase was extracted from fruiting bodies of Agrocybe aegirit and was purified by anion exchange. The ß-glucosidase was immobilized by SiO2 nanoparticles (nano-SiO2) crosslink with glutaraldehyde (GA). On the other hand, the ß-glucosidase firstly immobilized by process adsorption and then crosslink with genipin (GP) which was produced by the hydrolysis of geniposide by ß-glucosidase. Then we compared the properties of both immobilized GA-crosslinked ß-glucosidase and GP-crosslinked ß-glucosidase, their immobilization yields were 83.34% and 96.29% respectively. The GA-crosslinked ß-glucosidase and free ß-glucosidase revealed optimal pH at 6.0. The optimal pH of GP-crosslinked ß-glucosidase was ranging between 4.5 and 7.5. The GA-crosslinked ß-glucosidase and GP-crosslinked ß-glucosidase revealed the optimal temperature at 50 °C, 70 °C respectively. For the free ß-glucosidase, the optimal temperature was 55 °C. Furthermore, the GA-crosslinked ß-glucosidase and GP-crosslinked ß-glucosidase were characterized by scanning electron microscope (SEM). The geniposide consumption in the reaction system of GA-crosslinked ß-glucosidase was analyzed by high performance liquid chromatography (HPLC).

6.
Nat Commun ; 10(1): 4158, 2019 09 13.
Artigo em Inglês | MEDLINE | ID: mdl-31519886

RESUMO

The ubiquitination status of RIPK1 is considered to be critical for cell fate determination. However, the in vivo role for RIPK1 ubiquitination remains undefined. Here we show that mice expressing RIPK1K376R which is defective in RIPK1 ubiquitination die during embryogenesis. This lethality is fully rescued by concomitant deletion of Fadd and Ripk3 or Mlkl. Mechanistically, cells expressing RIPK1K376R are more susceptible to TNF-α induced apoptosis and necroptosis with more complex II formation and increased RIPK1 activation, which is consistent with the observation that Ripk1K376R/K376R lethality is effectively prevented by treatment of RIPK1 kinase inhibitor and is rescued by deletion of Tnfr1. However, Tnfr1-/- Ripk1K376R/K376R mice display systemic inflammation and die within 2 weeks. Significantly, this lethal inflammation is rescued by deletion of Ripk3. Taken together, these findings reveal a critical role of Lys376-mediated ubiquitination of RIPK1 in suppressing RIPK1 kinase activity-dependent lethal pathways during embryogenesis and RIPK3-dependent inflammation postnatally.


Assuntos
Sobrevivência Celular/fisiologia , Desenvolvimento Embrionário/fisiologia , Proteína Serina-Treonina Quinases de Interação com Receptores/metabolismo , Ubiquitinação/fisiologia , Animais , Apoptose/genética , Apoptose/fisiologia , Linhagem Celular , Sobrevivência Celular/genética , Desenvolvimento Embrionário/genética , Feminino , Citometria de Fluxo , Immunoblotting , Imunoprecipitação , Inflamação/genética , Inflamação/metabolismo , Camundongos , Proteínas Quinases/genética , Proteínas Quinases/metabolismo , Proteína Serina-Treonina Quinases de Interação com Receptores/genética , Receptores Tipo I de Fatores de Necrose Tumoral/genética , Receptores Tipo I de Fatores de Necrose Tumoral/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Ubiquitinação/genética
7.
Biochem Biophys Res Commun ; 519(4): 754-760, 2019 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-31547988

RESUMO

Wnt signaling is essential for the maintenance of adult stem cells and its aberrant activation is a stimulator of carcinogenesis. The transmembrane protein, Wntless, is an essential Wnt signaling component through regulating the secretion of Wnt ligands. Here, we generated a mouse model with specific Wntless knockout in intestinal epithelium to study its function in the intestinal epithelium. Wntless knockout exhibits no obvious defects in mice but significantly disrupted proliferation and differentiation of small intestinal organoids. We also discovered that these deficiencies could be partially rescued by Wnt3a supplement but not Wnt9b. To further investigate the role of Wntless in tumorigenesis, APC-deficient spontaneous intestinal tumors and chemical induced colorectal cancer mouse models were employed. To our surprise, intestinal epithelium-specific knockout of Wntless did not cause significant differences in tumor number and size. In summary, our data demonstrated that epithelial Wntless was required for the growth and differentiation of small intestinal organoids but not in live animals, suggesting the other tissues, such as mesenchymal tissue, play critical role for Wnt secretion in both intestinal homeostasis as well as tumorigenesis.

8.
Stem Cells Int ; 2019: 9271746, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31320907
9.
Mol Cell ; 75(4): 823-834.e5, 2019 08 22.
Artigo em Inglês | MEDLINE | ID: mdl-31302001

RESUMO

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.


Assuntos
Cisteína Endopeptidases/metabolismo , Mitocôndrias/metabolismo , Mutação , Obesidade/metabolismo , Transdução de Sinais , Sirtuína 3/metabolismo , Sumoilação , Acetilação , Animais , Cisteína Endopeptidases/genética , Gorduras na Dieta/efeitos adversos , Gorduras na Dieta/farmacologia , Células HEK293 , Humanos , Masculino , Camundongos , Camundongos Mutantes , Mitocôndrias/genética , Mitocôndrias/patologia , Obesidade/induzido quimicamente , Obesidade/genética , Obesidade/patologia , Sirtuína 3/genética
10.
Nat Commun ; 10(1): 2882, 2019 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-31253783

RESUMO

NLR Family CARD Domain Containing 5 (NLRC5), an important immune regulator in innate immunity, is involved in regulating inflammation and antigen presentation. However, the role of NLRC5 in vascular remodeling remains unknown. Here we report the role of NLRC5 on vascular remodeling and provide a better understanding of its underlying mechanism. Nlrc5 knockout (Nlrc5-/-) mice exhibit more severe intimal hyperplasia compared with wild-type mice after carotid ligation. Ex vivo data shows that NLRC5 deficiency leads to increased proliferation and migration of human aortic smooth muscle cells (HASMCs). NLRC5 binds to PPARγ and inhibits HASMC dedifferentiation. NACHT domain of NLRC5 is essential for the interaction with PPARγ and stimulation of PPARγ activity. Pioglitazone significantly rescues excessive intimal hyperplasia in Nlrc5-/- mice and attenuates the increased proliferation and dedifferentiation in NLRC5-deficient HASMCs. Our study demonstrates that NLRC5 regulates vascular remodeling by directly inhibiting SMC dysfunction via its interaction with PPARγ.


Assuntos
Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Túnica Íntima/metabolismo , Animais , Aorta , Apoptose , Pressão Sanguínea , Transplante de Medula Óssea , Proliferação de Células , Células Cultivadas , Citocinas/genética , Citocinas/metabolismo , Frequência Cardíaca , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Camundongos , Camundongos Knockout , Miócitos de Músculo Liso/metabolismo , Plasmídeos , Transcriptoma , Remodelação Vascular
12.
Protein Cell ; 10(9): 700, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31087276

RESUMO

In the original publication the grant number is incorrectly published. The correct grant number should be read as "17140901600". The corrected contents are provided in this correction article. This work was partially supported by grants from the National Natural Science Foundation of China (Nos. 81670470 and 81600149), a grant from the Shanghai Municipal Commission for Science and Technology (17140901600, 18411953500 and 15JC1400201) and a grant from National Key Research and Development Program (2016YFC0905100).

13.
Enzyme Microb Technol ; 122: 1-6, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30638504

RESUMO

Metal-organic framework (MOF) has generated a lot of research interests for it can be employed as immobilization material for enzyme. There are many small molecules in enzyme solution during the extraction process, such as NaCl. It is important to study the effects of the small molecules on MOFs. Here we report a facile and efficient method to encapsulate (R)-1-phenylethanol dehydrogenase ((R)-PEDH) into zeolitic imidazolate framework-8 (ZIF-8). In this work, the effects of NaCl on shape of ZIF-8 and enzyme encapsulation have been investigated. The scanning electron microscope (SEM) results showed that 0.1 M NaCl affect the morphology of ZIF-8 while the crystal structure was not changed analyzed by X-ray diffraction (XRD), and 0.1 M NaCl consisted in the encapsulation system of (R)-PEDH@ZIF-8 enhanced the activity up to 2.5 folds than no NaCl consisted during prepared (R)-PEDH@ZIF-8. The (R)-PEDH@ZIF-8 (prepared with 0.1 M NaCl) enhanced the storage stability and resist ability against trypsin, and the (R)-PEDH@ZIF-8 has been successfully reused.


Assuntos
Enzimas Imobilizadas/química , Imidazóis/química , Oxirredutases/química , Cloreto de Sódio/química , Zeolitas/química , Estabilidade Enzimática , Enzimas Imobilizadas/metabolismo , Estruturas Metalorgânicas/química , Oxirredutases/metabolismo , Tripsina/metabolismo
14.
Curr Mol Med ; 18(7): 436-447, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30539697

RESUMO

BACKGROUND: Primary hyperoxaluria type 1 (PH1) is an inherited disease caused by mutations in alanine-glyoxylate aminotransferase (AGXT). It is characterized by abnormal metabolism of glyoxylic acid in the liver leading to endogenous oxalate overproduction and deposition of oxalate in multiple organs, mainly the kidney. Patients of PH1 often suffer from recurrent urinary tract stones, and finally renal failure. There is no effective treatment other than combined liver-kidney transplantation. METHODS: Microinjection was administered to PH1 rats. Urine samples were collected for urine analysis. Kidney tissues were for Western blotting, quantitative PCR, AGT assays and histological evaluation. RESULTS: In this study, we generated a novel PH1 disease model through CRISPR/Cas9 mediated disruption of mitochondrial localized Agxt gene isoform in rats. Agxt-deficient rats excreted more oxalate in the urine than WT animals. Meanwhile, mutant rats exhibited crystalluria and showed a slight dilatation of renal tubules with mild fibrosis in the kidney. When supplied with 0.4% ethylene glycol (EG) in drinking water, mutant rats excreted greater abundance of oxalate and developed severe nephrocalcinosis in contrast to WT animals. Significantly elevated expression of inflammation- and fibrosisrelated genes was also detected in mutants. CONCLUSION: These data suggest that Agxt-deficiency in mitochondria impairs glyoxylic acid metabolism and leads to PH1 in rats. This rat strain would not only be a useful model for the study of the pathogenesis and pathology of PH1 but also a valuable tool for the development and evaluation of innovative drugs and therapeutics.


Assuntos
Sistemas CRISPR-Cas , Modelos Animais de Doenças , Hiperoxalúria Primária , Nefrocalcinose , Transaminases/deficiência , Animais , Glioxilatos/metabolismo , Hiperoxalúria Primária/genética , Hiperoxalúria Primária/patologia , Hiperoxalúria Primária/urina , Mitocôndrias/genética , Mitocôndrias/metabolismo , Nefrocalcinose/genética , Nefrocalcinose/patologia , Nefrocalcinose/urina , Oxalatos/urina , Ratos , Ratos Transgênicos
15.
EMBO J ; 37(18)2018 09 14.
Artigo em Inglês | MEDLINE | ID: mdl-30111536

RESUMO

Mammalian oocytes and zygotes have the unique ability to reprogram a somatic cell nucleus into a totipotent state. SUV39H1/2-mediated histone H3 lysine-9 trimethylation (H3K9me3) is a major barrier to efficient reprogramming. How SUV39H1/2 activities are regulated in early embryos and during generation of induced pluripotent stem cells (iPSCs) remains unclear. Since expression of the CRL4 E3 ubiquitin ligase in oocytes is crucial for female fertility, we analyzed putative CRL4 adaptors (DCAFs) and identified DCAF13 as a novel CRL4 adaptor that is essential for preimplantation embryonic development. Dcaf13 is expressed from eight-cell to morula stages in both murine and human embryos, and Dcaf13 knockout in mice causes preimplantation-stage mortality. Dcaf13 knockout embryos are arrested at the eight- to sixteen-cell stage before compaction, and this arrest is accompanied by high levels of H3K9me3. Mechanistically, CRL4-DCAF13 targets SUV39H1 for polyubiquitination and proteasomal degradation and therefore facilitates H3K9me3 removal and zygotic gene expression. Taken together, CRL4-DCAF13-mediated SUV39H1 degradation is an essential step for progressive genome reprogramming during preimplantation embryonic development.


Assuntos
Blastocisto/metabolismo , Desenvolvimento Embrionário , Regulação da Expressão Gênica no Desenvolvimento , Células-Tronco Pluripotentes Induzidas/metabolismo , Metiltransferases/metabolismo , Proteínas de Ligação a RNA/metabolismo , Proteínas Repressoras/metabolismo , Animais , Blastocisto/citologia , Estabilidade Enzimática , Histonas/genética , Histonas/metabolismo , Células-Tronco Pluripotentes Induzidas/citologia , Metiltransferases/genética , Camundongos , Camundongos Endogâmicos ICR , Camundongos Knockout , Oócitos/citologia , Oócitos/metabolismo , Proteólise , Proteínas de Ligação a RNA/genética , Proteínas Repressoras/genética , Complexos Ubiquitina-Proteína Ligase/genética , Complexos Ubiquitina-Proteína Ligase/metabolismo , Ubiquitinação/genética
17.
Clin Cancer Res ; 24(23): 5883-5894, 2018 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-30082477

RESUMO

PURPOSE: Pancreatic ductal adenocarcinoma (PDAC) is a heterogeneous disease with variable presentations and natural histories of disease. We hypothesized that different morphologic characteristics of PDAC tumors on diagnostic computed tomography (CT) scans would reflect their underlying biology. EXPERIMENTAL DESIGN: We developed a quantitative method to categorize the PDAC morphology on pretherapy CT scans from multiple datasets of patients with resectable and metastatic disease and correlated these patterns with clinical/pathologic measurements. We modeled macroscopic lesion growth computationally to test the effects of stroma on morphologic patterns, hypothesizing that the balance of proliferation and local migration rates of the cancer cells would determine tumor morphology. RESULTS: In localized and metastatic PDAC, quantifying the change in enhancement on CT scans at the interface between tumor and parenchyma (delta) demonstrated that patients with conspicuous (high-delta) tumors had significantly less stroma, higher likelihood of multiple common pathway mutations, more mesenchymal features, higher likelihood of early distant metastasis, and shorter survival times compared with those with inconspicuous (low-delta) tumors. Pathologic measurements of stromal and mesenchymal features of the tumors supported the mathematical model's underlying theory for PDAC growth. CONCLUSIONS: At baseline diagnosis, a visually striking and quantifiable CT imaging feature reflects the molecular and pathological heterogeneity of PDAC, and may be used to stratify patients into distinct subtypes. Moreover, growth patterns of PDAC may be described using physical principles, enabling new insights into diagnosis and treatment of this deadly disease.


Assuntos
Adenocarcinoma/diagnóstico por imagem , Adenocarcinoma/patologia , Carcinoma Ductal Pancreático/diagnóstico por imagem , Carcinoma Ductal Pancreático/patologia , Tomografia Computadorizada por Raios X , Adenocarcinoma/genética , Adenocarcinoma/terapia , Algoritmos , Biópsia , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/terapia , Linhagem Celular Tumoral , Terapia Combinada , Análise Mutacional de DNA , Humanos , Processamento de Imagem Assistida por Computador , Imuno-Histoquímica , Modelos Teóricos , Gradação de Tumores , Metástase Neoplásica , Estadiamento de Neoplasias , Carga Tumoral , Sequenciamento Completo do Exoma
18.
Aging (Albany NY) ; 10(4): 592-605, 2018 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-29676997

RESUMO

Glioblastoma (GBM) is one of the most deadly brain tumors. The convenient access to The Cancer Genome Atlas (TCGA) database allows for large-scale global gene expression profiling and database mining for potential correlation between genes and overall survival of a variety of malignancies including GBM. Previous reports have shown that tumor microenvironment cells and the extent of infiltrating immune and stromal cells in tumors contribute significantly to prognosis. Immune scores and stromal scores calculated based on the ESTIMATE algorithm could facilitate the quantification of the immune and stromal components in a tumor. To better understand the effects of genes involved in immune and stromal cells on prognosis, we categorized GBM cases in the TCGA database according to their immune/stromal scores into high and low score groups, and identified differentially expressed genes whose expression was significantly associated with prognosis in GBM patients. Functional enrichment analysis and protein-protein interaction networks further showed that these genes mainly participated in immune response, extracellular matrix, and cell adhesion. Finally, we validated these genes in an independent GBM cohort from the Chinese Glioma Genome Atlas (CGGA). Thus, we obtained a list of tumor microenvironment-related genes that predict poor outcomes in GBM patients.


Assuntos
Neoplasias Encefálicas/genética , Mineração de Dados/métodos , Glioblastoma/genética , Microambiente Tumoral/genética , Algoritmos , Neoplasias Encefálicas/imunologia , Neoplasias Encefálicas/patologia , Bases de Dados Genéticas , Feminino , Perfilação da Expressão Gênica/métodos , Glioblastoma/imunologia , Glioblastoma/patologia , Humanos , Estimativa de Kaplan-Meier , Masculino , Prognóstico , Transcriptoma , Microambiente Tumoral/imunologia
19.
J Biol Chem ; 293(18): 6883-6892, 2018 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-29507093

RESUMO

Hereditary tyrosinemia type I (HTI) is a metabolic genetic disorder caused by mutation of fumarylacetoacetate hydrolase (FAH). Because of the accumulation of toxic metabolites, HTI causes severe liver cirrhosis, liver failure, and even hepatocellular carcinoma. HTI is an ideal model for gene therapy, and several strategies have been shown to ameliorate HTI symptoms in animal models. Although CRISPR/Cas9-mediated genome editing is able to correct the Fah mutation in mouse models, WT Cas9 induces numerous undesired mutations that have raised safety concerns for clinical applications. To develop a new method for gene correction with high fidelity, we generated a Fah mutant rat model to investigate whether Cas9 nickase (Cas9n)-mediated genome editing can efficiently correct the Fah First, we confirmed that Cas9n rarely induces indels in both on-target and off-target sites in cell lines. Using WT Cas9 as a positive control, we delivered Cas9n and the repair donor template/single guide (sg)RNA through adenoviral vectors into HTI rats. Analyses of the initial genome editing efficiency indicated that only WT Cas9 but not Cas9n causes indels at the on-target site in the liver tissue. After receiving either Cas9n or WT Cas9-mediated gene correction therapy, HTI rats gained weight steadily and survived. Fah-expressing hepatocytes occupied over 95% of the liver tissue 9 months after the treatment. Moreover, CRISPR/Cas9-mediated gene therapy prevented the progression of liver cirrhosis, a phenotype that could not be recapitulated in the HTI mouse model. These results strongly suggest that Cas9n-mediated genome editing is a valuable and safe gene therapy strategy for this genetic disease.


Assuntos
Proteína 9 Associada à CRISPR/metabolismo , Desoxirribonuclease I/metabolismo , Edição de Genes , Terapia Genética/métodos , Tirosinemias/genética , Adenoviridae/genética , Animais , Modelos Animais de Doenças , Feminino , Vetores Genéticos , Células HEK293 , Hepatócitos/citologia , Humanos , Hidrolases/genética , Mutação INDEL , Cirrose Hepática/etiologia , Cirrose Hepática/prevenção & controle , Masculino , Ratos , Tirosinemias/complicações , Tirosinemias/imunologia , Tirosinemias/terapia
20.
Materials (Basel) ; 11(4)2018 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-29587358

RESUMO

Surface texture (ST) has been confirmed as an effective and economical surface treatment technique that can be applied to a great range of materials and presents growing interests in various engineering fields. Ti6Al4V which is the most frequently and successfully used titanium alloy has long been restricted in tribological-related operations due to the shortcomings of low surface hardness, high friction coefficient, and poor abrasive wear resistance. Ti6Al4V has benefited from surface texture-based surface treatments over the last decade. This review begins with a brief introduction, analysis approaches, and processing methods of surface texture. The specific applications of the surface texture-based surface treatments for improving surface performance of Ti6Al4V are thoroughly reviewed from the point of view of tribology and biology.

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