RESUMO
Cisplatin (CDDP) is a platinum-based drug with anti-cancer activity and is widely used as a standard therapy for bladder cancer. It is well known that CDDP causes cell death by increasing the generation of reactive oxygen species (ROS) and lipid peroxidation, but the mechanism of its anti-cancer effects has not been fully elucidated. There are still some problems such as chemoresistance in CDDP therapy. In the present study, we found the expression of Ca2+-independent phospholipase A2γ (iPLA2γ), which has been reported to regulate cellular redox homeostasis by inhibiting lipid peroxide accumulation, in human bladder cancer tissues. Thus, we investigated the effect of iPLA2γ knockdown on CDDP-induced bladder cancer cell death. As a result, we found that iPLA2γ knockdown significantly enhanced CDDP-induced apoptosis, intracellular and mitochondrial ROS production, cytochrome c release and caspase activation in bladder cancer cells. Moreover, mitochondrial membrane potential was decreased and peroxidation of mitochondrial phospholipids was increased by iPLA2γ knockdown. It was also shown that co-treatment of bromoenol lactone, an iPLA2 inhibitor, increased CDDP-induced apoptosis. These results indicated that iPLA2γ plays an important role in protecting bladder cancer cells from CDDP-induced apoptosis, and that iPLA2γ inhibitors might represent a novel strategy in CDDP-based multi-drug therapy.
Assuntos
Apoptose , Cisplatino , Fosfolipases A2 do Grupo VI , Peroxidação de Lipídeos , Mitocôndrias , Fosfolipídeos , Espécies Reativas de Oxigênio , Neoplasias da Bexiga Urinária , Humanos , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/patologia , Neoplasias da Bexiga Urinária/metabolismo , Neoplasias da Bexiga Urinária/tratamento farmacológico , Apoptose/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Cisplatino/farmacologia , Mitocôndrias/metabolismo , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/genética , Fosfolipases A2 do Grupo VI/metabolismo , Fosfolipases A2 do Grupo VI/genética , Peroxidação de Lipídeos/efeitos dos fármacos , Linhagem Celular Tumoral , Fosfolipídeos/metabolismo , Antineoplásicos/farmacologia , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Pironas/farmacologia , NaftalenosRESUMO
Neurodegeneration with brain iron accumulation (NBIA) encompasses a clinically and genetically heterogeneous group of rare disorders. Here, we report clinical, neuroimaging and genetic studies in twenty three Brazilian NBIA patients. In thirteen subjects, deleterious variants were detected in known NBIA-causing genes (PANK2, PLA2G6, C9ORF12, WDR45 and FA2H), including previously unreported variants in PANK2 and PLA2G6. Two patients carried rare, likely pathogenic variants in genes not previously associated with NBIA: KMT2A c.11785A > C (p.Ile3929Leu), and TIMM8A c.127T > C (p.Cys43Arg), suggesting an expansion of their associated phenotypes to include NBIA. In eight patients the etiology remains unsolved, suggesting variants undetectable by the adopted methods, or the existence of additional NBIA-causing genes.
Assuntos
Neuroimagem , Humanos , Brasil , Feminino , Masculino , Adulto , Adolescente , Adulto Jovem , Criança , Pré-Escolar , Imageamento por Ressonância Magnética , Encéfalo/diagnóstico por imagem , Encéfalo/metabolismo , Encéfalo/patologia , Distrofias Neuroaxonais/genética , Distrofias Neuroaxonais/diagnóstico por imagem , Fosfotransferases (Aceptor do Grupo Álcool)/genética , Ferro/metabolismo , Distúrbios do Metabolismo do Ferro/genética , Distúrbios do Metabolismo do Ferro/diagnóstico por imagem , Fosfolipases A2 do Grupo VIAssuntos
Lissencefalias Clássicas e Heterotopias Subcorticais em Banda , Distrofias Neuroaxonais , Humanos , Ferro , Imageamento por Ressonância Magnética/métodos , Distrofias Neuroaxonais/diagnóstico por imagem , Distrofias Neuroaxonais/genética , Encéfalo/diagnóstico por imagem , Fosfolipases A2 do Grupo VI/genéticaRESUMO
Infantile neuroaxonal dystrophy (INAD) is caused by recessive variants in PLA2G6 and is a lethal pediatric neurodegenerative disorder. Loss of the Drosophila homolog of PLA2G6, leads to ceramide accumulation, lysosome expansion, and mitochondrial defects. Here, we report that retromer function, ceramide metabolism, the endolysosomal pathway, and mitochondrial morphology are affected in INAD patient-derived neurons. We show that in INAD mouse models, the same features are affected in Purkinje cells, arguing that the neuropathological mechanisms are evolutionary conserved and that these features can be used as biomarkers. We tested 20 drugs that target these pathways and found that Ambroxol, Desipramine, Azoramide, and Genistein alleviate neurodegenerative phenotypes in INAD flies and INAD patient-derived neural progenitor cells. We also develop an AAV-based gene therapy approach that delays neurodegeneration and prolongs lifespan in an INAD mouse model.
Assuntos
Proteínas de Drosophila , Distrofias Neuroaxonais , Transtornos Parkinsonianos , Camundongos , Animais , Neurônios/metabolismo , Transtornos Parkinsonianos/metabolismo , Drosophila/metabolismo , Ceramidas/metabolismo , Distrofias Neuroaxonais/genética , Distrofias Neuroaxonais/metabolismo , Distrofias Neuroaxonais/patologia , Fosfolipases A2 do Grupo VI/metabolismo , Proteínas do Olho/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismoRESUMO
Polymorphisms of phospholipase A2VIA (iPLA2ß or PLA2G6) are associated with body weights and blood C-reactive protein. The role of iPLA2ß/PLA2G6 in non-alcoholic steatohepatitis (NASH) is still elusive because female iPla2ß-null mice showed attenuated hepatic steatosis but exacerbated hepatic fibrosis after feeding with methionine- and choline-deficient diet (MCDD). Herein, female mice with myeloid- (MPla2g6-/-) and hepatocyte- (LPla2g6-/-) specific PLA2G6 deletion were generated and phenotyped after MCDD feeding. Without any effects on hepatic steatosis, MCDD-fed MPla2g6-/- mice showed further exaggeration of liver inflammation and fibrosis as well as elevation of plasma TNFα, CCL2, and circulating monocytes. Bone-marrow-derived macrophages (BMDMs) from MPla2g6-/- mice displayed upregulation of PPARγ and CEBPα proteins, and elevated release of IL6 and CXCL1 under LPS stimulation. LPS-stimulated BMDMs from MCDD-fed MPla2g6-/- mice showed suppressed expression of M1 Tnfa and Il6, but marked upregulation of M2 Arg1, Chil3, IL10, and IL13 as well as chemokine receptors Ccr2 and Ccr5. This in vitro shift was associated with exaggeration of hepatic M1/M2 cytokines, chemokines/chemokine receptors, and fibrosis genes. Contrarily, MCDD-fed LPla2g6-/- mice showed a complete protection which was associated with upregulation of Ppara/PPARα and attenuated expression of Pparg/PPARγ, fatty-acid uptake, triglyceride synthesis, and de novo lipogenesis genes. Interestingly, LPla2g6-/- mice fed with chow or MCDD displayed an attenuation of blood monocytes and elevation of anti-inflammatory lipoxin A4 in plasma and liver. Thus, PLA2G6 inactivation specifically in myeloid cells and hepatocytes led to opposing phenotypes in female mice undergoing NASH. Hepatocyte-specific PLA2G6 inhibitors may be further developed for treatment of this disease.
Assuntos
Hepatopatia Gordurosa não Alcoólica , Feminino , Camundongos , Animais , Hepatopatia Gordurosa não Alcoólica/genética , Fosfolipases A2 Independentes de Cálcio , PPAR gama/genética , Interleucina-6 , Lipopolissacarídeos , Dieta , Hepatócitos , Fenótipo , Metionina , Colina , Racemetionina , PPAR alfa , Receptores de Quimiocinas , Fosfolipases A2 do Grupo VI/genéticaRESUMO
BACKGROUND: PLA2G6-Associated Neurodegeneration (PLAN) is a rare neurodegenerative disease with autosomal recessive inheritance, which belongs to the NBIA (Neurodegeneration with Brain Iron Accumulation) group. Although the pathogenesis of the disease remains largely unclear, lipid peroxidation seems to play a central role in the pathogenesis. Currently, there is no cure for the disease. OBJECTIVE: In this work, we examined the presence of lipid peroxidation, iron accumulation and mitochondrial dysfunction in two cellular models of PLAN, patients-derived fibroblasts and induced neurons, and assessed the effects of α-tocopherol (vitamin E) in correcting the pathophysiological alterations in PLAN cell cultures. METHODS: Pathophysiological alterations were examined in fibroblasts and induced neurons generated by direct reprograming. Iron and lipofuscin accumulation were assessed using light and electron microscopy, as well as biochemical analysis techniques. Reactive Oxygen species production, lipid peroxidation and mitochondrial dysfunction were measured using specific fluorescent probes analysed by fluorescence microscopy and flow cytometry. RESULTS: PLAN fibroblasts and induced neurons clearly showed increased lipid peroxidation, iron accumulation and altered mitochondrial membrane potential. All these pathological features were reverted with vitamin E treatment. CONCLUSIONS: PLAN fibroblasts and induced neurons reproduce the main pathological alterations of the disease and provide useful tools for disease modelling. The main pathological alterations were corrected by Vitamin E supplementation in both models, suggesting that blocking lipid peroxidation progression is a critical therapeutic target.
Assuntos
Distrofias Neuroaxonais , Doenças Neurodegenerativas , Fosfolipases A2 do Grupo VI/metabolismo , Humanos , Ferro/metabolismo , Peroxidação de Lipídeos , Mitocôndrias/metabolismo , Distrofias Neuroaxonais/metabolismo , Distrofias Neuroaxonais/patologia , Doenças Neurodegenerativas/metabolismo , Vitamina E/metabolismo , Vitamina E/farmacologiaRESUMO
PLA2G6 is the causative gene for a group of autosomal recessive neurodegenerative disorders known as PLA2G6-associated neurodegeneration (PLAN). We present a case with early-onset parkinsonism, ataxia, cognitive decline, cerebellar atrophy, and brain iron accumulation. Sequencing of PLA2G6 coding regions identified only a heterozygous nonsense variant, but mRNA analysis revealed the presence of an aberrant transcript isoform due to a novel deep intronic variant (c.2035-274G > A) leading to activation of an intronic pseudo-exon. These results expand the genotypic spectrum of PLAN, showing the paramount importance of detecting possible pathogenic variants in deep intronic regions in undiagnosed patients.
Assuntos
Encéfalo/patologia , Fosfolipases A2 do Grupo VI/genética , Mutação/genética , Doença de Parkinson/genética , Adulto , Idade de Início , Atrofia/patologia , Feminino , Humanos , Malformações do Sistema Nervoso/genética , Distrofias Neuroaxonais/genética , Doenças Neurodegenerativas/genética , Doenças Neurodegenerativas/patologia , Doença de Parkinson/diagnóstico , Doença de Parkinson/patologia , FenótipoRESUMO
Here, we identify iPLA2ß as a critical regulator for p53-driven ferroptosis upon reactive oxygen species (ROS)-induced stress. The calcium-independent phospholipase iPLA2ß is known to cleave acyl tails from the glycerol backbone of lipids and release oxidized fatty acids from phospholipids. We found that iPLA2ß-mediated detoxification of peroxidized lipids is sufficient to suppress p53-driven ferroptosis upon ROS-induced stress, even in GPX4-null cells. Moreover, iPLA2ß is overexpressed in human cancers; inhibition of endogenous iPLA2ß sensitizes tumor cells to p53-driven ferroptosis and promotes p53-dependent tumor suppression in xenograft mouse models. These results demonstrate that iPLA2ß acts as a major ferroptosis repressor in a GPX4-independent manner. Notably, unlike GPX4, loss of iPLA2ß has no obvious effect on normal development or cell viability in normal tissues but iPLA2ß plays an essential role in regulating ferroptosis upon ROS-induced stress. Thus, our study suggests that iPLA2ß is a promising therapeutic target for activating ferroptosis-mediated tumor suppression without serious toxicity concerns.
Assuntos
Ferroptose/fisiologia , Fosfolipases A2 do Grupo VI/metabolismo , Fosfolipídeo Hidroperóxido Glutationa Peroxidase/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Células A549 , Animais , Linhagem Celular Tumoral , Sobrevivência Celular , Modelos Animais de Doenças , Ácidos Graxos/metabolismo , Feminino , Ferroptose/genética , Fosfolipases A2 do Grupo VI/genética , Humanos , Camundongos , Camundongos Nus , Fosfolipídeo Hidroperóxido Glutationa Peroxidase/genética , Fosfolipídeos , Espécies Reativas de Oxigênio/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
The Ca2+-independent phospholipase A2ß (iPLA2ß) is a member of the PLA2 family that has been proposed to have roles in multiple biological processes including membrane remodeling, cell proliferation, bone formation, male fertility, cell death, and signaling. Such involvement has led to the identification of iPLA2ß activation in several diseases such as cancer, cardiovascular abnormalities, glaucoma, periodontitis, neurological disorders, diabetes, and other metabolic disorders. More recently, there has been heightened interest in the role that iPLA2ß plays in promoting inflammation. Recognizing the potential contribution of iPLA2ß in the development of autoimmune diseases, we review this issue in the context of an iPLA2ß link with macrophages and T-cells.
Assuntos
Fosfolipases A2 do Grupo VI/metabolismo , Imunidade Inata , Animais , Humanos , Inflamação , Linfócitos/imunologia , Macrófagos/imunologiaAssuntos
Cerebelo/anormalidades , Fosfolipases A2 do Grupo VI/genética , Mutação , Miopatias Congênitas Estruturais/genética , Adolescente , Alelos , Biópsia , Humanos , Masculino , Miopatias Congênitas Estruturais/enzimologia , Miopatias Congênitas Estruturais/patologia , Irmãos , Sequenciamento Completo do GenomaRESUMO
Eosinophils mediate pathological manifestations during tropical pulmonary eosinophilia (TPE), a potentially fatal complication of lymphatic filariasis, by mechanisms that are incompletely understood. Using two-dimensional gel electrophoresis, mass spectrometry, flow cytometry, and pharmacological and functional studies, we identified acidic calcium-independent phospholipase A2 (aiPLA2) as the master regulator of TPE pathogenesis. FACS-sorted lung eosinophils from TPE mice exhibited aiPLA2-dependent activation characterized by heavy calcium influx, F-actin polymerization, increased degranulation, and heightened reactive oxygen species generation. Interestingly, aiPLA2 also promoted alternative activation in lung macrophages and regulated the release of inflammatory intermediates from them. Treatment of TPE mice with MJ33, a nontoxic pharmacological inhibitor of aiPLA2, lowered eosinophil counts in the bronchoalveolar lavage fluid, reduced eosinophil peroxidase and ß-hexosaminidase activity, increased airway width, improved lung endothelial barrier, and lowered the production of inflammatory lipid intermediates, which significantly improved the pathological condition of the lungs. Importantly, ex vivo reconstitution of arachidonic acid to eosinophils from MJ33-treated TPE mice increased eosinophil degranulation and inflammatory lipid intermediates underlining the pivotal role of aiPLA2 in arachidonic acid metabolism. Mechanistically, phosphorylation of JNK-1 regulated phospholipase activity of aiPLA2, whereas IgG cross-linking mediated pathological activation of eosinophils. Taken together, ours is the first study, to our knowledge, to report hitherto undocumented role of aiPLA2 in regulating TPE pathogenesis.
Assuntos
Brugia Malayi/imunologia , Filariose Linfática/imunologia , Eosinófilos/imunologia , Fosfolipases A2 do Grupo VI/imunologia , Macrófagos/imunologia , Eosinofilia Pulmonar/imunologia , Animais , Modelos Animais de Doenças , Filariose Linfática/patologia , Eosinófilos/patologia , Macrófagos/patologia , Camundongos , Camundongos Endogâmicos BALB C , Eosinofilia Pulmonar/parasitologia , Eosinofilia Pulmonar/patologiaRESUMO
BACKGROUND: To elucidate the role of iPLA2/PLA2G6 in gingivobuccal squamous cell carcinoma (GB-SCC) and to ascertain the synthetic lethality-based chemoprevention role of aspirin in arachidonic acid metabolism (AAM) pathway down-regulated GB-SCC. METHODS: The in vitro efficacy of aspirin on GB-SCC cells (ITOC-03 and ITOC-04) was assessed by cell proliferation, colony formation, apoptosis, cell migration, cell cycle assay and RNA-seq, while inhibition of PLA2G6 and AAM pathway components was affirmed by qPCR, Western blot and immunofluorescence staining. The in vivo effect of aspirin was evaluated using NOD-SCID mice xenografts and immunohistochemical analysis. RESULTS: We found that aspirin, which has been reported to act through the COX pathway, is inhibiting PLA2G6, and thereby the COX and LOX components of the AAM pathway. The findings were validated using PLA2G6 siRNA and immunohistochemical marker panel. Moreover, a pronounced effect in ITOC-04 cells and xenografts implied aspirin-induced synthetic lethality in the AAM pathway down-regulated GB-SCC. CONCLUSIONS: This study reveals that aspirin induces the anti-tumor effect by a previously unrecognized mechanism of PLA2G6 inhibition. In addition, the effect of aspirin is influenced by the baseline AAM pathway status and could guide precision prevention clinical trials of AAM pathway inhibitors.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Aspirina/uso terapêutico , Carcinoma de Células Escamosas/tratamento farmacológico , Neoplasias Gengivais/tratamento farmacológico , Fosfolipases A2 do Grupo VI/efeitos dos fármacos , Mutações Sintéticas Letais/efeitos dos fármacos , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Aspirina/farmacologia , Linhagem Celular Tumoral , Proliferação de Células , Modelos Animais de Doenças , Regulação para Baixo , Humanos , Camundongos , Camundongos SCID , Prognóstico , TransfecçãoRESUMO
Inflammasomes have been implicated in the detection and clearance of a variety of bacterial pathogens, but little is known about whether this innate sensing mechanism has any regulatory effect on the expression of stimulatory ligands by the pathogen. During infection with Salmonella and many other pathogens, flagellin is a major activator of NLRC4 inflammasome-mediated macrophage pyroptosis and pathogen eradication. Salmonella switches to a flagellin-low phenotype as infection progresses to avoid this mechanism of clearance by the host. However, the host cues that Salmonella perceives to undergo this switch remain unclear. Here, we report an unexpected role of the NLRC4 inflammasome in promoting expression of its microbial ligand, flagellin, and identify a role for type 1 IFN signaling in switching of Salmonella to a flagellin-low phenotype. Early in infection, activation of NLRC4 by flagellin initiates pyroptosis and concomitant release of lysophospholipids which in turn enhance expression of flagellin by Salmonella thereby amplifying its ability to elicit cell death. TRIF-dependent production of type 1 IFN, however, later represses NLRC4 and the lysophospholipid biosynthetic enzyme iPLA2, causing a decline in intracellular lysophospholipids that results in down-regulation of flagellin expression by Salmonella These findings reveal a previously unrecognized immune-modulating regulatory cross-talk between endosomal TLR signaling and cytosolic NLR activation with significant implications for the establishment of infection with Salmonella.
Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Proteínas de Ligação ao Cálcio/metabolismo , Flagelina/metabolismo , Fosfolipases A2 do Grupo VI/metabolismo , Interferon Tipo I/metabolismo , Infecções por Salmonella/imunologia , Salmonella typhimurium/imunologia , Proteínas Adaptadoras de Transporte Vesicular/genética , Proteínas Adaptadoras de Transporte Vesicular/metabolismo , Animais , Proteínas Reguladoras de Apoptose/genética , Proteínas de Ligação ao Cálcio/genética , Células Cultivadas , Modelos Animais de Doenças , Regulação para Baixo , Flagelina/imunologia , Fosfolipases A2 do Grupo VI/antagonistas & inibidores , Humanos , Imunidade Inata , Inflamassomos/efeitos dos fármacos , Inflamassomos/imunologia , Inflamassomos/metabolismo , Cetonas/administração & dosagem , Lipopolissacarídeos/administração & dosagem , Lipopolissacarídeos/imunologia , Lisofosfolipídeos/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos , Camundongos Knockout , Naftalenos/administração & dosagem , Cultura Primária de Células , Piroptose/imunologia , Infecções por Salmonella/microbiologia , Salmonella typhimurium/isolamento & purificação , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologiaRESUMO
To examine the role of group VIA phospholipase A2 (iPLA2ß) in specific cell lineages in insulin secretion and insulin action, we prepared mice with a selective iPLA2ß deficiency in cells of myelomonocytic lineage, including macrophages (MØ-iPLA2ß-KO), or in insulin-secreting ß-cells (ß-Cell-iPLA2ß-KO), respectively. MØ-iPLA2ß-KO mice exhibited normal glucose tolerance when fed standard chow and better glucose tolerance than floxed-iPLA2ß control mice after consuming a high-fat diet (HFD). MØ-iPLA2ß-KO mice exhibited normal glucose-stimulated insulin secretion (GSIS) in vivo and from isolated islets ex vivo compared to controls. Male MØ-iPLA2ß-KO mice exhibited enhanced insulin responsivity vs. controls after a prolonged HFD. In contrast, ß-cell-iPLA2ß-KO mice exhibited impaired glucose tolerance when fed standard chow, and glucose tolerance deteriorated further when introduced to a HFD. ß-Cell-iPLA2ß-KO mice exhibited impaired GSIS in vivo and from isolated islets ex vivo vs. controls. ß-Cell-iPLA2ß-KO mice also exhibited an enhanced insulin responsivity compared to controls. These findings suggest that MØ iPLA2ß participates in HFD-induced deterioration in glucose tolerance and that this mainly reflects an effect on insulin responsivity rather than on insulin secretion. In contrast, ß-cell iPLA2ß plays a role in GSIS and also appears to confer some protection against deterioration in ß-cell functions induced by a HFD.
Assuntos
Fosfolipases A2 do Grupo VI/genética , Células Secretoras de Insulina/metabolismo , Fosfolipases A2/genética , Animais , Glicemia/genética , Dieta Hiperlipídica/efeitos adversos , Glucose/genética , Intolerância à Glucose/tratamento farmacológico , Intolerância à Glucose/genética , Teste de Tolerância a Glucose , Humanos , Insulina/genética , Insulina/metabolismo , Secreção de Insulina/genética , Ilhotas Pancreáticas/metabolismo , Ilhotas Pancreáticas/patologia , Macrófagos/efeitos dos fármacos , Camundongos , Camundongos Knockout , Fosfolipases A2/deficiênciaRESUMO
OBJECTIVE: To identify potential variant in a child diagnosed as infantile neuroaxonal dystrophy. METHODS: Genomic DNA was extracted from peripheral blood samples from the patient and his parents and subjected to next generation sequencing. Suspected variant was verified by PCR and Sanger sequencing. Pathogenicity of the mutation was predicted by using bioinformatic software including SIFT and PolyPhen-2. RESULTS: The child was found to carry compound heterozygous variations c.668C>A (p.Pro223Gln) and c.2266C>T (p.Gln756Ter) of the PLA2G6 gene, which were respectively inherited from his father and mother. c.2266C>T has changed codon 756 (glutamine) into a stop codon, resulting premature termination of peptide chain synthesis. c.2266C>T has not been reported previously and was predicted to be harmful. CONCLUSION: The compound variants of c.668C>A (p.Pro223Gln) and c.2266C>T (p.Gln756Ter) of the PLA2G6 gene probably underlies the disease in the child. Above finding has enriched the variant spectrum of the PLA2G6 gene.
Assuntos
Fosfolipases A2 do Grupo VI , Distrofias Neuroaxonais , Criança , Fosfolipases A2 do Grupo VI/genética , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Mutação , Distrofias Neuroaxonais/genéticaRESUMO
We investigated the relationship between the rs10798059 (BanI) and rs4375 polymorphisms in the phospholipase A2 (PLA2)G4A and PLA2G6 genes and the risk of nicotine dependence in 263 Croatian patients with schizophrenia. We also examined whether interactions between these polymorphisms and smoking contributed to schizophrenia onset and Positive and Negative Syndrome Scale (PANSS) psychopathology. We found no significant differences in the distribution of PLA2G4A genotypes and alleles according to smoking status, and no effect of the PLA2G4A genotype-smoking interaction on disease onset or PANSS. The PLA2G6-TT homozygous genotype was significantly overrepresented in male smokers compared to nonsmokers (34.7% vs. 17.1%, p < 0.05). These patients had â¼2.6-fold higher risk of becoming smokers than males with heterozygous PLA2G6-CT and homozygous PLA2G6-CC genotypes. In addition, male smokers without the PLA2G6-C allele (PLA2G6-TT homozygous) experienced earlier onset than nonsmoking homozygous PLA2G6-TT males. Thus, the PLA2G6 polymorphism affected the risk of nicotine dependence in male patients and the PLA2G6 genotype-smoking interaction was linked to the age of disease onset.
Assuntos
Fosfolipases A2 do Grupo VI/genética , Polimorfismo de Nucleotídeo Único , Esquizofrenia/genética , Tabagismo/epidemiologia , Adulto , Idade de Início , Croácia , Feminino , Estudos de Associação Genética , Fosfolipases A2 do Grupo IV/genética , Humanos , Masculino , Pessoa de Meia-Idade , Caracteres Sexuais , Tabagismo/genéticaRESUMO
NBIA (Neurodegeneration with brain iron accumulation) is a group of inherited neurologic disorders characterized by marked genetic heterogeneity, in which iron atypical accumulates in basal ganglia resulting in brain magnetic resonance imaging changes, histopathological abnormalities, and neuropsychiatric clinical symptoms. With the rapid development of high-throughput sequencing technologies, ten candidate genes have been identified, including PANK2, PLA2G6, C19orf12, WDR45, FA2H, ATP13A2, FTL, CP, C2orf37, and COASY. They are involved in seemingly unrelated cellular pathways, such as iron homeostasis (FTL, CP), lipid metabolism (PLA2G6, C19orf12, FA2H), Coenzyme A synthesis (PANK2, COASY), and autophagy (WDR45, ATP13A2). In particular, PANK2, COASY, PLA2G6, and C19orf12 are located on mitochondria, which associate with certain subtypes of NBIA showing mitochondria dysregulation. However, the relationships among those four genes are still unclear. Therefore, this review is specifically focused on dysregulation of mitochondria in NBIA and afore-mentioned four genes, with summaries of both pathological and clinical findings.
Assuntos
Fosfolipases A2 do Grupo VI/genética , Distúrbios do Metabolismo do Ferro/genética , Mitocôndrias/patologia , Proteínas Mitocondriais/genética , Distrofias Neuroaxonais/genética , Fosfotransferases (Aceptor do Grupo Álcool)/genética , Transferases/genética , Humanos , Distúrbios do Metabolismo do Ferro/metabolismo , Distúrbios do Metabolismo do Ferro/patologia , Metabolismo dos Lipídeos/genética , Potencial da Membrana Mitocondrial/genética , Distrofias Neuroaxonais/metabolismo , Distrofias Neuroaxonais/patologiaRESUMO
Increased activity of secretory phospholipases A2 (sPLA2) type-II was previously observed in ileum of Crohn's disease (CD). Our aims were to explore the involvement of calcium-independent (i)PLA2ß in the release of sPLA2s from the human mast cell (MC) line (HMC-1) and investigate expressions of cytosolic (c)PLA2α, iPLA2ß, sPLA2-IIA and sPLA2-V in MCs of CD ileum. The release of sPLA2 was investigated in HMC-1 by immunocytochemistry and ELISA. The expression intensities of PLA2s in mucosal MCs, and the proportion of PLA2-positive MCs, were investigated in normal ileum and in ileum from patients with CD by immunohistochemistry. The calcium ionophore-stimulated release of sPLA2-IIA and sPLA2-V from HMC-1 was reduced by the iPLA2-inhibitor bromoenol lactone. All four PLA2s were detectable in mucosal MCs, both in normal ileum and in CD, but the proportion of iPLA2ß-containing mucosal MCs and the expression intensity of sPLA2-IIA was increased in CD. Results indicate that iPLA2ß is involved in the secretion of sPLA2s from HMC-1, and suggest that iPLA2ß-mediated release of sPLA2 from intestinal MCs may contribute to CD pathophysiology. Ex vivo studies on isolated mucosal mast cells are however needed to clarify the precise role of MC PLA2s in the inflammatory processes of CD.
Assuntos
Doença de Crohn/imunologia , Fosfolipases A2 do Grupo VI/metabolismo , Mastócitos/imunologia , Fosfolipases A2 Secretórias/metabolismo , Adulto , Idoso , Ionóforos de Cálcio/farmacologia , Linhagem Celular Tumoral , Doença de Crohn/patologia , Feminino , Humanos , Íleo/citologia , Íleo/imunologia , Íleo/patologia , Mucosa Intestinal/imunologia , Mucosa Intestinal/patologia , Masculino , Mastócitos/efeitos dos fármacos , Mastócitos/metabolismo , Pessoa de Meia-IdadeRESUMO
INTRODUCTION: This study reports a novel mutation site of the phospholipase A2 group VI (PLA2G6) gene, and analyzes the information of 67 previously published cases to elucidate PLA2G6 phenotype-genotype variations. METHODS: We collected clinical data and examined gene mutation sites from one Chinese patient with adult-onset ataxia and her family. Next-generation sequencing (NGS) and Sanger sequencing were used to verify possible mutations. PolyPhen-2, SIFT, and MutationTaster were used to predict their pathogenicity. For analyzing the distribution frequency of the mutation, 597 healthy controls were recruited. We also analyzed the clinical and genetic information of 67 cases from 23 studies in Pubmed database. RESULTS: A novel compound heterozygous mutation of the PLA2G6 gene, c.1648delC and c.991Gâ¯>â¯T, was found in the Chinese patient, and classified as pathogenic. The c.1648delC variation was absent in ExAC, 1000G, dbSNP databases and the 597 healthy controls. Of the 67 cases, 29 presented ataxia. The signs of cerebellar atrophy appeared in the MRIs of most patients, while signs of iron accumulation were absent in older-aged patients with a compound heterozygous mutation. Thirty-eight patients showed no ataxia. A negative or mild extrapyramidal symptom accompanied by a low age, a homogenous mutation, while moderate or severe extrapyramidal symptoms were associated with an old age and a compound heterozygous mutation. CONCLUSION: A novel compound heterozygous mutation of the PLA2G6 gene, c.1648delC and c.991Gâ¯>â¯T, is associated with adult onset ataxia. Phenotype-genotype variations of PLA2G6 are predicted to be caused by the loss of protein or enzyme activity of phospholipase-2.
Assuntos
Ataxia/genética , Fosfolipases A2 do Grupo VI/genética , Doenças Neurodegenerativas/genética , Adolescente , Adulto , Fatores Etários , Idade de Início , Idoso , Idoso de 80 Anos ou mais , Ataxia/patologia , Ataxia/fisiopatologia , Atrofia/patologia , Cerebelo/diagnóstico por imagem , Cerebelo/patologia , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Doenças Neurodegenerativas/patologia , Doenças Neurodegenerativas/fisiopatologia , Linhagem , Fenótipo , Adulto JovemRESUMO
A recessive mutation in PLA2G6, which is known to cause a heterogeneous neurodegenerative clinical spectrum, has recently been shown to be responsible for autosomal-recessive familial forms of Parkinson's disease (PD). Here, we generated induced pluripotent stem cells (iPSCs) from the peripheral blood mononuclear cells of a female patient with a homozygous PLA2G6 c.991Gâ¯>â¯T (p.D331Y) mutation by using the Sendai-virus delivery system. The resulting iPSCs showed pluripotency confirmed by immunofluorescent staining for pluripotency markers and differentiated into the 3 germ layers in vivo. This cellular model will provide a good resource for further pathophysiological studies of PD.