RESUMO
BACKGROUND: Efforts to profile atopic dermatitis (AD) tissues have intensified, yet comprehensive analysis of systemic immune landscapes in severe AD remains crucial. METHODS: Employing single-cell RNA sequencing, we analyzed over 300,000 peripheral blood mononuclear cells from 12 severe AD patients (Eczema area and severity index (EASI) > 21) and six healthy controls. RESULTS: Results revealed significant immune cell shifts in AD patients, including increased Th2 cell abundance, reduced NK cell clusters with compromised cytotoxicity, and correlated Type 2 innate lymphoid cell proportions with disease severity. Moreover, unique monocyte clusters reflecting activated innate immunity emerged in very severe AD (EASI > 30). While overall dendritic cells (DCs) counts decreased, a distinct Th2-priming subset termed "Th2_DC" correlated strongly with disease severity, validated across skin tissue data, and flow cytometry with additional independent severe AD samples. Beyond the recognized role of Th2 adaptive immunity, our findings highlight significant innate immune cell alterations in severe AD, implicating their roles in disease pathogenesis and therapeutic potentials. CONCLUSION: Apart from the widely recognized role of Th2 adaptive immunity in AD pathogenesis, alterations in innate immune cells and impaired cytotoxic cells have also been observed in severe AD. The impact of these alterations on disease pathogenesis and the effectiveness of potential therapeutic targets requires further investigation.
Assuntos
Dermatite Atópica , RNA-Seq , Índice de Gravidade de Doença , Análise de Célula Única , Dermatite Atópica/imunologia , Humanos , Imunidade Inata , Masculino , Células Th2/imunologia , Células Th2/metabolismo , Feminino , Adulto , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Estudos de Casos e Controles , Análise da Expressão Gênica de Célula ÚnicaRESUMO
Fumaramic acid derivatives can be converted into their cis isomer maleamic acid derivatives under UV illumination, and these maleamic acid derivatives show pH-responsive degradability at acidic pH only after the preceding photoisomerization. The rate of the tandem photoisomerization-degradation of fumaramic acid derivatives can be finely controlled by changing the substituents on the double bond. Photoisomerization-based unlocking of the pH-responsive degradability of fumaramic acid derivatives has strong potential for the development of multisignal-responsive smart materials in biomedical applications.
Assuntos
Ácidos/química , Fumaratos/química , Concentração de Íons de Hidrogênio , Sistemas de Liberação de Medicamentos , Isomerismo , Nitrobenzenos/química , Processos Fotoquímicos , Succinatos/químicaRESUMO
The Ras effector NORE1 is frequently silenced in primary adenocarcinomas, although the significance of this silencing for tumorigenesis is unclear. Here we show that NORE1 induces polyubiquitination and proteasomal degradation of oncoprotein HIPK1 by facilitating its interaction with the Mdm2 E3 ubiquitin ligase. Endogenous HIPK1 is stabilized in Nore1-deficient mouse embryonic fibroblasts, and depletion of HIPK1 in NORE1-silenced lung adenocarcinoma cells inhibits anchorage-independent cell growth and tumour formation in nude mice. These findings indicate that the control of HIPK1 stability by Mdm2-NORE1 has a major effect on cell behaviour, and epigenetic inactivation of NORE1 enables adenocarcinoma formation in vivo through HIPK1 stabilization.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Proteínas Monoméricas de Ligação ao GTP/metabolismo , Proteínas Oncogênicas/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Proteólise , Proteínas Proto-Oncogênicas c-mdm2/metabolismo , Proteínas ras/metabolismo , Animais , Proteínas Reguladoras de Apoptose , Linhagem Celular Tumoral , Transformação Celular Neoplásica/metabolismo , Transformação Celular Neoplásica/patologia , Técnicas de Silenciamento de Genes , Humanos , Camundongos , Poliubiquitina/metabolismo , Ligação Proteica , Saccharomyces cerevisiae/metabolismo , UbiquitinaçãoRESUMO
We have developed a technique for isolating and culturing primary lung cancer cells extracted from patient tissue to facilitate anti-cancer drug development. Patient-derived lung cancer tissues were mechanically dissociated to 40-100 µm. Dispase was then used to isolate cultured lung cancer cell populations, which were re-plated on Matrigel-coated dishes containing N2-supplemented medium and growth factors. This method allows pure populations of primary non-small cell lung cancer cells to be grown in vitro. The isolated cells exhibited hallmark cancerous properties such as abnormal chromosomes and in vivo tumor formation. The cell lines generated through this procedure may help to advance our knowledge of certain forms of lung cancer and may also be useful for developing patient-specific anti-cancer drug screening procedures.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/patologia , Técnicas Citológicas/métodos , Técnicas de Cultura de Células/métodos , Células Cultivadas , Avaliação Pré-Clínica de Medicamentos/métodos , HumanosRESUMO
An increasing number of chromosomal aberrations is being identified in solid tumors providing novel biomarkers for various types of cancer and new insights into the mechanisms of carcinogenesis. We applied next generation sequencing technique to analyze the transcriptome of the non-small cell lung carcinoma (NSCLC) cell line H2228 and discovered a fusion transcript composed of multiple exons of ALK (anaplastic lymphoma receptor tyrosine kinase) and PTPN3 (protein tyrosine phosphatase, nonreceptor Type 3). Detailed analysis of the genomic structure revealed that a portion of genomic region encompassing Exons 10 and 11 of ALK has been translocated into the intronic region between Exons 2 and 3 of PTPN3. The key net result appears to be the null mutation of one allele of PTPN3, a gene with tumor suppressor activity. Consistently, ectopic expression of PTPN3 in NSCLC cell lines led to inhibition of colony formation. Our study confirms the utility of next generation sequencing as a tool for the discovery of somatic mutations and has led to the identification of a novel mutation in NSCLC that may be of diagnostic, prognostic, and therapeutic importance.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , Neoplasias Pulmonares/genética , Proteínas de Fusão Oncogênica/genética , Proteína Tirosina Fosfatase não Receptora Tipo 3/genética , RNA Neoplásico/genética , Receptores Proteína Tirosina Quinases/genética , Quinase do Linfoma Anaplásico , Sequência de Bases , Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Linhagem Celular Tumoral , Humanos , Neoplasias Pulmonares/diagnóstico , Dados de Sequência Molecular , RNA Neoplásico/química , Análise de Sequência de DNARESUMO
We determined the somatic mutations in the mitochondrial genomes of 70 lung cancer patients by pair-wise comparative analyses of the normal- and tumor-genome sequences acquired using Affymetrix Mitochondrial Resequencing Array 2.0. The overall mutation rates in lung cancers were Approximately 100 fold higher than those in normal cells, with significant statistical correlation with smoking (p=0.00088). Total of 532 somatic mutations were evenly distributed in 499 positions with very low overall frequency (1.07/bp), but the non-synonymous mutations causing amino acid substitution occurred more frequently (1.83/bp), particularly at two positions, 8701 and 10398 (10.5/bp) that code for ATPase6 and NADH dehydrogenase 3, respectively. Despite the randomness or even distribution of the mutations, these two mutations occurred together in 86% of the cases. The linkage between the two most frequent mutations suggests that they were selected together, possibly due to their cooperative role during cancer development. Indeed, the mutation at 10398 was shown by Canter, Pezzotti, and their colleagues in 2009, as a risk factor for breast cancer. In this study, we identified two potential biomarkers that might be functionally linked together during the development of cancer.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , Complexo I de Transporte de Elétrons/genética , Genoma Mitocondrial/genética , Mutação em Linhagem Germinativa , Neoplasias Pulmonares/genética , ATPases Mitocondriais Próton-Translocadoras/genética , Análise Mutacional de DNA , Feminino , Humanos , Masculino , Mutagênese , Polimorfismo Genético , República da Coreia , Fumar/genéticaRESUMO
AIMP2/p38 is a scaffolding protein required for the assembly of the macromolecular tRNA synthetase complex. Here, we describe a previously unknown function for AIMP2 as a positive regulator of p53 in response to genotoxic stresses. Depletion of AIMP2 increased resistance to DNA damage-induced apoptosis, and introduction of AIMP2 into AIMP2-deficient cells restored the susceptibility to apoptosis. Upon DNA damage, AIMP2 was phosphorylated, dissociated from the multi-tRNA synthetase complex, and translocated into the nuclei of cells. AIMP2 directly interacts with p53, thereby preventing MDM2-mediated ubiquitination and degradation of p53. Mutations in AIMP2, affecting its interaction with p53, hampered its ability to activate p53. Nutlin-3 recovered the level of p53 and the susceptibility to UV-induced cell death in AIMP2-deficient cells. This work demonstrates that AIMP2, a component of the translational machinery, functions as proapoptotic factor via p53 in response to DNA damage.
Assuntos
Aminoacil-tRNA Sintetases/metabolismo , Núcleo Celular/metabolismo , Dano ao DNA/efeitos da radiação , Complexos Multiproteicos/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Raios Ultravioleta , Transporte Ativo do Núcleo Celular/efeitos dos fármacos , Transporte Ativo do Núcleo Celular/genética , Transporte Ativo do Núcleo Celular/efeitos da radiação , Aminoacil-tRNA Sintetases/genética , Animais , Apoptose/genética , Apoptose/efeitos da radiação , Linhagem Celular Tumoral , Núcleo Celular/genética , Dano ao DNA/efeitos dos fármacos , Dano ao DNA/genética , Imidazóis/farmacologia , Camundongos , Complexos Multiproteicos/genética , Mutação , Fosforilação/efeitos dos fármacos , Fosforilação/efeitos da radiação , Piperazinas/farmacologia , Proteínas Proto-Oncogênicas c-mdm2/genética , Proteínas Proto-Oncogênicas c-mdm2/metabolismo , Proteína Supressora de Tumor p53/genética , Ubiquitinação/efeitos dos fármacos , Ubiquitinação/genética , Ubiquitinação/efeitos da radiaçãoRESUMO
The new allele, HLA-DQB1*05:247 differs from HLA-DQB1*05:02:01:01 by one nucleotide substitution at codon 35.
Assuntos
Sequenciamento de Nucleotídeos em Larga Escala , Alelos , Éxons/genética , Cadeias beta de HLA-DQ/genética , HumanosRESUMO
HLA-B*51:315 differs from B*51:01:01:01 by a single nucleotide polymorphism in codon 87 (CAG â CGG).
Assuntos
Antígenos HLA-B , Sequenciamento de Nucleotídeos em Larga Escala , Alelos , Códon , Antígenos HLA-B/genética , Teste de Histocompatibilidade , Humanos , Polimorfismo de Nucleotídeo Único , República da Coreia , Análise de Sequência de DNARESUMO
DQB1*06:344 differs from DQB1*06:02:01:01 by one nucleotide substitution at codon 211 in exon 3.
Assuntos
Sequenciamento de Nucleotídeos em Larga Escala , Alelos , Cadeias beta de HLA-DQ/genética , Humanos , República da CoreiaRESUMO
Flavins, comprising flavin mononucleotide (FMN), flavin adenine dinucleotide (FAD), and riboflavin (RF, vitamin B(2)), play important roles in numerous redox reactions such as those taking place in the electron-transfer chains of mitochondria in all eukaryotes and of plastids in plants. A selective chemosensor for flavins would be useful not only in the investigation of metabolic processes but also in the diagnosis of diseases related to flavins; such a sensor is presently unavailable. Herein, we report the first bifunctional chemosensor (PTZ-DPA) for flavins. PTZ-DPA consists of bis(Zn(2+)-dipicolylamine) and phenothiazine. Bis(Zn(2+)-dipicolylamine) (referred to here as XyDPA) was found to be an excellent catalyst in the conversion of FAD into cyclic FMN (riboflavin 4',5'-cyclic phosphate, cFMN) under physiological conditions, even at pH 7.4 and 27 degrees C, with less than 1 mol % of substrate. Utilizing XyDPA's superior function as an artificial FMN cyclase and phenothiazine as an electron donor able to quench the fluorescence of an isoalloxazine ring, PTZ-DPA enabled selective fluorescent discrimination of flavins (FMN, FAD, and RF): FAD shows ON(+), FMN shows OFF(-), and RF shows NO(0) fluorescence changes upon the addition of PTZ-DPA. With this selective sensing property, PTZ-DPA is applicable to real-time fluorescent monitoring of riboflavin kinase (RF to FMN), alkaline phosphatase (FMN to RF), and FAD synthetase (FMN to FAD).
Assuntos
Materiais Biomiméticos/química , Técnicas de Química Analítica/métodos , Flavinas/análise , Fluorescência , Compostos Organometálicos/química , Fósforo-Oxigênio Liases/metabolismo , Aminas/química , Materiais Biomiméticos/metabolismo , Mononucleotídeo de Flavina/análise , Mononucleotídeo de Flavina/metabolismo , Corantes Fluorescentes/análise , Corantes Fluorescentes/química , Medições Luminescentes , Estrutura Molecular , Ácidos Picolínicos/química , Zinco/químicaRESUMO
The new HLA-C*04:387 allele was found and characterized in a Korean hematopoietic stem cell donor.
Assuntos
Loci Gênicos/genética , Antígenos HLA-C/genética , Polimorfismo Genético , Recombinação Genética , Alelos , Povo Asiático/genética , Doadores de Sangue , Feminino , Transplante de Células-Tronco Hematopoéticas , Sequenciamento de Nucleotídeos em Larga Escala , Teste de Histocompatibilidade , Humanos , Pessoa de Meia-Idade , República da Coreia , Análise de Sequência de DNARESUMO
HLA-A*24:460 differs from HLA-A*24:02 by one nucleotide in codon 285.
Assuntos
Alelos , Antígeno HLA-A24/genética , Células-Tronco Hematopoéticas , Doadores de Tecidos , Transplantados , Éxons , Feminino , Genótipo , Transplante de Células-Tronco Hematopoéticas , Teste de Histocompatibilidade , Humanos , Polimorfismo Genético , República da CoreiaRESUMO
The new allele, HLA-B*40:405 differs from B*40:02:01:01 by one nucleotide substitution at codon 304.
Assuntos
Alelos , Antígeno HLA-B40/genética , Síndromes Mielodisplásicas/genética , Adulto , Códon , Éxons , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , República da CoreiaRESUMO
AIMP3 (previously known as p18) was shown to up-regulate p53 in response to DNA damage. Here, we show that AIMP3 couples oncogenic stresses to p53 activation to prevent cell transformation. Growth factor- or Ras-dependent induction of p53 was blocked by single allelic loss of AIMP3 as well as by suppression of AIMP3. AIMP3 heterozygous cells became susceptible to cell transformation induced by oncogenes such as Ras or Myc alone. The transformed AIMP3+/- cells showed severe abnormality in cell division and chromosomal structure. Thus, AIMP3 plays crucial roles in p53-mediated tumor-suppressive response against oncogenic stresses via differential activation of ATM and ATR, and in the maintenance of genomic stability.
Assuntos
Transformação Celular Neoplásica/genética , Inibidor de Quinase Dependente de Ciclina p18/genética , Regulação da Expressão Gênica/genética , Genes ras , Instabilidade Genômica/genética , Proteína Supressora de Tumor p53/genética , Alelos , Animais , Inibidor de Quinase Dependente de Ciclina p18/metabolismo , Embrião de Mamíferos , Fibroblastos , Camundongos , RNA Antissenso/genética , Transfecção , Proteína Supressora de Tumor p53/biossínteseRESUMO
The upper critical solution temperature (UCST) phase transition of halide salts of branched polyethylenimine (PEI) and methylated branched polyethylenimine (MPEI) is first reported in aqueous solutions. In particular, iodide counter-ions can introduce UCST properties in MPEI. The importance of the counter-ion composition of MPEI for UCST transition is discussed in detail.
RESUMO
Anaplastic lymphoma kinase (ALK) fusion is a common mechanism underlying pathogenesis of non-small cell lung carcinoma (NSCLC) where these rearrangements represent important diagnostic and therapeutic targets. In this study, we found a new ALK fusion gene, SEC31A-ALK, in lung carcinoma from a 53-year-old Korean man. The conjoined region in the fusion transcript was generated by the fusion of SEC31A exon 21 and ALK exon 20 by genomic rearrangement, which contributed to generation of an intact, in-frame open reading frame. SEC31A-ALK encodes a predicted fusion protein of 1,438 amino acids comprising the WD40 domain of SEC31A at the N-terminus and ALK kinase domain at the C-terminus. Fluorescence in situ hybridization studies suggested that SEC31A-ALK was generated by an unbalanced genomic rearrangement associated with loss of the 3'-end of SEC31A. This is the first report of SEC31A-ALK fusion transcript in clinical NSCLC, which could be a novel diagnostic and therapeutic target for patients with NSCLC.
Assuntos
Adenocarcinoma/genética , Neoplasias Pulmonares/genética , Fusão Oncogênica , Proteínas de Fusão Oncogênica/genética , Receptores Proteína Tirosina Quinases/genética , Proteínas de Transporte Vesicular/genética , Adenocarcinoma/tratamento farmacológico , Quinase do Linfoma Anaplásico , Éxons/genética , Humanos , Hibridização in Situ Fluorescente , Neoplasias Pulmonares/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Proteínas de Fusão Oncogênica/química , Receptores Proteína Tirosina Quinases/química , Proteínas de Transporte Vesicular/químicaRESUMO
INTRODUCTION: The aim of our analysis was to evaluate the prognostic effect of programmed cell death ligand-1 (PD-L1) expression in patients with non-small cell lung cancer (NSCLC). METHODS: PD-L1 expression among 1070 surgically resected NSCLC specimens was evaluated by immunohistochemical analysis. Data were analyzed using Cox proportional hazard models adjusting for age, sex, smoking status, histologic type, stage, and performance status. RESULTS: Sixty-eight patients (6%) were strongly PD-L1 positive and 410 (38%) were weakly PD-L1 positive. A significantly higher prevalence of PD-L1 positivity was observed among patients with squamous cell carcinoma and among stage IIIB and IV patients. PD-L1 expression may be associated with poorer overall survival, with an adjusted hazard ratio of 1.56 (95% confidence interval [CI]: 1.08-2.26, p = 0.02) for strong PD-L1 positivity, 1.18 (95% CI: 0.96-1.46; p = 0.12) for weak PD-L1 positivity, and 1.23 (95% CI: 1.00-1.51; p = 0.05) for the combined strongly and weakly positive groups compared with PD-L1 negativity. Negative prognostic effect of PD-L1 expression was not statistically significant after adjustment for postoperative chemotherapy or radiotherapy. Similar results were observed for progression-free survival. Among stage I patients, the disease recurrence rate was higher in the PD-L1-positive versus in the PD-L1-negative group (48% versus 27%, p < 0.001), with an adjusted hazard ratio for disease-free survival of 2.01 (95% CI, 1.08-3.73; p = 0.03) for strong PD-L1 positivity and 1.57 (95% CI, 1.17-2.11; p = 0.003) for weak PD-L1 positivity compared with PD-L1 negativity. CONCLUSIONS: Tumor PD-L1 expression may be associated with poor prognosis in patients with NSCLC, although its significance weakens when postoperative therapy is considered.
Assuntos
Antígeno B7-H1/análise , Carcinoma Pulmonar de Células não Pequenas/química , Neoplasias Pulmonares/química , Adulto , Idoso , Carcinoma Pulmonar de Células não Pequenas/mortalidade , Carcinoma Pulmonar de Células não Pequenas/cirurgia , Estudos de Coortes , Feminino , Humanos , Neoplasias Pulmonares/mortalidade , Neoplasias Pulmonares/cirurgia , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , PrognósticoRESUMO
Homeobox (hox) genes encode transcription factors which are critically involved in embryonic development. The 39 different hox genes are organized into four unlinked chromosomal gene clusters (hoxA-D) in mammals and a number of studies have suggested that hox genes play important roles in human leukemogenesis. Acute promyelocytic leukemia (APL) cells carrying the PML-RARa fusion protein, respond well by differentiating in their response to an all-trans-retinoic acid (ATRA) treatment. We examined the expression of the individual hox genes during this differentiation. Besides the constitutive expression of hoxA9 and no expression of hoxB7, the expression of hoxC4 increased significantly during differentiation of NB4 cells (PML-RAR(alpha)+). We also examined the expression of hoxC4 in bone marrow cells from APL patients and found that the hoxC4 expression was reproducibly induced during an ATRA treatment. To further examine this finding, HoxC4 was stably expressed in NB4 cells by retroviral transduction. The NB4 cells expressing HoxC4 showed differentiated phenotypes including a CD14 expression, which strongly suggests that upregulated hoxC4 is functionally involved in NB4 cell differentiation in response to ATRA. Upregulation of CD14 is at the transcription level and mediated by the homeodomain of the HoxC4.