RESUMEN
Although critical to T cell function, antigen specificity is often omitted in high-throughput multiomics-based T cell profiling due to technical challenges. We describe a high-dimensional, tetramer-associated T cell antigen receptor (TCR) sequencing (TetTCR-SeqHD) method to simultaneously profile cognate antigen specificities, TCR sequences, targeted gene expression and surface-protein expression from tens of thousands of single cells. Using human polyclonal CD8+ T cells with known antigen specificity and TCR sequences, we demonstrate over 98% precision for detecting the correct antigen specificity. We also evaluate gene expression and phenotypic differences among antigen-specific CD8+ T cells and characterize phenotype signatures of influenza- and Epstein-Barr virus-specific CD8+ T cells that are unique to their pathogen targets. Moreover, with the high-throughput capacity of profiling hundreds of antigens simultaneously, we apply TetTCR-SeqHD to identify antigens that preferentially enrich cognate CD8+ T cells in patients with type 1 diabetes compared to healthy controls and discover a TCR that cross-reacts with diabetes-related and microbiome antigens. TetTCR-SeqHD is a powerful approach for profiling T cell responses in humans and mice.
Asunto(s)
Antígenos/inmunología , Linfocitos T CD8-positivos/inmunología , Secuenciación de Nucleótidos de Alto Rendimiento , Receptores de Antígenos de Linfocitos T/genética , Análisis de la Célula Individual , Antígenos/metabolismo , Antígenos Virales/inmunología , Antígenos Virales/metabolismo , Autoantígenos/inmunología , Autoantígenos/metabolismo , Autoinmunidad , Linfocitos T CD8-positivos/metabolismo , Linfocitos T CD8-positivos/virología , Estudios de Casos y Controles , Separación Celular , Células Cultivadas , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/inmunología , Diabetes Mellitus Tipo 1/metabolismo , Herpesvirus Humano 4/inmunología , Herpesvirus Humano 4/patogenicidad , Humanos , Orthomyxoviridae/inmunología , Orthomyxoviridae/patogenicidad , Fenotipo , Receptores de Antígenos de Linfocitos T/inmunología , Receptores de Antígenos de Linfocitos T/metabolismoRESUMEN
Cell identity annotation for single-cell transcriptome data is a crucial process for constructing cell atlases, unraveling pathogenesis, and inspiring therapeutic approaches. Currently, the efficacy of existing methodologies is contingent upon specific data sets. Nevertheless, such data are often sourced from various batches, sequencing technologies, tissues, and even species. Notably, the gene regulatory relationship remains unaffected by the aforementioned factors, highlighting the extensive gene interactions within organisms. Therefore, we propose scHGR, an automated annotation tool designed to leverage gene regulatory relationships in constructing gene-mediated cell communication graphs for single-cell transcriptome data. This strategy helps reduce noise from diverse data sources while establishing distant cellular connections, yielding valuable biological insights. Experiments involving 22 scenarios demonstrate that scHGR precisely and consistently annotates cell identities, benchmarked against state-of-the-art methods. Crucially, scHGR uncovers novel subtypes within peripheral blood mononuclear cells, specifically from CD4+ T cells and cytotoxic T cells. Furthermore, by characterizing a cell atlas comprising 56 cell types for COVID-19 patients, scHGR identifies vital factors like IL1 and calcium ions, offering insights for targeted therapeutic interventions.
Asunto(s)
COVID-19 , Redes Reguladoras de Genes , RNA-Seq , Análisis de Expresión Génica de una Sola Célula , Humanos , Linfocitos T CD4-Positivos/metabolismo , COVID-19/genética , COVID-19/virología , Leucocitos Mononucleares/metabolismo , Anotación de Secuencia Molecular , RNA-Seq/métodos , SARS-CoV-2/genética , TranscriptomaRESUMEN
Our understanding of nerve regeneration can be enhanced by delineating its underlying molecular activities at single-neuron resolution in model organisms such as Caenorhabditis elegans. Existing cell isolation techniques cannot isolate neurons with specific regeneration phenotypes from C. elegans. We present femtosecond laser microdissection (fs-LM), a single-cell isolation method that dissects specific cells directly from living tissue by leveraging the micrometer-scale precision of fs-laser ablation. We show that fs-LM facilitates sensitive and specific gene expression profiling by single-cell RNA sequencing (scRNA-seq), while mitigating the stress-related transcriptional artifacts induced by tissue dissociation. scRNA-seq of fs-LM isolated regenerating neurons revealed transcriptional programs that are correlated with either successful or failed regeneration in wild-type and dlk-1 (0) animals, respectively. This method also allowed studying heterogeneity displayed by the same type of neuron and found gene modules with expression patterns correlated with axon regrowth rate. Our results establish fs-LM as a spatially resolved single-cell isolation method for phenotype-to-genotype mapping.
Asunto(s)
Proteínas de Caenorhabditis elegans , Caenorhabditis elegans , Animales , Caenorhabditis elegans/genética , Microdisección/métodos , Neuronas/fisiología , Rayos Láser , Análisis de Secuencia de ARN , Quinasas Quinasa Quinasa PAM , Proteínas de Caenorhabditis elegans/genéticaRESUMEN
Moiré superlattices have emerged as a new platform for studying strongly correlated quantum phenomena, but these systems have been largely limited to van der Waals layer two-dimensional materials. Here we introduce moiré superlattices leveraging ultrathin, ligand-free halide perovskites, facilitated by ionic interactions. Square moiré superlattices with varying periodic lengths are clearly visualized through high-resolution transmission electron microscopy. Twist-angle-dependent transient photoluminescence microscopy and electrical characterizations indicate the emergence of localized bright excitons and trapped charge carriers near a twist angle of ~10°. The localized excitons are accompanied by enhanced exciton emission, attributed to an increased oscillator strength by a theoretically predicted flat band. This research showcases the promise of two-dimensional perovskites as unique room-temperature moiré materials.
RESUMEN
Tin-based two-dimensional (2D) perovskites are emerging as lead-free alternatives in halide perovskite materials, yet their exciton dynamics and transport remain less understood due to defect scattering. Addressing this, we employed temperature-dependent transient photoluminescence (PL) microscopy to investigate intrinsic exciton transport in three structurally analogous Sn- and Pb-based 2D perovskites. Employing conjugated ligands, we synthesized high-quality crystals with enhanced phase stability at various temperatures. Our results revealed phonon-limited exciton transport in Sn perovskites, with diffusion constants increasing from 0.2 cm2 s-1 at room temperature to 0.6 cm2 s-1 at 40 K, and a narrowing PL line width. Notably, Sn-based perovskites exhibited greater exciton mobility than their Pb-based equivalents, which is attributed to lighter effective masses. Thermally activated optical phonon scattering was observed in Sn-based compounds but was absent in Pb-based materials. These findings, supported by molecular dynamics simulations, demonstrate that the phonon scattering mechanism in Sn-based halide perovskites can be distinct from their Pb counterparts.
RESUMEN
Carotid body tumor (CBT) is a rare neck tumor located at the adventitia of the common carotid artery bifurcation. The prominent pathological features of CBT are high vascularization and abnormal proliferation. However, single-cell transcriptome analysis of the microenvironment composition and molecular complexity in CBT has yet to be performed. In this study, we performed single-cell RNA sequencing (scRNA-seq) analysis on human CBT to define the cells that contribute to hypervascularization and chronic hyperplasia. Unbiased clustering analysis of transcriptional profiles identified 16 distinct cell populations including endothelial cells (ECs), smooth muscle cells (SMCs), neuron cells, macrophage cells, neutrophil cells, and T cells. Within the ECs population, we defined subsets with angiogenic capacity plus clear signs of later endothelial progenitor cells (EPCs) to normal ECs. Two populations of macrophages were detectable in CBT, macrophage1 showed enrichment in hypoxia-inducible factor-1 (HIF-1) and as well as an early EPCs cell-like population expressing CD14 and vascular endothelial growth factor. In addition to HIF-1-related transcriptional protein expression, macrophages1 also display a neovasculogenesis-promoting phenotype. SMCs included three populations showing platelet-derived growth factor receptor beta and vimentin expression, indicative of a cancer-associated fibroblast phenotype. Finally, we identified three types of neuronal cells, including chief cells and sustentacular cells, and elucidated their distinct roles in the pathogenesis of CBT and abnormal proliferation of tumors. Overall, our study provided the first comprehensive characterization of the transcriptional landscape of CBT at scRNA-seq profiles, providing novel insights into the mechanisms underlying its formation.
Asunto(s)
Tumor del Cuerpo Carotídeo , Células Progenitoras Endoteliales , Neovascularización Patológica , Humanos , Arterias Carótidas/patología , Tumor del Cuerpo Carotídeo/irrigación sanguínea , Análisis de la Célula Individual , Análisis de Expresión Génica de una Sola Célula , Transcriptoma/genética , Microambiente Tumoral/genética , Factor A de Crecimiento Endotelial Vascular , Neovascularización Patológica/diagnóstico , Neovascularización Patológica/genéticaRESUMEN
The circadian clock is entrained to daily environmental cues. Integrin-linked signaling via actin cytoskeleton dynamics transduces physical niche cues from the extracellular matrix to myocardin-related transcription factor (MRTF)/serum response factor (SRF)-mediated transcription. The actin cytoskeleton organization and SRF-MRTF activity display diurnal oscillations. By interrogating disparate upstream events in the actin cytoskeleton-MRTF-A/SRF signaling cascade, we show that this pathway transduces extracellular niche cues to modulate circadian clock function. Pharmacological inhibition of MRTF-A/SRF by disrupting actin polymerization or blocking the ROCK kinase induced period lengthening with augmented clock amplitude, and genetic loss of function of Srf or Mrtfa mimicked the effects of treatment with actin-depolymerizing agents. In contrast, actin polymerization shortened circadian clock period and attenuated clock amplitude. Moreover, interfering with the cell-matrix interaction through blockade of integrin, inhibition of focal adhesion kinase (FAK, encoded by Ptk2) or attenuating matrix rigidity reduced the period length while enhancing amplitude. Mechanistically, we identified that the core clock repressors Per2, Nr1d1 and Nfil3 are direct transcriptional targets of MRTF-A/SRF in mediating actin dynamics-induced clock response. Collectively, our findings defined an integrin-actin cytoskeleton-MRTF/SRF pathway in linking clock entrainment with extracellular cues that might facilitate cellular adaptation to the physical niche environment.
Asunto(s)
Relojes Circadianos , Factor de Respuesta Sérica , Citoesqueleto de Actina/metabolismo , Actinas/metabolismo , Relojes Circadianos/genética , Señales (Psicología) , Integrinas , Proteínas Nucleares , Factor de Respuesta Sérica/genética , Factor de Respuesta Sérica/metabolismo , Transactivadores , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Traditional alternating current filter based on aluminum electrolytic capacitors (AECs) suffer from abrupt drop of filtering capability at ultra-low temperatures (≤-30 °C), which greatly hinders the reliable working of electronics at extremely cold conditions. Herein, an ultra-low-temperature alternating current (AC) filter for the first time enabled by high-frequency supercapacitor based on covalently bonded hollow carbon onion-graphene hybrid structure is reported. It is found that the covalent bonding junctions enable high electronic conductivity and efficient ion adsorption/desorption behavior in the hybrid structure. Moreover, the hybrid structure owns positive curvature and shallows pores for fast ion diffusion kinetics. Consequently, the supercapacitor exhibits a record short resistor-capacitor time constant (τRC ) of 0.098 ms at 120 Hz at room temperature. Combining with low-melting-point electrolyte, the supercapacitor possesses excellent filtering capability and can output stable direct current signal with low fluctuation coefficients in a temperature range of -50 to 0 °C. More interestingly, the filter presents high negative phase angle, low dissipation factor, short τRC , and high capacitance retention below -30 °C, whereas AEC cannot work properly owing to its phase angle<45°. This work realizes the fabrication of an ultra-low-temperature AC filter, which presents a critical step forward for promoting the development of ultra-low-temperature electronics.
RESUMEN
The intricate processes that govern the interactions between peripatetic immune cells and distal renal injury in obesity are not fully understood. Employing transcriptomic analysis of circulating extracellular vesicles (EVs), a marked amplification of small RNA (miR-3960) is discerned within CD3-CD19+ B cells. This RNA is found to be preferentially augmented in kidney tissues, contrasting with its subdued expression in other organs. By synthesizing dual-luciferase reporter assay with co-immunoprecipitation analysis, it is pinpointed that miR-3960 specifically targets the nuclear gene TRMT5, a pivotal actor in the methylation of mitochondrial tRNA. This liaison instigates aberrations in the post-transcriptional modifications of mitochondrial tRNA, engendering deficiencies within the electron respiratory chain, primarily attributable to the diminution of the mitochondrial bioenergetic compound (NDUFA7) complex I. Such perturbations lead to a compromised mitochondrial respiratory capacity in renal tubular cells, thereby exacerbating tubular injury. In contrast, EV blockade or miR-3960 depletion markedly alleviates renal tubular injury in obesity. This investigation unveils a hitherto unexplored pathway by which obesity-induced circulating immune cells remotely manipulate mitochondrial metabolism in target organs. The strategic targeting of obese EVs or infiltrative immune cells and their specifically secreted RNAs emerges as a promising therapeutic avenue to forestall obesity-related renal afflictions.
RESUMEN
DNA methylation, an epigenetic regulatory mechanism dictating gene transcription, plays a critical role in the occurrence and development of cancer. However, the molecular underpinnings of LINC00987 methylation in the regulation of lung adenocarcinoma (LUAD) remain elusive. This study investigated LINC00987 expression in LUAD patients through analysis of The Cancer Genome Atlas data sets. Quantitative real-time polymerase chain reaction (RT-qPCR) and fluorescence in situ hybridization assays were used to assess LINC00987 expression in LUAD. The bisulfite genomic sequence PCR (BSP) assay was used to determine the methylation levels of the LINC00987 promoter. The interaction between LINC00987 and SND1 was elucidated via immunoprecipitation and RNA pull-down assays. The functional significance of LINC00987 and SND1 in Calu-3 and NCI-H1688 cells was evaluated in vitro through CCK-8, EdU, Transwell, flow cytometry, and vasculogenic mimicry (VM) tube formation assays. LINC00987 expression decreased in LUAD concomitant with hypermethylation of the promoter region, while hypomethylation of the LINC00987 promoter in LUAD tissues correlated with tumor progression. Treatment with 5-Aza-CdR augmented LINC00987 expression and inhibited tumor growth. Mechanistically, LINC00987 overexpression impeded LUAD progression and VM through direct binding with SND1, thereby facilitating its phosphorylation and subsequent degradation. Additionally, overexpression of SND1 counteracted the adverse effects of LINC00987 downregulation on cell proliferation, apoptosis, cell migration, invasion, and VM in LUAD in vitro. In conclusion, this pioneering study focuses on the expression and function of LINC00987 and reveals that hypermethylation of the LINC00987 gene may contribute to LUAD progression. LINC00987 has emerged as a potential tumor suppressor gene in tumorigenesis through its binding with SND1 to facilitate its phosphorylation and subsequent degradation.
Asunto(s)
Adenocarcinoma del Pulmón , Proliferación Celular , Metilación de ADN , Progresión de la Enfermedad , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica , Neoplasias Pulmonares , ARN Largo no Codificante , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adenocarcinoma del Pulmón/genética , Adenocarcinoma del Pulmón/patología , Adenocarcinoma del Pulmón/metabolismo , Apoptosis , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Endonucleasas/genética , Endonucleasas/metabolismo , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patología , Neoplasias Pulmonares/metabolismo , Fosforilación , Regiones Promotoras Genéticas , ARN Largo no Codificante/genéticaRESUMEN
We propose and design a multi-stage cascaded scanning laser ophthalmoscope (SLO) for ultra-wide field (UWF), which uses conicoid mirrors, constructed by conjugation of pupil plane. The vergence uniformity and the angular magnification of a cascaded conicoid mirrors (CCM) system are analyzed recursively and optimized preliminarily to achieve high quality imaging with UWF, and the optimal system with the model eye are obtained by simulation and optimization. Two-stage and three-stage cascaded systems are designed with this method, and the formulas of beam vergence and angular magnification are obtained by theoretical derivation. As compared to the two-stage CCM system, the proposed three-stage cascaded UWF SLO has superior performance in imaging quality. Its average RMS radius of spot diagram is calculated to be 26.372â µm, close to the diffractive limit resolution. The image resolution of human retina can be up to 30â µm with 135° FOV in theory. The three-stage cascaded SLO is more suitable for UWF fundus imaging. This study will be helpful for early screening and accurate diagnosis of various diseases in the peripheral retina.
Asunto(s)
Oftalmoscopios , Retina , Humanos , Oftalmoscopía/métodos , Fondo de Ojo , Retina/diagnóstico por imagen , Rayos LáserRESUMEN
Phase-selective organogelators (PSOGs) are considered as a prospective tool for their application in oil spill remediation. However, the number of reports on the PSOGs that can be used in powder form for prompt phase-selective gelation of crude oils is still limited. In this study, a series of compounds with l-mandelic acid as the scaffold bearing different amino acid fragments have been prepared. Also, the gelation behaviors and properties of these derivatives toward organic liquids, product oils, and a type of Chinese crude oil were investigated via heating-and-cooling process, stirring, or resting operation. Besides, the micromorphologies of the resulting gels and the driving forces for the gel formation have been studied by scanning electron microscopy, Fourier transform infrared, UV spectroscopy, concentration-dependent 1H NMR, and X-ray diffraction. Particularly, gelator C15-Phe-Mac-Nap was shown to have the capability of congealing the Chinese crude oil selectively from water in powder form with a relatively lower gelator dosage, as compared with the other gelators we reported in the current and previous works. Moreover, gelator C15-Phe-Mac-Nap displayed some advantageous behaviors such as the reusability of gelator, excellent mechanical and chemical stability of the crude oil gels, and nontoxicity of the gelator in the aquatic environment, indicating its great potential application value for marine oil spill remediation.
RESUMEN
Layered double hydroxides (LDH) are a class of functional anionic clays that typically consist of orthorhombic arrays of metal hydroxides with anions sandwiched between the layers. Due to their unique properties, including high chemical stability, good biocompatibility, controlled drug loading, and enhanced drug bioavailability, LDHs have many potential applications in the medical field. Especially in the fields of bioimaging and tumor therapy. This paper reviews the research progress of LDHs and their nanocomposites in the field of tumor imaging and therapy. First, the structure and advantages of LDH are discussed. Then, several commonly used methods for the preparation of LDH are presented, including co-precipitation, hydrothermal and ion exchange methods. Subsequently, recent advances in layered hydroxides and their nanocomposites for cancer imaging and therapy are highlighted. Finally, based on current research, we summaries the prospects and challenges of layered hydroxides and nanocomposites for cancer diagnosis and therapy.
Asunto(s)
Nanocompuestos , Neoplasias , Humanos , Hidróxidos/química , Neoplasias/diagnóstico por imagen , Neoplasias/tratamiento farmacológico , Nanocompuestos/uso terapéutico , Nanocompuestos/químicaRESUMEN
BACKGROUND: Accurate prediction of death is an unmet need in patients with acute decompensated heart failure (HF). Arachidonic acid (AA) metabolites play an important role in the multiple pathophysiological processes. We aimed to develop an AA score to accurately predict mortality in patients with acute decompensated HF and explore the causal relationship between the AA predictors and HF. METHODS: The serum AA metabolites was measured in patients with acute decompensated HF (discovery cohort n=419; validation cohort n=386) by mass spectroscopy. We assessed the prognostic importance of AA metabolites for 1-year death using Cox regression and machine learning approaches. A machine learning-based AA score for predicting 1-year death was created and validated. We explored the mechanisms using transcriptome and functional experiments in a mouse model of early ischemic cardiomyopathy. RESULTS: Among the 27 AA metabolites, elevated 14,15-DHET/14,15-EET ratio was the strongest predictor of 1-year death (hazard ratio, 2.10, P=3.1×10-6). Machine learning-based AA score using a combination of the 14,15-DHET/14,15-EET ratio, 14,15-DHET, PGD2, and 9-HETE performed best (area under the curve [AUC]: 0.85). The machine learning-based AA score provided incremental information to predict mortality beyond BNP (B-type natriuretic peptide; ΔAUC: 0.19), clinical score (ΔAUC: 0.09), and preexisting Acute Decompensated Heart Failure National Registry, Organized Program to Initiate Lifesaving Treatment in Hospitalized Patients With Heart Failure, and Get With The Guidelines Heart Failure scores (ΔAUC: 0.17, 0.17, 0.15, respectively). In the validation cohort, the AA score accurately predicted mortality (AUC:0.81). False-negative and false-positive findings, as classified by the BNP threshold, were correctly reclassified by the AA score (46.2% of false-negative and 84.5% of false-positive). In a murine model, the expression and enzymatic activity of sEH (soluble epoxide hydrolase) increased after myocardial infarction. Genetic deletion of sEH improved HF and the blockade of 14,15-EET abolished this cardioprotection. We mechanistically revealed the beneficial effect of 14,15-EET by impairing the activation of monocytes/macrophages. CONCLUSIONS: Our studies propose that the AA score predicts death in patients with acute decompensated HF and inhibiting sEH serves as a therapeutic target for treating HF. REGISTRATION: URL: https://www. CLINICALTRIALS: gov; Unique identifier: NCT04108182.
Asunto(s)
Insuficiencia Cardíaca , Péptido Natriurético Encefálico , Animales , Ácido Araquidónico , Insuficiencia Cardíaca/diagnóstico , Insuficiencia Cardíaca/terapia , Humanos , Ratones , Pronóstico , Sistema de RegistrosRESUMEN
OBJECTIVES: The focus of this research was to explore any potential link between nocturia and the risk of suicidal ideation. METHODS: Drawing from the National Health and Nutrition Survey, data relating to 25 241 participants was scrutinized. This included 13 421 individuals identifying as male and 11 820 individuals identifying as female. Participants provided information on nocturia and suicidal ideation via self-completed questionnaires. To determine if nocturia was independently related to suicidal ideation, a multivariable logistic regression analysis was employed. Analyses were also undertaken separately for adult males and females. RESULTS: It was found that around 3.5% of participants had experienced suicidal ideation. The results indicated that nocturia increased the risk for suicidal ideation in all adult groups (odds ratio [OR] = 1.67, 95% confidence interval[CI]: 1.37-2.03, p < 0.0001), including both males (OR = 1.91, 95% CI: 1.38-2.65, p < 0.001) and females (OR = 1.48, 95% CI: 1.158-1.90, p = 0.002). The risk for suicidal ideation increased with the severity of nocturia, with significant trends observed in adult males (p for trend = 0.04) and adult females (p for trend = 0.01). Additionally, subgroup examination showed a significant interaction between nocturia and educational level in adult males (p for interaction = 0.03). Among adult females, a noteworthy interaction was observed between nocturia and body mass index (p for interaction = 0.02). CONCLUSION: The research uncovered a connection between nocturia and an elevated risk of suicidal ideation.
RESUMEN
Interferon-inducible protein 204 (IFI204) is an intracellular DNA receptor that can recognize DNA viruses and intracellular bacteria. Extracellular traps (ETs) have been recognized as an indispensable antimicrobial barrier that play an indispensable role in bacterial, fungal, parasitic, and viral infections. However, how ETs form and the mechanisms by which IFI204 function in Staphylococcus aureus pneumonia are still unclear. Moreover, by in vitro experiments, we proved that IFI204 deficiency decreases the formation of ETs induced by Staphylococcus aureus in a NOX-independent manner. More importantly, Deoxyribonuclease I (DNase I) treatment significantly inhibited the formation of ETs. IFI204 contributed to ETs formation by promoting citrullination of histone H3 and the expression of PAD4 (peptidylarginine deiminase 4). Altogether, these findings highlight the potential importance of IFI204 for host defense against S. aureus USA300-TCH1516 infection.
Asunto(s)
Trampas Extracelulares , Neumonía Estafilocócica , Trampas Extracelulares/genética , Trampas Extracelulares/metabolismo , Interferones/metabolismo , Neutrófilos/metabolismo , Neumonía Estafilocócica/genética , Neumonía Estafilocócica/metabolismo , Staphylococcus aureus , Ratones , AnimalesRESUMEN
Histone deacetylase inhibitors (HDACis) are important drugs for cancer therapy, but the indistinct resistant mechanisms of solid tumor therapy greatly limit their clinical application. In this study we conducted HDACi-perturbated proteomics and phosphoproteomics analyses in HDACi-sensitive and -resistant cell lines using a tandem mass tag (TMT)-based quantitative proteomic strategy. We found that the ribosome biogenesis proteins MRTO4, PES1, WDR74 and NOP16 vital to tumorigenesis might regulate the tumor sensitivity to HDACi. By integrating HDACi-perturbated protein signature with previously reported proteomics and drug sensitivity data, we predicted and validated a series of drug combination pairs potentially to enhance the sensitivity of HDACi in diverse solid tumor. Functional phosphoproteomic analysis further identified the kinase PDK1 and ROCK as potential HDACi-resistant signatures. Overall, this study reveals the potential HDACi-resistant signatures and may provide promising drug combination strategies to attenuate the resistance of solid tumor to HDACi.
Asunto(s)
Resistencia a Antineoplásicos , Inhibidores de Histona Desacetilasas , Neoplasias , Proteómica , Humanos , Inhibidores de Histona Desacetilasas/farmacología , Resistencia a Antineoplásicos/efectos de los fármacos , Neoplasias/tratamiento farmacológico , Neoplasias/metabolismo , Línea Celular Tumoral , Antineoplásicos/farmacología , Antineoplásicos/uso terapéuticoRESUMEN
Autophagy is an intracellular degradation system that delivers cytoplasmic constituents to the lysosome, and loss of autophagy has been linked to increased genome instability. Here, we report that loss of autophagy is coupled to reduced histone H2A ubiquitination after DNA damage. p62/SQSTM1, which accumulates in autophagy-defective cells, directly binds to and inhibits nuclear RNF168, an E3 ligase essential for histone H2A ubiquitination and DNA damage responses. As a result, DNA repair proteins such as BRCA1, RAP80, and Rad51 cannot be recruited to the sites of DNA double-strand breaks (DSBs), which impairs DSB repair. Moreover, nuclear-localized p62 increased the sensitivity of tumor cells to radiation both in vitro and in vivo, and this required its interaction with RNF168. Our findings indicate that autophagy-deficiency-induced p62 accumulation results in inhibition of histone ubiquitination and highlight the complex relationship between autophagy and the DNA damage response.
Asunto(s)
Autofagia , Ensamble y Desensamble de Cromatina , Cromatina/metabolismo , Neoplasias Colorrectales/metabolismo , Roturas del ADN de Doble Cadena , Reparación del ADN , Proteína Sequestosoma-1/metabolismo , Ubiquitinación , Autofagia/efectos de la radiación , Proteínas Relacionadas con la Autofagia/genética , Proteínas Relacionadas con la Autofagia/metabolismo , Ensamble y Desensamble de Cromatina/efectos de la radiación , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Neoplasias Colorrectales/radioterapia , Reparación del ADN/efectos de la radiación , Células HCT116 , Histonas/metabolismo , Humanos , Interferencia de ARN , Tolerancia a Radiación , Proteína Sequestosoma-1/genética , Transducción de Señal , Transfección , Enzimas Ubiquitina-Conjugadoras/genética , Enzimas Ubiquitina-Conjugadoras/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Ubiquitinación/efectos de la radiaciónRESUMEN
PURPOSE: To explore whether implantable collamer lens implantation has any effect on the postoperative objective indicators of dry eye and to determine the severity of its influence on each indicator. METHODS: This prospective observational trial was performed in the Department of Ophthalmology of West China Hospital and was approved by the Biomedical Ethics Sub-Committee of the West China Hospital of Sichuan University. A total of 89 non-dry eye patients (178 eyes in total) who received ICL implantation surgery at West China Hospital of Sichuan University were enrolled. The noninvasive keratograph tear film breakup time (NIKBUT), noninvasive keratograph tear meniscus height (NIKTMH), score of lipid layer, score of meibomian gland function, and hyperemia index were obtained via the OCULUS Keratograph for all subjects before surgery and at 1 week, 1 month, and 3 months after surgery. The fluorescein tear film breakup time (FBUT), corneal fluorescein staining score (CFS), and Schirmer test I were also measured at the same time. RESULTS: A total of 178 eyes completed the 1-week and 1-month follow-up, and 40 eyes completed the 3-month follow-up. Compared with the preoperative baseline, there was no significant difference in the NIKBUT or the corneal fluorescein staining score at each follow-up time point (P > 0.05, P > 0.05, P > 0.05). The FBUT and Schirmer test I at 1 week, 1 month, and 3 months after surgery were significantly higher than the preoperative baseline (P < 0.01, P < 0.01, P < 0.01). The NIKTMH and the score of lipid layer were significantly lower than the preoperative baseline at 1 week, 1 month, and 3 months after surgery (P < 0.01, P < 0.01, P < 0.05). The score of meibomian gland function and hyperemia index were significantly lower than the preoperative baseline 1 week after surgery (P < 0.01). CONCLUSION: ICL implantation has no adverse effect on the occurrence of postoperative dry eye, but it reduces the basal tear secretion of patients after surgery and has adverse effects on the indices of meibomian gland function in the short term postoperatively.
Asunto(s)
Síndromes de Ojo Seco , Miopía , Lentes Intraoculares Fáquicas , Lágrimas , Humanos , Estudios Prospectivos , Síndromes de Ojo Seco/diagnóstico , Síndromes de Ojo Seco/metabolismo , Síndromes de Ojo Seco/etiología , Síndromes de Ojo Seco/fisiopatología , Femenino , Masculino , Lágrimas/metabolismo , Lágrimas/fisiología , Adulto , Estudios de Seguimiento , Miopía/cirugía , Miopía/fisiopatología , Implantación de Lentes Intraoculares , Adulto Joven , Persona de Mediana Edad , Complicaciones Posoperatorias/diagnóstico , Glándulas Tarsales/metabolismo , Córnea/cirugía , Córnea/metabolismo , Córnea/patologíaRESUMEN
Calcium ions (Ca2+) play crucial roles in sperm motility and fertilization. The copine (CPNE) family comprises several Ca2+-dependent phospholipid-binding proteins. Of these, CPNE1 is extensively expressed in mammalian tissues; however, its precise role in testicular development and spermatogenesis is yet to be fully characterized. In this study, we used proteomics to analyze testicular biopsies and found that levels of CPNE1 were significantly reduced in patients with non-obstructive azoospermia (defective spermatogenesis) compared to those in patients with obstructive azoospermia (physiological spermatogenesis). In mice, CPNE1 is expressed at various stages of germ cell development and is associated with the Golgi apparatus. Ultimately, CPNE1 is expressed in the flagella of mature sperms. To further examine the role of CPNE1, we developed a Cpne1 knockout mouse model. Analysis showed that the loss of Cpne1 did not impair testicular development, spermatogenesis, or sperm morphology and motility in physiological conditions. When treated with gadolinium (III) chloride or 2-aminoethoxydiphenyl borate, known inhibitors of store-operated Ca2+ entry, Ca2+ signals and sperm motility were significantly compromised in wild-type mice; however, both mechanisms were conserved in KO mice. These results suggested that CPNE1 is dispensable for testicular development, spermatogenesis or sperm motility in physiological conditions. In addition, CPNE1 may represent a target of Ca2+ channel inhibitors and may therefore be implicated in the regulation of Ca2+ signaling and sperm motility.