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BACKGROUND: Rheumatoid arthritis (RA) is a chronic autoimmune disorder characterized by exacerbated synovial inflammation and joint destruction. Recent studies suggest toll-like receptor 4 (TLR4) internalization facilitate inflammatory response of macrophage. The role of TLR4 internalization in the pathogenesis of RA is unknown. PURPOSE: To investigate the role and mechanism of TLR4 internalization in macrophage inflammatory response of RA and explore whether TLR4 internalization mediates the anti-arthritic effect of Xiaowugui (XWG) decoction, a patented herbal formula used in China. METHODS: The co-expression of TLR4 and the internalization marker, early endosome antigen 1 (EEA1), in the synovial samples of RA patients and joint tissue of collagen-induced arthritis (CIA) mice, were evaluated using immunofluorescence. The effect of Rab5a-mediated early internalization of TLR4 on the activation induced by lipopolysaccharide (LPS) in RAW264.7 cells was investigated using small interfering RNAs that act against Rab5a. CIA was induced in Rab5a-/- mice to evaluate the role of Rab5a in vivo. The disease progression and expression of Rab5a and TLR4 in the joint tissue were evaluated in CIA mice treated with XWG. Inflammatory factors production, TLR4 internalization, and activation of downstream signaling pathways were examined in RAW264.7 cells treated with XWG in vitro. RESULTS: The co-expression and co-localization of TLR4 and EEA1 were elevated in the synovial samples of RA patients and joint tissue of CIA mice. Pharmaceutical inhibition of TLR4 internalization reduced macrophages inflammatory responses induced by LPS. The co-expression and co-localization of Rab5a and TLR4 were significantly increased in macrophages treated with LPS. Silencing Rab5a reduced LPS-induced TLR4 internalization, inflammatory factors production, and phosphorylation of Jun N-terminal kinases (JNK) and p65. Genetic deletion of Rab5a inhibited TLR4 internalization and the development of arthritis in vivo. The co-expression of TLR4 and Rab5a was also elevated in the synovial samples of RA patients. XWG treatment of mice with CIA alleviated arthritis and reduced the co-expression of Rab5a and TLR4 in the joint tissue. XWG treatment of macrophage inhibited LPS-induced IL-6 and TNF-α production, co-expression of Rab5a and TLR4, and phosphorylation of JNK and p65. CONCLUSIONS: Our findings highlight the pathogenic role of TLR4 internalization in patients with RA and identify a novel Rab5a-dependent internalization pathway that promotes macrophage inflammatory response. XWG treatment demonstrated outstanding therapeutic effects in experimental arthritis, and targeting the Rab5a-mediated internalization of TLR4 may be the main underlying mechanism.
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Dermatomicoses , Mucor , Mucormicose , Humanos , Masculino , Antifúngicos/uso terapêutico , China , Dermatomicoses/microbiologia , Dermatomicoses/patologia , Dermatomicoses/diagnóstico , Dermatomicoses/tratamento farmacológico , DNA Fúngico/genética , População do Leste Asiático , Histocitoquímica , Microscopia , Mucor/isolamento & purificação , Mucormicose/diagnóstico , Mucormicose/microbiologia , Mucormicose/patologia , Análise de Sequência de DNA , IdosoRESUMO
OBJECTIVES: Although elevated levels of neutrophil extracellular traps (NETs) have been reported in patients with rheumatoid arthritis (RA), the role of NETs in RA and the relationship between NETs and macrophages in the pathogenesis of RA requires further research. Here, we sought to determine the role of NETs in RA pathogenesis and reveal the potential mechanism. METHODS: Neutrophil elastase (NE) and myeloperoxidase (MPO)-DNA were measured in human serum and synovium. NETs inhibitor GSK484 was used to examine whether NETs involved with RA progression. We stimulated macrophages with NETs and detected internalisation-related proteins to investigate whether NETs entry into macrophages and induced inflammatory cytokines secretion through internalisation. To reveal mechanisms mediating NETs-induced inflammation aggravation, we silenced GTPases involved in internalisation and inflammatory pathways in vivo and in vitro and detected downstream inflammatory pathways. RESULTS: Serum and synovium from patients with RA showed a significant increase in NE and MPO, which positively correlated to disease activity. Inhibiting NETs formation alleviated the collagen-induced arthritis severity. In vitro, NETs are internalised by macrophages and located in early endosomes. Rab 5a was identified as the key mediator of the NETs internalisation and inflammatory cytokines secretion. Rab 5a knockout mice exhibited arthritis alleviation. Moreover, we found that NE contained in NETs activated the Rab5a-nuclear factor kappa B (NF-κB) signal pathway and promoted the inflammatory cytokines secretion in macrophages. CONCLUSIONS: This study demonstrated that NETs-induced macrophages inflammation to aggravate RA in Rab 5a dependent manner. Mechanically, Rab5a mediated internalisation of NETs by macrophages and NE contained in NETs promoted macrophages inflammatory cytokines secretion through NF-κB-light-chain-enhancer of activated B cells signal pathway. Therapeutic targeting Rab 5a or NE might extend novel strategies to minimise inflammation in RA.
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Artrite Reumatoide , Armadilhas Extracelulares , Animais , Humanos , Camundongos , Artrite Reumatoide/patologia , Citocinas/metabolismo , Inflamação , Macrófagos/metabolismo , Neutrófilos/metabolismo , NF-kappa B/metabolismo , Proteínas rab5 de Ligação ao GTPRESUMO
BACKGROUND: This study investigated the overall survival (OS) of patients with distant metastasis of ovarian cancer after chemotherapy and surgery, and explored differences in the survival of these patients with single-site metastasis. METHODS: We selected only patients with stage IV ovarian cancer who had undergone chemotherapy. Using GraphPad Prism 7 software and Kaplan-Meier survival analysis, we confirmed the factors that were meaningful for OS. Survival-related risk factors were compared using the Cox proportional hazards model. RESULTS: Patients with lung, bone, and brain metastasis experienced shorter survival than patients with liver metastasis who underwent chemotherapy. Among patients with distant metastasis who underwent chemotherapy and surgery, patients with brain metastasis experienced longer survival than patients with liver metastasis, while patients with bone and lung metastasis presented shorter survival than patients with liver metastasis. Finally, surgery prolonged the survival of patients with stage IV ovarian cancer with single-site metastasis, who were treated with chemotherapy. CONCLUSIONS: Chemotherapy combined with surgery significantly prolonged OS and cancer-specific survival (CSS) of patients with distant metastatic stage IV ovarian cancer. Lung metastasis resulted in a shorter survival than liver metastasis in patients treated with chemotherapy alone or in combination with surgery.
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Background: The discovery and development of immune checkpoint inhibitors (ICIs) has significantly enhanced the arsenal of immunotherapy treatments available for cancer patients. The identification of biomarkers that are indicative of an individual's sensitivity to treatment with ICIs is useful for screening SCLC patients prior to commencement of any ICIs based immunotherapy. However, the relationship between GBP5 and the prognosis of SCLC immunotherapy is still unclear and requires further study. Methods: We downloaded two SCLC datasets, namely the George-SCLC and Jiang-SCLC cohorts. We used the TIDE algorithm to predict the efficacy of immunotherapy for SCLC patients. The QuanTIseq, MCPcounter, and EPIC algorithms are used to calculate the proportions of immune cells in SCLC patients. Additionally, we retrospectively collected 35 SCLC samples from the first affiliated hospital of the Hengyang Medical school. Results: Patients in each cohort were devided into two groups with high (GBP5-High) and low (GBP5-Low) expression of GBP5. In both cohorts, the GBP5-High population had a higher proportion of patients that responded well to immunotherapy (responders) (p < 0.05). In addition, both GBP5-High subgroups had significantly increased cytotoxicity, chemokines, antigen presenting, and TNF family related genes. We also determined that GBP5 was related to high-level infiltration of B cells, CD4+T cells, CD8+T cells and NK cells. Conclusion: In this study, we found that GBP5 has the potential to be used as a biomarker of ICIs efficacy for SCLC patients. GBP5 is related to the quantity of inflammatory molecules, a high level of immune infiltration, and a highly activated immune response pathway.
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BACKGROUND: Wearable devices may be useful for identification, quantification and characterization, and management of atrial fibrillation (AF). To date, consumer wrist-worn devices for AF detection using photoplethysmography-based algorithms perform only periodic checks when the user is stationary and are US Food and Drug Administration cleared for prediagnostic uses without intended use for clinical decision-making. There is an unmet need for medical-grade diagnostic wrist-worn devices that provide long-term, continuous AF monitoring. METHODS AND RESULTS: We evaluated the performance of a wrist-worn device with lead-I ECG and continuous photoplethysmography (Verily Study Watch) and photoplethysmography-based convolutional neural network for AF detection and burden estimation in a prospective multicenter study that enrolled 117 patients with paroxysmal AF. A 14-day continuous ECG monitor (Zio XT) served as the reference device to evaluate algorithm sensitivity and specificity for detection of AF in 15-minute intervals. A total of 91 857 intervals were contributed by 111 subjects with evaluable reference and test data (18.3 h/d median watch wear time). The watch was 96.1% sensitive (95% CI, 92.7%-98.0%) and 98.1% specific (95% CI, 97.2%-99.1%) for interval-level AF detection. Photoplethysmography-derived AF burden estimation was highly correlated with the reference device burden (R2=0.986) with a mean difference of 0.8% (95% limits of agreement, -6.6% to 8.2%). CONCLUSIONS: Continuous monitoring using a photoplethysmography-based convolutional neural network incorporated in a wrist-worn device has clinical-grade performance for AF detection and burden estimation. These findings suggest that monitoring can be performed with wrist-worn wearables for diagnosis and clinical management of AF. REGISTRATION INFORMATION: URL: https://www.clinicaltrials.gov; Unique identifier: NCT04546763.
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Fibrilação Atrial , Aprendizado Profundo , Humanos , Algoritmos , Fibrilação Atrial/diagnóstico , Eletrocardiografia , Estudos Prospectivos , PunhoRESUMO
Mucous membrane pemphigoid (MMP) is a type of subepithelial autoimmune bullous disease, affecting various mucosae, occasionally with skin lesions. Both diagnosis and treatment of MMP are difficult. Although multiple autoantigens have been identified for MMP, the pathogenesis of MMP is still unclear. In this study, we presented a female MMP case with extensive oral mucosal lesions and skin lesions, particularly on the extremities. IgG and IgA autoantibodies against multiple autoantigens including BP180, laminin 332, integrinα6ß4 and desmoglein 3, and IgM autoantibodies against BP180 were identified during the disease course. Compared with IgG autoantibodies, the levels of IgA autoantibodies against various autoantigens decreased more significantly with improvement of clinical features after the initiation of treatments. Our findings indicated the importance of comprehensive autoantibody screening for different immunoglobulin types and autoantigens at multiple time points for the precise diagnosis of various autoimmune bullous diseases, and the significant involvement of IgA autoantibodies into the pathogenesis of MMP.
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Doenças Autoimunes , Penfigoide Mucomembranoso Benigno , Penfigoide Bolhoso , Humanos , Feminino , Penfigoide Mucomembranoso Benigno/diagnóstico , Autoanticorpos , Imunoglobulina A , Colágenos não Fibrilares , Autoantígenos , Mucosa , Imunoglobulina GRESUMO
Mesenchymal stem cells (MSCs) have been proved to have anti-inflammatory capabilities, but the mechanisms are still under investigation. Recently, oxylipins have been identified as being related to the immuno-regulation function of MSCs, but the MSC-derived oxylipins, especially under the stimulation of versatile pro-inflammatory cytokines, have never been comprehensively analyzed. In the present research, a UPLC-MS/MS method was employed to identify and quantify the oxylipin profiles of adipose-derived mesenchymal stem cells (ADSCs) under cytokine stimulation (IL-1ß, TNF-α, IFN-ð¾ and TNF-α + IFN-ð¾). The differentially produced oxylipins between experimental groups were analyzed and compared. The elevated level of lipoxygenase-15 (LOX-15) mRNA was further verified by qRT-PCR analysis. From the targeted 71 oxylipins, we detected and quantified 57 oxylipins, while 14 were not detected. Distinctive from other cytokines, ADSCs activated by the combination of IFN-ð¾ and TNF-α up-regulated LOX-15 products 7-HDHA and 15-HEPE, which were metabolized from docosahexaenoic acid (DHA) and eicosapentaenoic acid, respectively, and involved in the pro-resolution phase of inflammation. The results reported here make a first step towards a comprehensive characterization of MSC-derived oxylipins under differential proinflammatory cytokine stimulation. The findings may lay a fundamental foundation for MSC-based therapies and further determine ways to optimize the therapeutic potential of MSCs.
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Citocinas , Oxilipinas , Anti-Inflamatórios , Cromatografia Líquida , Citocinas/metabolismo , Ácidos Docosa-Hexaenoicos , Ácido Eicosapentaenoico , Lipoxigenases , Oxilipinas/análise , Oxilipinas/metabolismo , Oxilipinas/farmacologia , RNA Mensageiro , Espectrometria de Massas em Tandem , Fator de Necrose Tumoral alfaRESUMO
The efficacy of Adoptive Cell Therapy (ACT) for solid tumor is still mediocre. This is mainly because tumor cells can hijack ACT T cells' immune checkpoint pathways to exert immunosuppression in the tumor microenvironment. Immune Checkpoint Inhibitors such as anti-PD-1 (aPD1) can counter the immunosuppression, but the synergizing effects of aPD1 to ACT was still not satisfactory. Here we demonstrate an approach to safely anchor aPD1-formed nanogels onto T cell surface via bio-orthogonal click chemistry before adoptive transfer. The spatial-temporal co-existence of aPD1 with ACT T cells and the responsive drug release significantly improved the treatment outcome of ACT in murine solid tumor model. The average tumor weight of the group treated by cell-surface anchored aPD1 was only 18 % of the group treated by equivalent dose of free aPD1 and T cells. The technology can be broadly applicable in ACTs employing natural or Chimeric Antigen Receptor (CAR) T cells.
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Neoplasias , Receptores de Antígenos Quiméricos , Animais , Terapia Baseada em Transplante de Células e Tecidos , Inibidores de Checkpoint Imunológico , Imunoterapia Adotiva , Camundongos , Nanogéis , Neoplasias/metabolismo , Receptores de Antígenos de Linfócitos T/metabolismo , Microambiente TumoralRESUMO
Psoriasis is an immune-mediated chronic inflammatory dermatosis influenced by hereditary and environmental factors. Human immunodeficiency virus (HIV) infection affects the immune system and exacerbates psoriatic lesions. We report the case of a 33-year-old male patient diagnosed with psoriasis vulgaris, psoriatic arthritis and HIV infection. Acitretin capsules, etanercept and high-active antiretroviral therapy (HAART) were effective. Two months after etanercept was discontinued, his condition worsened. After switching to secukinumab combined with HAART, the symptoms of psoriatic arthritis resolved rapidly after four weeks, with a Disease Activity Index for Psoriatic Arthritis score of 0. The time to achieve psoriasis area and severity index 40, 75, 90, and 100 were 2, 4, 8, and 29 weeks. The treatment was maintained for 1 year with no adverse reactions. Regarding the stable CD4+ T lymphocyte count and the viral load, administering anti-IL-17 monoclonal antibodies is an effective treatment option for psoriasis patients.
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BACKGROUND: This study aimed to investigate the correlation between the ratio of diastolic to systolic durations (D/S) and echocardiographic parameters of patients with chronic heart failure (CHF) and evaluate whether the D/S can be used as a supplementary biomarker for the classification of heart failure (HF) phenotypes. METHODS: In total, 122 CHF patients with a left ventricular ejection fraction (LVEF) <40%, 40%≤LVEF<50%, or ≥50% were categorized as having HF with a reduced ejection fraction (HFrEF) (N=32), HF with a mid-range ejection fraction (HFmrEF) (N=21) or HF with a preserved ejection fraction (HFpEF) (N=69), respectively. All patients underwent echocardiography for assessment of nineteen structural and functional echocardiographic parameters and digital phonocardiography for the measurement of D/S. Spearman correlation was used to analyse the associations between the D/S and echocardiographic parameters. Multivariate logistic regression analysis was performed to examine the associations between the D/S and HF phenotypes, and receiver operating characteristic (ROC) curve analysis was employed to evaluate the predictive value of the D/S in the classification of HF phenotypes. RESULTS: The D/S values of patients with HFrEF, HFmrEF and HFpEF were 1.32±0.06, 1.44±0.11 and 1.54±0.08, respectively, which were significantly different (All P<0.05). A close correlation between the D/S and LVEF was found (r=0.777, P<0.001). The multivariate analysis indicated that the D/S was an independent risk factor for CHF phenotypes (OR=4.927, 95% CI 2.532-9.587; P<0.001). The area under the ROC curve for distinguishing between HFmrEF and HFpEF using the D/S was 0.764 (95% CI 0.707-0.845; P < 0.001) and that for distinguishing between HFmrEF and HFrEF using the D/S was 0.821 (95% CI 0.755-0.882; P < 0.001). CONCLUSION: The D/S was significantly associated with LVEF, and as LVEF decreased, the D/S tended to decrease, which could also serve as a noninvasive supplementary indicator for detecting systolic and diastolic dysfunction.
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Tumor tissues/cells are the best sources of antigens to prepare cancer vaccines. However, due to the difficulty of solubilization and delivery of water-insoluble antigens in tumor tissues/cells, including water-insoluble antigens into cancer vaccines and delivering such vaccines efficiently to antigen-presenting cells (APCs) remain challenging. To solve these problems, herein, water-insoluble components of tumor tissues/cells are solubilized by 8 m urea and thus whole components of micrometer-sized tumor cells are reasssembled into nanosized nanovaccines. To induce maximized immunization efficacy, various antigens are loaded both inside and on the surface of nanovaccines. By encapsulating both water-insoluble and water-soluble components of tumor tissues/cells into nanovaccines, the nanovaccines are efficiently phagocytosed by APCs and showed better therapeutic efficacy than the nanovaccine loaded with only water-soluble components in melanoma and breast cancer. Anti-PD-1 antibody and metformin can improve the efficacy of nanovaccines. In addition, the nanovaccines can prevent lung cancer (100%) and melanoma (70%) efficiently in mice. T cell analysis and tumor microenvironment analysis indicate that tumor-specific T cells are induced by nanovaccines and both adaptive and innate immune responses against cancer cells are activated by nanovaccines. Overall, this study demonstrates a universal method to make tumor-cell-based nanovaccines for cancer immunotherapy and prevention.
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ImunoterapiaRESUMO
Lung cancer is still a main cause of cancer-related death worldwide. Non-small-cell lung cancer (NSCLC) accounts for the majority of lung cancers, and gefitinib is an effective targeted drug for NSCLC. It is important to explore the underlying molecular mechanisms of gefitinib resistance to provide new treatment strategies and to improve the prognosis of gefitinib-resistant NSCLC patients. This study aimed to examine the role of filamin A (FLNA) in acquired resistance to gefitinib in NSCLC, and identify ANXA2 (annexin A2), one of calcium-dependent phospholipid-binding proteins, as its corresponding regulatory factor. First, we established resistant cells via long-term exposure to gefitinib to analyse the association between FLNA and gefitinib resistance. Through quantitative real-time polymerase chain reaction (qRT-PCR), Cell Counting Kit-8 (CCK-8), western blotting (WB), and flow cytometry assays, we evaluated the role of FLNA. The effect of FLNA knockdown or overexpression was analysed not only in cell lines but also in mouse models. We verified the FLNA-interacting protein through coimmunoprecipitation (CoIP) experiments and found that the downstream signalling pathway was regulated by FLNA and its interacting protein. Finally, the upstream transcription factor was identified by chromatin immunoprecipitation (ChIP). Increased FLNA expression induced gefitinib resistance. Knockdown of FLNA restored gefitinib sensitivity and induced apoptosis in vivo and in vitro. FLNA and ANXA2 cooperatively led to the activation of the Wnt pathway, which was closely linked to gefitinib resistance. Subsequently, SP1 promoted transcriptional activation of FLNA to regulate gefitinib resistance. We determined that FLNA serves as a regulator of gefitinib resistance in NSCLC and found that FLNA and ANXA2 together induced gefitinib resistance by activating the Wnt pathway.
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Anexina A2/metabolismo , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Filaminas/metabolismo , Gefitinibe/farmacologia , Neoplasias Pulmonares/metabolismo , Proteínas de Neoplasias/metabolismo , Via de Sinalização Wnt/efeitos dos fármacos , Anexina A2/genética , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral , Filaminas/genética , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Proteínas de Neoplasias/genéticaRESUMO
To improve antitumor efficiency of chemotherapy and reduce side effect, according to the physiological characteristics of tumor tissues and tumor intracellular microenvironment, a multifunctional drug delivery system with properties of long circulation, active targeting, redox and pH triggered drug release was established based on the Generation 4 polyamidoamine dendrimer (PAMAM). First, the redox cleavable disulfide bonds (SS) were introduced for linking polyethylene glycol (PEG) with PAMAM to form PAMAM-S-S-PEG (PSSP). Then cRGD peptide was applied to the PEG end of PSSP to construct RGD-PSSP conjugates. Finally, encapsulating the antitumor chemotherapy drug doxorubicin (DOX) into the hydrophobic cavity of RGD-PSSP conjugates constructed the RGD-PSSP/DOX drug delivery system. The in vitro experiments displayed that RGD-PSSP/DOX NPs showed obviously redox and pH dual sensitive drug release profile. MTT and cell uptake observation elucidated cRGD modification could increase the cytotoxicity, and promote the uptake of B16 cells and HUVEC cells both overexpressing integrin ανß3on cell membrane. Cell uptake mechanism investigation further revealed that RGD-PSSP/DOX interacted with plasma membrane via specific recognition of cRGD peptide with integrin ανß3, and was subsequently internalized mainly through clathrin- and caveolin-mediated endocytosis. Remarkably, RGD-PSSP/DOX presented superior anticancer activity and lower heart and kidney toxicity in vivo, which could be regarded as a potential candidate for efficient antitumor chemotherapy drug delivery.
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Antineoplásicos , Dendrímeros , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Doxorrubicina/farmacologia , Portadores de Fármacos , Sistemas de Liberação de Medicamentos , Concentração de Íons de Hidrogênio , Oxirredução , Poliaminas , PolietilenoglicóisRESUMO
BACKGROUND: For real-time monitoring of hospital patients, high-quality inference of patients' health status using all information available from clinical covariates and lab test results is essential to enable successful medical interventions and improve patient outcomes. Developing a computational framework that can learn from observational large-scale electronic health records (EHRs) and make accurate real-time predictions is a critical step. In this work, we develop and explore a Bayesian nonparametric model based on multi-output Gaussian process (GP) regression for hospital patient monitoring. METHODS: We propose MedGP, a statistical framework that incorporates 24 clinical covariates and supports a rich reference data set from which relationships between observed covariates may be inferred and exploited for high-quality inference of patient state over time. To do this, we develop a highly structured sparse GP kernel to enable tractable computation over tens of thousands of time points while estimating correlations among clinical covariates, patients, and periodicity in patient observations. MedGP has a number of benefits over current methods, including (i) not requiring an alignment of the time series data, (ii) quantifying confidence regions in the predictions, (iii) exploiting a vast and rich database of patients, and (iv) inferring interpretable relationships among clinical covariates. RESULTS: We evaluate and compare results from MedGP on the task of online prediction for three patient subgroups from two medical data sets across 8,043 patients. We find MedGP improves online prediction over baseline and state-of-the-art methods for nearly all covariates across different disease subgroups and hospitals. CONCLUSIONS: The MedGP framework is robust and efficient in estimating the temporal dependencies from sparse and irregularly sampled medical time series data for online prediction. The publicly available code is at https://github.com/bee-hive/MedGP .
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Algoritmos , Modelos Estatísticos , Teorema de Bayes , Distribuição NormalRESUMO
Erlotinib, an epidermal growth factor receptor tyrosine kinase inhibitor (EGFRTKI), is widely applied as a firstline treatment for nonsmall cell lung cancer (NSCLC) and greatly improves the clinical outcomes of patients. However, acquired resistance to EGFRTKIs remains a major clinical challenge. Here, we identified guanylatebinding protein1 (GBP1) as a novel protein related to erlotinib resistance, and explored the specific mechanism by which GBP1 is involved in erlotinib resistance. First, the human NSCLC cells PC9ER and HCC827ER were generated by exposing cells to increasing concentrations of erlotinib over 6 months. We screened different genes between erlotinibsensitive and erlotinibresistant cells with data from the Gene Expression Omnibus database and detected the expression of these genes in erlotinibresistant and erlotinibsensitive cells by quantitative realtime polymerase chain reaction (qPCR). Moreover, we constructed GBP1knockdown and GBP1overexpressing cells to determine the IC50 value of erlotinib, to perform an apoptosis assay and to examine cell cycle distribution. A subcutaneous tumorigenesis test was used to analyze how GBP1 affects erlotinib resistance. Then, mass spectrometry analysis and coimmunoprecipitation were performed to verify the interaction between GBP1 and phosphoglycerate kinase 1 (PGK1). Changes in epithelialmesenchymal transition (EMT)related markers were observed following the upregulation and downregulation of PGK1 expression. Finally, a rescue experiment was used to determine whether GBP1 regulates EMT through PGK1. In the present study, GBP1 was significantly upregulated in erlotinibresistant cells, compared with erlotinibsensitive cells. In vitro and in vivo experiments showed that upregulated GBP1 expression contributed to erlotinib resistance, while decreased GBP1 expression had the opposite effect. As shown by performing survival analysis, high GBP1 expression predicted poor prognosis in patients with lung adenocarcinoma. Furthermore, the interaction between GBP1 and PGK1 was confirmed, and a rescue experiment revealed that GBP1 regulates EMT via PGK1. Finally, functional experiments showed that EMT is involved in erlotinib resistance. Our study suggests that GBP1 regulates erlotinib resistance via PGK1mediated EMT signaling, suggesting GBP1 as a potential therapeutic target in erlotinibresistant NSCLC.
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Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Cloridrato de Erlotinib/farmacologia , Proteínas de Ligação ao GTP/metabolismo , Neoplasias Pulmonares/tratamento farmacológico , Fosfoglicerato Quinase/metabolismo , Animais , Antineoplásicos/farmacologia , Carcinoma Pulmonar de Células não Pequenas/mortalidade , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Feminino , Proteínas de Ligação ao GTP/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Neoplasias Pulmonares/mortalidade , Neoplasias Pulmonares/patologia , Camundongos Nus , Transdução de Sinais/efeitos dos fármacos , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
INTRODUCTION: A multifunctional redox- and pH-responsive polymeric drug delivery system is designed and investigated for targeted anticancer drug delivery to liver cancer. METHODS: The nanocarrier (His-PAMAM-ss-PEG-Tf, HP-ss-PEG-Tf) is constructed based on generation 4 polyamidoamine dendrimer (G4 PAMAM). Optimized amount of histidine (His) residues is grafted on the surface of PAMAM to obtain enhanced pH-sensitivity and proton-buffering capacity. Disulfide bonds (ss) are introduced between PAMAM and PEG to reach accelerated intracellular drug release. Transferrin (Tf) was applied to achieve active tumor targeting. Doxorubicin (DOX) is loaded in the hydrophobic cavity of the nanocarrier to exert its anti-tumor effect. RESULTS: The results obtained from in vitro and in vivo evaluation indicate that HP-ss-PEG-Tf/DOX complex has pH and redox dual-sensitive properties, and exhibit higher cellular uptake and cytotoxicity than the other control groups. Flow cytometry and confocal microscopy display internalization of HP-ss-PEG-Tf/DOX via clathrin mediated endocytosis and effective endosomal escape in HepG2 cancer cells. Additionally, cyanine 7 labeled HP-ss-PEG-Tf conjugate could quickly accumulate in the HepG2 tumor. Remarkably, HP-ss-PEG-Tf/DOX present superior anticancer activity, enhanced apoptotic activity and lower heart and kidney toxicity in vivo. DISCUSSION: Thus, HP-ss-PEG-Tf is proved to be a promising candidate for effective targeting delivery of DOX into the tumor.
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Antineoplásicos/administração & dosagem , Dendrímeros/química , Portadores de Fármacos/administração & dosagem , Nylons/química , Transferrina/química , Animais , Antineoplásicos/química , Antineoplásicos/farmacocinética , Dendrímeros/administração & dosagem , Doxorrubicina/administração & dosagem , Doxorrubicina/química , Doxorrubicina/farmacocinética , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Liberação Controlada de Fármacos , Feminino , Células Hep G2 , Histidina/química , Humanos , Concentração de Íons de Hidrogênio , Camundongos Endogâmicos BALB C , Oxirredução , Polietilenoglicóis/química , Succinimidas/química , Transferrina/farmacocinética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Laboratory testing is an integral tool in the management of patient care in hospitals, particularly in intensive care units (ICUs). There exists an inherent trade-off in the selection and timing of lab tests between considerations of the expected utility in clinical decision-making of a given test at a specific time, and the associated cost or risk it poses to the patient. In this work, we introduce a framework that learns policies for ordering lab tests which optimizes for this trade-off. Our approach uses batch off-policy reinforcement learning with a composite reward function based on clinical imperatives, applied to data that include examples of clinicians ordering labs for patients. To this end, we develop and extend principles of Pareto optimality to improve the selection of actions based on multiple reward function components while respecting typical procedural considerations and prioritization of clinical goals in the ICU. Our experiments show that we can estimate a policy that reduces the frequency of lab tests and optimizes timing to minimize information redundancy. We also find that the estimated policies typically suggest ordering lab tests well ahead of critical onsets-such as mechanical ventilation or dialysis-that depend on the lab results. We evaluate our approach by quantifying how these policies may initiate earlier onset of treatment.
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Técnicas de Laboratório Clínico , Unidades de Terapia Intensiva , Injúria Renal Aguda/diagnóstico , Técnicas de Laboratório Clínico/estatística & dados numéricos , Biologia Computacional , Cuidados Críticos/estatística & dados numéricos , Técnicas de Apoio para a Decisão , Humanos , Unidades de Terapia Intensiva/organização & administração , Unidades de Terapia Intensiva/estatística & dados numéricos , Administração dos Cuidados ao Paciente/organização & administração , Administração dos Cuidados ao Paciente/estatística & dados numéricos , Reforço Psicológico , Recompensa , Sepse/diagnósticoRESUMO
Most human protein-coding genes can be transcribed into multiple distinct mRNA isoforms. These alternative splicing patterns encourage molecular diversity, and dysregulation of isoform expression plays an important role in disease etiology. However, isoforms are difficult to characterize from short-read RNA-seq data because they share identical subsequences and occur in different frequencies across tissues and samples. Here, we develop BIISQ, a Bayesian nonparametric model for isoform discovery and individual specific quantification from short-read RNA-seq data. BIISQ does not require isoform reference sequences but instead estimates an isoform catalog shared across samples. We use stochastic variational inference for efficient posterior estimates and demonstrate superior precision and recall for simulations compared to state-of-the-art isoform reconstruction methods. BIISQ shows the most gains for low abundance isoforms, with 36% more isoforms correctly inferred at low coverage versus a multi-sample method and 170% more versus single-sample methods. We estimate isoforms in the GEUVADIS RNA-seq data and validate inferred isoforms by associating genetic variants with isoform ratios.