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BACKGROUND AIMS: Hu8F4 is a T-cell receptor-like antibody with high affinity for the leukemia-associated antigen PR1/HLA-A2 epitope. Adapted into a chimeric antigen receptor (CAR) format, Hu8F4-CAR is composed of the Hu8F4 single-chain variable fragment, the human IgG1 CH2CH3 extracellular spacer domain, a human CD28 costimulatory domain and the human CD3ζ signaling domain. We have demonstrated high efficacy of Hu8F4-CAR-T cells against PR1/HLA-A2-expressing cell lines and leukemic blasts from patients with acute myeloid leukemia in vitro. Previous studies have shown that modification of the Fc domains of IgG4 CH2CH3 spacer regions can eliminate activation-induced cell death and off-target killing mediated by mouse Fc gamma receptor-expressing cells. METHODS: We generated Hu8F4-CAR(PQ) with mutated Fc receptor binding sites on the CH2 domain of Hu8F4-CAR to prevent unwanted interactions with Fc gamma receptor-expressing cells in vivo. RESULTS: The primary human T cells transduced with Hu8F4-CAR(PQ) can specifically lyse HLA-A2+ PR1-expressing leukemia cell lines in vitro. Furthermore, both adult donor-derived and cord blood-derived Hu8F4-CAR(PQ)-T cells are active and can eliminate U937 leukemia cells in NSG mice. CONCLUSIONS: Herein, we demonstrate that modification of the IgG1-based spacer can eliminate Fc receptor binding-induced adverse effects and Hu8F4-CAR(PQ)-T cells can kill leukemia in vivo.
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PURPOSE: Exercise-based cardiac rehabilitation (EBCR) improves functional capacity in heart failure (HF). However, data on the effect of EBCR in patients with advanced HF and left ventricular assist devices (LVADs) are limited. This meta-analysis aimed to evaluate the impact of EBCR on the functional ability of LVAD patients by comparing the corresponding outcome indicators between the EBCR and ST groups. METHODS: PubMed, Embase, Clinical Trials, and Cochrane Library databases were searched for studies assessing and comparing the effects of EBCR and standard therapy (ST) in patients following LVAD implantation. Using pre-defined criteria, appropriate studies were identified and selected. Data from selected studies were extracted in a standardized fashion, and a meta-analysis was performed using a fixed-effects model. The protocol was registered on INPLASY (202340073). RESULTS: In total, 12 trials involving 477 patients were identified. The mean age of the participants was 52.9 years, and 78.6% were male. The initiation of EBCR varied from LVAD implantation during the index hospitalization to 11 months post-LVAD implantation. The median rehabilitation period ranged from 2 weeks to 18 months. EBCR was associated with improved peak oxygen uptake (VO2) in all trials. Quantitative analysis was performed in six randomized studies involving 214 patients (EBCR: n = 130, ST: n = 84). EBCR was associated with a significantly high peak VO2 (weighted mean difference [WMD] = 1.64 mL/kg/min; 95% confidence interval [CI], 0.20-3.08; p = .03). Similarly, 6-min walk distance (6MWD) showed significantly greater improvement in the EBCR group than in the ST group (WMD = 34.54 m; 95% CI, 12.47-56.42; p = .002) in 266 patients (EBCR, n = 140; ST, n = 126). Heterogeneity was low among the included trials. None of the included studies reported serious adverse events related to EBCR, indicating the safety of EBCR after LVAD implantation. CONCLUSION: This study demonstrated that EBCR following LVAD implantation is associated with greater improvement in functional capacity compared with ST as reflected by the improved peak VO2 and 6MWD values. Considering the small number of patients in this analysis, further research on the clinical impact of EBCR in LVAD patients is warranted.
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Hu8F4 is a T cell receptor (TCR)-like antibody with high affinity for leukemia-associated antigen PR1/HLA-A2 epitope. Adapted into a chimeric antigen receptor (CAR) format, Hu8F4-CAR is comprised of the Hu8F4 scFv, the human IgG1 CH2CH3 extracellular spacer domain, a human CD28 costimulatory domain, and the human CD3ζ signaling domain. We have demonstrated high efficacy of Hu8F4-CAR-T cells against PR1/HLA-A2-expressing cell lines and leukemic blasts from AML patients in vitro. Previous studies have shown that modification of the Fc domains of IgG4 CH2CH3 spacer regions can eliminate activation-induced cell death and off-target killing mediated by mouse Fc gamma receptor (FcgR)-expressing cells. We generated Hu8F4-CAR(PQ) with mutated Fc receptor binding sites on the CH2 domain of Hu8F4-CAR to prevent unwanted interactions with FcgR-expressing cells in vivo. The primary human T cells transduced with Hu8F4-CAR(PQ) can specifically lyse HLA-A2+ PR1-expressing leukemia cell lines in vitro. Furthermore, both adult donor-derived and cord blood-derived Hu8F4-CAR(PQ)-T cells are active and can eliminate U937 leukemia cells in NSG mice. Herein, we demonstrate that modification of the IgG1-based spacer can eliminate Fc receptor-binding-induced adverse effects and Hu8F4-CAR(PQ)-T cells can kill leukemia in vivo.
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This article investigates the efficacy of a regularized multitask learning (MTL) framework based on SVM (M-SVM) to answer whether MTL always provides reliable results and how MTL outperforms independent learning. We first find that the M-SVM is Bayes risk consistent in the limit of a large sample size. This implies that despite the task dissimilarities, the M-SVM always produces a reliable decision rule for each task in terms of the misclassification error when the data size is large enough. Furthermore, we find that the task-interaction vanishes as the data size goes to infinity, and the convergence rates of the M-SVM and its single-task counterpart have the same upper bound. The former suggests that the M-SVM cannot improve the limit classifier's performance; based on the latter, we conjecture that the optimal convergence rate is not improved when the task number is fixed. As a novel insight into MTL, our theoretical and experimental results achieved an excellent agreement that the benefit of the MTL methods lies in the improvement of the preconvergence-rate (PCR) factor (to be denoted in Section III) rather than the convergence rate. Moreover, this improvement of PCR factors is more significant when the data size is small. In addition, our experimental results of five other MTL methods demonstrate the generality of this new insight.
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PURPOSE: A new type of visualized steerable sheath (Vizigo sheath; Biosense Webster Inc., Irvine, CA, USA) has been employed in clinical treatment. This study aimed to compare the effectiveness and safety of the Vizigo sheath to a fixed sheath (Swartz sheath; St. Jude Inc., St. Paul, MN, USA) for catheter ablation of paroxysmal atrial fibrillation (PAF). METHODS: We analyzed the procedural time, fluoroscopy time, contact force (CF), and initial pulmonary vein isolation (PVI) rate. After 6 months of follow-up, the success rate of ablation between the two groups was compared. RESULTS: Compared to the Swartz sheath, using the Vizigo sheath can significantly reduce the total procedural time and fluoroscopy time and increase the overall average CF, especially in the anterior left pulmonary vein (LPV), superior LPV, posterior right pulmonary vein (RPV), and superior RPV. The proportion of CF within a reasonable range in the Vizigo group was significantly higher than that in the Swartz group, especially in the anterior LPV, posterior RPV, and superior RPV. Besides, the left, right, and bilateral initial PVI rates in the Vizigo group were significantly higher. CONCLUSIONS: The visualized steerable sheath for PAF catheter ablation not only reduced radiation exposure but also significantly improved CF and initial PVI rate, all of which indicated an increased rate of successful ablation.
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Fibrilación Atrial , Ablación por Catéter , Venas Pulmonares , Fibrilación Atrial/diagnóstico por imagen , Fibrilación Atrial/cirugía , Fluoroscopía , Humanos , Venas Pulmonares/cirugía , Resultado del TratamientoRESUMEN
BACKGROUND AIMS: Human myeloperoxidase has been shown to be overexpressed in many types of leukemia, such as chronic myeloid leukemia, acute myeloid leukemia and myelodysplastic syndrome. The authors identified two myeloperoxidase-derived HLA-A2-restricted peptides, MY4 and MY8, as novel leukemia-associated antigens. METHODS: Ex vivo-elicited MY4- and MY8-specific cytotoxic T lymphocytes were generated, and tested for leukemia cell lysis in vitro and in NOD/SCID AML xenograft model. RESULTS: These MY4- and MY8-specific cytotoxic T lymphocytes killed leukemic blasts while sparing healthy donor bone marrow cells. In addition, co-injection of MY4- and MY8-specific cytotoxic T lymphocytes into nonobese diabetic/severe combined immunodeficiency mice with acute myeloid leukemia drastically reduced tumor burden in vivo. The authors also found that MY4- and MY8-specific T cells could be detected in the peripheral blood mononuclear cells of allogeneic stem cell transplant recipients. CONCLUSIONS: These antigen-specific T cells were significantly increased in blood samples from patients compared with healthy donors, suggesting that both MY4 and MY8 are immunogenic and that MY4- and MY8-specific cytotoxic T lymphocytes may play a role in reducing leukemia in vivo. Thus, the discovery of MY4 and MY8 as novel leukemia-associated antigens paves the way for targeting these antigens in immunotherapy against myeloid leukemia.
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Antígeno HLA-A2 , Leucemia Mieloide Aguda , Animales , Humanos , Leucemia Mieloide Aguda/terapia , Leucocitos Mononucleares , Ratones , Ratones Endogámicos NOD , Ratones SCID , Péptidos , Peroxidasa , Linfocitos T CitotóxicosRESUMEN
BACKGROUND AND OBJECTIVE: Systematically review the current published literature on the impact of exercise training (ET) in chronic heart failure (CHF) patients who were conducted cardiac resynchronization therapy (CRT). METHODS: PubMed, EMBASE, and the Cochrane Library of Controlled Trails databases were searched for trials comparing the additional effects of ET in CHF patients after CRT implantation with no exercise or usual care control up until 2020.03.07. We independently screened the literature, extracted data, employed the tool for the assEssment of Study qualiTy and reporting in EXercise (TESTEX) to evaluate study quality and risk of bias, and performed meta-analysis with Revman 5.3 software. RESULTS: Eight trials were identified for qualitative analysis and 7 randomized controlled trails (RCTs) included 235 participants (120 ET; 115 controls) for quantitative analysis. The results showed that the maximal workload (mean difference [MD] 26.32âW, 95% CI 19.41-33.23; Pâ<â.00001, I2â=â0%) and the exercise duration (MD 68.95âseconds, 95% CI 15.41-122.48; Pâ=â.01, I2â=â76%) had significant improvement in the ET group versus control. Subgroup analysis showed that compared with control, the change in peak oxygen uptake (VO2) (MD 3.05âml/kg/minute, 95% CI 2.53-3.56; Pâ<â.00001, I2â=â0%), left ventricular ejection fraction (LVEF) (MD 4.97%, 95% CI 1.44-8.49; Pâ=â.006, I2â=â59%), and health related quality of life (HRQoL) (the change in Minnesota living with heart failure questionnaire [MLHFQ]: MD -19.96, 95% CI -21.57 to -18.34; Pâ<â.00001, I2â=â0%) were significantly improved in the light to moderate intensity training (non-HIT) group, while there seemed no statistical difference of above endpoints in the high intensity training (HIT) group. CONCLUSION: During the short term (up to 6âmonths), non-HIT could improve exercise capacity, cardiac function, and HRQoL in CHF patients with CRT. However, due to the small number of participants, a high-quality large-sample multicenter trial is demanded.
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Rehabilitación Cardiaca/métodos , Terapia de Resincronización Cardíaca/métodos , Terapia por Ejercicio/métodos , Ejercicio Físico/fisiología , Insuficiencia Cardíaca/rehabilitación , Anciano , Enfermedad Crónica , Tolerancia al Ejercicio , Femenino , Insuficiencia Cardíaca/fisiopatología , Humanos , Masculino , Persona de Mediana Edad , Calidad de Vida , Ensayos Clínicos Controlados Aleatorios como Asunto , Volumen Sistólico/fisiología , Resultado del Tratamiento , Función Ventricular Izquierda/fisiologíaRESUMEN
INTRODUCTION: Hemodynamically-instable ventricular arrhythmias (VAs) are rare in patients with pulmonary hypertension (PH). To the best of our knowledge, only 1 case has been reported so far. Moreover, the pathogenesis of this kind of arrhythmia remains obscured and its treatment is challenging. Here we report another case and presented the substrate for VAs initiation and therapeutic effect of radiofrequency ablation. PATIENT CONCERNS: This is a 57-year-old man who presented paroxysmal palpitation associated with presyncope at rest. Surface electrocardiogram (ECG) revealed frequent ventricular premature contractions and non-sustained ventricular tachycardia when symptoms occurred. He also had a history of severe PH which was secondary to atrial septal defect and partial anomalous pulmonary venous drainage and suffered from obvious dyspnea when climbing stairs World Health Organization Class III (WHO Class III). DIAGNOSIS: Hemodynamically-instable VAs associated with severe PH. INTERVENTION: Echocardiography revealed enlargement of right ventricle (right ventricle [RV]: 43âmm). Electrophysiological examination showed the origin of VAs is next to a small low-voltage zone of RV. Radiofrequency delivery at the origin successfully terminated VAs without occurrence of complication. OUTCOME: The patient was free from arrhythmias and got an improvement of exercise tolerance, just with mild dyspnea when climbing stairs World Health Organization Class II (WHO class II), during six-month follow up. LESSONS: This case suggests the low-voltage zone of remodeled RV, which may be secondary to increased pulmonary artery pressure, serves as the substrate for VAs initiation in patient with PH. Radiofrequency ablation can successfully terminate VAs and the termination of VAs can significantly improve the patient's impaired exercise tolerance.
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Hipertensión Pulmonar/complicaciones , Ablación por Radiofrecuencia/métodos , Complejos Prematuros Ventriculares/complicaciones , Ecocardiografía , Técnicas Electrofisiológicas Cardíacas/métodos , Ventrículos Cardíacos/diagnóstico por imagen , Ventrículos Cardíacos/fisiopatología , Humanos , Hipertensión Pulmonar/fisiopatología , Masculino , Persona de Mediana Edad , Complejos Prematuros Ventriculares/fisiopatología , Complejos Prematuros Ventriculares/terapia , Remodelación VentricularRESUMEN
The driving process and explanatory factors regulating the transformation and migration of arsenic (As) mediated by dissimilatory iron reducing bacteria (DFeRB) and sulfate reducing bacteria (SRB) remain poorly understood. The novelty of this study is to explore the driving process and key environmental factors governing As mobilization mediated by DFeRB and SRB based on continuous As speciation and environmental parameter monitoring in a sediment-water system. The results illustrate the reduction process mediated by DFeRB and SRB significantly promotes the reduction of As(V) and the endogenous release of As. However, in the DFeRB and SRB mediated reductions, the main driving process and key explanatory factors that dominate the As mobility are significantly different. DFeRB has significant effects on the reductive dissolution and re-distribution of Fe(III) oxyhydroxides and As-containing Fe(III) minerals and on adsorption-desorption, which in turn influenced the transformation of iron species and the release and ecotoxicity of As. Meanwhile, the environmental factors that affect As mobility depend on Fe2+ and Fe3+ in DFeRB-induced reduction, presenting two main pathways: the process of As mobilization mediated by DFeRB, and the process influenced by the inorganic phosphorus involved in the competitive adsorption and anion exchange. Significantly different from DFeRB, the effects of SRB on As behavior mainly occur by influencing the adsorbed As, pyrite, and As sulfides in the sediments and through the formation of sulfides during the sulfate reduction. The main pathways of As mobilization reflect the direct effects of SRB, S2-, and Fe2+. In addition, the role of NH4+-N in the driving process of As mobility is more pronounced in SRB-induced reduction. NO3--N is an essential factor affecting As mobility, but the effects of NO3--N on As lead to non-significant pathways. This work provides insights into the environmental effects of DFeRB and SRB on the biogeochemical cycle of As.
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PURPOSE: In radiofrequency ablation near coronary arteries (CA), coronary angiography is traditionally recommended to estimate distance between catheter and CA. This study aimed to investigate the feasibility of an alternative approach for intuitively demonstrating spatial location of catheter and CA during ablation of ventricular arrhythmias (VAs) originating from aortic root (AR) and great cardiac vein (GCV). METHODS: During mapping and ablation, 3D-reconstructed cardiac CT and electroanatomic mapping were merged, and distance between CA and catheter was monitored. Coronary angiography, for distance verification, was used when the distance was less than 5 mm in image integration model (IIM). RESULTS: Twenty-three patients (52.26 ± 17.89 years, 12 men) with ablation originating in left cusp (LCC, n = 8), right cusp (n = 2), and left-right cusp junction (LCC-RCC, n = 12) and GCV (n = 1) were enrolled. In IIM, the distance between origin and CA was less than 5 mm in 2 VAs originating in LCC and one in GCV (3/23), whereas distance for ablation was always safe (12.3-22.3 mm) for VAs of LCC-RCC origin. IIM avoided angiography use in 20 patients, reducing radiation exposure by 80.6% (650.18 ± 624.31 vs 3356.97 ± 1529.46uGycm2, P = 0.088). VA termination failed in two cases of LCC origin due to proximity to CA, and was achieved in all other patients (91.3%). No CA damage occurred during the procedures. CONCLUSION: Mapping and ablation under IIM guidance of VAs of AR and GCV origin appears feasible and safe, while avoiding angiography use particularly in VAs of LCC-RCC origin.
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Ablación por Catéter , Taquicardia Ventricular , Aorta , Arritmias Cardíacas/cirugía , Vasos Coronarios , Electrocardiografía , Técnicas Electrofisiológicas Cardíacas , Femenino , Humanos , Imagenología Tridimensional , Masculino , Persona de Mediana Edad , Taquicardia Ventricular/diagnóstico por imagen , Taquicardia Ventricular/cirugía , Tomografía Computarizada por Rayos X , Resultado del TratamientoRESUMEN
Immunotherapy targeting leukemia-associated antigens has shown promising results. Because of the heterogeneity of leukemia, vaccines with a single peptide have elicited only a limited immune response. Targeting several peptides together elicited peptide-specific cytotoxic T lymphocytes (CTLs) in leukemia patients, and this was associated with clinical responses. Thus, the discovery of novel antigens is essential. In the current study, we investigated cyclin E as a novel target for immunotherapy. Cyclin E1 and cyclin E2 were found to be highly expressed in hematologic malignancies, according to reverse transcription polymerase chain reaction and western blot analysis. We identified two HLA-A*0201 binding nonameric peptides, CCNE1M from cyclin E1 and CCNE2L from cyclin E2, which both elicited the peptide-specific CTLs. The peptide-specific CTLs specifically kill leukemia cells. Furthermore, CCNE1M and CCNE2L CTLs were increased in leukemia patients who underwent allogeneic hematopoietic stem cell transplantation, and this was associated with desired clinical outcomes. Our findings suggest that cyclin E1 and cyclin E2 are potential targets for immunotherapy in leukemia.
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Antígenos de Neoplasias/metabolismo , Vacunas contra el Cáncer/inmunología , Ciclina E/inmunología , Ciclinas/inmunología , Antígeno HLA-A2/inmunología , Leucemia/inmunología , Proteínas Oncogénicas/inmunología , Adulto , Anciano , Antígenos de Neoplasias/inmunología , Vacunas contra el Cáncer/farmacología , Epítopos de Linfocito T/inmunología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Linfocitos T Citotóxicos/inmunología , Adulto JovenRESUMEN
Purpose: PR1 is a human leukocyte antigen (HLA)-A2 nonameric peptide derived from neutrophil elastase (NE) and proteinase 3 (P3). We have previously shown that PR1 is cross-presented by solid tumors, leukemia, and antigen-presenting cells, including B cells. We have also shown that cross-presentation of PR1 by solid tumors renders them susceptible to killing by PR1-targeting immunotherapies. As multiple myeloma is derived from B cells, we investigated whether multiple myeloma is also capable of PR1 cross-presentation and subsequently capable of being targeted by using PR1 immunotherapies.Experimental Design: We tested whether multiple myeloma is capable of cross-presenting PR1 and subsequently becomes susceptible to PR1-targeting immunotherapies, using multiple myeloma cell lines, a xenograft mouse model, and primary multiple myeloma patient samples.Results: Here we show that multiple myeloma cells lack endogenous NE and P3, are able to take up exogenous NE and P3, and cross-present PR1 on HLA-A2. Cross-presentation by multiple myeloma utilizes the conventional antigen processing machinery, including the proteasome and Golgi, and is not affected by immunomodulating drugs (IMiD). Following PR1 cross-presentation, we are able to target multiple myeloma with PR1-CTL and anti-PR1/HLA-A2 antibody both in vitro and in vivoConclusions: Collectively, our data demonstrate that PR1 is a novel tumor-associated antigen target in multiple myeloma and that multiple myeloma is susceptible to immunotherapies that target cross-presented antigens. Clin Cancer Res; 24(14); 3386-96. ©2018 AACR.
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Antineoplásicos Inmunológicos/farmacología , Antígeno HLA-A2/inmunología , Mieloma Múltiple/inmunología , Fragmentos de Péptidos/antagonistas & inhibidores , Fragmentos de Péptidos/inmunología , Animales , Presentación de Antígeno/efectos de los fármacos , Presentación de Antígeno/inmunología , Células Presentadoras de Antígenos/inmunología , Transporte Biológico , Línea Celular Tumoral , Activación de Complemento , Reactividad Cruzada/efectos de los fármacos , Reactividad Cruzada/inmunología , Citotoxicidad Inmunológica , Modelos Animales de Enfermedad , Antígeno HLA-A2/química , Antígeno HLA-A2/metabolismo , Humanos , Factores Inmunológicos/farmacología , Inmunomodulación/efectos de los fármacos , Ratones , Mieloma Múltiple/tratamiento farmacológico , Mieloma Múltiple/metabolismo , Mieloma Múltiple/patología , Complejo de la Endopetidasa Proteasomal/metabolismo , Linfocitos T Citotóxicos/inmunología , Linfocitos T Citotóxicos/metabolismo , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Despite substantial advances in the treatment of acute myeloid leukemia (AML), only 30% of patients survive more than 5 years. Therefore, new therapeutics are much needed. Here, we present a novel therapeutic strategy targeting PR1, an HLA-A2 restricted myeloid leukemia antigen. Previously, we have developed and characterized a novel T-cell receptor-like monoclonal antibody (8F4) that targets PR1/HLA-A2 and eliminates AML xenografts by antibody-dependent cellular cytotoxicity (ADCC). To improve the potency of 8F4, we adopted a strategy to link T-cell cytotoxicity with a bi-specific T-cell-engaging antibody that binds PR1/HLA-A2 on leukemia and CD3 on neighboring T-cells. The 8F4 bi-specific antibody maintained high affinity and specific binding to PR1/HLA-A2 comparable to parent 8F4 antibody, shown by flow cytometry and Bio-Layer Interferometry. In addition, 8F4 bi-specific antibody activated donor T-cells in the presence of HLA-A2+ primary AML blasts and cell lines in a dose dependent manner. Importantly, activated T-cells lysed HLA-A2+ primary AML blasts and cell lines after addition of 8F4 bi-specific antibody. In conclusion, our studies demonstrate the therapeutic potential of a novel bi-specific antibody targeting the PR1/HLA-A2 leukemia-associated antigen, justifying further clinical development of this strategy.
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Anticuerpos Biespecíficos/inmunología , Antígenos de Neoplasias/inmunología , Antígeno HLA-A2/inmunología , Leucemia Mieloide Aguda/inmunología , Linfocitos T/inmunología , Animales , Anticuerpos Biespecíficos/farmacología , Especificidad de Anticuerpos/inmunología , Antígenos de Neoplasias/metabolismo , Células CHO , Línea Celular , Cricetulus , Citotoxicidad Inmunológica , Antígeno HLA-A2/metabolismo , Humanos , Inmunoterapia Adoptiva , Leucemia Mieloide Aguda/metabolismo , Leucemia Mieloide Aguda/patología , Leucemia Mieloide Aguda/terapia , Activación de Linfocitos , Unión Proteica , Linfocitos T/metabolismoRESUMEN
PURPOSE: The aim of this analysis is to compare the pharmacokinetics of drug substrates in healthy Chinese and European subjects of aligned CYP2C9, CYP2C19, or CYP2D6 enzyme activity, providing further insight into drivers of interethnic differences in pharmacokinetics. METHODS: Following identification of appropriate drug substrates, a comprehensive and structured literature search was conducted to identify single-dose pharmacokinetic data in healthy Chinese or European subjects with reported CYP2C9, CYP2C19, or CYP2D6 activity (genotype or phenotype). The ratio of drug AUC in the Chinese and European subjects classified with aligned enzyme activity was calculated (ethnicity ratio (ER)). RESULTS: For 22/25 drugs identified, the ERs calculated indicated no or only limited interethnic differences in exposure (Asunto(s)
Citocromo P-450 CYP2C19/metabolismo
, Citocromo P-450 CYP2C9/metabolismo
, Citocromo P-450 CYP2D6/metabolismo
, Farmacocinética
, Polimorfismo Genético
, Algoritmos
, Área Bajo la Curva
, Pueblo Asiatico
, China
, Estudios de Cohortes
, Citocromo P-450 CYP2C19/genética
, Citocromo P-450 CYP2C9/genética
, Citocromo P-450 CYP2D6/genética
, Bases de Datos Genéticas
, Bases de Datos Farmacéuticas
, Femenino
, Humanos
, Internet
, Masculino
, Preparaciones Farmacéuticas/sangre
, Farmacogenética/métodos
, Especificidad por Sustrato
, Población Blanca
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Neutrophil elastase (NE) can be rapidly taken up by tumor cells that lack endogenous NE expression, including breast cancer, which results in cross-presentation of PR1, an NE-derived HLA-A2-restricted peptide that is an immunotherapy target in hematological and solid tumor malignancies. The mechanism of NE uptake, however, remains unknown. Using the mass spectrometry-based approach, we identify neuropilin-1 (NRP1) as a NE receptor that mediates uptake and PR1 cross-presentation in breast cancer cells. We demonstrated that soluble NE is a specific, high-affinity ligand for NRP1 with a calculated Kd of 38.7 nm Furthermore, we showed that NRP1 binds to the RRXR motif in NE. Notably, NRP1 knockdown with interfering RNA or CRISPR-cas9 system and blocking using anti-NRP1 antibody decreased NE uptake and, subsequently, susceptibility to lysis by PR1-specific cytotoxic T cells. Expression of NRP1 in NRP1-deficient cells was sufficient to induce NE uptake. Altogether, because NRP1 is broadly expressed in tumors, our findings suggest a role for this receptor in immunotherapy strategies that target cross-presented antigens.
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Absorción Fisiológica , Neoplasias de la Mama/metabolismo , Reactividad Cruzada , Elastasa de Leucocito/metabolismo , Proteínas de Neoplasias/metabolismo , Neuropilina-1/metabolismo , Secuencias de Aminoácidos , Anticuerpos Bloqueadores/metabolismo , Neoplasias de la Mama/inmunología , Neoplasias de la Mama/patología , Sistemas CRISPR-Cas , Línea Celular Tumoral , Femenino , Humanos , Cinética , Elastasa de Leucocito/química , Elastasa de Leucocito/inmunología , Ligandos , Proteínas de Neoplasias/antagonistas & inhibidores , Proteínas de Neoplasias/química , Proteínas de Neoplasias/genética , Neuropilina-1/antagonistas & inhibidores , Neuropilina-1/química , Neuropilina-1/genética , Fragmentos de Péptidos/química , Fragmentos de Péptidos/genética , Fragmentos de Péptidos/metabolismo , Dominios y Motivos de Interacción de Proteínas , Interferencia de ARN , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Solubilidad , Linfocitos T Citotóxicos/inmunología , Linfocitos T Citotóxicos/metabolismoRESUMEN
In our preliminary studies, we observed zolmitriptan (ZOL) treatment led to induction of CYP3A2 in male not female rats. To figure out the reason is of great significance for drug-drug interactions and personalized administration. Since growth hormone (GH) is known as the major mechanistic determinant of sexually-dimorphic gene expression like CYP3A2 in rat liver, the impacts of ZOL on both plasma GH levels in non monosodium glutamate (MSG)-treated rats and CYP3A2 expression in GH depleted MSG-treated rats were studied. ZOL was shown to partially suppress GH levels in both genders. Furthermore, CYP3A2 protein and mRNA level declined in male not female MSG-treated rats. In order to study the possible molecular events involved in the depression of GH and gender-selective induction on rat CYP3A2 by ZOL, the mRNA and protein level(whole protein and nuclear protein) of hepatocyte nuclear factor 4α (HNF4α) was investigated. Nuclear accumulation of HNF4α was observed in the normal male not female rat liver tissue following ZOL treatment. However, this kind of nuclear translocation did not occur in rat hepatocytes and MSG-treated rats. These findings demonstrated CYP3A2 inducibility by ZOL was gender-selective. GH and HNF4α may play an important role in CYP3A2 induction.
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Inductores del Citocromo P-450 CYP3A/farmacología , Citocromo P-450 CYP3A/metabolismo , Hormona del Crecimiento/antagonistas & inhibidores , Oxazolidinonas/farmacología , Factores Sexuales , Triptaminas/farmacología , Animales , Factor Nuclear 4 del Hepatocito/metabolismo , Masculino , ARN Mensajero , Ratas , Glutamato de SodioRESUMEN
BACKGROUND AIMS: The PR1 peptide, derived from the leukemia-associated antigens proteinase 3 and neutrophil elastase, is overexpressed on HLA-A2 in acute myeloid leukemia (AML). We developed a T-cell receptor (TCR)-like monoclonal antibody (8F4) that binds the PR1/HLA-A2 complex on the surface of AML cells, efficiently killing them in vitro and eliminating them in preclinical models. Humanized 8F4 (h8F4) with high affinity for the PR1/HLA-A2 epitope was used to construct an h8F4- chimeric antigen receptor (CAR) that was transduced into T cells to mediate anti-leukemia activity. METHODS: Human T cells were transduced to express the PR1/HLA-A2-specific CAR (h8F4-CAR-T cells) containing the scFv of h8F4 fused to the intracellular signaling endo-domain of CD3 zeta chain through the transmembrane and intracellular costimulatory domain of CD28. RESULTS: Adult human normal peripheral blood (PB) T cells were efficiently transduced with the h8F4-CAR construct and predominantly displayed an effector memory phenotype with a minor population (12%) of central memory cells in vitro. Umbilical cord blood (UCB) T cells could also be efficiently transduced with the h8F4-CAR. The PB and UCB-derived h8F4-CAR-T cells specifically recognized the PR1/HLA-A2 complex and were capable of killing leukemia cell lines and primary AML blasts in an HLA-A2-dependent manner. CONCLUSIONS: Human adult PB and UCB-derived T cells expressing a CAR derived from the TCR-like 8F4 antibody rapidly and efficiently kill AML in vitro. Our data could lead to a new treatment paradigm for AML in which targeting leukemia stem cells could transfer long-term immunity to protect against relapse.
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Sangre Fetal , Antígeno HLA-A2/inmunología , Leucemia Mieloide Aguda/terapia , Leucocitos Mononucleares/metabolismo , Mieloblastina/inmunología , Receptores de Antígenos de Linfocitos T/metabolismo , Linfocitos T/metabolismo , Adulto , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/metabolismo , Línea Celular , Epítopos/inmunología , Sangre Fetal/citología , Sangre Fetal/inmunología , Terapia Genética , Humanos , Fragmentos Fc de Inmunoglobulinas/química , Fragmentos Fc de Inmunoglobulinas/inmunología , Fragmentos Fc de Inmunoglobulinas/metabolismo , Inmunoterapia Adoptiva/métodos , Leucemia Mieloide Aguda/inmunología , Leucemia Mieloide Aguda/patología , Leucocitos Mononucleares/inmunología , Mieloblastina/química , Fragmentos de Péptidos/química , Fragmentos de Péptidos/inmunología , Receptores de Antígenos de Linfocitos T/genética , Receptores de Antígenos de Linfocitos T/inmunología , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Proteínas Recombinantes de Fusión/metabolismo , Especificidad del Receptor de Antígeno de Linfocitos T , Linfocitos T/inmunologíaRESUMEN
A novel series of N-(4-aryl-5-aryloxy-thiazol-2-yl)-amides as RORγt inverse agonists was discovered. Binding mode analysis of a RORγt partial agonist (2c) revealed by co-crystal structure in RORγt LBD suggests that the inverse agonists do not directly interfere with the interaction between H12 and the RORγt LBD. Detailed SAR exploration led to identification of potent RORγt inverse agonists such as 3m with a pIC50 of 8.0. Selected compounds in the series showed reasonable activity in Th17 cell differentiation assay as well as low intrinsic clearance in mouse liver microsomes.
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Amidas/química , Amidas/farmacología , Agonismo Inverso de Drogas , Miembro 3 del Grupo F de la Subfamilia 1 de Receptores Nucleares/agonistas , Células Th17/efectos de los fármacos , Tiazoles/química , Tiazoles/farmacología , Animales , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Humanos , Ratones , Simulación del Acoplamiento Molecular , Miembro 3 del Grupo F de la Subfamilia 1 de Receptores Nucleares/metabolismo , Células Th17/citologíaRESUMEN
A novel series of biaryl amides was identified as RORγt inhibitors through core replacement of a starting hit 1. Structure-activity relationship exploration on the biaryl moiety led to discovery of potent RORγt inhibitors with good oral bioavailability and CNS penetration. Compounds 9a and 9g demonstrated excellent in vivo efficacy in EAE mice dose dependently with once daily oral administration.
RESUMEN
Organic anion-transporting polypeptide (OATP) 1A2 has the potential to be a target for central nervous system drug delivery due to its luminal localization at the human blood-brain barrier and broad substrate specificity. We found OATP1A2 mRNA expression in the human brain to be comparable to breast cancer resistance protein and OATP2B1 and much higher than P-glycoprotein (P-gp), and confirmed greater expression in the brain relative to other tissues. The goal of this study was to establish a model system to explore OATP1A2-mediated transcellular transport of substrate drugs and the interplay with P-gp. In vitro (human embryonic kidney 293 cells stably expressing Oatp1a4, the closest murine isoform) and in vivo (naïve and Oatp1a4 knock-out mice) studies with OATP1A2 substrate triptan drugs demonstrated that these drugs were not Oatp1a4 substrates. This species difference demonstrates that the rodent is not a good model to investigate the active brain uptake of potential OATP1A2 substrates. Thus, we constructed a novel OATP1A2 expressing Madin-Darby canine kidney (MDCK) II wild type and an MDCKII-multidrug resistance protein 1 (MDR1) system using BacMam virus transduction. The spatial expression pattern of OATP1A2 after transduction in MDCKII-MDR1 cells was superimposed to P-gp, confirming apical membrane localization. OATP1A2-mediated uptake of zolmitriptan, rosuvastatin, and fexofenadine across monolayers increased with increasing OATP1A2 protein expression. OATP1A2 counteracted P-gp efflux for cosubstrates zolmitriptan and fexofenadine. A three-compartment model incorporating OATP1A2-mediated influx was used to quantitatively describe the time- and concentration-dependent apical-to-basolateral transcellular transport of rosuvastatin across OATP1A2 expressing the MDCKII monolayer. This novel, simple and versatile experimental system is useful for understanding the contribution of OATP1A2-mediated transcellular transport across barriers, such as the blood-brain barrier.