Expression of PD-1/PD-L1 and PD-L2 in peripheral T-cells from non-small cell lung cancer patients.

Binding of programmed death-1 (PD-1) with its ligands (PD-L1/2) transmits a co-inhibitory signal in activated T-cells that promotes T-cell exhaustion, leading to tumor immune evasion. The efficacy of antibodies targeting PD-1 and PD-L1 has led to a paradigm shift in lung cancer treatment but the prognostic and predictive value of tumor PD-L1 expression remains controversial. Evaluating PD-1, PD-L1/2 expression in peripheral blood cells may serve as a potential biomarker for prognosis and response to therapy. In this prospective observational study, plasma cytokine levels and PD-1, PD-L1 and PD-L2 expression was evaluated in circulating CD3+, CD3+CD4+ and CD3+CD8+ cells from 70 treatment-naïve patients with advanced NSCLC (Stage IIIB and IV) and from 10 healthy donors. The primary objective was to assess OS according to PD-1, PD-L1, PD-L2 expression status on PBMCs and lymphocyte subsets. Our results indicate that the percentage of PD-L1+CD3+, PD-L1+CD3+CD8+ PD-L2+PBMCs, PD-L2+CD3+, PD-L2+CD3+CD4+ cells was higher in patients than in healthy donors. Survival was decreased among patients with a high percentage of either PD-1+PBMCs, PD-1+CD3+, PD-L1+CD3+, PD-L1+CD3+CD8+, PD-L2+CD3+, PD-L2+CD3+CD4+, or PD-L2+CD3+CD8+ cells. IL-2 and TNF-α showed the strongest association with PD-L1 and PD-L2 expression on specific subsets of T-lymphocytes. Our findings suggest that increased PD-1/PD-L1/PDL-2 expression in PBMCs, particularly in T-cells, may be an additional mechanism leading to tumor escape from immune control. This study is registered with ClinicalTrials.gov, number NCT02758314.

Apolipoprotein B100 is required for hepatitis C infectivity and Mipomersen inhibits hepatitis C.

AIM: To characterize the role of apolipoprotein B100 (apoB100) in hepatitis C viral (HCV) infection. METHODS: In this study, we utilize a gene editing tool, transcription activator-like effector nucleases (TALENs), to generate human hepatoma cells with a stable genetic deletion of APOB to assess of apoB in HCV. Using infectious cell culture-competent HCV, viral pseudoparticles, replicon models, and lipidomic analysis we determined the contribution of apoB to each step of the viral lifecycle. We further studied the effect of mipomersen, an FDA-approved antisense inhibitor of apoB100, on HCV using in vitro cell-culture competent HCV and determined its impact on viral infectivity with the TCID50 method. RESULTS: We found that apoB100 is indispensable for HCV infection. Using the JFH-1 fully infectious cell-culture competent virus in Huh 7 hepatoma cells with TALEN-mediated gene deletion of apoB (APOB KO), we found a significant reduction in HCV RNA and protein levels following infection. Pseudoparticle and replicon models demonstrated that apoB did not play a role in HCV entry or replication. However, the virus produced by APOB KO cells had significantly diminished infectivity as measured by the TCID-50 method compared to wild-type virus. Lipidomic analysis demonstrated that these virions have a fundamentally altered lipidome, with complete depletion of cholesterol esters. We further demonstrate that inhibition of apoB using mipomersen, an FDA-approved anti-sense oligonucleotide, results in a potent anti-HCV effect and significantly reduces the infectivity of the virus. CONCLUSION: ApoB is required for the generation of fully infectious HCV virions, and inhibition of apoB with mipomersen blocks HCV. Targeting lipid metabolic pathways to impair viral infectivity represents a novel host targeted strategy to inhibit HCV.

Hepatic Injury in Nonalcoholic Steatohepatitis Contributes to Altered Intestinal Permeability.

BACKGROUND & AIMS: Emerging data suggest that changes in intestinal permeability and increased gut microbial translocation contribute to the inflammatory pathway involved in nonalcoholic steatohepatitis (NASH) development. Numerous studies have investigated the association between increased intestinal permeability and NASH. Our meta-analysis of this association investigates the underlying mechanism. METHODS: A meta-analysis was performed to compare the rates of increased intestinal permeability in patients with NASH and healthy controls. To further address the underlying mechanism of action, we studied changes in intestinal permeability in a diet-induced (methionine-and-choline-deficient; MCD) murine model of NASH. In vitro studies were also performed to investigate the effect of MCD culture medium at the cellular level on hepatocytes, Kupffer cells, and intestinal epithelial cells. RESULTS: Nonalcoholic fatty liver disease (NAFLD) patients, and in particular those with NASH, are more likely to have increased intestinal permeability compared with healthy controls. We correlate this clinical observation with in vivo data showing mice fed an MCD diet develop intestinal permeability changes after an initial phase of liver injury and tumor necrosis factor-α (TNFα) induction. In vitro studies reveal that MCD medium induces hepatic injury and TNFα production yet has no direct effect on intestinal epithelial cells. Although these data suggest a role for hepatic TNFα in altering intestinal permeability, we found that mice genetically resistant to TNFα-myosin light chain kinase (MLCK)-induced intestinal permeability changes fed an MCD diet still develop increased permeability and liver injury. CONCLUSIONS: Our clinical and experimental results strengthen the association between intestinal permeability increases and NASH and also suggest that an early phase of hepatic injury and inflammation contributes to altered intestinal permeability in a fashion independent of TNFα and MLCK.

Noninvasive Biomarkers of Liver Fibrosis: Clinical Applications and Future Directions.

Chronic liver disease is a significant cause of morbidity and mortality worldwide. Current strategies for assessing prognosis and treatment rely on accurate assessment of disease stage. Liver biopsy is the gold standard for assessing fibrosis stage but has many limitations. Noninvasive biomarkers of liver fibrosis have been extensively designed, studied, and validated in a variety of liver diseases. With the advent of direct acting antivirals and the rise in obesity-related liver disease, there is a growing need to establish these noninvasive methods in the clinic. In addition, it has become increasingly clear over the last few years that noninvasive biomarkers can also be used to monitor response to antifibrotic therapies and predict liver outcomes, including hepatocellular carcinoma development. This review highlights the most well-established noninvasive biomarkers to-date, with a particular emphasis on serum and imaging-based methodologies.

[Association between overweight, glucocorticoids and metabolic syndrome in cancer patients under chemotherapy]. / Asociación de sobrepeso y uso de glucocorticoides con componentes del síndrome metabólico en pacientes oncológicos en quimioterapia.

UNLABELLED: Metabolic syndrome components like overweight, obesity, insulin resistance, and hyperglycemia are common findings in patients with cancer diagnosis under chemotherapy treatment. These factors have been associated with higher recurrence rates. This study associates Body Mass index, steroids treatment and tumor site with metabolic syndrome (MS) components in patients with cancer diagnosis under chemotherapy treatment. METHODS: In this retrospective study, files from patients under chemotherapy treatment treated in a university oncology center from 2008 to 2010 where reviewed. Anthropometric data and ATP III MS criteria were reviewed. RESULTS: 158 patients were included, 75.9% female. Most common tumors were breast, gastrointestinal and lung cancer. 56.3% presented =3 component of MS; 43.6% of patients received Dexametasone as part of chemotherapy treatment. Mean BMI was 25.3 kg/m(2). Breast cancer diagnosis was associated with presence of 3 or more components of metabolic syndrome. Glococorticoid treatment was not significantly associated with MS diagnosis. CONCLUSIONS: patients with IMC>25 presented 12.6 more risk of MS, independently of glucocorticoids treatment. Weight maintenance is important to reduce MS.

En pacientes con diagnostico de cáncer en tratamiento con quimioterapia es común encontrar componentes del síndrome metabólico (SM) como sobrepeso, obesidad, resistencia a la insulina e hiperglicemia. Estos componentes se han asociado a mayor recurrencia de la enfermedad. Objetivos: describir la relación entre el IMC, el uso de glucocorticoides y el sitio de tumor con los factores del SM en pacientes tratados con quimioterapia de un centro hospitalario universitario. Métodos: Estudio retrospectivo donde se revisaron expedientes de pacientes tratados en el centro oncológico del 2008 al 2010, con diagnóstico de cáncer y en tratamiento con quimioterapia sistémica. Se recopilo información acerca de datos antropométricos y criterios de SM definidos por ATP III. Resultados: Se incluyeron 158 pacientes, 75,9% genero femenino, los tumores mas comunes fueron mama, gastrointestinal y pulmón. Más de la mitad de los pacientes presentaron >3 componentes del SM (56,3%); El 43,6% de pacientes recibieron dexametasona como parte del tratamiento. El IMC promedio fue de 25,3 kg/m2. El diagnostico de cáncer de mama en tratamiento con quimioterapia se asoció con la presencia de 3 o más componentes del SM. La administración de glucocorticoides no se asoció a la presencia de SM. Conclusiones: los sujetos con IMC>25 tienen 12,6 veces más el riesgo de padecer SM, independientemente del uso de glucocorticoides en el tratamiento. El mantenimiento de un peso adecuado en el paciente oncológico es importante para reducir los factores de riesgo del SM.

Oligomerization and regulated proteolytic processing of angiopoietin-like protein 4.

Angiopoietin-like protein 4 (Angptl4) is a recently identified circulating protein expressed primarily in adipose tissue and liver. Also known as peroxisome proliferator-activated receptor (PPAR)-gamma angiopoietin-related, fasting induced adipose factor, and hepatic fibrinogen/angiopoietin-related protein, recombinant Angptl4 causes increase of plasma very low density lipoprotein levels by inhibition of lipoprotein lipase activity. Similar to angiopoietins and other angiopoietin-like proteins, Angptl4 contains an amino-terminal coiled-coil domain and a carboxyl-terminal fibrinogen-like domain. We report here that Angptl4 is evolutionarily conserved among several mammalian species and that full-length Angptl4 protein is an oligomer containing intermolecular disulfide bonds. Oligomerized Angptl4 undergoes proteolytic processing to release its carboxyl fibrinogen-like domain, which circulates as a monomer. Angptl4's N-terminal coiled-coil domain mediates its oligomerization, which by itself is sufficient to form higher order oligomeric structure. Adenovirus-mediated overexpression of Angptl4 in 293 cells shows that conversion of full-length, oligomerized Angptl4 is mediated by a cell-associated protease activity induced by serum. These findings demonstrate a novel property of angiopoietin-like proteins and suggest that oligomerization and proteolytic processing of Angptl4 may regulate its biological activities in vivo.

Perfución miocárdica valorada con técnica de dos isótopos. Experiencia de los primeros 1600 casos en México / Myocardial perfusion assesed by two isotope technique

El SPECT tiene alta sensibilidad para la detección de enfermedad coronaria. Hace cuatro años se introdujo en México el estudio con dos isótopos (Talio reposo/MIBI esfuerzo), que reúne las ventajas de ambos radiotrazadores para el estudio de la perfusión miocárdica. Se presenta nuestra experiencia de los primeros tres años. Se estudiaron 1600 pacientes con sospecha de isquemia miocárdica; se excluyeron 288 por falta de un seguimiento adecuado. De los 1312 incluidos, 895 tenían coronariografía. Las imágenes obtenidas fueron evaluadas dividiendo el corazón en 20 segmentos y para cada segmento se utilizó una escala de 5 puntos: 0 = normal a 4 = ausencia de perfusión. Se consideró anormal si dos o más segmentos tuvieron puntuación en esfuerzo de MIBI igual o mayor a 2. La sensibilidad global para diagnóstico de isquemia fue de 96.28 por ciento. El método de dos isótopos es adecuado para el diagnóstico del paciente con cardiopatía isquémica. Tiene alta sensibilidad y especificidad para el reconocimiento de enfermedad coronaria global y por territorios coronarios específicos. Este trabajo constituye la serie más grande en América Latina que utiliza este método diagnóstico.