Inmuno-checkpoints: PD-L1, una diana terapéutica más allá del cáncer

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Farmacología. Fecha de lectura: 06-03-2020Los inmuno-chekpoints (ICs) son moléculas responsables de modular la duración y la intensidad de la respuesta inmunológica. El bloqueo de ICs inhibitorios con anticuerpos monoclonales ha sido aplicado con gran éxito en el cáncer, destacando la terapia frente a la interacción PD-1/PD-L1 por su rango de acción y eficacia. En este trabajo, hemos evaluado la implicación de PD-L1 en la tolerancia a endotoxinas (TE), así como el potencial terapéutico del bloqueo de su interacción con PD-1, en el contexto de tres patologías asociadas a infecciones: sepsis, fibrosis quística (FQ) e infección por virus de la inmunodeficiencia humana (VIH). Los monocitos humanos en estado de TE presentaron una elevada expresión de PD-L1 debida a la acción del factor de transcripción HIF-1α. Este hecho explica parte del agotamiento y la disminución de respuesta de los linfocitos T durante este fenómeno. En pacientes con sepsis, la expresión de PD-L1 correlacionó inversamente con la saturación de oxígeno (SaO2). Consecuentemente, los pacientes con hipoxemia (SaO2 ≤92 %) presentaron mayores niveles de PD-L1 y una mayor respuesta a la terapia antiPD-1 comparada con el resto de pacientes. Por su parte, en sujetos con FQ, se observó una sobre-expresión de PD-L1 en aquellos con infección por Pseudomonas aeruginosa. De acuerdo a lo observado en pacientes y en modelos in vitro, este efecto se reduce con la polimixina colistimetato, y puede explicarse por la translocación de fragmentos de lipopolisacárido de esta bacteria al torrente sanguíneo. Finalmente, en individuos infectados por el VIH, se encontró una correlación positiva entre los niveles de PD-L1 y la carga viral en el plasma. Asimismo, los pacientes con mayores niveles de esta molécula manifestaron fracaso virológico en la terapia antirretroviral. Globalmente, todos estos datos proponen un nuevo rol para PD-L1 clave en la fisiopatología de la TE. Consecuentemente durante la TE, no sólo se vería afectada la respuesta innata, sino también la adaptativa a través de la acción de este ligando. Los resultados aquí presentados, permiten identificar grupos de pacientes susceptibles de ser tratados con inmunoterapia antiPD-1/PD-L1 que serían aquellos con hipoxémia en el caso de la sepsis, los colonizados por Pseudomonas aeruginosa en el caso de la FQ y los pacientes con alta carga viral y/o fracaso virológico en el caso de los infectados por VIHThe Immuno-Checkpoints (ICs) are molecules responsible for the modulation of the immune response. The blockade of inhibitory ICs by monoclonal antibodies has been successfully applied in cancer, in which, the therapy against PD-1/PD-L1 interaction stands out due to its range of action and efficacy. In this work, we have evaluated the involvement of PD-L1 in endotoxin tolerance (ET) as well as the therapeutic potential of blocking its interaction with PD-1, in the context of three infection associated pathologies: sepsis, cystic fibrosis (CF) and human immunodeficiency virus (HIV) infection. Human monocytes under ET had an overexpression of PD-L1 triggered by the HIF-1α transcription factor. This fact explains part of the T lymphocytes depletion and their decreased responses during this phenomenon. In patients with sepsis, PD-L1 expression correlated inversely with oxygen saturation (SaO2). Consequently, those with hypoxemia (SaO2 ≤92%) had higher levels of PD-L1 and a greater response to an anti-PD-1 therapy compared to the rest of the patients. In subjects with CF, we observed an overexpression of PD-L1 in those with Pseudomonas aeruginosa infection. In in vitro models, this effect was reduced by polymyxin colistimethate and could be explained by the translocation of the lipopolysaccharide of this pathogen into the bloodstream. Finally, in HIV-infected individuals, a positive correlation was found between plasmatic PD-L1 levels and viral load. Likewise, patients with higher levels of this molecule manifested virological failure in antiretroviral therapy. Globally, all these data propose a new key role for PD-L1 in the pathophysiology of ET. Consequently, during ET, not only the innate response would be affected, but also the adaptive response through the action of this ligand. The results presented here allow the identification of groups of individuals likely to be treated with anti-PD/1-PD-L1 immunotherapy. These patients are those with hypoxemia in the case of sepsis, those colonized by Pseudomonas aeruginosa in the case of CF and those with high viral load and/or virological failure in the case of HIV-infected individualsEste trabajo fue apoyado por los siguientes proyectos y ayudas: • Ayuda a la contratación de ayudantes de investigación PEJ15/BIO/AI-0021 de la Comunidad de Madrid a José Avendaño Ortiz • Proyecto PI14/01234 del Instituto de Salud Carlos III” (ISCIII) “Fondos de Investigación Sanitarias” (FIS) y Fondos FEDER a Eduardo López Collazo • Proyecto Proyecto PIE15/00065) del Instituto de Salud Carlos III” (ISCIII) “Fondos de Investigación Sanitarias” (FIS) y Fondos FEDER a Eduardo López Collaz

The prognostic impact of SIGLEC5-induced impairment of CD8+ T cell activation in sepsis

Immune checkpoint; Sepsis; T-cell exhaustionPunto de control inmunológico; Septicemia; Agotamiento de células TPunt de control immunitari; Sèpsia; Esgotament de cèl·lules TBackground Sepsis is associated with T-cell exhaustion, which significantly reduces patient outcomes. Therefore, targeting of immune checkpoints (ICs) is deemed necessary for effective sepsis management. Here, we evaluated the role of SIGLEC5 as an IC ligand and explored its potential as a biomarker for sepsis. Methods In vitro and in vivo assays were conducted to both analyse SIGLEC5's role as an IC ligand, as well as assess its impact on survival in sepsis. A multicentre prospective cohort study was conducted to evaluate the plasmatic soluble SIGLEC5 (sSIGLEC5) as a mortality predictor in the first 60 days after admission in sepsis patients. Recruitment included sepsis patients (n = 346), controls with systemic inflammatory response syndrome (n = 80), aneurism (n = 11), stroke (n = 16), and healthy volunteers (HVs, n = 100). Findings SIGLEC5 expression on monocytes was increased by HIF1α and was higher in septic patients than in healthy volunteers after ex vivo LPS challenge. Furthermore, SIGLEC5-PSGL1 interaction inhibited CD8+ T-cell proliferation. Administration of sSIGLEC5r (0.8 mg/kg) had adverse effects in mouse endotoxemia models. Additionally, plasma sSIGLEC5 levels of septic patients were higher than HVs and ROC analysis revealed it as a mortality marker with an AUC of 0.713 (95% CI, 0.656–0.769; p < 0.0001). Kaplan–Meier survival curve showed a significant decrease in survival above the calculated cut-off (HR of 3.418, 95% CI, 2.380–4.907, p < 0.0001 by log-rank test) estimated by Youden Index (523.6 ng/mL). Interpretation SIGLEC5 displays the hallmarks of an IC ligand, and plasma levels of sSIGLEC5 have been linked with increased mortality in septic patients.This work was supported by grants from Instituto de Salud Carlos III (ISCIII) and “Fondos FEDER” to ELC (PIE15/00065, PI18/00148, PI14/01234, PI21/00869), to PP (20859/PI/18) and to CdF (PI21/01178), and received funding from the European Union's Horizon 2020 research and innovation program under the Marie Sklodowaska-Curie grant agreement to KMH (No. 713673; “laCaixa”). R.L.-R. was supported by “Predoctotales de formación en Investigación” (PFIS) grant FI19/00334 and J.A.-O. by Sara Borrell grant CD21/00059 from ISCIII. The Vall d'Hebron University Hospital and Vall d’Hebron Research Institute were supported by Plan Nacional de I+D+i 2013–2016, the ISCIII and Spanish Network for Research in Infectious Diseases (REIPI RD16/0016/0003)—co-financed by European Development Regional Fund “A way to achieve Europe”, and by the European Union’s Horizon 2020 Research and Innovation Program (JCRR, RF, JJGL, AF). Authors thank Emilio Llanos for his technical assistance

Oxygen Saturation on Admission Is a Predictive Biomarker for PD-L1 Expression on Circulating Monocytes and Impaired Immune Response in Patients With Sepsis

Sepsis is a pathology in which patients suffer from a proinflammatory response and a dysregulated immune response, including T cell exhaustion. A number of therapeutic strategies to treat human sepsis, which are different from antimicrobial and fluid resuscitation treatments, have failed in clinical trials, and solid biomarkers for sepsis are still lacking. Herein, we classified 85 patients with sepsis into two groups according to their blood oxygen saturation (SaO2): group I (SaO2 ≤ 92%, n = 42) and group II (SaO2 &gt; 92%, n = 43). Blood samples were taken before any treatment, and the immune response after ex vivo LPS challenge was analyzed, as well as basal expression of PD-L1 on monocytes and levels of sPD-L1 in sera. The patients were followed up for 1 month. Taking into account reinfection and exitus frequency, a significantly poorer evolution was observed in patients from group I. The analysis of HLA-DR expression on monocytes, T cell proliferation and cytokine profile after ex vivo LPS stimulation confirmed an impaired immune response in group I. In addition, these patients showed both, high levels of PD-L1 on monocytes and sPD-L1 in serum, resulting in a down-regulation of the adaptive response. A blocking assay using an anti-PD-1 antibody reverted the impaired response. Our data indicated that SaO2 levels on admission have emerged as a potential signature for immune status, including PD-L1 expression. An anti-PD-1 therapy could restore the T cell response in hypoxemic sepsis patients with SaO2 ≤ 92% and high PD-L1 levels

A system dynamics model to predict the human monocyte response to endotoxins

System dynamics is a powerful tool that allows modeling of complex and highly networked systems such as those found in the human immune system. We have developed a model that reproduces how the exposure of human monocytes to lipopolysaccharides (LPSs) induces an inflammatory state characterized by high production of tumor necrosis factor alpha (TNFα), which is rapidly modulated to enter into a tolerant state, known as endotoxin tolerance (ET). The model contains two subsystems with a total of six states, seven flows, two auxiliary variables, and 14 parameters that interact through six differential and nine algebraic equations. The parameters were estimated and optimized to obtain a model that fits the experimental data obtained from human monocytes treated with various LPS doses. In contrast to publications on other animal models, stimulation of human monocytes with super-low-dose LPSs did not alter the response to a second LPSs challenge, neither inducing ET, nor enhancing the inflammatory response. Moreover, the model confirms the low production of TNFα and increased levels of C-C motif ligand 2 when monocytes exhibit a tolerant state similar to that of patients with sepsis. At present, the model can help us better understand the ET response and might offer new insights on sepsis diagnostics and prognosis by examining the monocyte response to endotoxins in patients with sepsisThis work was supported by grants from the “Instituto de Salud Carlos III” (ISCiii), “Fondo de Investigación Sanitaria” (FIS), and Fondos FEDER (PI14/01234, PIE15/00065) to EL-C. EA work contract is supported by the Torres Quevedo program from “Ministerio de Economía y Competitividad” (SPTQ1300X006175XV0). VT work contract is supported by the “Ministerio de Economía y Competitividad” (PTA2013-8265-I

Experiences in flipped learning

El presente trabajo describe unas experiencias de innovación docente, de igual perfil y metodología, desarrolladas por un conjunto de profesores noveles participantes bajo la tutela experta de profesorado senior. El tipo de experiencia seleccionada ha sido el diseño y ejecución de una flipped classroom, o clase invertida. Además, se incluye la correspondiente evaluación de la acción con una valoración académica o impacto en los aprendizajes, y una valoración de la experiencia por parte del alumnado y profesorado responsable. Los resultados de cinco experiencias con impacto a mas de 270 alumnos de distinto perfil académico (tres asignaturas de titulo de Grado y dos de Máster) y las evaluaciones cuantificadas indican una valoracion muy positiva de esta metodología flipped learning (FL) por los alumnos y su deseo de aplicabilidad. Por su parte, el profesorado constata que las experiencias FL han incrementado satisfactoriamente el aprendizaje de sus alumnos y declaran una alta valoración de la experiencia para la mejora de sus competencia docentes.The present work describes experiences of teaching innovation, of the same profile and methodology, developed by a group of participating junior teachers under the expert guidance of senior teaching staff. The type of experience selected has been the design and execution of a flipped classroom, or inverted class. In addition, the corresponding evaluation of the action is included with an academic assessment or impact on learning, and an assessment of the experience by the students and teaching staff responsible. The results of five experiences with impact to more than 270 students of different academic profiles (three degrees and two Masters) and the quantified evaluations indicate a very positive assessment of this flipped learning methodology by the students and their desire for applicability. On the other hand, the teachers note that the FL experiences have successfully increased the learning of their students and declare a high evaluation of the experience for the improvement of their teaching competences

Cellular and humoral functional responses after BNT162b2 mRNA vaccination differ longitudinally between naive and subjects recovered from COVID-19

We have analyzed BNT162b2 vaccine-induced immune responses in naive subjects and individuals recovered from coronavirus disease 2019 (COVID-19), both soon after (14 days) and later after (almost 8 months) vaccination. Plasma spike (S)-specific immunoglobulins peak after one vaccine shot in individuals recovered from COVID-19, while a second dose is needed in naive subjects, although the latter group shows reduced levels all along the analyzed period. Despite how the neutralization capacity against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mirrors this behavior early after vaccination, both groups show comparable neutralizing antibodies and S-specific B cell levels late post-vaccination. When studying cellular responses, naive individuals exhibit higher SARS-CoV-2-specific cytokine production, CD4+ T cell activation, and proliferation than do individuals recovered from COVID-19, with patent inverse correlations between humoral and cellular variables early post-vaccination. However, almost 8 months post-vaccination, SARS-CoV-2-specific responses are comparable between both groups. Our data indicate that a previous history of COVID-19 differentially determines the functional T and B cell-mediated responses to BNT162b2 vaccination over time.C.d.F., J.G.-P., and J.A. are supported by Instituto de Salud Carlos III (ISCII). We thank JM Ligos and Cytek Biosciences for their technical support. Research in E.L.-C.’s lab was supported by Fundación Familia Alonso, Santander Bank, Real Seguros, Fundación Mutua Madrileña, Fundación Uria, Fundación La Caixa, and Ayuntamiento de Madrid.S

Inflammatory cytokines and organ dysfunction associate with the aberrant DNA methylome of monocytes in sepsis

Sepsis, a life-threatening organ dysfunction caused by a dysregulated systemic immune response to infection, associates with reduced responsiveness to subsequent infections. How such tolerance is acquired is not well understood but is known to involve epigenetic and transcriptional dysregulation. Bead arrays were used to compare global DNA methylation changes in patients with sepsis, non-infectious systemic inflammatory response syndrome, and healthy controls. Bioinformatic analyses were performed to dissect functional reprogramming and signaling pathways related to the acquisition of these specific DNA methylation alterations. Finally, in vitro experiments using human monocytes were performed to test the induction of similar DNA methylation reprogramming. Here, we focused on DNA methylation changes associated with sepsis, given their potential role in stabilizing altered phenotypes. Tolerized monocytes from patients with sepsis display changes in their DNA methylomes with respect to those from healthy controls, affecting critical monocyte-related genes. DNA methylation profiles correlate with IL-10 and IL-6 levels, significantly increased in monocytes in sepsis, as well as with the Sequential Organ Failure Assessment score; the observed changes associate with TFs and pathways downstream to toll-like receptors and inflammatory cytokines. In fact, in vitro stimulation of toll-like receptors in monocytes results in similar gains and losses of methylation together with the acquisition of tolerance. We have identified a DNA methylation signature associated with sepsis that is downstream to the response of monocytes to inflammatory signals associated with the acquisition of a tolerized phenotype and organic dysfunction

Impact of vaccination against COVID-19 on patients with cancer in ACHOC-C19 study: Real world evidence from one Latin American country

Introducción: Durante la pandemia, se ha recomendado que la vacunación contra COVID-19 sea una prioridad para los pacientes con cáncer; sin embargo, estos pacientes no se incluyeron en los estudios iniciales de evaluación de las vacunas disponibles. Objetivo: Definir el impacto de la vacunación contra COVID-19 en la prevención del riesgo de complicaciones asociadas a la infección en una cohorte de pacientes con cáncer en Colombia. Métodos: Se realizó un estudio observacional analítico de cohorte, basado en el registro nacional de pacientes con cáncer e infección por COVID 19 ACHOC-C19. Los datos se recolectaron desde junio de 2021, hasta octubre de 2021. Los criterios de inclusión fueron: Pacientes mayores de 18 años con diagnóstico de cáncer e infección confirmada por COVID-19. Se compararon los datos de las cohortes no vacunadas y vacunadas. Los resultados evaluados incluyeron mortalidad por todas las causas en los 30 días siguientes al diagnóstico de COVID-19, hospitalización y necesidad de ventilación mecánica. La estimación del efecto se realizó mediante el riesgo relativo (RR), la reducción absoluta del riesgo (RRA) y el número necesario a tratar (NNT). El análisis multivariante se realizó mediante modelos lineales generalizados. Resultados: Se incluyeron 896 pacientes, de los cuales 470 eran mayores de 60 años (52,4%) y el 59% eran mujeres (n=530). Se reclutaron 172 pacientes en la cohorte vacunada y 724 en la cohorte no vacunada (ratio: 1 a 4,2). La incidencia acumulada de resultados clínicos entre los pacientes no vacunados frente a los vacunados fue: para hospitalización 42% (IC 95%: 38,7%-46,1%) frente a 29%; (IC 95%: 22,4%-36,5%); para requerimiento de ventilación mecánica invasiva 8,4% (n=61) frente a 4,6% (n=8) y para mortalidad por todas las causas 17% (n=123) frente a 4,65% (n=8). Conclusiones: En nuestra población, los pacientes con cáncer no vacunados tienen un mayor riesgo de complicaciones por infección por COVID -19, como hospitalización, ventilación mecánica y mortalidad. Es muy recomendable promover activamente la vacunación entre esta población. El autor (es). Este es un artículo de acceso abierto distribuido bajo los términos de la Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). Consulte los términos y condicionesIntroduction: During the pandemic, it has been recommended that vaccination against COVID-19 be a priority for patients with cancer; however, these patients were not included in the initial studies evaluating the available vaccines. Objective: To define the impact of vaccination against COVID-19 in preventing the risk of complications associated with the infection in a cohort of patients with cancer in Colombia. Methods: An analytical observational cohort study, based on national registry of patients with cancer and COVID 19 infection ACHOC-C19, was done. The data was collected from June 2021, until October 2021. Inclusion criteria were: Patients older than 18 years with cancer diagnosis and confirmed COVID-19 infection. Data from the unvaccinated and vaccinated cohorts were compared. Outcomes evaluated included all-cause mortality within 30 days of COVID-19 diagnosis, hospitalization, and need for mechanical ventilation. The estimation of the effect was made through the relative risk (RR), the absolute risk reduction (ARR) and the number needed to treat (NNT). Multivariate analysis was performed using generalized linear models. Results: 896 patients were included, of whom 470 were older than 60 years (52.4%) and 59% were women (n=530). 172 patients were recruited in the vaccinated cohort and 724 in the non-vaccinated cohort (ratio: 1 to 4.2). The cumulative incidence of clinical outcomes among the unvaccinated vs vaccinated patients were: for hospitalization 42% (95% CI: 38.7%-46.1%) vs 29%; (95% CI: 22.4%-36.5%); for invasive mechanical ventilation requirement 8.4% (n=61) vs 4.6% (n=8) and for mortality from all causes 17% (n=123) vs 4.65% (n=8). Conclusion: In our population, unvaccinated patients with cancer have an increased risk of complications for COVID -19 infection, as hospitalization, mechanical ventilation, and mortality. It is highly recommended to actively promote the vaccination among this population. © The author(s). This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). See http://ivyspring.com/terms for full terms and conditions

Gestión del conocimiento. Perspectiva multidisciplinaria. Volumen 17

El libro “Gestión del Conocimiento. Perspectiva Multidisciplinaria”, Volumen 17 de la Colección Unión Global, es resultado de investigaciones. Los capítulos del libro, son resultados de investigaciones desarrolladas por sus autores. El libro es una publicación internacional, seriada, continua, arbitrada, de acceso abierto a todas las áreas del conocimiento, orientada a contribuir con procesos de gestión del conocimiento científico, tecnológico y humanístico. Con esta colección, se aspira contribuir con el cultivo, la comprensión, la recopilación y la apropiación social del conocimiento en cuanto a patrimonio intangible de la humanidad, con el propósito de hacer aportes con la transformación de las relaciones socioculturales que sustentan la construcción social de los saberes y su reconocimiento como bien público