Application limits of Q-Switched Nd: YAG laser irradiation for stone cleaning based on colour measurements

The application limits of the laser-cleaning technique for different types of building stones have been investigated by measuring colour variations. The selected stones differ in their chemical and mineralogical composition, colour, texture and crystallinity degree. The experimentation was carried out with a Q-switched Nd:YAG laser. The colour variations on stones associated with different operative fluences were measured using a colorimeter. Further, surface morphological changes were examined under SEM. From the calculation of colour differences, a damage threshold fluence was established for each stone type. The response of the stones to laser radiation at a particular fluence was found to be mainly conditioned by their chemical and mineralogical composition and, to a less extent, by their textural characteristics

A novel [89Zr]-anti-PD-1-PET-CT to assess response to PD-1/PD-L1 blockade in lung cancer

BackgroundHarnessing the anti-tumor immune system response by targeting the program cell death protein (PD-1) and program cell death ligand protein (PD-L1) axis has been a major breakthrough in non-small cell lung cancer (NSCLC) therapy. Nonetheless, conventional imaging tools cannot accurately assess response in immunotherapy-treated patients. Using a lung cancer syngeneic mouse model responder to immunotherapy, we aimed to demonstrate that [89Zr]-anti-PD-1 immuno-PET is a safe and feasible imaging modality to assess the response to PD-1/PD-L1 blockade in NSCLC.Materials and methodsA syngeneic mouse model responder to anti-PD-1 therapy was used. Tumor growth and response to PD-1 blockade were monitored by conventional 2-deoxy-2-[18F]fluoro-D-glucose ([18F]-FDG) PET scans. Additionally, tumor lymphocyte infiltration was analyzed by the use of an [89Zr]-labeled anti-PD-1 antibody and measured as 89Zr tumor uptake.ResultsConventional [18F]-FDG-PET scans failed to detect the antitumor activity exerted by anti-PD-1 therapy. However, [89Zr]-anti-PD-1 uptake was substantially higher in mice that responded to PD-1 blockade. The analysis of tumor-infiltrating immune cell populations and interleukins demonstrated an increased anti-tumor effect elicited by activation of effector immune cells in PD-1-responder mice. Interestingly, a positive correlation between [89Zr]-anti-PD-1 uptake and the proportion of tumor-infiltrating lymphocytes (TILs) was found (Cor = 0.8; p = 0.001).ConclusionOur data may support the clinical implementation of immuno-PET as a promising novel imaging tool to predict and assess the response of PD-1/PD-L1 inhibitors in patients with NSCLC

Overcoming PLK1 inhibitor resistance by targeting mevalonate pathway to impair AXL-TWIST axis in colorectal cancer

© 2021 The Author(s).New therapeutic targets are revolutionizing colorectal cancer clinical management, opening new horizons in metastatic patients’ outcome. Polo Like Kinase1 (PLK1) inhibitors have high potential as antitumoral agents, however, the emergence of drug resistance is a major challenge for their use in clinical practice. Overcoming this challenge represents a hot topic in current drug discovery research. BI2536-resistant colorectal cancer cell lines HT29R, RKOR, SW837R and HCT116R, were generated in vitro and validated by IG50 assays and xenografts models by the T/C ratio. Exons 1 and 2 of PLK1 gene were sequenced by Sanger method. AXL pathway, Epithelial-to-Mesenchymal transition (EMT) and Multidrug Resistance (MDR1) were studied by qPCR and western blot in resistant cells. Simvastatin as a re-sensitizer drug was tested in vitro and the drug combination strategies were validated in vitro and in vivo. PLK1 gene mutation R136G was found for RKOR. AXL pathway trough TWIST1 transcription factor was identified as one of the mechanisms involved in HT29R, SW837R and HCT116R lines, inducing EMT and upregulation of MDR1. Simvastatin was able to impair the mechanisms activated by adaptive resistance and its combination with BI2536 re-sensitized resistant cells in vitro and in vivo. Targeting the mevalonate pathway contributes to re-sensitizing BI2536-resistant cells in vitro and in vivo, raising as a new strategy for the clinical management of PLK1 inhibitors.This study has been funded by Instituto de Salud Carlos III (ISCIII) -Fondos FEDER proyects PI16/01468 and PI19/01231

Del color de los ojos al interior del genoma. Nuevas tecnologías aplicadas a la educación: una experiencia en la enseñanza de la Genética

Depto. de Genética, Fisiología y MicrobiologíaFac. de Ciencias BiológicasFALSEsubmitte

UNR/CDSE1 expression as prognosis biomarker in resectable pancreatic ductal adenocarcinoma patients: A proof-of-concept

Pancreatic ductal adenocarcinoma is an aggressive form of pancreatic cancer and the fourth leading cause of cancer-related death. When possible, curative approaches are based on surgical resection, though not every patient is a candidate for surgery. There are clinical guidelines for the management of these patients that offer different treatment options depending on the clinical and pathologic characteristics. However, the survival rates seen in this kind of patients are still low. The CDSE1 gene is located upstream of NRAS and encodes an RNA-binding protein termed UNR. The aim of this study was to analyze UNR expression and its correlation with outcome in patients with resectable pancreatic ductal adenocarcinoma (PDAC). For this, samples from resectable PDAC patients who underwent duodenopancreatectomy were used to evaluate UNR protein expression by immunohistochemistry using a tissue microarray. Here, we observed that low UNR expression was significantly associated with shorter progression-free survival after surgery (P = 0.010). Moreover, this prognostic marker remained significant after Cox proportional hazards model (P = 0.036). We further studied the role of CDSE1 expression in patient’s prognosis using data from public repositories (GEO and TGCA), confirming our results. Interestingly, CDSE1 expression correlated with that of genes characteristic of an immunogenic molecular subtype of pancreatic cancer. Based on these findings, UNR may be considered a potential prognostic biomarker for resectable PDAC and may serve to guide subsequent adjuvant treatment decisionsThis work has been carried out with the support of the RNA-Reg CONSOLIDER Network CSD2009-00080 (J.M.-U. and J.G.-F.), and Spanish Health Research Project Funds PI16/ 01468 from ªInstituto de Salud Carlos IIIº (A.C. and J.G.-F.), both of the Spanish Ministry of Economy, Industry and Competitivenes

Herramientas de aprendizaje para estudiantes de secundaria: Aplicación de la Genética y la Genómica

Depto. de Genética, Fisiología y MicrobiologíaFac. de Ciencias BiológicasFALSEsubmitte

Un examen actualizado de la percepción de las barreras para la implementación de la farmacogenómica y la utilidad de los pares fármaco/gen en América Latina y el Caribe

La farmacogenómica (PGx) se considera un campo emergente en los países en desarrollo. La investigación sobre PGx en la región de América Latina y el Caribe (ALC) sigue siendo escasa, con información limitada en algunas poblaciones. Por lo tanto, las extrapolaciones son complicadas, especialmente en poblaciones mixtas. En este trabajo, revisamos y analizamos el conocimiento farmacogenómico entre la comunidad científica y clínica de ALC y examinamos las barreras para la aplicación clínica. Realizamos una búsqueda de publicaciones y ensayos clínicos en este campo en todo el mundo y evaluamos la contribución de ALC. A continuación, realizamos una encuesta regional estructurada que evaluó una lista de 14 barreras potenciales para la aplicación clínica de biomarcadores en función de su importancia. Además, se analizó una lista emparejada de 54 genes/fármacos para determinar una asociación entre los biomarcadores y la respuesta a la medicina genómica. Esta encuesta se comparó con una encuesta anterior realizada en 2014 para evaluar el progreso en la región. Los resultados de la búsqueda indicaron que los países de América Latina y el Caribe han contribuido con el 3,44% del total de publicaciones y el 2,45% de los ensayos clínicos relacionados con PGx en todo el mundo hasta el momento. Un total de 106 profesionales de 17 países respondieron a la encuesta. Se identificaron seis grandes grupos de obstáculos. A pesar de los continuos esfuerzos de la región en la última década, la principal barrera para la implementación de PGx en ALC sigue siendo la misma, la "necesidad de directrices, procesos y protocolos para la aplicación clínica de la farmacogenética/farmacogenómica". Las cuestiones de coste-eficacia se consideran factores críticos en la región. Los puntos relacionados con la reticencia de los clínicos son actualmente menos relevantes. Según los resultados de la encuesta, los pares gen/fármaco mejor clasificados (96%-99%) y percibidos como importantes fueron CYP2D6/tamoxifeno, CYP3A5/tacrolimus, CYP2D6/opioides, DPYD/fluoropirimidinas, TMPT/tiopurinas, CYP2D6/antidepresivos tricíclicos, CYP2C19/antidepresivos tricíclicos, NUDT15/tiopurinas, CYP2B6/efavirenz y CYP2C19/clopidogrel. En conclusión, aunque la contribución global de los países de ALC sigue siendo baja en el campo del PGx, se ha observado una mejora relevante en la región. La percepción de la utilidad de las pruebas PGx en la comunidad biomédica ha cambiado drásticamente, aumentando la concienciación entre los médicos, lo que sugiere un futuro prometedor en las aplicaciones clínicas de PGx en ALC.Pharmacogenomics (PGx) is considered an emergent field in developing countries. Research on PGx in the Latin American and the Caribbean (LAC) region remains scarce, with limited information in some populations. Thus, extrapolations are complicated, especially in mixed populations. In this paper, we reviewed and analyzed pharmacogenomic knowledge among the LAC scientific and clinical community and examined barriers to clinical application. We performed a search for publications and clinical trials in the field worldwide and evaluated the contribution of LAC. Next, we conducted a regional structured survey that evaluated a list of 14 potential barriers to the clinical implementation of biomarkers based on their importance. In addition, a paired list of 54 genes/drugs was analyzed to determine an association between biomarkers and response to genomic medicine. This survey was compared to a previous survey performed in 2014 to assess progress in the region. The search results indicated that Latin American and Caribbean countries have contributed 3.44% of the total publications and 2.45% of the PGx-related clinical trials worldwide thus far. A total of 106 professionals from 17 countries answered the survey. Six major groups of barriers were identified. Despite the region’s continuous efforts in the last decade, the primary barrier to PGx implementation in LAC remains the same, the “need for guidelines, processes, and protocols for the clinical application of pharmacogenetics/pharmacogenomics”. Cost-effectiveness issues are considered critical factors in the region. Items related to the reluctance of clinicians are currently less relevant. Based on the survey results, the highest ranked (96%–99%) gene/drug pairs perceived as important were CYP2D6/tamoxifen, CYP3A5/tacrolimus, CYP2D6/opioids, DPYD/fluoropyrimidines, TMPT/thiopurines, CYP2D6/tricyclic antidepressants, CYP2C19/tricyclic antidepressants, NUDT15/thiopurines, CYP2B6/efavirenz, and CYP2C19/clopidogrel. In conclusion, although the global contribution of LAC countries remains low in the PGx field, a relevant improvement has been observed in the region. The perception of the usefulness of PGx tests in biomedical community has drastically changed, raising awareness among physicians, which suggests a promising future in the clinical applications of PGx in LAC

Evaluación preclínica y caracterización molecular de terapias antiangiogénicas en carcinomas neuroendocrinos. Papel de Galectina-1 como mediador de respuesta

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Bioquímica. Fecha de lectura: 24-04-2019Esta tesis tiene embargado el acceso al texto completo hasta el 24-10-2020Los carcinomas neuroendocrinos (CNE), son considerados una enfermedad rara, suponen aproximadamente entre 10-20% de todas las neoplasias neuroendocrinas (NNE) diagnosticadas y su supervivencia media está alrededor de los 10 meses. Son tumores que, al diagnóstico, presentan enfermedad avanzada y metástasis. A día de hoy el único tratamiento aprobado es la quimioterapia basada en cisplatino/etopósido en primera línea, con una segunda línea no definida. Los CNE presentan una elevada actividad angiogénica caracterizada por una alta expresión de factores proangiogénicos, como son los miembros de la familia del factor de crecimiento vascular (VEGF). Por este motivo, este trabajo plantea la evaluación in vivo de dos tratamientos antiangiogénicos, aflibercept y bevacizumab, en dos modelos de xenoinjerto murinos desarrollados a partir de células humanas de CNE de colon y pulmón. Nuestros resultados demuestran que ambos fármacos tienen una actividad antitumoral elevada en ambos modelos, siendo especialmente llamativo el efecto denominado de “estabilidad tumoral” inducido por aflibercept en el modelo de pulmón. En el estudio de los mecanismos relacionados con la actividad antitumoral asociada a las terapias antiangiogénicas estudiadas, los resultados muestran que las diferencias de inhibición de crecimiento tumoral con aflibercept entre ambos modelos son debidas, al menos en parte, a los diferentes niveles de expresión de la proteína Galectina-1 (Gal-1). Los tumores generados a partir de la línea de CNE de pulmón presentaban altos niveles de Gal-1 que eran significativamente reducidos con aflibercept. Dicha reducción afectó directamente a procesos vitales para el desarrollo del tumor tales como la invasión celular, la transición epitelio-mesenquima y la desestructuración de la matriz extracelular. Estos resultados no se observaron en el modelo derivado de la línea de CNE de colon cuyos tumores presentaban niveles apenas detectables de Gal-1, sugiriendo que esta proteína puede convertirse en un potencial marcador predictivo para el tratamiento con aflibercept. El análisis de Gal-1 en un pequeño grupo de pacientes con CNE confirmó que tumores con origen pancreático o pulmonar presentaban niveles elevados de esta proteína mientras que los CNE de colon mostraban niveles similares a sus correspondientes tejidos normales. Estos resultados junto la alta eficacia de aflibercept demostrada en el modelo preclínico con elevados niveles de Gal-1 abren la puerta al uso de este antiangiogénico en pacientes cuyos tumores sobreexpresen Gal-1.Neuroendocrine carcinomas (NEC) are considered a rare disease. Approximately, they represent 10- 20% of all diagnosed neuroendocrine neoplasms (NEN), and their median survival is around 10 months. These tumors debut with advanced disease and metastasis at diagnosis. So far, chemotherapy scheme based on cisplatin/etoposide is the unique therapy as first line, and after relapse, the second treatment line is not well defined. NECs show a high angiogenic activity characterized by an over expression of proangiogenic factors, such as vascular growth factor (VEGF) family. For this reason, this work proposes the in vivo evaluation of two antiangiogenic treatments, aflibercept and bevacizumab, in two murine xenograft models developed from human colon and lung CNE cell lines. Our results show that both drugs exert a high antitumor activity in both models, being especially striking the "tumor stability" induced by aflibercept in the lung model. Regarding to the potential mechanisms associated with the antitumoral activity of antiangiogenic therapies analyzed in this study, the results show that the differences of tumor growth inhibition by aflibercept between both xenograft models are due, at least partially, to the different protein levels of Galectin-1 (Gal-1). Tumors derived from lung NEC cell line showed high levels of Gal-1 that were significantly reduced by aflibercept. This reduction directly affects essential processes for tumor development such as cell invasion, transition epithelium-mesenchyma and the extracellular matrix remodeling. These effects were not observed in xenograft model from colon CNE cell line whose tumors show almost undetectable Gal-1 levels, suggesting that this protein could be a potential predictive marker of aflibercept treatment. Analysis of Gal-1 in a set of patients with NEC confirmed that tumors with pancreatic or pulmonary origin showed high levels of this protein whereas colon NECs showed similar levels as their matched normal tissues. These results together with the high efficacy of aflibecept demonstrated in the preclinical model with high levels of Gal-1 open the door to the use of this antiangiogenic as therapy in patients whose tumors overexpress Gal-1

Id1 and PD-1 Combined Blockade Impairs Tumor Growth and Survival of KRAS-mutant Lung Cancer by Stimulating PD-L1 Expression and Tumor Infiltrating CD8+ T Cells

The use of PD-1/PD-L1 checkpoint inhibitors in advanced NSCLC is associated with longer survival. However, many patients do not benefit from PD-1/PD-L1 blockade, largely because of immunosuppression. New immunotherapy-based combinations are under investigation in an attempt to improve outcomes. Id1 (inhibitor of differentiation 1) is involved in immunosuppression. In this study, we explored the potential synergistic effect of the combination of Id1 inhibition and pharmacological PD-L1 blockade in three different syngeneic murine KRAS-mutant lung adenocarcinoma models. TCGA analysis demonstrated a negative and statistically significant correlation between PD-L1 and Id1 expression levels. This observation was confirmed in vitro in human and murine KRAS-driven lung cancer cell lines. In vivo experiments in KRAS-mutant syngeneic and metastatic murine lung adenocarcinoma models showed that the combined blockade targeting Id1 and PD-1 was more effective than each treatment alone in terms of tumor growth impairment and overall survival improvement. Mechanistically, multiplex quantification of CD3+/CD4+/CD8+ T cells and flow cytometry analysis showed that combined therapy favors tumor infiltration by CD8+ T cells, whilst in vivo CD8+ T cell depletion led to tumor growth restoration. Co-culture assays using CD8+ cells and tumor cells showed that T cells present a higher antitumor effect when tumor cells lack Id1 expression. These findings highlight that Id1 blockade may contribute to a significant immune enhancement of antitumor efficacy of PD-1 inhibitors by increasing PD-L1 expression and harnessing tumor infiltration of CD8+ T lymphocytes