Análisis de los factores pronósticos en el cáncer de colon estadios II y III

ANÁLISIS DE LOS FACTORES PRONOSTICOS EN EL CANCER DE COLON ESTADIOS II Y IIILa tesis versa sobre un análisis retrospectivo, que incluye 213 paciente que padecieron cáncer de colon en estadio patológico II o III entre el año 2007 y 2010. Todos los sujetos fueron intervenidos de su neoplasia y posteriormente completaron tratamiento adyuvante con quimioterapia, dado que eran estadios III o estadios II con algún factor de mal pronóstico. El objetivo primario de la tesis era determinar si alguno de los factores clinicos, histopatológicos y moleculares estudiados se verificaba como factor pronóstico independiente, a los 5 años de seguimientos, en términos de superviencia; supervivencia libre enfermedad [SLE], supervivencia global [SG] y/o supervivencia cáncer específica [SCE].<br /

Correlación radio-patológica de criterios morfológicos en la valoración de metástasis hepáticas de adenocarcinoma de colon

El cáncer colorrectal (CCR) es una de las neoplasias más comunes en el mundo occidental, habiendo aumentado su incidencia durante las últimas décadas. En España es la segunda causa de muerte por cáncer en la población general, solamente superada por el cáncer de pulmón. Cerca del 50% de las metástasis a distancia del CCR se encuentran exclusivamente en el hígado y dos tercios de los pacientes con CCR diseminado fallecen a consecuencia de las metástasis hepáticas. La evaluación de la respuesta al tratamiento oncológico es una tarea crucial que comparten hoy en día radiólogos y patólogos. La respuesta histológica a la terapia preoperatoria ha demostrado ser un factor predictivo de la supervivencia en los pacientes con tumores sólidos. Una evaluación radiológica confiable, reproducible y de aplicación universal es fundamental en el abordaje del CCR, tanto en la valoración diagnóstica y de extensión de la enfermedad como en la respuesta al tratamiento, ya que dicha respuesta proporciona información pronóstica de gran importancia. Clásicamente la respuesta al tratamiento en las metástasis hepáticas de CCR ha sido evaluada mediante la variación radiológica del tamaño tumoral, no obstante el advenimiento de nuevos fármacos quimioterápicos con predominio de la actividad citostática sobre la citotóxica ha significado que la sola medición de la reducción del tamaño tumoral no sea suficiente, dado que no estima la necrosis o la fibrosis tumoral secundaria al tratamiento. Además, la determinación del porcentaje de células tumorales viables en el seno de la metástasis analizada puede considerarse un factor pronóstico en la supervivencia. Dada la amplia disponibilidad de la TAC en la práctica clínica habitual, aunado a las mejoras en la resolución de las imágenes introducidas por la tecnología multidetector y la capacidad de cuantificación de los valores de atenuación en las estructuras estudiadas, nos hemos propuesto determinar el grado de correlación entre los parámetros de caracterización histomorfológica y los hallazgos morfodensitométricos objetivables por TAC en las metástasis hepáticas de adenocarcinoma de colon, en lo relativo a la posible correlación radiológica con los fenómenos histopatológicos de desvitalización de las lesiones, concretamente en forma de necrosis o fibrosis y la morfología de la interfase tumor/parénquima hepático. Para ello hemos realizamos un estudio retrospectivo de carácter descriptivo, respecto a los parámetros radio-patológicos de una base de datos de 150 pacientes intervenidos de hepatectomía con intención curativa por parte de la Unidad de Cirugía Hepatobiliopancreática del Servicio de Cirugía General y del Aparato Digestivo del Hospital Universitario Miguel Servet de Zaragoza. Los casos debían disponer de imágenes de TAC que valorasen la presencia, extensión y características morfológicas de las metástasis hepáticas. De los 150 casos disponibles, sólo 60 pacientes cumplieron con los criterios de inclusión en el estudio. Se observó una distribución por sexo de 57% hombres (34/60) y 43% de mujeres (26/60), con una media de edad de 64,4 ± 10,2 años. Hemos demostrado una asociación negativa entre los valores de atenuación tumoral y la respuesta histológica, interpretada como la suma de los porcentajes de necrosis y fibrosis (-0,256 con p = 0,05), tal que los valores de atenuación disminuyen a mayor respuesta histológica. Respecto a la celularidad viable, mayormente observable en la periferia tumoral, hemos demostrado asociación estadística entre el Anillo de Hiperrealce Periférico (equivalente radiológico al Casquete Tumoral o Tumor Normal Interface - TNI) y la respuesta histológica estimada mediante los porcentajes de necrosis y fibrosis tumoral, tal que los valores de fibrosis tumoral son mayores en los casos con ausencia de anillo de hiperrealce, mientras que la necrosis tumoral es mayor en los casos con presencia del mencionado anillo (p valor de 0,038 y 0,007 respectivamente). Si se considera la respuesta histológica de predominio fibroso como un indicador de buen pronóstico, nuestros resultados abren la posibilidad de sugerir la valoración del anillo de hiperrealce periférico como un indicador pronóstico prequirúrgico. No hemos demostrado correlación radio-patológica en lo referente a las determinaciones de los contornos y márgenes tumorales, en concreto, la estimación radiológica del contorno (liso vs. irregular) con respecto a la presencia de pseudocápsula fibrosa (p = 0,173); la valoración histológica del margen tumoral (infiltrante vs. expansivo) respecto a la estimación radiológica del contorno (liso vs. irregular) (p = 0,144); el patrón vascular (hipóxico vs. no hipóxico) frente a la estimación radiológica del contorno (liso vs. irregular) (x2=0,356) y frente a la presencia de anillo de hiperrealce periférico (x2=1,000); o la estimación histológica del margen tumoral (infiltrante vs. expansivo) con respecto a la presencia de anillo de hiperrealce periférico (x2=0,301). Tampoco hemos demostrado asociación estadística entre la valoración radiológica del contorno tumoral (liso vs. irregular) y los porcentajes de necrosis (x2=0,389) o fibrosis (x2=1,000). En conclusión, la tomografía axial computada (TAC) proporciona información valiosa en la valoración prequirúrgica y de respuesta al tratamiento en las metástasis hepáticas de adenocarcinoma de colon, con correlación radio-patológica en lo relativo a parámetros macroscópicos y de variaciones densitométricas relacionadas con la respuesta histopatológica. Los patrones de respuesta histopatológica considerados como porcentajes de fibrosis y necrosis tumoral guardan asociación estadística con respecto a la atenuación global objetivada por TAC, y la presencia de Anillo de Hiperrealce Periférico (equivalente radiológico del Casquete Tumoral) se asocia a respuestas histológicas de predominio necrótico, mientras que su ausencia guarda relación con respuestas histológicas de tipo fibroso, de mejor pronóstico. En tal sentido, se propone el estudio del Anillo de Hiperrealce Periférico como un indicador radiológico pronóstico, de utilidad prequirúrgica

Identification of novel targets in adipose tissue involved in non-alcoholic fatty liver disease progression

Obesity is a major risk factor for the development of Nonalcoholic fatty liver disease (NAFLD). We hypothesize that a dysfunctional subcutaneous white adipose tissue (scWAT) may lead to an accumulation of ectopic fat in the liver. Our aim was to investigate the molecular mechanisms involved in the causative role of scWAT in NALFD progression. We performed a RNA-sequencing analysis in a discovery cohort (n = 45) to identify genes in scWAT correlated with fatty liver index, a qualitative marker of liver steatosis. We then validated those targets in a second cohort (n = 47) of obese patients who had liver biopsies available. Finally, we obtained scWAT mesenchymal stem cells (MSCs) from 13 obese patients at different stages of NAFLD and established in vitro models of human MSC (hMSC)-derived adipocytes. We observed impaired adipogenesis in hMSC-derived adipocytes as liver steatosis increased, suggesting that an impaired adipogenic capacity is a critical event in the development of NAFLD. Four genes showed a differential expression pattern in both scWAT and hMSC-derived adipocytes, where their expression paralleled steatosis degree: SOCS3, DUSP1, SIK1, and GADD45B. We propose these genes as key players in NAFLD progression. They could eventually constitute potential new targets for future therapies against liver steatosis

Flow cytometry multiplexed method for the detection of neutralizing human antibodies to the native SARS-CoV-2 spike protein

A correct identification of seropositive individuals for the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection is of paramount relevance to assess the degree of protection of a human population to present and future outbreaks of the COVID-19 pandemic. We describe here a sensitive and quantitative flow cytometry method using the cytometer-friendly non-adherent Jurkat T-cell line that stably expresses the full-length native spike “S” protein of SARS-CoV-2 and a truncated form of the human EGFR that serves a normalizing role. S protein and huEGFRt coding sequences are separated by a T2A self-cleaving sequence, allowing to accurately quantify the presence of anti-S immunoglobulins by calculating a score based on the ratio of fluorescence intensities obtained by double-staining with the test sera and anti-EGFR. The method allows to detect immune individuals regardless of the result of other serological tests or even repeated PCR monitoring. As examples of its use, we show that as much as 28% of the personnel working at the CBMSO in Madrid is already immune. Additionally, we show that anti-S antibodies with protective neutralizing activity are long-lasting and can be detected in sera 8 months after infectionThis work was funded by intramural grant CSIC-COVID19-004: 202020E081 (to B.A.) and CSIC-COVID19-004: 202020E165 (to MF). L.H has been supported by an FPI fellowship from the Spanish Ministry of Science and Innovation. I.B. has been supported by an H2020-MSCA-ITN-2016 training network grant of the European Union (GA 721358

Decreased breadth of the antibody response to the spike protein of SARS-CoV-2 after repeated vaccination

Introduction: The rapid development of vaccines to prevent COVID-19 has raised the need to compare the capacity of different vaccines in terms of developing a protective humoral response. Previous studies have shown inconsistent results in this area, highlighting the importance of further research to evaluate the efficacy of different vaccines. Methods: This study utilized a highly sensitive and reliable flow cytometry method to measure the titers of IgG1 isotype antibodies in the blood of healthy volunteers after receiving one or two doses of various vaccines administered in Spain. The method was also used to simultaneously measure the reactivity of antibodies to the S protein of the original Wuhan strain and variants B.1.1.7 (Alpha), B.1.617.2 (Delta), and B.1.617.1 (Kappa). Results: Significant differences were observed in the titer of anti-S antibodies produced after a first dose of the vaccines ChAdOx1 nCov-19/AstraZeneca, mRNA-1273/Moderna, BNT162b2/Pfizer-BioNTech, and Ad26.COV.S/Janssen. Furthermore, a relative reduction in the reactivity of the sera with the Alpha, Delta, and Kappa variants, compared to the Wuhan strain, was observed after the second boosting immunization. Discussion: The findings of this study provide a comparison of different vaccines in terms of anti-S antibody generation and cast doubts on the convenience of repeated immunization with the same S protein sequence. The multiplexed capacity of the flow cytometry method utilized in this study allowed for a comprehensive evaluation of the efficacy of various vaccines in generating a protective humoral response. Future research could focus on the implications of these findings for the development of effective COVID-19 vaccination strategies.This work was funded by intramural grant CSIC-COVID19- 004: 202020E081 (to BA) and CSIC-COVID19-004: 202020E165 (to MF). LH has been supported by an FPI fellowship from the Spanish Ministry of Science and Innovation. IB has been supported by an H2020-MSCA-ITN-2016 training network grant of the European Union (GA 721358)

Diversity of immune responses in children highly exposed to SARS-CoV-2

Background: Children are less susceptible than adults to symptomatic COVID‐19 infection, but very few studies addressed their underlying cause. Moreover, very few studies analyzed why children highly exposed to the virus remain uninfected. Methods: We analyzed the serum levels of ACE2, angiotensin II, anti-spike and anti-N antibodies, cytokine profiles, and virus neutralization in a cohort of children at high risk of viral exposure, cohabiting with infected close relatives during the lockdown in Spain. Results: We analyzed 40 children who were highly exposed to the virus since they lived with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2)-infected relatives during the lockdown for several months without taking preventive measures. Of those, 26 reported mild or very mild symptoms. The induced immune response to the virus was analyzed 3 months after the household infection. Surprisingly, only 15 children had IgG anti-S (IgG+) determined by a sensitive method indicative of a past infection. The rest, negative for IgG anti-N or S in various tests, could be further subdivided, according to IgM antibodies, into those having IgM anti-S and IgM anti-N (IgG−IgMhigh) and those having only IgM anti-N (IgG−IgMlow). Interestingly, those two subgroups of children with IgM antibodies have strikingly different patterns of cytokines. The IgMhigh group had significantly higher IFN-α2 and IFN-γ levels as well as IL-10 and GM-CSF than the IgMlow group. In contrast, the IgMlow group had low levels of ACE2 in the serum. Both groups have a weaker but significant capacity to neutralize the virus in the serum than the IgG+ group. Two children were negative in all immunological antibody tests. Conclusions: A significant proportion of children highly exposed to SARS-CoV-2 did not develop a classical adaptive immune response, defined by the production of IgG, despite being in close contact with infected relatives. A large proportion of those children show immunological signs compatible with innate immune responses (as secretion of natural antibodies and cytokines), and others displayed very low levels of the viral receptor ACE2 that may have protected them from the virus spreading in the body despite high and constant viral exposure.This research was funded by grants from “Ministerio de Ciencia e Innovación” (PID-2019104760RB-100); “Comunidad de Madrid” (S2017/BMD-3671. INFLAMUNE-CM; FEDER and COTRAVI-19-CM) to MF; Consejo Superior de Investigaciones Científica, CSIC (CSIC-COV19-108, SGL210235) to MF and UB; CRUE-Supera COVID; the European Development Regional Fund “A way to achieve Europe” (ERDF); Merck, Sharp and Dohme Investigator Studies Program (code MISP# IIS 60257); and Fondo Supera COVID-19 (2020–001) to SSV. The study also received institutional grants from “Fundación Ramón Areces” and “Banco de Santander.”Peer reviewe

ACE2 Serum Levels as Predictor of Infectability and Outcome in COVID-19

Background: COVID‐19 can generate a broad spectrum of severity and symptoms. Many studies analysed the determinants of severity but not among some types of symptoms. More importantly, very few studies analysed patients highly exposed to the virus that nonetheless remain uninfected. Methods: We analysed serum levels of ACE2, Angiotensin II and anti-Spike antibodies in 2 different cohorts at high risk of viral exposure, highly exposed but uninfected subjects, either high risk health care workers or persons cohabiting with infected close relatives and seropositive patients with symptoms. We tested the ability of the sera of these subjects to neutralize lentivirus pseudotyped with the Spike-protein. Results: We found that the serum levels of ACE2 are significantly higher in highly exposed but uninfected subjects. Moreover, sera from this seronegative persons can neutralize SARS-CoV-2 infection in cellular assays more strongly that sera from non-exposed negative controls eventhough they do not have anti-CoV-2 IgG antibodies suggesting that high levels of ACE2 in serum may somewhat protect against an active infection without generating a conventional antibody response. Finally, we show that among patients with symptoms, ACE2 levels were significantly higher in infected patients who developed cutaneous as compared with respiratory symptoms and ACE2 was also higher in those with milder symptoms. Conclusions: These findings suggest that soluble ACE2 could be used as a potential biomarker to predict SARS-CoV-2 infection risk and to discriminate COVID-19 disease subtypes.This research was funded by grants from “Ministerio de Ciencia e Innovación” (SAF2013-42850-R, SAF2016-75988-R and PID-2019104760RB-100; FEDER), “Comunidad de Madrid (S2017/BMD-3671. INFLAMUNE-CM; FEDER) to MF, Consejo Superior de Investigaciones Científica, CSIC (CSIC-COV19-108, SGL210235) to MF and UB, CRUE-Supera COVID, the European Development Regional Fund ‘‘A way to achieve Europe’’ (ERDF), Merck, Sharp & Dohme Investigator Studies Program (code MISP# IIS 60257), and Fondo Supera COVID-19 (2020-001) to SS-V. Institutional grants from “Fundación Ramón Areces” and “Banco de Santander”. This research work was also funded by the European Commission – NextGenerationEU (Regulation EU 2020/2094), through CSIC's Global Health Platform (PTI Salud Global).Peer reviewe

Decreased breadth after boost

The rapid development of vaccines to prevent infection by SARS-CoV-2 virus causing COVID-19 makes necessary to compare the capacity of the different vaccines in terms of development of a protective humoral response. Here, we have used a highly sensitive and reliable flow cytometry method to measure the titers of antibodies of the IgG1 isotype in blood of volunteers after receiving one or two doses of the vaccines being administered in Spain. We took advantage of the multiplexed capacity of the method to measure simultaneously the reactivity of antibodies with the S protein of the original strain Wuhan-1 and the variant B.1.1.7 (Alpha). We found significant differences in the titer of anti-S antibodies produced after a first dose of the vaccines ChAdOx1 nCov-19/AstraZeneca, mRNA-1273/Moderna, BNT162b2/Pfizer-BioNTech and Ad26.COV.S/Janssen. Most important, we found a relative reduction in the reactivity of the sera with the B.1.1.7 versus the Wuhan-1 variant after the second boosting immunization. These data allow to make a comparison of different vaccines in terms of anti-S antibody generation and cast doubts about the convenience of repeatedly immunizing with the same S protein sequence.This work was funded by intramural grant CSIC-COVID19-004: 202020E081 (to B.A.) and CSIC-COVID19-004: 202020E165 (to MF). L.H has been supported by an FPI fellowship from the Spanish Ministry of Science and Innovation. I.B. has been supported by an H2020-MSCA-ITN2016 training network grant of the European Union (GA 721358)Peer reviewe

Decreased breadth of the antibody response to the spike protein of SARS-CoV-2 after repeated vaccination

IntroductionThe rapid development of vaccines to prevent COVID-19 has raised the need to compare the capacity of different vaccines in terms of developing a protective humoral response. Previous studies have shown inconsistent results in this area, highlighting the importance of further research to evaluate the efficacy of different vaccines.MethodsThis study utilized a highly sensitive and reliable flow cytometry method to measure the titers of IgG1 isotype antibodies in the blood of healthy volunteers after receiving one or two doses of various vaccines administered in Spain. The method was also used to simultaneously measure the reactivity of antibodies to the S protein of the original Wuhan strain and variants B.1.1.7 (Alpha), B.1.617.2 (Delta), and B.1.617.1 (Kappa).ResultsSignificant differences were observed in the titer of anti-S antibodies produced after a first dose of the vaccines ChAdOx1 nCov-19/AstraZeneca, mRNA-1273/Moderna, BNT162b2/Pfizer-BioNTech, and Ad26.COV.S/Janssen. Furthermore, a relative reduction in the reactivity of the sera with the Alpha, Delta, and Kappa variants, compared to the Wuhan strain, was observed after the second boosting immunization.DiscussionThe findings of this study provide a comparison of different vaccines in terms of anti-S antibody generation and cast doubts on the convenience of repeated immunization with the same S protein sequence. The multiplexed capacity of the flow cytometry method utilized in this study allowed for a comprehensive evaluation of the efficacy of various vaccines in generating a protective humoral response. Future research could focus on the implications of these findings for the development of effective COVID-19 vaccination strategies