Dynamic epigenetic regulation of the microRNA-200 family mediates epithelial and mesenchymal transitions in human tumorigenesis

Epithelial-mesenchymal (EMT) and mesenchymal-epithelial (MET) transitions occur in the development of human tumorigenesis and are part of the natural history of the process to adapt to the changing microenvironment. In this setting, the miR-200 family is recognized as a master regulator of the epithelial phenotype by targeting ZEB1 and ZEB2, two important transcriptional repressors of the cell adherence (E-cadherin) and polarity (CRB3 and LGL2) genes. Recently, the putative DNA methylation associated inactivation of various miR-200 members has been described in cancer. Herein, we show that the miR-200ba429 and miR-200c141 transcripts undergo a dynamic epigenetic regulation linked to EMT or MET phenotypes in tumor progression. The 5′-CpG islands of both miR-200 loci were found unmethylated and coupled to the expression of the corresponding miRNAs in human cancer cell lines with epithelial features, such as low levels of ZEB1/ZEB2 and high expression of E-cadherin, CRB3 and LGL2, while CpG island hypermethylation-associated silencing was observed in transformed cells with mesenchymal characteristics. The recovery of miR-200ba429 and miR-200c141 expression by stable transfection in the hypermethylated cells restored the epithelial markers and inhibited migration in cell culture and tumoral growth and metastasis formation in nude mice. We also discovered, using both cell culture and animal models, that the miR-200 epigenetic silencing is not an static and fixed process but it can be shifted to hypermethylated or unmethylated 5′-CpG island status corresponding to the EMT and MET phenotypes, respectively. In fact, careful laser microdissection in human primary colorectal tumorigenesis unveiled that in normal colon mucosa crypts (epithelia) and stroma (mesenchyma) already are unmethylated and methylated at these loci, respectively; and that the colorectal tumors undergo selective miR-200 hypermethylation of their epithelial component. These findings indicate that the epigenetic silencing plasticity of the miR-200 family contributes to the evolving and adapting phenotypes of human tumors

Maternal care boosted by paternal imprinting in mammals.

In mammals, mothers are the primary caregiver, programmed, in part, by hormones produced during pregnancy. High-quality maternal care is essential for the survival and lifelong health of offspring. We previously showed that the paternally silenced imprinted gene pleckstrin homology-like domain family A member 2 (Phlda2) functions to negatively regulate a single lineage in the mouse placenta called the spongiotrophoblast, a major source of hormones in pregnancy. Consequently, the offspring's Phlda2 gene dosage may influence the quality of care provided by the mother. Here, we show that wild-type (WT) female mice exposed to offspring with three different doses of the maternally expressed Phlda2 gene-two active alleles, one active allele (the extant state), and loss of function-show changes in the maternal hypothalamus and hippocampus during pregnancy, regions important for maternal-care behaviour. After birth, WT dams exposed in utero to offspring with the highest Phlda2 dose exhibit decreased nursing and grooming of pups and increased focus on nest building. Conversely, 'paternalised' dams, exposed to the lowest Phlda2 dose, showed increased nurturing of their pups, increased self-directed behaviour, and a decreased focus on nest building, behaviour that was robustly maintained in the absence of genetically modified pups. This work raises the intriguing possibility that imprinting of Phlda2 contributed to increased maternal care during the evolution of mammals

Assessment of variability sources in grape ripening parameters by using FTIR and multivariate modelling

The variability in grape ripening is associated with the fact that each grape berry undergoes its own biochemical processes. Traditional viticulture manages this by averaging the physicochemical values of hundreds of grapes to make decisions. However, to obtain accurate results it is necessary to evaluate the different sources of variability, so exhaustive sampling is essential. In this article, the factors “grape maturity over time” and “position of the grape” (both in the grapevine and in the bunch/cluster) were considered and studied by analyzing the grapes with a portable ATR-FTIR instrument and evaluating the spectra obtained with ANOVA–simultaneous component analysis (ASCA). Ripeness over time was the main factor affecting the characteristics of the grapes. Position in the vine and in the bunch (in that order) were also significantly important, and their effect on the grapes evolves over time. In addition, it was also possible to predict basic oenological parameters (TSS and pH with errors of 0.3 °Brix and 0.7, respectively). Finally, a quality control chart was built based on the spectra obtained in the optimal state of ripening, which could be used to decide which grapes are suitable for harvest

Unified behavioral scoring for preclinical models

Preclinical mental health research relies upon animal models, and whilst many encouraging advances are being made, reproducibility and translational relevance may be limited by sub-optimal testing or model choices. Animal behaviors are complex and test batteries should be designed to include their multifaceted nature. However, multiple behavioral testing is often avoided due to cost, availability or statistical rigor. Additionally, despite the disparity in the incidence of mental health problems between the sexes, a move toward reducing animal numbers could be a deterrent to including both male and female animals. The current study introduces a unified scoring system for specific behavioral traits with the aim of maximizing the use of all data generated whilst reducing the incidence of statistical errors. Female and male mice from two common background strains were tested on behavior batteries designed to probe multiple aspects of anxiety-related and social behavioral traits. Results for every outcome measure were normalized to generate scores for each test and combined to give each mouse a single unified score for each behavioral trait. The unified behavioral scores revealed clear differences in the anxiety and stress-related, and sociability traits of mice. Principle component analysis of data demonstrated significant clustering of animals into their experimental groups. In contrast, individual tests returned an ambiguous mixture of non-significant trends and significant effects for various outcome measures. Utilizing a range of behavioral measures and combining all outcome measure data to produce unified scores provides a useful tool for detecting subtle behavioral traits in preclinical models

Prevention of diet-induced obesity by apple polyphenols in Wistar rats through regulation of adipocyte gene expression and DNA methylation patterns

This study was conducted to determine the mechanisms implicated in the beneficial effects of apple polyphenols (APs) against diet-induced obesity in Wistar rats, described in a previous study from our group. Supplementation of high-fat sucrose diet with AP prevented adiposity increase by inhibition of adipocyte hypertrophy. Rats supplemented with AP exhibited improved glucose tolerance while adipocytes isolated from these rats showed an enhanced lipolytic response to isoproterenol. AP intake led to reduced Lep, Plin, and sterol regulatory element binding transcription factor 1 (Srebf1) mRNA levels and increased aquaporin 7 (Aqp7), adipocyte enhancer binding protein 1 (Aebp1), and peroxisome proliferator-activated receptor gamma co-activator 1 alpha (Ppargc1a) mRNA levels in epididymal adipocytes. In addition, we found different methylation patterns of Aqp7, Lep, Ppargc1a, and Srebf1 promoters in adipocytes from apple-supplemented rats compared to high-fat sucrose fed rats. The administration of AP protects against body weight gain and fat deposition and improves glucose tolerance in rats. We propose that AP exerts the antiobesity effects through the regulation of genes involved in adipogenesis, lipolysis, and fatty acid oxidation, in a process that could be mediated in part by epigenetic mechanisms

Placental endocrine insufficiency programs anxiety, deficits in cognition and atypical social behaviour in offspring

Abnormally elevated expression of the imprinted PHLDA2 gene has been reported in the placenta of human babies that are growth restricted in utero in several studies. We previously modelled this gene alteration in mice and found that just two-fold increased expression of Phlda2 resulted in placental endocrine insufficiency. In addition, elevated Phlda2 was found to drive fetal growth restriction (FGR) of transgenic offspring and impaired maternal care by their wild type mothers. Being born small and being exposed to suboptimal maternal care have both been associated with the increased risk of mental health disorders in human populations. In the current study we probed behavioural consequences of elevated Phlda2 for the offspring. We discovered increased anxiety-like behaviours, deficits in cognition and atypical social behaviours, with the greatest impact on male offspring. Subsequent analysis revealed alterations in the transcriptome of the adult offspring hippocampus, hypothalamus, and amygdala, regions consistent with these behavioural observations. The inclusion of a group of fully wild type controls raised in a normal maternal environment allowed us to attribute behavioural and molecular alterations to the adverse maternal environment induced by placental endocrine insufficiency rather than the specific gene change of elevated Phlda2. Our work demonstrates that a highly common alteration reported in human fetal growth restriction is associated with negative behavioural outcomes later in life. Importantly, we also establish the experimental paradigm that placental endocrine insufficiency can program atypical behaviour in offspring highlighting the under-appreciated role of placental endocrine insufficiency in driving disorders of later life behaviour

Plan de formación y docencia en un centro integral de cáncer: estrategia e implementación

Introducción: El Institut Català d'Oncologia (ICO) puso en marcha la Unidad de Formación y Docencia (UFiD), en el año 2006, con el objetivo de potenciar el desarrollo profesional, la difusión del conocimiento y la calidad de la atención oncológica. En el presente artículo se describe el proceso de puesta en marcha de la UFiD, los resultados del estudio de las necesidades formativas realizada por los profesionales y colectivos de la institución, las acciones formativas organizadas, y en el área de docencia, los resultados de la reorganización de las trayectorias docentes de los diferentes servicios, así como el apoyo de nuevas iniciativas. Materiales y métodos: Durante el periodo 2008-2009, se implantaron acciones formativas y docentes en concordancia con los resultados del estudio de necesidades y con las líneas estratégicas de la institución. Se realizaron un total de 71 actividades formativas, con 898 participantes de todas las categorías profesionales, con una tasa de satisfacción de los participantes de 7,8 sobre 10. En el apartado de docencia, durante el mismo periodo se realizaron 3 másteres, 3 posgrados, 59 cursos de actualización con metodología semipresencial y 19 cursos con metodología online, todos ellos con acreditación académica. La satisfacción media fue de 8 sobre 10. Resultados y conclusiones: La actividad llevada a cabo por la UFiD ha permitido, por una parte, articular la formación continuada en base al estudio de las necesidades de los profesionales, lo que ha contribuido a promover el desarrollo profesional y la calidad asistencial de éstos, y por otra parte, dar soporte a las actividades docentes ya existentes y promover nuevas iniciativas para favorecer la difusión de conocimientos en el área oncológica, lo que ha posicionado al ICO como un centro absolutamente comprometido con la formación oncológica

Suspensión del tratamiento con inhibidores de BCR-ABL en los pacientes con leucemia mieloide crónica en respuesta molecular profunda dentro de la práctica clínica asistencial: Experiencia española en un total de 236 casos

Oral Presentation [CO-092] Introducción: La mitad de los pacientes con leucemia mieloide crónica (LMC) en respuesta molecular profunda no pierde la respuesta molecular mayor (RMM) tras la suspensión del tratamiento con inhibidores de BCR-ABL (ITC). Esta estrategia ha demostrado ser segura en ensayos clínicos pero hay poca información acerca de su aplicabilidad en la práctica clínica asistencial. El objetivo del estudio fue analizar la experiencia con la suspensión del tratamiento fuera de ensayo clínico en España. Métodos: Se analizan los resultados de 236 pacientes con LMC en fase crónica que suspendieron el tratamiento fuera de ensayo clínico en 33 hospitales. Criterios de inclusión: a) tratamiento con ITC >3 años, b) respuesta molecular grado 4.5 durante >2 años (se permitió una única determinación de RM4 durante ese período). Se excluyeron los pacientes trasplantados. Resultados: Las características de la serie se muestran en la tabla 1. Los motivos principales para suspender el tratamiento fueron los efectos secundarios (n=66), lograr la remisión libre de tratamiento (n=166) y el embarazo (n=4). La mediana de seguimiento tras la suspensión fue de 21, 5 meses y 5 pacientes fallecieron por causas no relacionadas con la LMC. Durante este periodo, 67 pacientes reiniciaron el tratamiento por recaída molecular (pérdida de RMM: n=52, aumento de tránscritos >1 log en dos controles sucesivos sin pérdida de RMM: n=12), decisión del paciente (n=2) o síndrome de discontinuación (n=1). Un paciente perdió la RMM a los 20 meses y decidió no tratarse, recuperando la RMM espontáneamente. Cuarenta y nueve recaídas (75% del total) ocurrieron en los primeros 6 meses, 8 entre los meses 7-12, y 8 tras los 12 meses, produciéndose la pérdida de RMM más tardía a los 30 meses. La supervivencia libre de reinicio del tratamiento (figura 1) y de recaída molecular fue del 66, 8% y del 67, 5% a los 3 años, respectivamente. Los factores asociados a mayor supervivencia libre de recaída fueron la duración del tratamiento con ITC >5 años (p=0.01) y la RM4.5 >4 años antes de la suspensión (p=0.017). Un total de 51 pacientes (22%) desarrollaron dolor osteomuscular tras la suspensión. No se registró ningún caso de progresión a fases avanzadas. El valor mediano de la carga de BCR-ABL al reinicio del tratamiento fue del 0, 3% (>5% en 7 casos). La mediana de seguimiento tras reinicio del tratamiento fue de 20 meses; 46 de 52 casos (88%) recuperaron la RMM tras una mediana de tiempo de 3 meses; 50 de 64 recuperaron la RM4 (mediana 3, 5 meses) y 47 de 64 recuperaron la MR4.5 (mediana 5 meses). En el último control, el estado de la respuesta fue: RM4.5 (n=195), RM4 (n=15), RMM (n=14), respuesta citogenética completa (n=10), otros (n=2). Conclusiones: los resultados confirman que la suspensión del tratamiento es factible en la práctica clínica asistencial en España. La duración del tratamiento y de la respuesta molecular profunda se asociaron con la supervivencia libre de recaída. Esta información puede ser útil para establecer recomendaciones generales acerca de la discontinuación del tratamiento de la LMC en nuestro medio

RICORS2040 : The need for collaborative research in chronic kidney disease

Chronic kidney disease (CKD) is a silent and poorly known killer. The current concept of CKD is relatively young and uptake by the public, physicians and health authorities is not widespread. Physicians still confuse CKD with chronic kidney insufficiency or failure. For the wider public and health authorities, CKD evokes kidney replacement therapy (KRT). In Spain, the prevalence of KRT is 0.13%. Thus health authorities may consider CKD a non-issue: very few persons eventually need KRT and, for those in whom kidneys fail, the problem is 'solved' by dialysis or kidney transplantation. However, KRT is the tip of the iceberg in the burden of CKD. The main burden of CKD is accelerated ageing and premature death. The cut-off points for kidney function and kidney damage indexes that define CKD also mark an increased risk for all-cause premature death. CKD is the most prevalent risk factor for lethal coronavirus disease 2019 (COVID-19) and the factor that most increases the risk of death in COVID-19, after old age. Men and women undergoing KRT still have an annual mortality that is 10- to 100-fold higher than similar-age peers, and life expectancy is shortened by ~40 years for young persons on dialysis and by 15 years for young persons with a functioning kidney graft. CKD is expected to become the fifth greatest global cause of death by 2040 and the second greatest cause of death in Spain before the end of the century, a time when one in four Spaniards will have CKD. However, by 2022, CKD will become the only top-15 global predicted cause of death that is not supported by a dedicated well-funded Centres for Biomedical Research (CIBER) network structure in Spain. Realizing the underestimation of the CKD burden of disease by health authorities, the Decade of the Kidney initiative for 2020-2030 was launched by the American Association of Kidney Patients and the European Kidney Health Alliance. Leading Spanish kidney researchers grouped in the kidney collaborative research network Red de Investigación Renal have now applied for the Redes de Investigación Cooperativa Orientadas a Resultados en Salud (RICORS) call for collaborative research in Spain with the support of the Spanish Society of Nephrology, Federación Nacional de Asociaciones para la Lucha Contra las Enfermedades del Riñón and ONT: RICORS2040 aims to prevent the dire predictions for the global 2040 burden of CKD from becoming true