Intratumor genetic heterogeneity and clonal evolution to decode endometrial cancer progression

Analyzing different tumor regions by next generation sequencing allows the assessment of intratumor genetic heterogeneity (ITGH), a phenomenon that has been studied widely in some tumor types but has been less well explored in endometrial carcinoma (EC). In this study, we sought to characterize the spatial and temporal heterogeneity of 9 different ECs using whole-exome sequencing, and by performing targeted sequencing validation of the 42 primary tumor regions and 30 metastatic samples analyzed. In addition, copy number alterations of serous carcinomas were assessed by comparative genomic hybridization arrays. From the somatic mutations, identified by whole-exome sequencing, 532 were validated by targeted sequencing. Based on these data, the phylogenetic tree reconstructed for each case allowed us to establish the tumors’ evolution and correlate this to tumor progression, prognosis, and the presence of recurrent disease. Moreover, we studied the genetic landscape of an ambiguous EC and the molecular profile obtained was used to guide the selection of a potential personalized therapy for this patient, which was subsequently validated by preclinical testing in patient-derived xenograft models. Overall, our study reveals the impact of analyzing different tumor regions to decipher the ITGH in ECs, which could help make the best treatment decisionWe thank all those at the Translational Research Laboratory of the MD Anderson Cancer Center Madrid for their invaluable help with this study. Tissue samples were obtained with the support of the MD Anderson Foundation Biobank (recordnumber B.0000745, ISCIII National Biobank Record), the “Xarxa Catalana de Bancs de Tumors” and “Plataforma de Biobancos” ISCIII (PT13/0010/0014, B.000609). This study has been supported by the Spanish Ministry of Economy and Innovation (PID2019-104644RB-I00 (GMB), the Instituto de Salud Carlos III (ISCIII, CIBERONC, CB16/12/00295 - GMB-, CB16/12/00328 -EC, AGM- and CB16/12/00231 -XMG- [all partly supported by FEDER funds]) and by the AECC Scientific Foundation (FC_AECC PROYE19036MOR -GMB- and Coordinated groups 2018 -XMG, AGM, GMB-). SO is funded by an AECC-postdoctoral grant (2020). JSR-F and BW are funded in part by the Breast Cancer Research Foundation and in part by the NIH/ NCI P50 CA247749 01 grant. Research reported in this publication was supported in part by a Cancer Center Support Grant of the NIH/NCI (Grant No. P30CA008748; MSK). We thank the Eurofins Megalab laboratory for helping us to perform the analysis of DNA HPV detection

Deciphering intratumor molecular heterogeneity in gynecological cancer progression

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Bioquímica. Fecha de lectura: 12-05-2017Esta tesis tiene embargado el acceso al texto completo hasta el 12-11-201

Mecanismos para abordar la resiliencia urbana desde la escala local.

Planificar en la incertidumbre es uno de los mayores retos de la humanidad en sus dimensiones políticas, sociales y medio ambientales, la crisis de la incertidumbre a los fenómenos climáticos y sus impactos han modificado y resignificado el concepto de vulnerabilidad a lo largo y ancho de Latinoamérica y el Caribe.Es erróneo y peligroso pensar que la culpa de los desequilibrios ambientales y sus repercusiones es solo atribuible al Cambio Climático, es evidente que nuestra forma actual de habitar es social, ambiental y económicamente obsoleta, sumados al claro desinterés de los medios de difusión masiva en mostrar la cruda realidad social y la necesidad de asumir nuestras responsabilidades en el cambio de paradigma en cuanto a la convivencia simbiótica y armónica con el medio físico.Según el informe monitor de vulnerabilidad climática de la asociación climática DARA y el Climate Vulnerable Forum, alrededor de 350.000 personas pierden la vida actualmente por asuntos relacionados con el cambio climático, pero el mundo podría además sufrir un millón de muertes anuales a partir de 2030, si no se ponen en marcha acciones correctivas; los costes económicos alcanzarían cerca de un tercio de un billón de dólares al año, superior a los apenas 100.000 millones actuales.Estas premisas presuponen un cambio de ruta en la manera de pensar estrategias resilientes de acción local con repercusiones globales, de re-pensar lo complejo y la operatividad concreta de la investigación científica hacia la pro-actividad social, la participación ciudadana en la toma de decisiones y la recuperación de la memoria ambiental de los pueblos, esta nueva visión transdisciplinar, multisectorial y de fuertes arraigos en la cultura local permiten dar respuestas adaptativas a territorios únicos.Este artículo es fruto de la experiencia del curso CELFI, del Nodo de Sustentabilidad y Desarrollo “Resiliencia urbana y cambio climático global en ciudades latinoamericanas” y explora tres dimensiones para abordar la resiliencia urbana en ciudades latinoamericanas.“La influencia humana en el sistema climático es clara. Es evidente a tenor de las crecientes concentraciones de gases de efecto invernadero en la atmósfera, el forzamiento radiactivo positivo y el calentamiento observado, y gracias a la comprensión del sistema climático.

The use of SIGS (Spray-Induced Gene Silencing) strategy in the control of Botrytis cinerea in horticultural crops.

The causal agent of the gray mold disease, Botrytis cinerea, is one of the main limiting factors of horticultural crops production worldwide. Its control is very dependent on the use of fungicides; however, due to the ability of this pathogen to develop resistance and the new European legislation regarding the reduction of fungicide diversity available for grower in the next years, new advances and technologies are needed to control this important harvest and postharvest disease. In this study, the potential of the RNA interference (RNAi) technology known as "spray-induced gene silencing" (SIGS) as an alternative for the management of B. cinerea was explored. For this purpose, a double-stranded RNA (dsRNA) molecule was designed to target the transcription factor mrr1, which is involved in the overexpression of the ABC transporters (drug efflux transporters) and the resistance to several commercial fungicides. To test the efficacy of mrr1-dsRNA, dsRNA uptake by the fungus, sensitivity assays (effect on conidia germination, detached leaf, and fruit assays), fungal biomass analysis and gene expression studies were carried out. The results showed that the fungus was able to uptake it, reducing the conidia germination of the pathogen. Subsequently, it was shown, in in vivo assays performed on tomato leaves and apple fruit, that the application of mrr1-dsRNA downregulated mrr1 gene expression and its silencing produced a significantly reduction of B. cinerea development, resulting in a reduction of the fungal biomass. These results demonstrated the potential of the SIGS strategy for the control of B. cinerea and mrr1-dsRNA as a new molecule with fungicide activity that could be included into the several strategies carried out for sustainable plant protection control programs in the field.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Inflammatory fibro-epithelial hyperplasia related to a fixed implant-supported prosthesis : a case report

The gingival overgrowth is a common finding in the clinical practice with a diverse etiology. There are no treatment guidelines defined for this oral lesions. These can provoke discomfort to the patient and often, can alter the function of the stomatologic system. This article presents a case report of a bilateral gingival overgrowth in a 68 years old woman wearing a fixed upper-arch implant-supported prosthesis placed five years ago. The clinical exam after removing the prosthesis showed an intense accumulation of plaque around the intermediate abutments associated to a mucosal enlargement with suppuration on touching the buccal area of the implant in position 1.5 and a probing depth of 8mm. The 2.4 and 2.5 implants also showed vestibular mucosal enlargement and a probing depth of 6mm. No changes were observed in the peri-implant bone level measured in the periapical radiographs. An incisional biopsy was made on second quadrant and sent for the histopathological study. The definitive diagnosis was inflammatory fibro-epithelial hyperplasia. No recurrence has been reported after a 6 month follow-up

Low abundance of NDUFV2 and NDUFS4 subunits of the hydrophilic complex I domain and VDAC1 predicts mammalian longevity

Mitochondrial reactive oxygen species (ROS) production, specifically at complex I (Cx I), has been widely suggested to be one of the determinants of species longevity. The present study follows a comparative approach to analyse complex I in heart tissue from 8 mammalian species with a longevity ranging from 3.5 to 46 years. Gene expression and protein content of selected Cx I subunits were analysed using droplet digital PCR (ddPCR) and western blot, respectively. Our results demonstrate: 1) the existence of species-specific differences in gene expression and protein content of Cx I in relation to longevity; 2) the achievement of a longevity phenotype is associated with low protein abundance of subunits NDUFV2 and NDUFS4 from the matrix hydrophilic domain of Cx I; and 3) long-lived mammals show also lower levels of VDAC (voltage-dependent anion channel) amount. These differences could be associated with the lower mitochondrial ROS production and slower aging rate of long-lived animals and, unexpectedly, with a low content of the mitochondrial permeability transition pore in these species

Intra-tumor heterogeneity in TP53 null high grade serous ovarian carcinoma progression

[Background]: High grade serous ovarian cancer is characterised by high initial response to chemotherapy but poor outcome in the long term due to acquired resistance. One of the main genetic features of this disease is TP53 mutation. The majority of TP53 mutated tumors harbor missense mutations in this gene, correlated with p53 accumulation. TP53 null tumors constitute a specific subgroup characterised by nonsense, frameshift or splice-site mutations associated to complete absence of p53 expression. Different studies show that this kind of tumors may have a worse prognosis than other TP53 mutated HGSC. [Methods]: In this study, we sought to characterise the intra-tumor heterogeneity of a TP53 null HGSC consisting of six primary tumor samples, two intra-pelvic and four extra-pelvic recurrences using exome sequencing and comparative genome hybridisation. [Results]: Significant heterogeneity was found among the different tumor samples, both at the mutational and copy number levels. Exome sequencing identified 102 variants, of which only 42 were common to all three samples; whereas 7 of the 18 copy number changes found by CGH analysis were presented in all samples. Sanger validation of 20 variants found by exome sequencing in additional regions of the primary tumor and the recurrence allowed us to establish a sequence of the tumor clonal evolution, identifying those populations that most likely gave rise to recurrences and genes potentially involved in this process, like GPNMB and TFDP1. Using functional annotation and network analysis, we identified those biological functions most significantly altered in this tumor. Remarkably, unexpected functions such as microtubule-based movement and lipid metabolism emerged as important for tumor development and progression, suggesting its potential interest as therapeutic targets. [Conclusions]: Altogether, our results shed light on the clonal evolution of the distinct tumor regions identifying the most aggressive subpopulations and at least some of the genes that may be implicated in its progression and recurrence, and highlights the importance of considering intra-tumor heterogeneity when carrying out genetic and genomic studies, especially when these are aimed to diagnostic procedures or to uncover possible therapeutic strategies.This work has been supported by grants from the AECC network-2012, Telemarató 2013, Instituto de Salud Carlos III (ISCIII) (PI13/00132 and RETIC-RD12/0036/0007), GEIS award 2013, and by the Community of Madrid (S2010/BMD-2303). AM is a predoctoral student supported by FPU fellowship (Spanish Education Ministry). PGS is founded by postdoc contracts from the AECC Scientific Foundation.Peer Reviewe

Intratumor genetic heterogeneity and clonal evolution to decode endometrial cancer progression

Endometrial cancer; Clonal evolution; MutationCáncer endometrial; Evolución clonal; MutaciónCàncer d'endometri; Evolució clonal; MutacióAnalyzing different tumor regions by next generation sequencing allows the assessment of intratumor genetic heterogeneity (ITGH), a phenomenon that has been studied widely in some tumor types but has been less well explored in endometrial carcinoma (EC). In this study, we sought to characterize the spatial and temporal heterogeneity of 9 different ECs using whole-exome sequencing, and by performing targeted sequencing validation of the 42 primary tumor regions and 30 metastatic samples analyzed. In addition, copy number alterations of serous carcinomas were assessed by comparative genomic hybridization arrays. From the somatic mutations, identified by whole-exome sequencing, 532 were validated by targeted sequencing. Based on these data, the phylogenetic tree reconstructed for each case allowed us to establish the tumors’ evolution and correlate this to tumor progression, prognosis, and the presence of recurrent disease. Moreover, we studied the genetic landscape of an ambiguous EC and the molecular profile obtained was used to guide the selection of a potential personalized therapy for this patient, which was subsequently validated by preclinical testing in patient-derived xenograft models. Overall, our study reveals the impact of analyzing different tumor regions to decipher the ITGH in ECs, which could help make the best treatment decision.We thank all those at the Translational Research Laboratory of the MD Anderson Cancer Center Madrid for their invaluable help with this study. Tissue samples were obtained with the support of the MD Anderson Foundation Biobank (record number B.0000745, ISCIII National Biobank Record), the “Xarxa Catalana de Bancs de Tumors” and “Plataforma de Biobancos” ISCIII (PT13/0010/0014, B.000609). This study has been supported by the Spanish Ministry of Economy and Innovation (PID2019-104644RB-I00 (GMB), the Instituto de Salud Carlos III (ISCIII, CIBERONC, CB16/12/00295 - GMB-, CB16/12/00328 -EC, AGM- and CB16/12/00231 -XMG- [all partly supported by FEDER funds]) and by the AECC Scientific Foundation (FC_AECC PROYE19036MOR -GMB- and Coordinated groups 2018 -XMG, AGM, GMB-). SO is funded by an AECC-postdoctoral grant (2020). JSR-F and BW are funded in part by the Breast Cancer Research Foundation and in part by the NIH/NCI P50 CA247749 01 grant. Research reported in this publication was supported in part by a Cancer Center Support Grant of the NIH/NCI (Grant No. P30CA008748; MSK). We thank the Eurofins Megalab laboratory for helping us to perform the analysis of DNA HPV detection