The Phosphorylation of Kv1.3: A Modulatory Mechanism for a Multifunctional Ion Channel

The voltage-gated potassium channel Kv1.3 plays a pivotal role in a myriad of biological processes, including cell proliferation, differentiation, and apoptosis. Kv1.3 undergoes fine-tuned regulation, and its altered expression or function correlates with tumorigenesis and cancer progression. Moreover, posttranslational modifications (PTMs), such as phosphorylation, have evolved as rapid switch-like moieties that tightly modulate channel activity. In addition, kinases are promising targets in anticancer therapies. The diverse serine/threonine and tyrosine kinases function on Kv1.3 and the effects of its phosphorylation vary depending on multiple factors. For instance, Kv1.3 regulatory subunits (KCNE4 and Kvβ) can be phosphorylated, increasing the complexity of channel modulation. Scaffold proteins allow the Kv1.3 channelosome and kinase to form protein complexes, thereby favoring the attachment of phosphate groups. This review compiles the network triggers and signaling pathways that culminate in Kv1.3 phosphorylation. Alterations to Kv1.3 expression and its phosphorylation are detailed, emphasizing the importance of this channel as an anticancer target. Overall, further research on Kv1.3 kinase-dependent effects should be addressed to develop effective antineoplastic drugs while minimizing side effects. This promising field encourages basic cancer research while inspiring new therapy development

A Novel NFIX-STAT6 Gene Fusion in Solitary Fibrous Tumor: A Case Report

Solitary fibrous tumor is a rare subtype of soft-tissue sarcoma with a wide spectrum of histopathological features and clinical behaviors, ranging from mildly to highly aggressive tumors. The defining genetic driver alteration is the gene fusion NAB2–STAT6, resulting from a paracentric inversion within chromosome 12q, and involving several different exons in each gene. STAT6 (signal transducer and activator of transcription 6) nuclear immunostaining and/or the identification of NAB2–STAT6 gene fusion is required for the diagnostic confirmation of solitary fibrous tumor. In the present study, a new gene fusion consisting of Nuclear Factor I X (NFIX), mapping to 19p13.2 and STAT6, mapping to 12q13.3 was identified by targeted RNA-Seq in a 74-year-old female patient diagnosed with a deep-seated solitary fibrous tumor in the pelvis. Histopathologically, the neoplasm did not display nuclear pleomorphism or tumor necrosis and had a low proliferative index. A total of 378 unique reads spanning the NFIXexon8–STAT6exon2 breakpoint with 55 different start sites were detected in the bioinformatic analysis, which represented 59.5% of the reads intersecting the genomic location on either side of the breakpoint. Targeted RNA-Seq results were validated by RT-PCR/ Sanger sequencing. The identification of a new gene fusion partner for STAT6 in solitary fibrous tumor opens intriguing new hypotheses to refine the role of STAT6 in the sarcomatogenesis of this entity

Pharmacogenetic Profiling in High-Risk Soft Tissue Sarcomas Treated with Neoadjuvant Chemotherapy

Neoadjuvant chemotherapy based on anthracyclines and ifosfamide for high-risk soft tissue sarcomas (STS) of the extremities and trunk is a controversial treatment option. There are sub-stantial interindividual differences in clinical outcomes in patients treated with neoadjuvant chem-otherapy. The aim of this study was to evaluate, as biomarkers, polymorphisms in genes encoding drug-metabolizing enzymes, drug transporters, or drug targets and their association with toxicity and survival in STS patients treated with neoadjuvant chemotherapy. We analysed variants in genes involved in anthracycline metabolism (ABCB1, ABCC2, NQO1, CBR3, and SLC22A16) and in ifosfamide catabolism (ALDH1A1) in 79 treated patients. Two genes showed significant association after adjusted multivariate analysis: ABCC2 and ALDH1A1. In patients treated with anthracyclines, ABCC2 rs3740066 was associated with risk of febrile neutropenia (p = 0.031), and with decreased overall survival (OS) (p = 0.024). ABCC2 rs2273697 was associated with recurrence-free survival (RFS) (p = 0.024). In patients treated with ifosfamide, ALDH1A1 rs3764435 was associated with RFS (p = 0.046). Our pharmacogenetic study shows for the first time that variants in genes regulating the metabolism of neoadjuvant chemotherapy may be helpful to predict toxicity and survival benefit in high-risk STS treated with neoadjuvant chemotherapy. Further validation studies are needed to es-tablish their clinical utility

Antibody conversion rates to SARS-CoV-2 in saliva from children attending summer schools in Barcelona, Spain.

Background: Surveillance tools to estimate viral transmission dynamics in young populations are essential to guide recommendations for school opening and management during viral epidemics. Ideally, sensitive techniques are required to detect low viral load exposures among asymptomatic children. We aimed to estimate SARS-CoV-2 infection rates in children and adult populations in a school-like environment during the initial COVID-19 pandemic waves using an antibody-based field-deployable and non-invasive approach. Methods: Saliva antibody conversion defined as ≥ 4-fold increase in IgM, IgA, and/or IgG levels to five SARS-CoV-2 antigens including spike and nucleocapsid constructs was evaluated in 1509 children and 396 adults by high-throughput Luminex assays in samples collected weekly in 22 summer schools and 2 pre-schools in 27 venues in Barcelona, Spain, from June 29th to July 31st, 2020. Results: Saliva antibody conversion between two visits over a 5-week period was 3.22% (49/1518) or 2.36% if accounting for potentially cross-reactive antibodies, six times higher than the cumulative infection rate (0.53%) assessed by weekly saliva RT-PCR screening. IgG conversion was higher in adults (2.94%, 11/374) than children (1.31%, 15/1144) (p=0.035), IgG and IgA levels moderately increased with age, and antibodies were higher in females. Most antibody converters increased both IgG and IgA antibodies but some augmented either IgG or IgA, with a faster decay over time for IgA than IgG. Nucleocapsid rather than spike was the main antigen target. Anti-spike antibodies were significantly higher in individuals not reporting symptoms than symptomatic individuals, suggesting a protective role against COVID-19. Conclusion: Saliva antibody profiling including three isotypes and multiplexing antigens is a useful and user-friendlier tool for screening pediatric populations to detect low viral load exposures among children, particularly while they are not vaccinated and vulnerable to highly contagious variants, and to recommend public health policies during pandemics

Shells and humans: molluscs and other coastal resources from the earliest human occupations at the Mesolithic shell midden of El Mazo (Asturias, Northern Spain)

Human populations exploited coastal areas with intensity during the Mesolithic in Atlantic Europe, resulting in the accumulation of large shell middens. Northern Spain is one of the most prolific regions, and especially the so-called Asturian area. Large accumulations of shellfish led some scholars to propose the existence of intensification in the exploitation of coastal resources in the region during the Mesolithic. In this paper, shell remains (molluscs, crustaceans and echinoderms) from stratigraphic units 114 and 115 (dated to the early Mesolithic c. 9 kys cal BP) at El Mazo cave (Asturias, northern Spain) were studied in order to establish resource exploitation patterns and environmental conditions. Species representation showed that limpets, top shells and sea urchins were preferentially exploited. One-millimetre mesh screens were crucial in establishing an accurate minimum number of individuals for sea urchins and to determine their importance in exploitation patterns. Environmental conditions deduced from shell assemblages indicated that temperate conditions prevailed at the time of the occupation and the morphology of the coastline was similar to today (rocky exposed shores). Information recovered relating to species representation, collection areas and shell biometry reflected some evidence of intensification (reduced shell size, collection in lower areas of exposed shores, no size selection in some units and species) in the exploitation of coastal resources through time. However, the results suggested the existence of changes in collection strategies and resource management, and periods of intense shell collection may have alternated with times of shell stock recovery throughout the Mesolithic.This research was performed as part of the project “The human response to the global climatic change in a littoral zone: the case of the transition to the Holocene in the Cantabrian coast (10,000–5000 cal BC) (HAR2010-22115-C02-01)” funded by the Spanish Ministry of Economy and Competitiveness. AGE was funded by the University of Cantabria through a predoctoral grant and IGZ was funded by the Spanish Ministry of Economy and Competitiveness through a Juan de la Cierva grant. We also would like to thank the University of Cantabria and the IIIPC for providing support, David Cuenca-Solana, Alejandro García Moreno and Lucia Agudo Pérez for their help. We also thank Jennifer Jones for correcting the English. Comments from two anonymous reviewers helped to improve the paper

New insights into the genetic etiology of Alzheimer's disease and related dementias

Characterization of the genetic landscape of Alzheimer's disease (AD) and related dementias (ADD) provides a unique opportunity for a better understanding of the associated pathophysiological processes. We performed a two-stage genome-wide association study totaling 111,326 clinically diagnosed/'proxy' AD cases and 677,663 controls. We found 75 risk loci, of which 42 were new at the time of analysis. Pathway enrichment analyses confirmed the involvement of amyloid/tau pathways and highlighted microglia implication. Gene prioritization in the new loci identified 31 genes that were suggestive of new genetically associated processes, including the tumor necrosis factor alpha pathway through the linear ubiquitin chain assembly complex. We also built a new genetic risk score associated with the risk of future AD/dementia or progression from mild cognitive impairment to AD/dementia. The improvement in prediction led to a 1.6- to 1.9-fold increase in AD risk from the lowest to the highest decile, in addition to effects of age and the APOE ε4 allele

Multiancestry analysis of the HLA locus in Alzheimer’s and Parkinson’s diseases uncovers a shared adaptive immune response mediated by HLA-DRB1*04 subtypes

Across multiancestry groups, we analyzed Human Leukocyte Antigen (HLA) associations in over 176,000 individuals with Parkinson’s disease (PD) and Alzheimer’s disease (AD) versus controls. We demonstrate that the two diseases share the same protective association at the HLA locus. HLA-specific fine-mapping showed that hierarchical protective effects of HLA-DRB1*04 subtypes best accounted for the association, strongest with HLA-DRB1*04:04 and HLA-DRB1*04:07, and intermediary with HLA-DRB1*04:01 and HLA-DRB1*04:03. The same signal was associated with decreased neurofibrillary tangles in postmortem brains and was associated with reduced tau levels in cerebrospinal fluid and to a lower extent with increased Aβ42. Protective HLA-DRB1*04 subtypes strongly bound the aggregation-prone tau PHF6 sequence, however only when acetylated at a lysine (K311), a common posttranslational modification central to tau aggregation. An HLA-DRB1*04-mediated adaptive immune response decreases PD and AD risks, potentially by acting against tau, offering the possibility of therapeutic avenues

Estudio de las mutaciones de los genes c-kit y pdgfra y la expresión de mirnas en una serie de tumores estromales del tracto gastrointestinal (gist) de la cspt. Correlación entre los hallazgos moleculares y la evolución clínica

Los tumores estromales del tracto gastrointestinal (GISTs) son las neoplasias estromales más frecuentes en esta localización. Expresan c-kit (CD117) y CD34 de forma característica. Los GIST presentan mutaciones (“gain-of-function”) en uno de los dos genes de receptores tirosinquinasa (c-KIT y PDGFRA), aunque en el 10% no se detectan mutaciones (GIST “wild-type”). En los GISTs, destaca la implicación de los microRNA-221, 222 y 494. Estos microRNA se unen al mRNA del gen c-KIT y su infraregulación implica un aumento de expresión proteica de c-kit. Los objetivos planteados en este estudio son los siguientes, aplicados a una serie de casos de GIST estudiados en Parc Taulí Sabadell Hospital Universitari: determinar la asociación entre parámetros clínico-patológicos, inmunohistoquímicos e índice de proliferación celular Ki-67; determinar las mutaciones de los genes c-KIT y PDGFRA y niveles de expresión de miRNA-221, 222 y 494 y correlacionar con los grupos de riesgo pronóstico (criterios AFIP-Miettinen 2006), parámetros clínico-patológicos e inmunohistoquímicos y progresión tumoral; establecer un patrón molecular diferencial entre los diferentes grupos de riesgo y aplicar y determinar el “nomograma” del SKCC para predecir la probabilidad de supervivencia libre de recurrencia a los 2 y 5 años y comparar con las probabilidades observadas. Las conclusiones obtenidas en nuestro estudio son las siguientes: La presencia de necrosis, positividad de intensidad marcada (3+) para c-kit, positividad para c-kit con patrón de Golgi, expresión elevada de Ki-67 y elevada tendencia a presentar metástasis, se asocian con más frecuencia a GISTs del grupo de alto riego. Los GISTs con morfología fusocelular y mixta se asocian a peor comportamiento biológico que los GISTs con morfología epitelioide. La mutación en el exón 11 se asocia con más frecuencia a pacientes de edad más avanzada, mujeres, localización en intestino delgado, morfología fusocelular y positividad con intensidad marcada (3+) para c-kit. Los tipos de mutación deleción/indel tienen más tendencia a distribuirse en GISTs localizados en intestino delgado, de alto riesgo y muestran menor supervivencia global que los GISTs con mutaciones puntuales. La mutación en el exón 9 se asocia con más frecuencia a GISTs localizados en intestino delgado, de morfología de tipo mixto y con valores de expresión de Ki-67 elevados. Los casos con mutación en el exón 9 de c-KIT y en el 18 de PDGFRA tienen más tendencia a expresar CD34 que el resto de mutaciones. La mutación en el exón 18 se asocia con más frecuencia a sexo masculino, localización gástrica, morfología epitelioide, matriz mixoide y grupo de muy bajo riesgo. Los GISTs wild-type se localizan con más frecuencia en estómago, presentan morfología fusocelular y vacuolización citoplasmática. En relación a los niveles de expresión de miRNAs: la mayoría de casos presentan niveles de expresión inferiores a 1. Diferencias de expresión en los distintos miRNAs se han asociado a parámetros clínico-patológicos y evolutivos como: localización, positividad para CD34, grupos de riesgo, tipo histológico, intensidad y patrón de c-kit y supervivencia específica. Todos los casos de GISTs wild-type de nuestra serie han mostrado niveles de expresión de miRNAs inferiores a 0,7. Aplicado el nomograma se observa que las predicciones tienden a cumplirse en los casos con alta probabilidad de supervivencia libre de recurrencia a los 2 años (80-100%), aunque no se demuestran diferencias estadísticamente significativas.Gastrointestinal stromal tumors (GISTs) are the most common stromal malignancies in this location. GISTs express c-kit (CD117) and CD34 characteristically. Mutations (gain-of-function) are detected in one of two receptor tyrosine kinase genes (c-KIT and PDGFRA), although in 10% of GISTs mutations are not detected (GIST "wild-type"). In these tumors, involvement of microRNA-221, 222 and 494 has been described. These miRNAs bind to mRNA of c-KIT gene and its downregulation implies increased expression of c-kit protein. The aims of this study, applied in a series of GIST cases studied in Parc Taulí Sabadell Hospital Universitari, are: to determine the association between clinicopathological and immunohistochemical parameters and cell proliferation index (Ki-67); to determine the mutations of c-KIT and PDGFRA genes and expression levels of miRNA-221, 222 and 494 and correlate with prognostic risk groups (criteria AFIP-Miettinen 2006), clinicopathological and immunohistochemical parameters, and tumor progression; to establish a differential molecular signature between different risk groups; and to determine the "nomogram" of the SKCC for predicting the probability of recurrence-free survival at 2 and 5 years and compare with the observed probabilities. Conclusions obtained from our study are: Presence of necrosis, strong intensity (3+) for c-kit, c-kit-positive Golgi pattern, high expression of Ki-67 and high tendency to metastasize, are most often associated with GISTs of high-risk group than the other risk groups. GISTs with spindle and mixed cell morphology are associated with worse biological behaviour than GISTs with epithelioid morphology. Mutation in exon 11 is most often associated with older patients, women, small bowel localization, spindle cell morphology and strong intensity (3 +) for c-kit. Delection/indel types of mutation are more likely to be distributed in GISTs located in small bowel and high-risk group, and show lower overall survival that GISTs with punctual mutations. Mutation in exon 9 is more often associated with small bowel location, mixed morphology and high expression values of Ki-67. GIST with mutation in exon 9 of c-KIT and exon 18 of PDGFRA are more likely to express CD34 than other mutations. Mutation in exon 18 is more often associated with male gender, gastric location, epithelioid morphology, myxoid matrix and very low-risk group. The wild-type GISTs are found more often in stomach, with spindle cell morphology and cytoplasmic vacuolization. In relation with the expression levels of miRNAs, most cases present expression levels below 1. Differences in expression of the different miRNAs have been associated with some clinicopathological and biological behaviour parameters as: location, CD34 positivity, risk groups, histological type, intensity and pattern of c-kit and specific survival. All cases of wild-type GISTs in our series have shown miRNAs expression levels less than 0.7. When applying the nomogram, predictions and real values tend to coincide in cases with high probability of recurrence-free survival at 2 years (80-100%), although no statistically significant differences were found

Estudio de las mutaciones de los genes c-kit y pdgfra y la expresión de mirnas en una serie de tumores estromales del tracto gastrointestinal (gist) de la cspt. Correlación entre los hallazgos moleculares y la evolución clínica

Los tumores estromales del tracto gastrointestinal (GISTs) son las neoplasias estromales más frecuentes en esta localización. Expresan c-kit (CD117) y CD34 de forma característica. Los GIST presentan mutaciones ("gain-of-function") en uno de los dos genes de receptores tirosinquinasa (c-KIT y PDGFRA), aunque en el 10% no se detectan mutaciones (GIST "wild-type"). En los GISTs, destaca la implicación de los microRNA-221, 222 y 494. Estos microRNA se unen al mRNA del gen c-KIT y su infraregulación implica un aumento de expresión proteica de c-kit. Los objetivos planteados en este estudio son los siguientes, aplicados a una serie de casos de GIST estudiados en Parc Taulí Sabadell Hospital Universitari: determinar la asociación entre parámetros clínico-patológicos, inmunohistoquímicos e índice de proliferación celular Ki-67; determinar las mutaciones de los genes c-KIT y PDGFRA y niveles de expresión de miRNA-221, 222 y 494 y correlacionar con los grupos de riesgo pronóstico (criterios AFIP-Miettinen 2006), parámetros clínico-patológicos e inmunohistoquímicos y progresión tumoral; establecer un patrón molecular diferencial entre los diferentes grupos de riesgo y aplicar y determinar el "nomograma" del SKCC para predecir la probabilidad de supervivencia libre de recurrencia a los 2 y 5 años y comparar con las probabilidades observadas. Las conclusiones obtenidas en nuestro estudio son las siguientes: La presencia de necrosis, positividad de intensidad marcada (3+) para c-kit, positividad para c-kit con patrón de Golgi, expresión elevada de Ki-67 y elevada tendencia a presentar metástasis, se asocian con más frecuencia a GISTs del grupo de alto riego. Los GISTs con morfología fusocelular y mixta se asocian a peor comportamiento biológico que los GISTs con morfología epitelioide. La mutación en el exón 11 se asocia con más frecuencia a pacientes de edad más avanzada, mujeres, localización en intestino delgado, morfología fusocelular y positividad con intensidad marcada (3+) para c-kit. Los tipos de mutación deleción/indel tienen más tendencia a distribuirse en GISTs localizados en intestino delgado, de alto riesgo y muestran menor supervivencia global que los GISTs con mutaciones puntuales. La mutación en el exón 9 se asocia con más frecuencia a GISTs localizados en intestino delgado, de morfología de tipo mixto y con valores de expresión de Ki-67 elevados. Los casos con mutación en el exón 9 de c-KIT y en el 18 de PDGFRA tienen más tendencia a expresar CD34 que el resto de mutaciones. La mutación en el exón 18 se asocia con más frecuencia a sexo masculino, localización gástrica, morfología epitelioide, matriz mixoide y grupo de muy bajo riesgo. Los GISTs wild-type se localizan con más frecuencia en estómago, presentan morfología fusocelular y vacuolización citoplasmática. En relación a los niveles de expresión de miRNAs: la mayoría de casos presentan niveles de expresión inferiores a 1. Diferencias de expresión en los distintos miRNAs se han asociado a parámetros clínico-patológicos y evolutivos como: localización, positividad para CD34, grupos de riesgo, tipo histológico, intensidad y patrón de c-kit y supervivencia específica. Todos los casos de GISTs wild-type de nuestra serie han mostrado niveles de expresión de miRNAs inferiores a 0,7. Aplicado el nomograma se observa que las predicciones tienden a cumplirse en los casos con alta probabilidad de supervivencia libre de recurrencia a los 2 años (80-100%), aunque no se demuestran diferencias estadísticamente significativas.Gastrointestinal stromal tumors (GISTs) are the most common stromal malignancies in this location. GISTs express c-kit (CD117) and CD34 characteristically. Mutations (gain-of-function) are detected in one of two receptor tyrosine kinase genes (c-KIT and PDGFRA), although in 10% of GISTs mutations are not detected (GIST "wild-type"). In these tumors, involvement of microRNA-221, 222 and 494 has been described. These miRNAs bind to mRNA of c-KIT gene and its downregulation implies increased expression of c-kit protein. The aims of this study, applied in a series of GIST cases studied in Parc Taulí Sabadell Hospital Universitari, are: to determine the association between clinicopathological and immunohistochemical parameters and cell proliferation index (Ki-67); to determine the mutations of c-KIT and PDGFRA genes and expression levels of miRNA-221, 222 and 494 and correlate with prognostic risk groups (criteria AFIP-Miettinen 2006), clinicopathological and immunohistochemical parameters, and tumor progression; to establish a differential molecular signature between different risk groups; and to determine the "nomogram" of the SKCC for predicting the probability of recurrence-free survival at 2 and 5 years and compare with the observed probabilities. Conclusions obtained from our study are: Presence of necrosis, strong intensity (3+) for c-kit, c-kit-positive Golgi pattern, high expression of Ki-67 and high tendency to metastasize, are most often associated with GISTs of high-risk group than the other risk groups. GISTs with spindle and mixed cell morphology are associated with worse biological behaviour than GISTs with epithelioid morphology. Mutation in exon 11 is most often associated with older patients, women, small bowel localization, spindle cell morphology and strong intensity (3 +) for c-kit. Delection/indel types of mutation are more likely to be distributed in GISTs located in small bowel and high-risk group, and show lower overall survival that GISTs with punctual mutations. Mutation in exon 9 is more often associated with small bowel location, mixed morphology and high expression values of Ki-67. GIST with mutation in exon 9 of c-KIT and exon 18 of PDGFRA are more likely to express CD34 than other mutations. Mutation in exon 18 is more often associated with male gender, gastric location, epithelioid morphology, myxoid matrix and very low-risk group. The wild-type GISTs are found more often in stomach, with spindle cell morphology and cytoplasmic vacuolization. In relation with the expression levels of miRNAs, most cases present expression levels below 1. Differences in expression of the different miRNAs have been associated with some clinicopathological and biological behaviour parameters as: location, CD34 positivity, risk groups, histological type, intensity and pattern of c-kit and specific survival. All cases of wild-type GISTs in our series have shown miRNAs expression levels less than 0.7. When applying the nomogram, predictions and real values tend to coincide in cases with high probability of recurrence-free survival at 2 years (80-100%), although no statistically significant differences were found