25 research outputs found
Tumour mutations in long noncoding RNAs enhance cell fitness
Long noncoding RNAs (lncRNAs) are linked to cancer via pathogenic changes
in their expression levels. Yet, it remains unclear whether lncRNAs can also
impact tumour cell fitness via function-altering somatic “driver” mutations. To
search for such driver-lncRNAs, we here perform a genome-wide analysis of
fitness-altering single nucleotide variants (SNVs) across a cohort of 2583 primary
and 3527 metastatic tumours. The resulting 54 mutated and positivelyselected
lncRNAs are significantly enriched for previously-reported cancer
genes and a range of clinical and genomic features. A number of these lncRNAs
promote tumour cell proliferation when overexpressed in in vitro models. Our
results also highlight a dense SNV hotspot in the widely-studied NEAT1 oncogene.
To directly evaluate the functional significance of NEAT1 SNVs, we use in
cellulo mutagenesis to introduce tumour-like mutations in the gene and
observe a significant and reproducible increase in cell fitness, both in vitro and
in a mouse model. Mechanistic studies reveal that SNVs remodel the NEAT1
ribonucleoprotein and boost subnuclear paraspeckles. In summary, this work
demonstrates the utility of driver analysis for mapping cancer-promoting
lncRNAs, and provides experimental evidence that somatic mutations can act
through lncRNAs to enhance pathological cancer cell fitness.Swiss National Science Foundation through the National Centre of Competence in Research (NCCR) "RNA amp; Disease" 51NF40-182880The elements of long noncoding RNA function 31003A_182337Sinergia project "Regenerative strategies for heart disease via targeting the long noncoding transcriptome" 173738Medical Faculty of the University and University Hospital of BernHelmut Horten Stiftung, Swiss Cancer Research Foundation 4534-08-2018Science Foundation Ireland 18/FRL/6194
18/CRT/621
Deportes en los Juegos Olímpicos: Su reflejo en los medios de comunicación y buscadores
Los Juegos Olímpicos modernos están cambiando, y lo hacen con los tiempos y nuevas generaciones. Esto ha supuesto también un importante papel de los medios de comunicación, cuya repercusión afecta de manera activa a incentivar las opciones de que una disciplina se incluya en el programa olímpico por parte del Comité
Olímpico Internacional.
En este trabajo resaltaremos la importancia que juegan los medios de comunicación una vez las distintas disciplinas y deportes son aceptadas para ser olímpicas en las próximas citas de Tokio 2020 y París 2024. Lo haremos con una
perspectiva popular, ya que elegiremos deportes que tienen un pasado en las calles, barrios o playas de diversas partes del mundo. Para ello analizando las noticias editadas en los medios de comunicación y las búsquedas en los distintos motores de
internet llegaremos a las conclusiones sobre si afectan y de qué manera a la popularidad de los mismos.
Este trabajo pone de manifiesto la relevancia de los medios de comunicación y su interpolación a los lectores y consumidores de prensa no solo deportiva sino
también generalista y el impacto que esta tiene en el interés de los mismos a la hora de buscar e indagar de manera proactiva en internet.The Olympic Games are changing, and it comes with the times and new
generations. Mass Medias have had an important role in those changes and how they
could affect to th
e sports, as they could even be key for the decision taken by the CIO
to include them into the Olympic program for the next Olympic Games.
The following work tries to highlight the importance of the media once the
different disciplines and sports were ac
cepted to be part of the Olympic program for the
nexts Olympic Games in Tokyo 2021 and Paris 2024. Also we’ll use a popular
perspective, using sports that were born in the streets, beaches or neighborhoods from cities around the world. To analyze this, we
took some of the most important
newspapers in Spain and checked the number of news published since the CIO
announced they will be an Olympic Sport and also will check the different searching
volumes for those keywords to see how this affects the searches o
n Google.
Importance of the mass media and its interpolation to readers and consumers,
no only on specialized press but also the generic one. We show the impact of them on
the search volume on the Internet.Universidad de Sevilla. Grado en Periodism
Supplementing nursery pig feed with seaweed extracts increases final body weight of pigs
Probiotics have been used as an alternative method to antibiotic treatment. In this study, we evaluated the effect of seaweed extracts, commercially named OceanFeed Swine® (OFS), on body weight, average daily weight gain (ADG) and feed efficiency (FE) in nursery and fattening pigs of terminal lines. The study was conducted with 1809 piglets from weaning age until the end of the fattening phase on a commercial farm, divided into two groups (control and OFS; 905 and 904 pigs, respectively). Three replicates were made during the 6 weeks of production. Food was elaborated in the same productive unit and was given to the animals in the form of flour. OceanFeed Swine® (5 g/kg of diet) was added to the diet of the OFS groups during the nursery period, from 21 to 55 days of age. The faecal microbial population was evaluated at 35 and 60 days of life. In this paper, we observed a positive effect of administering OceanFeed Swine® on the production parameters analysed in the fattening phase, showing an improvement of 26 g in ADG and FE of 0.07. It can be concluded that the use of OceanFeed Swine® in the diets of the piglets, aged between 21 to 55 days old, improves FE (Control 2.36, OFS 2.29) and ADG (control 0.798 kg, OFS 0.824 kg). Moreover, its use exhibited an increase in the slaughter weight, a reduction of E coli CFU, and an increase in Lactobacillus sp. in fattening animals. All these improvements have a major positive economic impact per pig at time of slaughter
The value of desmosomal plaque-related markers to distinguish squamous cell carcinoma and adenocarcinoma of the lung
An antibody panel is needed to definitively differentiate between adenocarcinoma (AC)
and squamous cell carcinoma (SCC) in order to meet more stringent requirements for the histologic
classification of lung cancers. Staining of desmosomal plaque-related proteins may be useful in the
diagnosis of lung SCC. The specificity for SCC of membrane staining for PKP1, KRT15, and DSG3 was 97.4%, 94.6%,
and 100%, respectively, and it was 100% when the markers were used together and in combination
with the conventional markers (AUCs of 0.7619 for Panel 1 SCC, 0.7375 for Panel 2 SCC, 0.8552 for
Panel 1 AC, and 0.8088 for Panel 2 AC). In a stepwise multivariate logistic regression model, the combination
of CK5/6, p63, and PKP1 in membrane was the optimal panel to differentiate between SCC
and AC, with a percentage correct classification of 96.2% overall (94.6% of ACs and 97.6% of SCCs).
PKP1 and DSG3 are related to the prognosis. PKP1, KRT15, and DSG3 are highly specific for SCC, but they were more useful to differentiate
between SCC and AC when used together and in combination with conventional markers.
PKP1 and DSG3 expressions may have prognostic value.MEFV was supported by PAIDI programme, Group BIO309, Junta de
Andalucía
Multi‑omic alterations of the SWI/SNF complex define a clinical subgroup in lung adenocarcinoma
PPM's lab is funded by the Ministry of Economy of Spain (SAF2015-67919-R), Junta de Andalucia (P20-00688, PI-0135-2020, PIGE-0213-2020, PIGE-04402019, PI-0245-2017), University of Granada (B-CTS-480-UGR20), International Association for the Study of Lung Cancer (IASLC), and Spanish Association for Cancer Research (LAB-AECC-2018). PP is supported by a PhD "La Caixa Foundation"LCF/BQ/DE15/10360019 Fellowship. AA is supported by an FPU17/00067 fellowship. IFC was supported by a PhD FPI-fellowship (BES-2013-064596). DJG was supported by a "Fundacion Benefica Anticancer Santa Candida y San Francisco Javier"predoctoral fellowship. MSBC and CC's work is supported by the project DPI2017-84439-R Ministry of Economy of Spain and FEDER and by the fellowship "Beca de Iniciacion a la Investigacion del Plan Propio de Investigacion 2019" by University of Granada. MSBC is supported by an FPU19/00576 predoctoral fellowship. CNIO Proteomics Unit is a member of Proteored PRB3 and is supported by grant PT17/0019, of the PE I + D + i 2013-2016, funded by ISCIII and ERDF.SWI/SNF complexes are major targets of mutations in cancer. Here, we combined multiple “-omics” methods to assess
SWI/SNF composition and aberrations in LUAD. Mutations in lung SWI/SNF subunits were highly recurrent in our
LUAD cohort (41.4%), and over 70% of the mutations were predicted to have functional impact. Furthermore, SWI/
SNF expression in LUAD suffered an overall repression that could not be explained exclusively by genetic alterations.
Finally, SWI/SNF mutations were associated with poorer overall survival in TCGA-LUAD. We propose SWI/SNF-mutant
LUAD as a separate clinical subgroup with practical implications.Spanish Government SAF2015-67919-R
DPI2017-84439-RJunta de Andalucia P20-00688
PI-0135-2020
PIGE-0213-2020
PIGE-0440-2019
PI-0245-2017University of Granada B-CTS-480-UGR20International Association for the Study of Lung Cancer (IASLC)Spanish Association for Cancer Research LAB-AECC-2018La Caixa Foundation LCF/BQ/DE15/10360019PhD FPI-fellowship BES-2013-064596"Fundacion Benefica Anticancer Santa Candida y San Francisco Javier" predoctoral fellowshipEuropean Commissionfellowship "Beca de Iniciacion a la Investigacion del Plan Propio de Investigacion 2019" by University of Granada
Instituto de Salud Carlos III PT17/0019European Commission PT17/0019
FPU17/00067
FPU19/0057
ROS-MATLAB HMI for industrial robot teleoperation
[Resumen] ROS (Robot Operating System) es un framework para el desarrollo de sistemas robóticos de código abierto ampliamente utilizado en la industria y la investigación. Los sistemas robóticos asistidos requieren una comunicación fluida y fiable entre ROS y el controlador externo, a fin de lograr un control y supervisión eficaz del mismo. En este artículo, se presenta el procedimiento necesario para establecer la comunicación a través de una red local entre un robot móvil que implementa ROS y un dispositivo remoto desde el que enviar y recibir información del robot. Además, se propone un ejemplo de HMI (Interfaz Hombre-Máquina) desarrollado en MATLAB, y que puede ser instalada en equipos Windows, Linux o MacOSX, para la teleoperación de un robot en un entorno industrial. La comunicación bidireccional en tiempo real, las capacidades de procesamiento de datos y su versatilidad la convierten en una herramienta completa para la gestión de datos robóticos en entornos industriales.[Abstract] ROS (Robot Operating System) is an open-source robotics development framework widely used in industry and research. Human assisted robotic systems require of robust and constant communication between ROS and the remote controller in order to achieve effective control and monitoring of the system. In this paper, we propose a methodology to connect a mobile ROS with a remote device, using a wireless local network. This device serves as link between the robot and the rest of the system. Additionally, an HMI (Human-Machine Interface) developed using MATLAB is presented. The HMI can be implemented in Windows, Linux or MacOSX computers and provides teleoperation capabilities for a robot in an industrial environment. Real-time bidirectional communication, data processing capabilities and intrinsic versatility make this HMI a robust tool for robotic data management in industrial applications.Ministerio Ciencia e Innovación; PLEC2021-007819Comunidad de Madrid; S2018/NMT-433
Comprehensive Analysis of SWI/SNF Inactivation in Lung Adenocarcinoma Cell Models
Simple Summary: Mammalian SWI/SNF complexes regulate gene expression by reorganizing the
way DNA is packaged into chromatin. SWI/SNF subunits are recurrently altered in tumors at multiple
levels, including DNA mutations as well as alteration of the levels of RNA and protein. Cancer cell
lines are often used to study SWI/SNF function, but their patterns of SWI/SNF alterations can be
complex. Here, we present a comprehensive characterization of DNA mutations and RNA and protein
expression of SWI/SNF members in 38 lung adenocarcinoma (LUAD) cell lines. We show that over
85% of our cell lines harbored at least one alteration in one SWI/SNF subunit. In addition, over 75% of
our cell lines lacked expression of at least one SWI/SNF subunit at the protein level. Our catalog will
help researchers choose an appropriate cell line model to study SWI/SNF function in LUAD.
Abstract: Mammalian SWI/SNF (SWitch/Sucrose Non-Fermentable) complexes are ATP-dependent
chromatin remodelers whose subunits have emerged among the most frequently mutated genes
in cancer. Studying SWI/SNF function in cancer cell line models has unveiled vulnerabilities
in SWI/SNF-mutant tumors that can lead to the discovery of new therapeutic drugs. However,
choosing an appropriate cancer cell line model for SWI/SNF functional studies can be challenging
because SWI/SNF subunits are frequently altered in cancer by various mechanisms, including genetic
alterations and post-transcriptional mechanisms. In this work, we combined genomic, transcriptomic,
and proteomic approaches to study the mutational status and the expression levels of the SWI/SNF subunits in a panel of 38 lung adenocarcinoma (LUAD) cell lines. We found that the SWI/SNF
complex was mutated in more than 76% of our LUAD cell lines and there was a high variability in the
expression of the di erent SWI/SNF subunits. These results underline the importance of the SWI/SNF
complex as a tumor suppressor in LUAD and the di culties in defining altered and unaltered cell
models for the SWI/SNF complex. These findings will assist researchers in choosing the most suitable
cellular models for their studies of SWI/SNF to bring all of its potential to the development of novel
therapeutic applications.Ministry of Economy of Spain
SAF2015-67919-RJunta de Andalucía
CS2016-3
P12-BIO1655
PIGE-0440-2019
Pl-0245-2017
PI-0135-2020University of Granada
PPJIA2019-0
B-CTS-126-UGR18International Association for the Study of Lung Cancer (IASLC)Spanish Association for Cancer Research (LAB-AECC)PhD "La Caixa Foundation"
LCF/BQ/DE15/10360019"Fundacion Benefica Anticancer Santa Candida y San Francisco Javier" predoctoral fellowshipEuropean Commission
837897Spanish Ministry of Education, Culture and Sports FPU fellowship
FPU17/00067
FPU17/01258
FPU18/03709PhD FPI-fellowship
BES-2013-064596Fundación Científica de la Asociación Española Contra el Cáncer
GCB14-2170Fundación Ramon ArecesInstituto de Salud Carlos III-Fondo de Investigación Sanitaria-Fondo Europeo de Desarrollo Regional `Una manera de hacer Europa' (FEDER)
PI19/0009
Estado del arte de la medición de la productividad y la eficiencia técnica en América Latina: Caso Nicaragua
In this article were presented the results of a literature review from textbooks and scientific papers that address theory of "productivity" and "technical efficiency" as two key economic indicators to determine the economic growth reflected a productive unit, an economic sector or a nation. The study found that countries with a higher contribution of technical progress to the change in productivity in the period of 50 years (1960-2010) were analyzed recorded the Argentina, Brazil, Colombia and Ecuador, with indexes of around 0.3%. Furthermore, it is evident that the 19 countries analyzed in this study showed a decreasing technical efficiency, which means that the contribution of TFP that efficiency was negative in all countries. In the case of Nicaragua it was noted that some authors TFP estimated between 0.016 and 0.018 of growth.Rev. iberoam. bioecon. cambio clim. Vol. 1 num 2, 2015, pág. 76-100 En este artículo se plasman los resultados de una revisión bibliográfica de textos y artículos científicos que abordan la teoría de la “Productividad” y de la “Eficiencia técnica” como dos magnitudes económicas claves para determinar el crecimiento económico de una unidad productiva, un sector económico o de una nación. En el estudio se encontró que los países donde se registró una mayor contribución del progreso técnico a la variación de la productividad en el período de 50 años (de 1960-2010) analizado fueron la Argentina, el Brasil, Colombia y el Ecuador, con índices de alrededor del 0,3%. Además, se evidencia que los 19 países analizados en este trabajo registraron una eficiencia técnica decreciente, que supone que el aporte de dicha eficiencia a la PTF fue negativo en todos los países. En el caso de Nicaragua se notó que la PTF algunos autores la estimaron entre un 0.08 y 0.016 de ritmo de crecimiento interanual.Rev. iberoam. bioecon. cambio clim. Vol. 1 num 2, 2015, pág. 76-10
SMARCA4 deficient tumours are vulnerable to KDM6A/UTX and KDM6B/JMJD3 blockade
The authors thank Isabel Bartolessis (Cancer Genetics Group) at IJC for technical assistance. This work was supported by the Spanish Ministry of Economy and CompetitivityMINECO (grant number SAF-2017-82186R, to M.S.-C., and grant PI19/01320 to A. Villanueva) and from the Fundacion Cientifica of the Asociacion Espanola Contra el Cancer (AECC) (grant number GCB14142170MONT) to M.S.-C. A. Villanueva is also funded by the Department of Health of the Generalitat de Catalunya (2014SGR364). O.A. R. received a Juan de la Cierva postdoctoral contract (grant No. IJCI-2016-28201, until November 2019) and an AECC research contract (INVES19045ROME from December 2019). A. Vilarrubi, P.L. and A.A. are supported by pre-doctoral contracts from the Spanish MINECO (FPI-fellowship: PRE2018-084624, BES-2015-072204 and FPU17/00067). M.S. was supported by a Rio Hortega contract from the Instituto de Salud Carlos III (CM17/00180). L.F. received a European Union Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie Actions grant agreement, number 799850.Despite the genetic inactivation of SMARCA4, a core component of the SWI/SNF-complex
commonly found in cancer, there are no therapies that effectively target SMARCA4-deficient
tumours. Here, we show that, unlike the cells with activated MYC oncogene, cells with
SMARCA4 inactivation are refractory to the histone deacetylase inhibitor, SAHA, leading to
the aberrant accumulation of H3K27me3. SMARCA4-mutant cells also show an impaired
transactivation and significantly reduced levels of the histone demethylases KDM6A/UTX
and KDM6B/JMJD3, and a strong dependency on these histone demethylases, so that its
inhibition compromises cell viability. Administering the KDM6 inhibitor GSK-J4 to mice
orthotopically implanted with SMARCA4-mutant lung cancer cells or primary small cell
carcinoma of the ovary, hypercalcaemic type (SCCOHT), had strong anti-tumour effects. In
this work we highlight the vulnerability of KDM6 inhibitors as a characteristic that could be
exploited for treating SMARCA4-mutant cancer patients.Spanish Ministry of Economy and Competitivity-MINECO SAF-2017-82186R
PI19/01320Fundacion Cientifica of the Asociacion Espanola Contra el Cancer (AECC) GCB14142170MONTDepartment of Health of the Generalitat de Catalunya 2014SGR364Juan de la Cierva postdoctoral contract IJCI-2016-28201AECC research contract INVES19045ROMESpanish MINECO PRE2018-084624
BES-2015-072204
FPU17/00067Instituto de Salud Carlos III
European Commission CM17/00180European Union Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie Actions grant agreement 79985
Bioinformatic approaches for the discovery of non-coding alterations in cancer
Introduction. Cancer is one of the main causes of premature death
worldwide. Cancers arise when cell genomes accumulate driver mutations,
which are mutations that improve cell fitness. Driver mutations are a minority
among the thousands of mutations present in a typical cancer genome.
Although major efforts have been made to identify driver mutations in various
cancers, most of them have focused on the protein coding genome, which only
represents ~1.1% of the human genome. Part of the ~98.9% of the human
genome that does not code for protein contains functional elements, such as
regulatory DNA elements, intronic splice regions, untranslated regions of
protein coding genes, and non-coding RNA genes. Among non-coding RNAs,
microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) may
participate in the regulation of gene expression and their expression is often
altered in cancer. However, efforts to identify non-coding driver mutations
have been rare, and sample sizes in lung adenocarcinoma (LUAD) have been
low. In addition, the largest mutational study in diffuse large B-cell lymphoma
(DLBCL) to date omitted mutations in intronic splice regions.
Objectives. We aimed to computationally identify and characterize novel
driver mutations in non-coding DNA in in-house and external LUAD cohorts,
with special focus on miRNA genes, lncRNA genes, and intronic splice
regions. In addition, we aimed to identify and characterize previously missed
mutations in intronic splice sites in external DLBCL datasets.
Methods. We performed targeted sequencing of genomic DNA in an in-house
cohort of 70 LUAD primary tumors, 27 matched normal samples, and 37
LUAD cell lines. Our design included all human miRNA genes (n = 1881), as
well as exons of cancer-related lncRNA genes (n = 908) and protein coding
genes (n = 1307). We developed computational pipelines to identify highconfidence
somatic variants by combining multiple variant calling tools, and
we also applied them to external whole-genome sequencing data of LUAD
samples from The Cancer Genome Atlas (N = 59 tumor-normal pairs). In
addition, we applied state-of-the-art driver discovery tools to find putative
drivers in coding sequences, lncRNAs, miRNAs, intronic splice regions,
proximal promoters, and untranslated regions. We assessed the functional
relevance of the identified candidate drivers using external genomic, gene
expression, and clinical data as well as functional impact scores. Furthermore,
we developed a novel pipeline to annotate variants in a miRNA-centric
manner, identifying variants that affect seeds and those that disrupt or create
sequence motifs that mediate the processing of miRNA primary transcripts.
Finally, in DLBCL, we reanalyzed external datasets (combined N = 1711) to
identify previously missed recurrent mutations at intronic splice sites, we
analyzed the impact of the splice site mutations on RNA processing, and we
functionally characterized the most recurrent splice site mutation, which
affected CD79B.
Results. We successfully detected high-confidence somatic variants in all
analyzed datasets. However, driver discovery tools did not perform adequately
in our targeted sequencing cohorts of limited size, as one based on functional
impact predictions lacked sensitivity in non-coding regions whereas one based
on mutation clustering had a high false positive rate. Still, we identified three
candidate driver lncRNAs that accumulated mutations so that at least one
mutation had high predicted functional impact: TUSC7, SOX2-OT, and
ZEB2-AS1. However, the affected lncRNAs had very low expression in
external LUAD datasets. This, together with their mutational patterns and the
genomic characteristics of their loci, argued against an RNA sequencedependent
effect of their mutations. In miRNAs, a mutation in the seed of
miR-133b was predicted to have high functional impact and to prevent it from
targeting the oncogene EGFR. In addition, we identified mutations that
disrupted or created processing motifs in miRNA primary transcripts, such as
mutations that disrupted mismatched GHG motifs in mir-7-1, mir-7-2, and
mir-139. In intronic splice regions, we found mutations that altered RNA
splicing in LUAD driver genes such as MET and RBM10. In promoters and
untranslated regions, we detected no high-confidence drivers. In DLBCL,
intronic splice site mutations recurrently affected cancer driver genes and
caused major RNA aberrations in cis. The most recurrent RNA alteration was
intron 4 retention in CD79B. The alteration was caused by recurrent
mutations at the fourth splice donor site of CD79B, and it was associated with
an increase in the number of B cell receptors in the cell surface and a
subsequent increase in oncogenic signaling.
Conclusions. Non-coding variants with high predicted functional impact
were rare in our LUAD datasets. In addition, it was unclear whether the
candidate driver non-coding RNAs in LUAD had RNA sequence-dependent
functions. Experimental work will be necessary to confirm whether the
candidate driver non-coding RNAs have biological activity in LUAD and
whether their activity is altered by the observed mutations. In DLBCL, intronic
splice site mutations are recurrent and they can cause major cancer-promoting
aberrations in driver genesIntroducción. El cáncer es una de las principales causas de muerte prematura
mundialmente. El cáncer se origina cuando los genomas celulares acumulan
mutaciones conductoras, que son mutaciones que confieren ventaja selectiva
a la célula. Las mutaciones conductoras son una minoría entre las miles de
mutaciones que contiene un genoma tumoral promedio. Aunque se han
llevado a cabo grandes esfuerzos para identificar mutaciones conductoras en
una gran variedad de cánceres, la mayoría de los esfuerzos se han centrado en
el genoma codificante de proteína, que tan solo supone ~1,1% del genoma
humano. Parte del ~98,9% del genoma humano no codificante de proteína
contiene elementos funcionales, tales como ADN regulador, regiones
intrónicas de corte y empalme, regiones no traducidas de genes codificantes
de proteína y genes de ARN no codificante. Entre los ARNs no codificantes,
los microARNs (miARNs) y los ARNs largos no codificantes (ARNlncs)
pueden participar en la regulación de la expresión génica y su expresión está a
menudo alterada en cáncer. Sin embargo, los esfuerzos para identificar
mutaciones conductoras en secuencias no codificantes han sido escasos, y los
tamaños de muestra para adenocarcinoma de pulmón (ADC) han sido bajos.
Además, el mayor estudio hasta la fecha en linfoma difuso de células B grandes
(LDCBG) omitió las mutaciones en regiones intrónicas de corte y empalme.
Objetivos. Nuestro objetivo principal fue analizar y caracterizar
computacionalmente nuevas mutaciones conductoras en secuencias no
codificantes en cohortes propias y externas de ADC, con especial interés en
miARNs, ARNlncs y regiones intrónicas de corte y empalme. Un objetivo
adicional fue identificar y caracterizar mutaciones previamente no descritas
en sitios intrónicos de corte y empalme en datos externos de LDCBG.
Métodos. Realizamos secuenciación de DNA genómico dirigida a todos los
genes de miARNs humanos (n = 1881), así como a exones de genes de
ARNlncs relacionados con cáncer (n = 908) y de genes codificantes de proteína
relacionados con cáncer (n = 1307) en una cohorte propia de 70 tumores
primarios de LUAD, 27 muestras normales pareadas y 37 líneas celulares de
LUAD. Desarrollamos métodos computacionales para identificar variantes
somáticas con alta confianza mediante la combinación de múltiples
herramientas. Además, aplicamos dichos métodos para analizar datos de
secuenciación de genoma completo de muestras de ADC de The Cancer
Genome Atlas (N = 59 parejas tumor-normal). Asimismo, aplicamos
herramientas de descubrimiento de mutaciones conductoras en secuencias
codificantes, ARNlncs, miARNs, regiones intrónicas de corte y empalme,
promotores proximales y regiones no traducidas. Determinamos la relevancia
funcional de los elementos candidatos a conductores utilizando datos externos
genómicos, transcriptómicos y clínicos. Además, desarrollamos una nueva
metodología para anotar variantes de una forma miARN-céntrica, pudiendo
identificar variantes que afectan a secuencias semilla y aquellas que crean o
destruyen motivos de secuencia que median el procesamiento de los
transcritos primarios de miARNs. Finalmente, en LDCBG, reanalizamos
conjuntos de datos externos (N combinada = 1711) para identificar
mutaciones recurrentes en sitios intrónicos de corte y empalme no detectadas
en estudios anteriores. Analizamos el impacto de las mutaciones halladas en
el procesamiento del ARN afectado, y caracterizamos funcionalmente la
mutación más recurrente, que afectaba a CD79B.
Resultados. Detectamos exitosamente variantes somáticas con alta confianza
en todos los conjuntos de datos analizados. Sin embargo, las herramientas de
descubrimiento de mutaciones conductoras no tuvieron un rendimiento
adecuado en nuestras cohortes de secuenciación dirigida de tamaño limitado:
una herramienta basada en predicciones de impacto funcional tuvo baja
sensibilidad en regiones no codificantes, mientras que otra basada en el
agrupamiento de mutaciones tuvo una tasa elevada de falsos positivos. No
obstante, identificamos tres ARNlncs candidatos a conductores que
acumulaban mutaciones tal que al menos una de ellas tenía un alto impacto
funcional predicho: TUSC7, SOX2-OT y ZEB2-AS1. Sin embargo, los ARNlncs
afectados tenían una expresión extremadamente baja en datos externos de
ADC. Esto, unido a sus patrones mutacionales y a las características genómicas
de sus loci, hizo improbable que el efecto de sus mutaciones fuese dependiente
de la secuencia de ARN. En miARNs, una mutación en la semilla de miR-133b
tenía un alto impacto funcional predicho, impidiendo la unión de miR-133b
al oncogén EGFR. Asimismo, identificamos mutaciones que destruían o creaban motivos de procesamiento en los transcritos primarios de miARNs,
destacando las mutaciones que afectaban a motivos GHG desapareados en
mir-7-1, mir-7-2 y mir-139. En regiones intrónicas de corte y empalme,
hallamos mutaciones que alteraban el corte y empalme de genes conductores
de ADC como MET y RBM10. En promotores y en regiones no traducidas, no
encontramos ninguna mutación conductora con alto nivel de confianza. En
LDCBG, las mutaciones en sitios intrónicos de corte y empalme afectaban
recurrentemente a genes conductores de la enfermedad y causaban grandes
aberraciones a nivel de ARN en cis. La aberración más recurrente a nivel de
ARN fue la retención del intrón 4 de CD79B. La alteración estaba causada por
mutaciones recurrentes en el cuarto sitio intrónico donador de corte y
empalme de CD79B, y estaba asociada a un incremento en el número de
receptores de células B en la superficie celular y un consiguiente aumento en
la señalización oncogénica.
Conclusiones. Las variantes en ARNs no codificantes con impacto funcional
predicho elevado fueron infrecuentes en nuestros conjuntos de datos.
Asimismo, no se pudo determinar de manera concluyente que los ARNs no
codificantes candidatos a conductores de ADC tuviesen funciones
dependientes de la secuencia de ARN. Se requerirá trabajo experimental para
confirmar si los ARNs no codificantes candidatos a conductores tienen
actividad biológica en ADC y si las mutaciones detectadas en los mismos
alteran dicha actividad. En LDCBG, las mutaciones en sitios intrónicos de
corte y empalme son recurrentes y pueden causar grandes aberraciones en los
principales genes conductores de la enfermedad.Tesis Univ. Granada