Identificación de nuevos factores endocrinos-metabólicos y de la maquinaria de splicing con potencial diagnóstico, pronóstico y/o terapéutico en cáncer de próstata

Prostate cancer (PCa) is the tumor pathology with the highest incidence among men in developed countries and represents one of the main causes of cancer-related death in this group. Despite the advances achieved during the last years in PCa diagnosis and treatment, the diagnostic and therapeutic tools currently available are not sufficiently efficient. Specifically, prostate specific antigen (PSA) is the current “gold standard” used for PCa screening. Unfortunately, the PSA test has controversial specificity, since certain physiological conditions or non-tumor pathologies (e.g. benign prostatic hyperplasia, prostatitis) are also associated with high circulating PSA levels. Furthermore, the diagnostic capacity and clinical utility of the PSA test is especially low in patients with PSA levels between 3-10 ng/mL (the so-called grey zone). For these reasons, PSA test is associated with PCa overdiagnosis and unnecessary biopsies. Moreover, this invasive method required to subsequently confirm or discard the presence of PCa can cause side effects such as pain, bleeding and infections, which significantly impact patient’s quality of live. Therefore, there is an urgent need to identify new diagnostic tools that complement or replace PSA in clinical practice. Similarly, the therapeutic armamentarium in PCa is limited and clearly insufficient. Specifically, the pharmacological treatment used as the first option for the treatment of advanced-stage PCa or tumor recurrences consists of androgen deprivation (e.g. LHRH antagonists, abiraterone), due to the marked dependence of androgenic signaling of PCa cells. However, about 80% of patients treated with androgen deprivation therapy will develop resistance to this treatment, generating the so-called Castration Resistant PCa (CRPC), the most aggressive phenotype of this disease, which remains lethal nowadays. Therefore, the search for alternative therapeutic targets useful for the development of new, more efficient, treatments than those currently available continues to be one of the main objectives of the biomedical scientific community. PCa develops mainly in older patients, who suffer, with a high frequency, endocrinemetabolic pathologies (i.e. obesity, diabetes), which can influence the development and/or progression of PCa, increasing the complexity of this pathology. Specifically, obesity is associated with an increased risk of developing PCa, and with higher PCa aggressiveness. In this context, it has been described that metformin and statins (drugs commonly used in patients with endocrine-metabolic pathologies such as obesity or diabetes) exert antitumor effects in numerous tumor types, both in vitro and in vivo. However, although the combination of both drugs has been shown to exert additive antitumor actions in some cancer cell types, this possibility has not been systematically studied in the case of PCa. An important limitation in this field is that the information available regarding the exact molecular mechanisms underlying the pathophysiological interaction between obesity and PCa is very limited and fragmentary. In this sense, it is well-known that numerous components of different hormonal systems (i.e. androgen, ghrelin, etc.) are clearly dysregulated in PCa and obesity, which may constitute a source of biomarkers in these pathologies. The ghrelin system is a clear example in this context, since it is a pleiotropic hormonal system consisting of various ligands (e.g. Ghrelin, In1-ghrelin), enzymes (GOAT), and receptors (GHSR1a, GHSR1b). Specifically, In1-ghrelin (an alternative splicing variant derived as the result of the intron-1 retention) is overexpressed and plays an oncogenic role in PCa. Likewise, GOAT levels are higher in PCa tissues compared to nontumor prostate tissue, but the putative pathophysiological role of this enzyme in PCa is still unknown nowadays. Furthermore, it has been reported that circulating levels of both In1- ghrelin and GOAT are higher in PCa patients compared to healthy patients, suggesting that both elements could represent non-invasive diagnostic biomarkers of PCa. However, to date, it is unknown whether In1-ghrelin or GOAT can be detected and whether they are dysregulated in urine (which constitutes an enriched source of proteins derived from the prostate) in patients with PCa compared with patients without PCa. Finally, it is important to mention that the alteration of the splicing process has become a common hallmark of a large number of tumor pathologies, including PCa. Splicing is a physiological mechanism involved in the maturation of pre-mRNAs, which can generate different mature mRNAs from a single gene, thereby increasing the transcriptomic complexity of eukaryotic cells. However, increasing evidence demonstrates the existence of different splicing variants with oncogenic potential which are associated to the development, progression and/or resistance of pharmacological treatments of multiples tumor pathologies, including PCa. These dysregulations of the splicing process may be due to an alteration of the machinery that regulate this cellular process (spliceosome) and/or of the proteins that regulate this mechanism (splicing factors), suggesting that they could play a relevant role in the development and/or progression of PCa. Although certain studies have shown the specific dysregulation of some of these elements in PCa, the possible involvement of the alteration of this machinery and the putative implication in the development and/or progression of PCa has not been systematically explored yet. Based on the all the information mentioned above, the general aim of this Doctoral Thesis was to determine the potential role that certain splicing-related elements (spliceosome components and splicing factors) and metabolic modulators (endogenous elements and exogenous treatments) play and/or exert in the development, progression and/or aggressiveness of PCa, with the ultimate goal of discovering novel biomarkers (diagnostic and/or prognostic) and therapeutic tools that could improve the diagnosis, treatment and the clinical management of PCa patients. The first section of this Doctoral Thesis was focused on exploring the putative dysregulation and the clinical and functional implications of the splicing machinery in PCa. Results presented herein indicate that the splicing machinery is drastically dysregulated in PCa, including pivotal elements of this machinery such as SNRNP200, SRRM1, SRSF3, RBM22 or SF3B1. Among all the dysregulated spliceosome components and splicing factors, SNRNP200, SRRM1 and SRSF3 were selected initially to be further explored because they were associated with all the relevant aggressiveness features available in the cohort of patients used in this study. Mechanistically, silencing of SNRNP200, SRRM1 or SRSF3 inhibited oncogenic pathways and altered the splicing process of genes previously reported to be involved in tumor aggressiveness and/or CRPC development. Consistently, SNRNP200-, SRRM1- and SRSF3-silencing sensitized the CRPC cells to an androgen receptor (AR)-inhibitor (enzalutamide). Therefore, these data demonstrate that SNRNP200, SRRM1 and SRSF3 could represent attractive novel diagnostic and prognostic biomarkers as well as therapeutic targets for PCa and CRPC. This Doctoral Thesis was also focused on exploring the potential dysregulation and role of RBM22 in PCa, a splicing factor tightly involved in the activation of the spliceosome. Specifically, we found that RBM22 levels were lower (at mRNA and protein levels) in PCa tissues compared to non-tumor prostate tissues, and that these levels were inversely associated to key clinical parameters of aggressiveness in patients and in a preclinical mouse model. Moreover, we observed that the aggressiveness features of PCa cells were reduced in response to RBM22 overexpression in vitro and in vivo (i.e. reduced tumor growth, proliferation, migration, etc.) Mechanistically, results from high-throughput techniques (i.e. RNAseq, CLIP-seq and signaling array) performed in samples from in vitro and in vivo PCa models pointed out that the inhibition of MYC, MYCN and E2F, as well as a profound dysregulation of the splicing process could be the molecular mechanisms associated to the relevant findings previously observed after the RBM22 overexpression in PCa. Therefore, these results suggest that RBM22 plays a relevant antitumor role in PCa through the regulation of the splicing process and the reduction of MYC, MYCN and E2F levels/activity. In the case of SF3B1, a key component involved in the spliceosome assembly, we demonstrated that SF3B1 is overexpressed in PCa and that its levels were directly associated with important aggressiveness features of PCa. Moreover, the downregulation of SF3B1 and the inhibition of its activity (using the selective inhibitor pladienolide-B) significantly reduced functional aggressiveness features and increased the apoptotic rate of PCa cells. From a molecular perspective, SF3B1 inhibition altered PI3K/AKT and JNK oncogenic signaling pathways, down-regulated key oncogenic splicing variants (AR-v7 and In1-ghrelin), and dysregulated the expression of several elements involved in mRNA metabolism [splicing and non-sense mediated mRNA decay (NMD)]. Taken together, these results suggest that SF3B1 could represent a new prognostic biomarker and a therapeutic target in PCa, providing convincing evidence for the putative utility of pladienolide-B as novel therapeutic tool for the treatment of this devastating pathology. Once demonstrated the key role of several splicing-regulating factors in PCa, and since metformin has been found to exert antitumoral effects in various cancer types and that alter the expression of specific splicing factors in certain cell types, we next aimed to explore the potential involvement of the splicing dysregulation in the antitumor actions of metformin in PCa. Our data indicate that metformin produced a profound downregulation in the expression of different spliceosome components and splicing factors in PCa cells, being some of these changes (i.e. downregulation of SF3B1, SRRM1 and NOVA1) further validated in vivo using a PCa xenograft model. Moreover, the antiproliferative response of metformin treatment observed in PCa cells was completely blocked when the expression of SF3B1, SRRM1 or NOVA1 were silenced. Therefore, these results suggest that the dysregulation of some components of the splicing machinery, especially SF3B1, SRRM1 and NOVA1, might represent an additional molecular mechanism underlying the wellknown antitumor effects of metformin in PCa. The second section of this Doctoral Thesis was focused on the analysis of the antitumor actions of metformin, statins and their combination, as well as the underlying molecular mechanisms in PCa. Specifically, PCa patients treated with metformin and statins in combination were associated to more beneficial effects in key clinical parameters of aggressiveness compared to those receiving individual treatments. Consistently, although biguanides and statins significantly decreased the aggressiveness of PCa cells in vitro, these antitumor effects were more pronounced when combining both treatments. Mechanistically, we found that the treatment with the combination of metformin and simvastatin in vitro resulted in a hyper-inactivation of AR and mTOR, as well as in an upregulation of the expression of different cyclin-dependent kinase inhibitors (CKIs; CDKN1A, CDKN1B, CDKN2A and CDKN2D), being some of those changes further validated in samples from patients with PCa (i.e. upregulation of CDKN1B and CDKN2A). Altogether, this study demonstrates that metformin and simvastatin exert additive antitumor effects in PCa, thus suggesting that this combination might be a novel and attractive therapeutic approach to tackle PCa. Finally, in the third section of this Doctoral Thesis, we analysed specific components of the ghrelin system (specifically, GOAT enzyme and In1-ghrelin variant) to unveil their potential role as diagnostic, prognostic and/or therapeutic tools in PCa. These results demonstrated that GOAT could be detected in urine samples, wherein its levels outperformed the capacity of plasma PSA levels to diagnose PCa, especially clinically significant PCa (SigPCa; defined as Gleason score ≥7) in patients with PSA in the grey zone. Moreover, high urine GOAT levels were associated to increased risk of developing PCa and SigPCa, and were positively correlated to key clinical and molecular aggressiveness parameters in PCa patients. Furthermore, we demonstrated that GOAT overexpression increased PCa aggressiveness in vitro and in vivo, while its silencing and the blockade of its activity (using a specific GOAT inhibitor) significantly decreased the aggressiveness features of PCa cells in vitro. Therefore, these results demonstrated that GOAT enzyme may represent an additional non-invasive diagnostic biomarker, as well as a potential prognostic and therapeutic tool for PCa. Moreover, the analysis of In1-ghrelin in urine from patients with PSA in the grey zone indicated that its levels were able to distinguish between patients with and without PCa, were associated to higher risk of PCa, obesity and diabetes, and correlated with metabolic and aggressiveness parameters. In fact, we found that the capacity of urine In1-ghrelin levels to diagnose PCa was especially elevated when only obese patients were considered, suggesting its potential utility as a noninvasive diagnostic biomarker from a more personalized perspective. Altogether, the general conclusion derived from this Doctoral Thesis is that the alteration of the splicing process and the dysregulation of certain endocrine-metabolic systems could contribute to the development, progression and/or aggressiveness of PCa, representing a source of novel diagnostic and prognostic biomarkers, as well as therapeutic targets that could be exploited to improve the diagnosis of PCa patients, and to develop more effective prognostic and therapeutic tools to tackle this devastating pathology

Finding Multiple Solutions in Nonlinear Integer Programming with Algebraic Test-Sets

We explain how to compute all the solutions of a nonlinear integer problem using the algebraic test-sets associated to a suitable linear subproblem. These test-sets are obtained using Gröbner bases. The main advantage of this method, compared to other available alternatives, is its exactness within a quite good efficiency.Ministerio de Economía y Competitividad MTM2016-75024-PMinisterio de Economía y Competitividad MTM2016-74983-C2- 1-RJunta de Andalucía P12-FQM-269

Extending FuzAtAnalyzer to approach the management of classical negation

FuzAtAnalyzer was conceived as a Java framework which goes beyond of classical tools in formal concept analysis. Specifically, it successfully incorporated the management of uncertainty by means of methods and tools from the area of fuzzy formal concept analysis. One limitation of formal concept analysis is that they only consider the presence of properties in the objects (positive attributes) as much in fuzzy as in crisp case. In this paper, a first step in the incorporation of negations is presented. Our aim is the treatment of the absence of properties (negative attributes). Specifically, we extend the framework by including specific tools for mining knowledge combining crisp positive and negative attributes.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Cooperative Games on Antimatroids

AMS classification: 90D12;game theory;cooperative games;antimatroids

On the computation of Bernstein–Sato ideals

AbstractIn this paper we compare the approach of Briançon and Maisonobe for computing Bernstein–Sato ideals—based on computations in a Poincaré–Birkhoff–Witt algebra—with the readily available method of Oaku and Takayama. We show that it can deal with interesting examples that have proved intractable so far

Desórdenes de la locomoción, a dos velocidades de paso, en pacientes con fibromialgia

El objetivo del presente estudio es analizar si hay diferencias en las variables espaciotemporales de locomoción entre las pacientes afectadas con fibromialgia (FM) y un grupo de mujeres sanas, analizadas a velocidad confortable y rápida. Se evaluaron a 55 mujeres con FM y 44 sanas. El análisis de la locomoción se realizó con una plataforma de presiones (GaitRite system) y los sujetos realizaron una prueba a velocidad confortable y otra a velocidad rápida. Diferencias significativas (p<0.001) fueron encontradas entre el grupo control y el grupo FM en las variables de velocidad, longitud de paso, cadencia, fase de apoyo monopodal, fase de apoyo bipodal, fase de oscilación y fase de apoyo en las dos condiciones analizadas. No encontramos diferencias significativas entre las pacientes con FM caminando rápido y el grupo control a velocidad confortable, excepto en la cadencia. Los parámetros de locomoción en mujeres con FM están severamente afectados al compararlo con el grupo control, debido a diversos factores como la bradicinesia, la falta de actividad física, el sobrepeso, la fatiga y el dolor, que junto con una menor fuerza isométrica de las extremidades inferiores, pueden ser las responsables de estas alteraciones en la locomoción y en el descenso de la calidad de vida de estas pacientes.The aim of the present study was to determine if there are differences in spatiotemporal parameters among patients affected by fibromyalgia (FM) and healthy subjects, assess at comfortable and quickly speed of locomotion. We studied 55 women with FM and 44 controls. Gait analysis was performed using an instrumented walkway for measurement of the kinematic parameters of gait (GAITRite system), and the subjects was assessment in two conditions, comfortable and fast gait speed. Significant differences (p<0.001) between FM and control groups were found in velocity, stride length, cadence, single support ratio, double support ratio, stance phase ratio, and swing phase ratio at both analysis conditions. There aren’t statistical differences between FM group walking at quick speed and control group walking at comfortable speed, except in cadence variable. Gait parameters of women affected by FM were severely impaired when compared to those of healthy women. Different factors such as lack of physical activity, bradikinesia, overweight, fatigue, and pain together with a lower isometric force in the legs can be responsible for the alterations in gait and poorer life quality of women with FM.Peer Reviewe

Propiedades sorbentes de nanopartículas de carbono: nanotubos, nanoconos y nanocuernos

III Encuentro sobre Nanociencia y Nanotecnología de Investigadores y Tecnólogos Andaluce

Carbon nanocones/disks as new coating for solid-phase microextraction

II Encuentro sobre nanociencia y nanotecnología de investigadores y tecnólogos de la Universidad de Córodoba. NANOUC

Intervalo entre partos en el vacuno de carne extensivo de raza retinta

La eficacia reproductiva en el vacuno depende, entre otros parámetros, del intervalo entre partos. Esta variable está considerada una de las más importantes para la rentabilidad de la explotación en el caso del vacuno de carne en régimen extensivo. En este trabajo se estudia la influencia de distintos factores sobre el intervalo entre partos en la raza retinta mediante el análisis de 7356 partos de 1803 vacas pertenecientes al Núcleo de Control de Rendimientos de la Asociación de Ganaderos de esta raza. Nuestros resultados muestran que este parámetro se ve afectado principalmente por la ganadería, la campaña ganadera, época, sus interacciones y el tipo de parto, no existiendo relación significativa entre este período y el peso al destete de los terneros El valor medio obtenido para todo el intervalo analizado ha sido de 16,144 ± 0,086 meses y la heredabilidad estimada para este carácter fue de 0,1104 ± 0,08

Contribución al conocimiento del género Anthyllis L. (Fabaceae) en la Península Ibérica A. plumosa sp. nov

Se realiza un estudio morfológico, palinológico y cariológico de tres especies del género Anthyllis (sect. Oreanthyllis), describiéndose una nueva especie, A. plumosa E. Domínguez procedente de las arenas dolomíticas de la Sierra de Almijara (Málaga).In this paper a morphological, palynological and caryological study of three species of Anthyllis (sect. Oreanthyllis) is included. As a consecuence a new species A. plumosa E. Domínguez from the dolomitics sands of Sierra de Almijara (Málaga) is described