Topographic analysis of pancreatic cancer by TMA and digital spatial profiling reveals biological complexity with potential therapeutic implications.

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal human malignancies. Tissue microarrays (TMA) are an established method of high throughput biomarker interrogation in tissues but may not capture histological features of cancer with potential biological relevance. Topographic TMAs (T-TMAs) representing pathophysiological hallmarks of cancer were constructed from representative, retrospective PDAC diagnostic material, including 72 individual core tissue samples. The T-TMA was interrogated with tissue hybridization-based experiments to confirm the accuracy of the topographic sampling, expression of pro-tumourigenic and immune mediators of cancer, totalling more than 750 individual biomarker analyses. A custom designed Next Generation Sequencing (NGS) panel and a spatial distribution-specific transcriptomic evaluation were also employed. The morphological choice of the pathophysiological hallmarks of cancer was confirmed by protein-specific expression. Quantitative analysis identified topography-specific patterns of expression in the IDO/TGF-ß axis; with a heterogeneous relationship of inflammation and desmoplasia across hallmark areas and a general but variable protein and gene expression of c-MET. NGS results highlighted underlying genetic heterogeneity within samples, which may have a confounding influence on the expression of a particular biomarker. T-TMAs, integrated with quantitative biomarker digital scoring, are useful tools to identify hallmark specific expression of biomarkers in pancreatic cancer.

Evaluation of deep oscillation therapy for the treatment of lumbar pain syndrome using motion capture systems: A systematic review.

Low back pain is a painful disorder that prevents normal mobilization, increases muscle tension and whose first-line treatment is usually non-steroidal anti-inflammatory drugs, together with non-invasive manual therapies, such as deep oscillation therapy. This systematic review aims to investigate and examine the scientific evidence of the effectiveness of deep oscillation therapy in reducing pain and clinical symptomatology in patients with low back pain, through the use of motion capture technology. To carry out this systematic review, the guidelines of the PRISMA guide were followed. A literature search was performed from 2013 to March 2022 in the PubMed, Elsevier, Science Director, Cochrane Library, and Springer Link databases to collect information on low back pain, deep oscillation, and motion capture. The risk of bias of the articles was assessed using the Cochrane risk of bias tool. Finally, they were included 16 articles and 5 clinical trials which met the eligibility criteria. These articles discussed the effectiveness of deep oscillation therapy in reducing pain, eliminating inflammation, and increasing lumbar range of motion, as well as analyzing the use of motion capture systems in the analysis, diagnosis, and evaluation of a patient with low back pain before, during and after medical treatment. There is no strong scientific evidence that demonstrates the high effectiveness of deep oscillation therapy in patients with low back pain, using motion capture systems. This review outlines the background for future research directed at the use of deep oscillation therapy as a treatment for other types of musculoskeletal injuries.

Classification of the Pathological Range of Motion in Low Back Pain Using Wearable Sensors and Machine Learning.

Low back pain (LBP) is a highly common musculoskeletal condition and the leading cause of work absenteeism. This project aims to develop a medical test to help healthcare professionals decide on and assign physical treatment for patients with nonspecific LBP. The design uses machine learning (ML) models based on the classification of motion capture (MoCap) data obtained from the range of motion (ROM) exercises among healthy and clinically diagnosed patients with LBP from Imbabura-Ecuador. The following seven ML algorithms were tested for evaluation and comparison: logistic regression, decision tree, random forest, support vector machine (SVM), k-nearest neighbor (KNN), multilayer perceptron (MLP), and gradient boosting algorithms. All ML techniques obtained an accuracy above 80%, and three models (SVM, random forest, and MLP) obtained an accuracy of >90%. SVM was found to be the best-performing algorithm. This article aims to improve the applicability of inertial MoCap in healthcare by making use of precise spatiotemporal measurements with a data-driven treatment approach to improve the quality of life of people with chronic LBP.

BackMov: Individualized Motion Capture-Based Test to Assess Low Back Pain Mobility Recovery after Treatment.

Low back pain (LBP) is a common issue that negatively affects a person's quality of life and imposes substantial healthcare expenses. In this study, we introduce the (Back-pain Movement) BackMov test, using inertial motion capture (MoCap) to assess lumbar movement changes in LBP patients. The test includes flexion-extension, rotation, and lateralization movements focused on the lumbar spine. To validate its reproducibility, we conducted a test-retest involving 37 healthy volunteers, yielding results to build a minimal detectable change (MDC) graph map that would allow us to see if changes in certain variables of LBP patients are significant in relation to their recovery. Subsequently, we evaluated its applicability by having 30 LBP patients perform the movement's test before and after treatment (15 received deep oscillation therapy; 15 underwent conventional therapy) and compared the outcomes with a specialist's evaluations. The test-retest results demonstrated high reproducibility, especially in variables such as range of motion, flexion and extension ranges, as well as velocities of lumbar movements, which stand as the more important variables that are correlated with LBP disability, thus changes in them may be important for patient recovery. Among the 30 patients, the specialist's evaluations were confirmed using a low-back-specific Short Form (SF)-36 Physical Functioning scale, and agreement was observed, in which all patients improved their well-being after both treatments. The results from the specialist analysis coincided with changes exceeding MDC values in the expected variables. In conclusion, the BackMov test offers sensitive variables for tracking mobility recovery from LBP, enabling objective assessments of improvement. This test has the potential to enhance decision-making and personalized patient monitoring in LBP management.

What makes a word a good representative of the category of "emotion"? The role of feelings and interoception.

The words we use to describe emotions vary in terms of prototypicality; that is, some of these words may be more representative of the semantic category of emotion than others (e.g., anger refers more clearly to an emotion than boredom). Based on a multicomponential conception of emotions, the aim of the present study was to examine the contribution of several variables to emotion prototypicality. Some of those variables are related to the distinct components of emotions: evaluation, action, body expression, internal body sensations (interoception), and feelings. Other variables are related to the concreteness/abstractness distinction: sensory experience, social interaction, thought, and morality. We collected ratings for these variables for a large set of words (1,286) which varied in emotion prototypicality. A regression analysis revealed that the variables that most contributed to emotion prototypicality were feelings and interoception. Furthermore, a factor analysis identified two underlying factors: socioemotional polarity and emotional experience. The scores of each word in both factors were used to create a two-dimensional space and a density plot which provides relevant information about the organization of emotion concepts in memory. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

KeepRunning: A MoCap-Based Rapid Test to Prevent Musculoskeletal Running Injuries.

The worldwide popularisation of running as a sport and recreational practice has led to a high rate of musculoskeletal injuries, usually caused by a lack of knowledge about the most suitable running technique for each runner. This running technique is determined by a runner's anthropometric body characteristics, dexterity and skill. Therefore, this study aims to develop a motion capture-based running analysis test on a treadmill called KeepRunning to obtain running patterns rapidly, which will aid coaches and clinicians in assessing changes in running technique considering changes in the study variables. Therefore, a review and proposal of the most representative events and variables of analysis in running was conducted to develop the KeepRunning test. Likewise, the minimal detectable change (MDC) in these variables was obtained using test-retest reliability to demonstrate the reproducibility and viability of the test, as well as the use of MDC as a threshold for future assessments. The test-retest consisted of 32 healthy volunteer athletes with a running training routine of at least 15 km per week repeating the test twice. In each test, clusters of markers were placed on the runners' body segments using elastic bands and the volunteers' movements were captured while running on a treadmill. In this study, reproducibility was defined by the intraclass correlation coefficient (ICC) and MDC, obtaining a mean value of ICC = 0.94 ± 0.05 for all variables and MDC = 2.73 ± 1.16° for the angular kinematic variables. The results obtained in the test-retest reveal that the reproducibility of the test was similar or better than that found in the literature. KeepRunning is a running analysis test that provides data from the involved body segments rapidly and easily interpretable. This data allows clinicians and coaches to objectively provide indications for runners to improve their running technique and avoid possible injury. The proposed test can be used in the future with inertial motion capture and other wearable technologies.

Spatial transcriptomics map of the embryonic mouse brain - a tool to explore neurogenesis.

The developing brain has a well-organized anatomical structure comprising different types of neural and non-neural cells. Stem cells, progenitors and newborn neurons tightly interact with their neighbouring cells and tissue microenvironment, and this intricate interplay ultimately shapes the output of neurogenesis. Given the relevance of spatial cues during brain development, we acknowledge the necessity for a spatial transcriptomics map accessible to the neurodevelopmental community. To fulfil this need, we generated spatially resolved RNA sequencing (RNAseq) data from embryonic day 13.5 mouse brain sections immunostained for mitotic active neural and vascular cells. Unsupervised clustering defined specific cell type populations of diverse lineages and differentiation states. Differential expression analysis revealed unique transcriptional signatures across specific brain areas, uncovering novel features inherent to particular anatomical domains. Finally, we integrated existing single-cell RNAseq datasets into our spatial transcriptomics map, adding tissue context to single-cell RNAseq data. In summary, we provide a valuable tool that enables the exploration and discovery of unforeseen molecular players involved in neurogenesis, particularly in the crosstalk between different cell types.

Evolutionary conservation of embryonic DNA methylome remodelling in distantly related teleost species.

Methylation of cytosines in the CG context (mCG) is the most abundant DNA modification in vertebrates that plays crucial roles in cellular differentiation and identity. After fertilization, DNA methylation patterns inherited from parental gametes are remodelled into a state compatible with embryogenesis. In mammals, this is achieved through the global erasure and re-establishment of DNA methylation patterns. However, in non-mammalian vertebrates like zebrafish, no global erasure has been observed. To investigate the evolutionary conservation and divergence of DNA methylation remodelling in teleosts, we generated base resolution DNA methylome datasets of developing medaka and medaka-zebrafish hybrid embryos. In contrast to previous reports, we show that medaka display comparable DNA methylome dynamics to zebrafish with high gametic mCG levels (sperm: ∼90%; egg: ∼75%), and adoption of a paternal-like methylome during early embryogenesis, with no signs of prior DNA methylation erasure. We also demonstrate that non-canonical DNA methylation (mCH) reprogramming at TGCT tandem repeats is a conserved feature of teleost embryogenesis. Lastly, we find remarkable evolutionary conservation of DNA methylation remodelling patterns in medaka-zebrafish hybrids, indicative of compatible DNA methylation maintenance machinery in far-related teleost species. Overall, these results suggest strong evolutionary conservation of DNA methylation remodelling pathways in teleosts, which is distinct from the global DNA methylome erasure and reestablishment observed in mammals.

The glycolytic enzymes glyceraldehyde-3-phosphate dehydrogenase and hexokinase interact with cell cycle proteins in maize.

Cyclin/cyclin-dependent kinase (CDK) heterodimers have multiple phosphorylation targets and may alter the activity of these targets. Proteins from different metabolic processes are among the phosphorylation targets, that is, enzymes of central carbon metabolism. This work explores the interaction of Cyc/CDK complex members with the glycolytic enzymes hexokinase 7 (HXK7) and glyceraldehyde-3-phosphate dehydrogenase (GAP). Both enzymes interacted steadily with CycD2;2, CycB2;1 and CDKA;1 but not with CDKB1;1. However, Cyc/CDKB1;1 complexes phosphorylated both enzymes, decreasing their activities. Treatment with a CDK-specific inhibitor (RO-3306) or with lambda phosphatase after kinase assay restored total HXK7 activity, but not GAP activity. In enzymatic assays, increasing concentrations of CDKB1;1, but not of CycD2;2, CycB2;1 or CycD2;2/CDKB1;1 complex, decreased GAP activity. Cell cycle regulators may modulate carbon channeling in glycolysis by two different mechanisms: Cyc/CDK-mediated phosphorylation of targets (e.g., HXK7; canonical mechanism) or by direct and transient interaction of the metabolic enzyme (e.g., GAP) with CDKB1;1 without a Cyc partner (alternative mechanism).

A Yap-dependent mechanoregulatory program sustains cell migration for embryo axis assembly.

The assembly of the embryo's primary axis is a fundamental landmark for the establishment of the vertebrate body plan. Although the morphogenetic movements directing cell convergence towards the midline have been described extensively, little is known on how gastrulating cells interpret mechanical cues. Yap proteins are well-known transcriptional mechanotransducers, yet their role in gastrulation remains elusive. Here we show that the double knockout of yap and its paralog yap1b in medaka results in an axis assembly failure, due to reduced displacement and migratory persistence in mutant cells. Accordingly, we identified genes involved in cytoskeletal organization and cell-ECM adhesion as potentially direct Yap targets. Dynamic analysis of live sensors and downstream targets reveal that Yap is acting in migratory cells, promoting cortical actin and focal adhesions recruitment. Our results indicate that Yap coordinates a mechanoregulatory program to sustain intracellular tension and maintain the directed cell migration for embryo axis development.

Temporal and clonal characterization of neural stem cell niche recruitment in the medaka neuromast.

Stem cell populations are defined by their capacity to self-renew and to generate differentiated progeny. These unique characteristics largely depend on the stem cell micro-environment, the so-called stem cell niche. Niches were identified for most adult stem cells studied so far, but we know surprisingly little about how somatic stem cells and their niche come together during organ formation. Using the neuromasts of teleost fish, we have previously reported that neural stem cells recruit their niche from neighboring epithelial cells, which go through a morphological and molecular transformation. Here, we tackle quantitative, temporal, and clonal aspects of niche formation in neuromasts by using 4D imaging in transgenic lines, and lineage analysis in mosaic fish. We show that niche recruitment happens in a defined temporal window during the formation of neuromasts in medaka, and after that, the niche is enlarged mainly by the proliferation of niche cells. Niche recruitment is a non-clonal process that feeds from diverse epithelial cells that do not display a preferential position along the circumference of the forming neuromast. Additionally, we cover niche formation and expansion in zebrafish to show that distant species show common features during organogenesis in the lateral line system. Overall, our findings shed light on the process of niche formation, fundamental for the maintenance of stem cells not only in medaka but also in many other multicellular organisms.

Molecular dynamics simulations reveal the impact of NUDT15 R139C and R139H variants in structural conformation and dynamics.

NUDT15, also known as MTH2, is a member of the NUDIX protein family that catalyzes the hydrolysis of nucleotides and deoxynucleotides, as well as thioguanine analogues. NUDT15 has been reported as a DNA sanitizer in humans, and more recent studies have shown that some genetic variants are related to a poor prognosis in neoplastic and immunologic diseases treated with thioguanine drugs. Despite this, the role of NUDT15 in physiology and molecular biology is quite unclear, as is the mechanism of action of this enzyme. The existence of clinically relevant variants has prompted the study of these enzymes, whose capacity to bind and hydrolyze thioguanine nucleotides is still poorly understood. By using a combination of biomolecular modeling techniques and molecular dynamics, we have studied the monomeric wild type NUDT15 as well as two important variants, R139C and R139H. Our findings reveal not only how nucleotide binding stabilizes the enzyme but also how two loops are responsible for keeping the enzyme in a packed, close conformation. Mutations in α2 helix affect a network of hydrophobic and π-interactions that enclose the active site. This knowledge contributes to the understanding of NUDT15 structural dynamics and will be valuable for the design of new chemical probes and drugs targeting this protein.Communicated by Ramaswamy H. Sarma.

Toxicity of Asciminib in Real Clinical Practice: Analysis of Side Effects and Cross-Toxicity with Tyrosine Kinase Inhibitors.

(1) Background: Despite the prognostic improvements achieved with tyrosine kinase inhibitors (TKIs) in chronic myeloid leukemia (CML), a minority of patients still fail TKIs. The recent introduction of asciminib may be a promising option in intolerant patients, as it is a first-in-class inhibitor with a more selective mechanism of action different from the ATP-competitive inhibition that occurs with TKIs. Therefore, our goal was to analyze toxicities shown with asciminib as well as to study cross-toxicity with previous TKIs. (2) Methods: An observational, multicenter, retrospective study was performed with data from 77 patients with CML with therapeutic failure to second-generation TKIs who received asciminib through a managed-access program (MAP) (3) Results: With a median follow-up of 13.7 months, 22 patients (28.5%) discontinued treatment: 32% (7/22) due to intolerance and 45% (10/22) due to resistance. Fifty-five percent of the patients reported adverse effects (AEs) with asciminib and eighteen percent grade 3-4. Most frequent AEs were: fatigue (18%), thrombocytopenia (17%), anemia (12%), and arthralgias (12%). None of the patients experienced cardiovascular events or occlusive arterial disease. Further, 26%, 25%, and 9% of patients required dose adjustment, temporary suspension, or definitive discontinuation of treatment, respectively. Toxicities under asciminib seemed lower than with prior TKIs for anemia, cardiovascular events, pleural/pericardial effusion, diarrhea, and edema. Cross-toxicity risk was statistically significant for thrombocytopenia, anemia, neutropenia, fatigue, vomiting, and pancreatitis. (4) Conclusion: Asciminib is a molecule with a good safety profile and with a low rate of AEs. However, despite its new mechanism of action, asciminib presents a risk of cross-toxicity with classical TKIs for some AEs.

Prepandemic viral community-acquired pneumonia: Diagnostic sensitivity and specificity of nasopharyngeal swabs and performance of clinical severity scores.

The objectives of this work were to assess the diagnostic sensitivity and specificity of nasopharyngeal (NP) swabs for viral community-acquired pneumonia (CAP) and the performance of pneumonia severity index (PSI) and CURB-65 severity scores in the viral CAP in adults. A prospective observational cohort study of consecutive 341 hospitalized adults with CAP was performed between January 2018 and March 2020. Demographics, comorbidities, symptoms/signs, analytical data, severity scores, antimicrobials, and outcomes were recorded. Blood, NP swabs, sputum, and urine samples were collected at admission and assayed by multiplex real time-PCR, bacterial cultures, and Streptococcus pneumoniae and Legionella pneumophila antigens detection, to determine the etiologies and quantify the viral load. The etiology was identified in 174 (51.0%) patients, and in 85 (24.9%) it was viral, the most frequent rhinovirus and influenza virus. The sensitivity of viral detection in sputum (50.7%) was higher than in NP swabs (20.9%). Compared with sputum, the positive predictive value and specificity of NP swabs for viral diagnosis were 95.8% and 96.9%, respectively. Performance of PSI and CURB-65 scores in all CAP with etiologic diagnosis were as expected, with mortality associated with higher values, but they were not associated with mortality in patients with viral pneumonia. NP swabs have lower sensitivity but high specificity for the diagnosis of viral CAP in adults compared with sputum, reinforcing the use NP swabs for the diagnostic etiology work-up. The PSI and CURB-65 scores did not predict mortality in the viral CAP, suggesting that they need to be updated scores based on the identification of the etiological agent.

A reinforcement learning approach to improve the performance of the Avellaneda-Stoikov market-making algorithm.

Market making is a high-frequency trading problem for which solutions based on reinforcement learning (RL) are being explored increasingly. This paper presents an approach to market making using deep reinforcement learning, with the novelty that, rather than to set the bid and ask prices directly, the neural network output is used to tweak the risk aversion parameter and the output of the Avellaneda-Stoikov procedure to obtain bid and ask prices that minimise inventory risk. Two further contributions are, first, that the initial parameters for the Avellaneda-Stoikov equations are optimised with a genetic algorithm, which parameters are also used to create a baseline Avellaneda-Stoikov agent (Gen-AS); and second, that state-defining features forming the RL agent's neural network input are selected based on their relative importance by means of a random forest. Two variants of the deep RL model (Alpha-AS-1 and Alpha-AS-2) were backtested on real data (L2 tick data from 30 days of bitcoin-dollar pair trading) alongside the Gen-AS model and two other baselines. The performance of the five models was recorded through four indicators (the Sharpe, Sortino and P&L-to-MAP ratios, and the maximum drawdown). Gen-AS outperformed the two other baseline models on all indicators, and in turn the two Alpha-AS models substantially outperformed Gen-AS on Sharpe, Sortino and P&L-to-MAP. Localised excessive risk-taking by the Alpha-AS models, as reflected in a few heavy dropdowns, is a source of concern for which possible solutions are discussed.

Descifrando el mecanismo de inhibición de una pseudoquinasa: la quinasa ligada a integrinas y el cpd22 / Deciphering the inhibition mechanism of a pseudokinase: integrin-bound kinase and cpd22

Según la Organización Mundial de la Salud (OMS), el cáncer es una de las principales causas de muerte no infecciosas en todo el mundo. En este contexto, la búsqueda de nuevos fármacos potentes y seguros contra el cáncer es un área de gran interés para la industria farmacéutica y la investigación académica. A pesar de que las proteínas quinasas han sido una de las dianas anticancerosas más exploradas, sus parientes evolutivamente degenerado, las pseudoquinasas, han atraído la atención de la comunidad científica en la última década. La quinasa ligada a integrinas (ILK) es un miembro del pseudokinoma humano y ha sido reivindicada y validada como una diana terapéutica prometedora para las enfermedades neoplásicas. El único inhibidor de ILK conocido, CPD22, ha probado su actividad en ensayos fenotípicos; sin embargo, se dispone de muy poca información acerca de su mecanismo de acción a nivel molecular. Usando técnicas quimioinformáticas y de modelado molecular se ha dilucidado si el CPD22 es capaz de unirse a ILK y cómo lo hace. Además, nuestro modelo computacional explica el SAR generado en la campaña de optimización del cabeza de serie y los experimentos de SPR han probado, por primera vez, que la molécula se une a ILK, lo que respalda la hipótesis de inhibición por unión directa y el modelo generado. (AU)

According to World Health Organization (WHO) cancer is one of top non- infectious death causes worldwide. In this context, as such the searching of new potent and safe anticancer drugs is a hot point for pharmacy industry and academic investigation. Despite protein kinases have been one of the most explored anticancer targets, its evolutionary degenerated parents, pseudokinases, have attracted the attention of scientific community during the last decade. Integrin Linked Kinase (ILK) is a member of the human pseudokinome that has been claimed and validated as a prommissing target for neoplastic diseases. The only well-known ILK inhibitor, CPD22, has probed its activity in phenotypic assays, however very few knowledge regarding its mechanism of action at molecular level is available. Using chemoinformatic and molecular modelling techniques we were able to elucidate if this molecule is able to bind to ILK and how. Additionally, our model explains the SAR raised from the optimization campaign, and SPR experiments have probed, by first time, that this molecule binds to ILK, thus supporting the hypothesis of inhibition by direct binding and the computational model as well. (AU)

Propuesta de un modelo computacional para el estudio de la selectividad de los inhibidores alostéricos de PTP1B / Proposal for a computational model to study the selectivity between PTP1B and its allosteric inhibitors

La diabetes mellitus tipo II ha emergido como una pandemia mundial, convirtiéndose en un grave problema de salud pública para muchos países. Los tratamientos actuales presentan efectos adversos indeseables, falta de eficacia o simplemente solo tratan los síntomas, no las causas. La proteína tirosina fosfatasa 1B se ha reconocido como una diana prometedora para el tratamiento de esta enfermedad. A pesar de todos los estudios realizados sobre esta enzima, apenas existe conocimiento acerca de la selectividad entre esta y la tirosina fosfatasa de linfocitos T, su homólogo más cercano, responsable de complicados efectos adversos. En este estudio, las aproximaciones computacionales aplicadas a un inhibidor alostérico previamente descrito de esta enzima, han permitido identificar un residuo de fenilalanina como crucial para esta selectividad. Los resultados han permitido explicar esta selectividad desde el punto de vista bioquímico y guiarán el diseño de nuevos fármacos más potentes y selectivos. (AU)

Diabetes mellitus type II has spread as a problematic pandemia for countries all over the world. Despite of the current available treatments, approved drugs still show undesirable side effects, loss of efficacy or they target symptoms instead of causes. Since its discovery, protein tyrosine phosphatase 1B has emerged as a very promising target against this disease. Despite of the information about the enzyme, there is no knowledge regarding the selectivity between this enzyme and its closest homologue, lymphocyte T tyrosine phosphatase, responsible of complicate side effects. In this study different computational approaches have let us to highlight the importance of a phenylalanine residue located in protein tyrosine phosphatase 1B, but no in its homologue, as a crucial hotspot that causes selectivity. These results let to explain the observed selectivity and they can be used as a guide for the design of new inhibitors. (AU)

Diazepam binding inhibitor governs neurogenesis of excitatory and inhibitory neurons during embryonic development via GABA signaling.

Of the neurotransmitters that influence neurogenesis, gamma-aminobutyric acid (GABA) plays an outstanding role, and GABA receptors support non-synaptic signaling in progenitors and migrating neurons. Here, we report that expression levels of diazepam binding inhibitor (DBI), an endozepine that modulates GABA signaling, regulate embryonic neurogenesis, affecting the long-term outcome regarding the number of neurons in the postnatal mouse brain. We demonstrate that DBI is highly expressed in radial glia and intermediate progenitor cells in the germinal zones of the embryonic mouse brain that give rise to excitatory and inhibitory cells. The mechanism by which DBI controls neurogenesis involves its action as a negative allosteric modulator of GABA-induced currents on progenitor cells that express GABAA receptors containing γ2 subunits. DBI's modulatory effect parallels that of GABAA-receptor-mediating signaling in these cells in the proliferative areas, reflecting the tight control that DBI exerts on embryonic neurogenesis.

Insight into the mechanism of molecular recognition between human Integrin-Linked Kinase and Cpd22 and its implication at atomic level.

ABSRACT: Pseudokinases have received increasing attention over the past decade because of their role in different physiological phenomena. Although pseudokinases lack several active-site residues, thereby hindering their catalytic activity, recent discoveries have shown that these proteins can play a role in intracellular signaling thanks to their non-catalytic functions. Integrin-linked kinase (ILK) was discovered more than two decades ago and was subsequently validated as a promising target for neoplastic diseases. Since then, only a few small-molecule inhibitors have been described, with the V-shaped pyrazole Cpd22 being the most interesting and characterized. However, little is known about its detailed mechanism of action at atomic level. In this study, using a combination of computational chemistry methods including PELE calculations, docking, molecular dynamics and experimental surface plasmon resonance, we were able to prove the direct binding of this molecule to ILK, thus providing the basis of its molecular recognition by the protein and the effect over its architecture. Our breakthroughs show that Cpd22 binding stabilizes the ILK domain by binding to the pseudo-active site in a similar way to the ATP, possibly modulating its scaffolding properties as pseudokinase. Moreover, our results explain the experimental observations obtained during Cpd22 development, thus paving the way to the development of new chemical probes and potential drugs.

Descifrando el mecanismo de inhibición de una pseudoquinasa: la quinasa ligada a integrinas y el cpd22 / Deciphering the inhibition mechanism of a pseudokinase: integrin-bound kinase and cpd22

Según la Organización Mundial de la Salud (OMS), el cáncer es una de las principales causas de muerte no infecciosas en todo el mundo. En este contexto, la búsqueda de nuevos fármacos potentes y seguros contra el cáncer es un área de gran interés para la industria farmacéutica y la investigación académica. A pesar de que las proteínas quinasas han sido una de las dianas anticancerosas más exploradas, sus parientes evolutivamente degenerado, las pseudoquinasas, han atraído la atención de la comunidad científica en la última década. La quinasa ligada a integrinas (ILK) es un miembro del pseudokinoma humano y ha sido reivindicada y validada como una diana terapéutica prometedora para las enfermedades neoplásicas. El único inhibidor de ILK conocido, CPD22, ha probado su actividad en ensayos fenotípicos; sin embargo, se dispone de muy poca información acerca de su mecanismo de acción a nivel molecular. Usando técnicas quimioinformáticas y de modelado molecular se ha dilucidado si el CPD22 es capaz de unirse a ILK y cómo lo hace. Además, nuestro modelo computacional explica el SAR generado en la campaña de optimización del cabeza de serie y los experimentos de SPR han probado, por primera vez, que la molécula se une a ILK, lo que respalda la hipótesis de inhibición por unión directa y el modelo generado.(AU)

According to World Health Organization (WHO) cancer is one of top non- infectious death causes worldwide. In this context, as such the searching of new potent and safe anticancer drugs is a hot point for pharmacy industry and academic investigation. Despite protein kinases have been one of the most explored anticancer targets, its evolutionary degenerated parents, pseudokinases, have attracted the attention of scientific community during the last decade. Integrin Linked Kinase (ILK) is a member of the human pseudokinome that has been claimed and validated as a prommissing target for neoplastic diseases. The only well-known ILK inhibitor, CPD22, has probed its activity in phenotypic assays, however very few knowledge regarding its mechanism of action at molecular level is available. Using chemoinformatic and molecular modelling techniques we were able to elucidate if this molecule is able to bind to ILK and how. Additionally, our model explains the SAR raised from the optimization campaign, and SPR experiments have probed, by first time, that this molecule binds to ILK, thus supporting the hypothesis of inhibition by direct binding and the computational model as well.(AU)