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Rodriguez, S.

Publications and source records attributed to Rodriguez, S..

At least 19 recordsLinked to original sources

RNA Editing of DEGS1 Links Genetic Risk to Ceramide Dysregulation and Astrocyte Toxicity in ALS

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease with ~90% of cases being sporadic. Although genome-wide association studies have identified numerous genetic risk loci, these variants account for only a portion of ALS heritability, suggesting that additional genetic and regulatory mechanisms contribute to disease risk. Furthermore, the mechanisms linking common variants to disease pathogenesis remain poorly understood. Adenosine-to-inosine (A-to-I) RNA editing is dysregulated in ALS, yet systematic identification of disease-relevant editing events and their functional consequences has been limited. We performed comprehensive RNA editing analysis across eight central nervous system regions from a Target ALS cohort, identifying 752 differentially edited sites in 304 genes. Global RNA editing was significantly reduced in ALS tissues, particularly in spinal cord and choroid plexus, with altered editing in genes involved in immune signaling, stress response and RNA processing. Through genetic colocalization analysis integrating editing quantitative trait loci (edQTLs) with ALS GWAS data, we identified common variants at the DEGS1 locus associated with both increased 3' UTR editing and elevated ALS risk (posterior probability >0.88). DEGS1 encodes dihydroceramide desaturase 1, a key enzyme converting dihydroceramide to ceramide, the precursor to all sphingolipid species. Using CRISPR-generated DEGS1 knockout iPSCs and lipidomic profiling, we demonstrated that DEGS1 is essential for ceramide synthesis and downstream sphingolipid production in astrocytes. Functional validation revealed that ALS-associated editing of the DEGS1 3' UTR significantly increased DEGS1 protein expression. Critically, astrocytes expressing the fully edited DEGS1 3' UTR increased ceramide levels and reduced cell viability compared to those expressing the unedited construct. These findings establish a novel regulatory link between RNA editing, sphingolipid metabolism and ALS pathogenesis. Our results demonstrate that common genetic variants can influence disease risk through post-transcriptional mechanisms that alter lipid homeostasis in astrocytes, a cell type increasingly recognized as central to ALS progression.

neuroscience↗

Changes in the internal genitalia of Hermetia illucens (Diptera: Stratiomyidae) during reproductive maturation under a coffee pulp-based diet

The global expansion of Black Soldier Fly (BSF), Hermetia illucens (L.), production systems for organic waste management has increased the need to better understand its reproductive biology in order to optimize mass-rearing programs. In this study, we macroscopically characterized chronological morphological changes in the internal genitalia of adult H. illucens reared on a sustainable alternative larval diet based on coffee pulp and corn bran. Morphological assessments and dissections were conducted on female and male reproductive tracts at 3, 6, and 9 days after adult emergence. Overall, the general organization of both reproductive systems was consistent with previous descriptions. A notable observation was the presence of a tripartite fertilization chamber in females, composed of three distinct compartments apparently associated with the three spermathecae. The functional significance of this anatomical organization remains to be determined. Females showed progressively advanced ovarian development and reached the clearest morphological indicators of reproductive maturity at 9 days, while males showed the greatest testicular distention and opacity at the same age. Compared with maturation times reported in previous studies using conventional larval diets, these observations suggest a later pattern of reproductive maturation under the coffee pulp-based diet. The observed differences may be associated with the nutritional composition and carbohydrate-to-protein balance of the larval diet. These results provide chronological and iconographic information that may contribute to the optimization of laboratory rearing protocols and the use of coffee by-products in H. illucens bioconversion systems.

zoology↗

SOX9 regulates endothelial tip cell specification to promote cerebral and neuroretinal vascularization

BackgroundTissue vascularization relies on the organotypic specification of endothelial tip cells to interpret local cues and guide angiogenic sprouts into specific tissue compartments. However, the molecular regulators controlling brain and retinal endothelial tip cell identity remain poorly understood. MethodsEndothelial-specific Sox9 loss-of-function mouse was combined with single-cell, spatial, and bulk RNA sequencing analyses of developing mouse brain and retinal vasculature, as well as experimental ischemic stroke. Transcriptomic findings were validated using in situ hybridization, immunofluorescence, and functional angiogenesis assays. ResultsTranscription factor activity analysis of single-cell RNA-sequencing datasets identified SOX9 as a candidate regulator selectively enriched in developing brain and retinal endothelial tip cells. Endothelial-specific deletion of Sox9 impaired brain and neuroretina vascularization and disrupted the tip cell transcriptomic program, resulting in reduced sprouting angiogenesis and matrix-remodeling pathways. Conversely, SOX9 overexpression in HUVECs promoted neuro-tip-like signatures and enhanced endothelial invasion and sprouting. Following ischemic stroke, single-cell and spatial transcriptomic analyses identified a transient angiogenic endothelial population within the ischemic area. However, these cells failed to express Sox9 and lacked key developmental brain tip cell features. ConclusionsSOX9 is a key regulator of endothelial tip cell identity and neuronal angiogenesis. These findings reveal fundamental differences between developmental and injury-induced vascular responses and identify SOX9 as a potential therapeutic target to promote functional vascular regeneration.

developmental biology↗

LMNA Haploinsufficiency in Human iPSC-Derived Cardiac Organoids Reveals Early Fibrotic Signaling as a Therapeutically Targetable Process

LMNA mutations are a major cause of dilated cardiomyopathy (DCM), with haploinsufficiency representing a common pathogenic mechanism. Yet the earliest disease-initiating events remain poorly defined. Here, we identify a novel intronic splice-site variant, c.937-1G>A, that disrupts pre-mRNA processing and induces nonsense-mediated mRNA decay, resulting in LMNA haploinsufficiency. Using induced pluripotent stem cells (iPSCs) generated from the patients peripheral blood mononuclear cells (PBMCs), we differentiated into self-patterning human cardiac organoids to model early LMNA-DCM in a multicellular human context. Single-nucleus transcriptomics revealed unexpectedly broad remodeling across major cardiac cell types beyond cardiomyocytes. Functionally, LMNA-mutant organoids exhibited impaired contractility, altered calcium handling, and increased arrhythmic activity. These changes were accompanied by early profibrotic activation, including increased reactive oxygen species (ROS), periostin (POSTN) secretion, and CTGF expression. Treatment with the antifibrotic drug nintedanib attenuated this response. Together, these findings show that LMNA haploinsufficiency initiates global pathogenic remodeling at an unexpectedly early developmental stage.

cell biology↗

In silico drug repurposing and in vitro validation of cestode fatty acid binding proteins

Echinococcosis is a Neglected Tropical Disease (NTD) caused by Echinococcus granulosus and Echinococcus multilocularis, the etiological agents of cystic and alveolar echinococcosis, respectively. These infections pose a significant public health burden, particularly in endemic regions. Cestodes lack key enzymes involved in lipid metabolism and must acquire lipids from their hosts. Fatty Acid Binding Proteins (FABPs), which mediate lipid trafficking and intracellular transport, have therefore emerged as essential and potentially druggable targets. In this study, we implemented an integrated virtual screening strategy combining ligand-based and structure-based approaches to identify novel FABP binders as potential therapeutic agents against Echinococcus spp. High-specificity screening of approximately 435,000 compounds yielded a limited number of prioritized in silico hits. Four compounds--hydrochlorothiazide, naratriptan, fenticonazole, and montelukast--were selected for experimental validation, prioritizing repurposing candidates. Fluorescence displacement assays confirmed that hydrochlorothiazide binds to three cestode FABPs (EgFABP1, EmFABP1, and EmFABP3), validating the predictive performance of the computational workflow. These findings support the value of parallel in silico screening strategies and drug repurposing approaches for the discovery of new therapeutic candidates against neglected tropical diseases.

bioinformatics↗

Conformational ensembles of flexible multidomain proteins: How close are we to accurate and reliable predictions?

Multidomain proteins connected by flexible linkers populate conformational ensembles that are challenging to characterize using conventional structural biology methods. In domain-linker-domain (DLD) proteins, linker-mediated inter-domain relative positions and orientations are functionally relevant, yet their dynamical behavior in solution normally remain poorly described. Small-angle X-ray scattering (SAXS) provides ensemble-averaged structural information for such systems; however, coupling with computational modeling is required to accurately describe the dynamic behavior of this family of proteins in solution. Here, we present a systematic evaluation of five ensemble-generation strategies applied to a set of eighteen proteins sharing the same two globular domains, connected by naturally occurring linkers of varying length and composition. Modeling methods based on different underlying principles are compared by assessing their agreement to experimental SAXS data, showing a large disparity and systematic structural biases among them. Furthermore, for each approach, we examine the effect of refinement against SAXS restraints and assess its capacity to describe the experimental data, as well as the induced biases in global dimensions and inter-domain distance distributions. This analysis underlines the importance of the initial conformational pool for deriving experimentally compatible ensembles. Overall, this work provides a high-quality benchmark for SAXS-driven ensemble modeling of flexible, multidomain proteins and establishes a framework for the critical interpretation of solution scattering data in systems with pronounced conformational heterogeneity.

biophysics↗

Oral immune priming modulates microbiota composition and supports pathogen control in the Manila clam (Ruditapes philippinarum)

Immunological memory was long considered an exclusive feature of vertebrates. However, extensive evidence now shows that invertebrates possess forms of innate immune memory--known as immune priming-- where previous exposure to a pathogen enhances subsequent immune responses and host protection. Immune priming has been proposed as a promising strategy for disease prevention in shellfish aquaculture. However, immune priming remains largely unexplored in marine bivalves, particularly in the Manila clam (Ruditapes philippinarum), a top-ten global aquaculture species. Here, we investigated for the first time the effects of oral immune priming on host survival, pathogen dynamics, and microbiota composition in R. philippinarum against the emergent bivalve pathogen V. europaeus. Priming was induced using the live bacterial pathogen at a sublethal dose, followed by a lethal secondary exposure. Primed clams exhibited a significant survival following the second challenge (87% survival vs. 0% in non-primed clams), demonstrating robust protection against reinfection. Quantitative PCR (qPCR) revealed that primed clams rapidly reduced pathogen loads after 48 h during the second challenge, reaching concentrations below the mortality threshold observed in non-primed clams ([~]105 copies mg-{superscript 1}). Interestingly, the pathogen was able to persist at low and non-harmful concentration ([~]102 copies mg-{superscript 1}) in primed clams along both challenges. Full-length 16S rRNA metabarcoding analyses showed that immune priming shifts the host microbiota. Alpha and beta diversity indicated a progressive reduction in diversity and the establishment of a specific and resilient bacterial community in primed clams. Clustering analyses identified a priming-associated microbiota dominated by Acinetobacter, Brevundimonas, Sphingobium, and Psychrobacter, which persisted through the secondary challenge but was absent or depleted in non-primed clams. Conversely, members of the Arcobacteraceae (e.g., Arcobacter, Poseidonibacter) were absent after priming and emerged only during second infection, decreasing in primed clams but increasing in non-primed clams coinciding with high mortalities. Our findings provide the first phenotypic and microbiome-level evidence of oral immune priming in Manila clam. Here we demonstrate that priming enhances pathogen control and promotes the establishment of a protective microbiota that may interact with the host immune system to confer resistance against bacterial infection. These results open new avenues for immune-priming and microbiota-based strategies to improve disease resistance in bivalve aquaculture.

microbiology↗

Deciphering the pangenome of the shellfish pathogen Vibrio europaeus: Evolutionary history and functional impact of core and accessory genes in aquaculture.

Vibrio europaeus is an important pathogen in shellfish aquaculture, yet its genomic diversity and adaptive potential remain poorly understood. Here, we present the first comprehensive analysis of the V. europaeus pangenome, integrating genomic data from all strains available to the date sequenced specifically for this study. Those were isolated from different aquaculture facilities (shellfish hatcheries) associated to mass mollusks mortalities from different geographical locations, years and host species. Our findings revealed an open pangenome with the 61% of the genes associated to the accessory genome that contributes to environmental and host adaptations. Phylogenomic analyses of the core-genome (39% of the pangenome size) allowed to evaluate the evolutionary history and intraspecific diversity of V. europaeus and revealed that Spanish strains displayed a much lower genetic variability than French, Chilean or American strains, probably due to a monophyletic radiation event. Functional annotation of core and accessory genes revealed the key virulence factors of the species while it also disclosed that those are located mainly into the core genes. The high number of anti-phage defense systems encoded in the accessory genome explained almost all the variability of the species. The results provide important insights into the evolutionary history and ecological versatility of V. europaeus, with potential implications for diagnostics, epidemiological surveillance, and disease management strategies in aquaculture. Impact statementThis study presents the first comprehensive pangenome analysis of Vibrio europaeus, an emergent pathogen responsible for severe economic losses in shellfish aquaculture, the second most important sector of global aquaculture. Here, we characterized for the first time the V. europaeus pangenome, integrating genomic data from all strains isolated to date, sequenced specifically for this study using NGS and/or third-generation (PacBio) technologies. This work achieved the most complete species pangenome to date and is among the first studies on aquaculture-related bacterial pathogens. Beyond a descriptive framework, the pangenome was critically examined to identify key traits, including virulence factors, secondary metabolite biosynthesis, and antimicrobial resistance genes, essential for host infection and adaptation. Moreover, the study of anti-phage defense systems was shown to account for much of the species genomic variability. The genomic resources and insights generated here substantially expand our understanding of V. europaeus biology and provide valuable information that can be applied for diagnostics, epidemiological surveillance, and sustainable management of this pathogen in aquaculture industry. Data summaryAll genome assemblies have been uploaded to the National Center for Biotechnology Information. The GenBank accession numbers for each of the 39 strains used in this study and detailed information can be found in Table S1. All bioinformatics tools used for comparative genomics have been listed in the Methods section including references, associated databases and analysis parameters.

microbiology↗

Structure-Guided Development of a Potent BioA Inhibitor ValidatesBiotin Synthesis Inhibition as a Therapeutic Strategy forTuberculosis

Conditionally essential pathways - such as the biotin biosynthesis - represent promising targets for new antibiotics. However, the chemical interrogation of the biotin pathway with an orally effective lead remains elusive, and the preclinical development of biotin inhibitors for mycobacterial infections in vivo is challenging due to the unusually high concentration of biotin in standard mouse models. Structure-guided optimization was applied to develop the first oral lead targeting aminotransferase BioA, a key enzyme in bacterial biotin biosynthesis, resulting in C48, a picomolar inhibitor displaying sub-micromolar MICs against Mycobacterium tuberculosis (Mtb). Mechanism of action was confirmed by biochemical, structural, and genetic studies. C48 demonstrated favorable pharmacokinetics and excellent oral bioavailability resulting in over 39,000-fold improved exposure. We next developed an easy-to-operate, low-biotin mouse model that recapitulates human biotin physiology. C48 significantly reduced Mtb burden in this low-biotin mouse model, providing the first in vivo proof-of-concept for targeting biotin biosynthesis in Mtb.

microbiology↗

SIRPγ modulates effector differentiation of human CD8 T Cells under suboptimal TCR stimulation: implications for immune homeostasis and autoimmunity

BackgroundAberrant CD8 T-cell differentiation contributes to the pathogenesis of autoimmune diseases, and immune-mediated tissue damage. However, the molecular mechanisms that prevent premature effector T cell programming in humans remain incompletely defined. Signal regulatory protein gamma (SIRP{gamma}) is selectively expressed on T-cells in the human immune system. Notably, genetic variants associated with reduced SIRP{gamma} expression have been linked to increased risk of immune-mediated diseases, including type 1 diabetes and multiple sclerosis, but the contribution of SIRP{gamma} to CD8 T-cell dysregulation in these contexts remains unclear. ObjectiveTo determine how inter-individual variation in SIRP{gamma} expression influences immune homeostasis and CD8 T-cell effector programming. MethodsPeripheral blood CD8 T-cells from healthy donors were analyzed for SIRP{gamma} expression and associated differentiation phenotypes. Naive CD8 T-cells were purified and subjected to siRNA-mediated knockdown of SIRPG, followed by suboptimal TCR stimulation. Differentiation status, transcription factor expression, and effector cytokine production were measured using flow cytometry. CD47 blockade was used to assess ligand dependency. ResultsLow SIRP{gamma} expression on CD8 T-cells was associated with increased frequencies of CD27-CD45RO effector-like and CD27-CD45RO- terminally differentiated CD8 T-cells. SIRPG knockdown induced effector-like differentiation, with increased CD45RO and T-bet expression and elevated TNF-, IFN-{gamma}, and Granzyme B production. This effect was not recapitulated by CD47 blockade, suggesting a CD47-independent regulatory mechanism. ConclusionSIRP{gamma} serves as a negative regulator of CD8 T-cell effector differentiation under suboptimal stimulation. Inter-individual variation in its expression may influence susceptibility to immune dysregulation, positioning it as a potential biomarker and therapeutic target.

immunology↗

Interplay Between Intrinsically Disordered Proteins and Atomically Precise Gold Nanoclusters Modulates their Optical Properties

Understanding how structural and optical properties of metallic nanoclusters can be tuned by proteins is crucial for the use of these hybrid molecules in biomedical applications. The interaction of proteins with ultrasmall, atomically-precise gold nanoclusters (Au-NCs) has been mainly investigated in the context of structured proteins, while their behavior with intrinsically disordered proteins (IDPs) remains unexplored. This work examines the structural and optical properties of Au-NCs interacting with bioengineered IDPs containing up to three cysteines. We show that, by exploiting the conformational flexibility of cysteine-containing IDPs, we can anchor proteins to Au-NCs in a position-specific manner, leading to new bioconjugates with properties that differ from those of the individual components. We observed an up to 15-fold photoluminescence enhancement depending on the number of cysteines anchored. By combining mass spectrometry, small-angle X-ray scattering (SAXS), and computational modelling, the ensemble structures of nine bioconjugates with different stoichiometries were elucidated, indicating their overall compactness. Our results suggest that the interface between these atomically-precise species and the conformationally fluctuating protein is responsible for the optical properties of these nanobioconjugates. This research improves our understanding of Au-NC- protein interactions, paving the way to novel nano-molecular hybrid conjugates with tunable properties for bioimaging and therapeutic applications.

biochemistry↗

Protein language models accelerate the discovery of Plastic-Degrading Enzymes

Plastic pollution presents a critical environmental challenge, necessitating innovative and sustainable solutions. In this context, biodegradation using microorganisms and enzymes offers an environmentally friendly alternative. This work introduces an AI-driven frame-work that integrates machine learning (ML) and generative models to accelerate the discovery and design of plastic-degrading enzymes. By leveraging pre-trained protein language models and curated datasets, we developed seven ML-based binary classification models to identify enzymes targeting specific plastic substrates, achieving an average accuracy of 89%. The framework was applied to over 6,000 enzyme sequences from the RemeDB to classify enzymes targeting diverse plastics, including PET, PLA, and Nylon. Besides, generative learning strategies combined with trained classification models in this work were applied for de novo generation of PET-degrading enzymes. Structural bioinformatics validated potential candidates through in-silico analysis, highlighting differences in physicochemical properties between generated and experimentally validated enzymes. Moreover, generated sequences exhibited lower molecular weights and higher aliphatic indices, features that may enhance interactions with hydrophobic plastic substrates. These findings highlight the utility of AI-based approaches in enzyme discovery, providing a scalable and efficient tool for addressing plastic pollution. Future work will focus on experimental validation of promising candidates and further refinement of generative strategies to optimize enzymatic performance.

bioengineering↗

The inflammasome adaptor protein ASC triggers immunometabolic dysregulation induced by dysbiosis during obesity

The inflammasomes are a component of the innate immune system that induce inflammation after activation by danger and pathogen signals, this process is of extreme importance in the gut, where high-saturated fat diets modulate inflammasome activity resulting in dysbiosis and inflammation. In this work, we found that the common inflammasome adaptor protein, apoptosis speck-like protein with a caspase activation domain (ASC), is essential for the development of metabolic dysregulation during high fat diet. ASC-deficient mice (Pycard-/-) presented lower weight gain and improved glucose tolerance, enhancing metabolism, and reducing inflammation in both intestine and liver. The extracellular administration of ASC oligomers to Pycard-/- mice partially rescued dysbiosis, intestinal metabolism and inflammation. Therefore, ASC presents intracellular functions activating the inflammasome, as well as extracellular function as an oligomer during metabolic dysregulation, emerging ASC as a potential drug target beneficial to treat obesity and metabolism-associated liver diseases.

immunology↗

MicroRNA-502-3p modulates the GABA A subunits, synaptic proteins and mitochondrial morphology in hippocampal neurons

MicroRNA-502-3p (MiR-502-3p), a synapse enriched miRNA is considerably implicated in Alzheimers disease (AD). Our previous study found the high expression level of miR-502-3p in AD synapses relative to controls. Further, miR-502-3p was found to modulate the GABAergic synapse function via modulating the GABA A receptor subunit -1 (GABRA1) protein. The current study is attempted to examine the impact of miR-502-3p on other GABA subunit proteins, synaptic proteins, mitochondrial morphology and other hippocampal neuron genes. Mouse hippocampal neuronal (HT22) cells were transfected with miR-502-3p overexpression (OE) vector, miR-502-3p sponge (suppression) vector and scramble control vector. MiR-502-3p vectors transfection was confirmed by fluorescence microscopy. MiR-502-3p expression and GABRA1 expression was confirmed by qRT-PCR and miRNAScope in-situ hybridization. GABA A subunit and synaptic proteins were studied by immunoblotting analysis and mitochondrial morphology was analyzed by transmission electron microscopy (TEM) analysis. Further, Affymetrix gene array analysis was conducted in miR-502-3p overexpressed and suppressed cells. Our results observed that elevated miR-502-3p, negatively modulates the GABRA1 level. The levels of GABA A subunit and synaptic proteins were reduced by ectopic expression of miR-502-3p and increase by miR-502-3p suppression. The mitochondrial morphology was found to be improved in-terms of their number and length in miR-502-3p suppressed cells. Further, Gene array analysis unveiled the deregulation of several genes by miR-502-3p, which are associated with oxidative stress, immune response and synaptic function. These results provide new insights and an update to understand the biological roles of miR-502-3p in regulation of neuron function and synaptic activity.

neuroscience↗

Local alkalinity enhancement using artificial substrates increases survivorship of early-stage coral recruits

Efforts to restore coral reefs using sexually derived coral recruits are often hindered by their low survivorship and growth, hence scalable interventions to improve these parameters are urgently needed. Here, we developed novel settlement substrates that modify the local chemical and hydrodynamic environments to provide local alkalinity enhancement (AE) within the laminar boundary layer to aid in coral restoration. Cement tiles with four different chemistries and two different surface topographies were tested in a novel flume system to quantify their ability to change local pH under reef-like conditions and their effect on larval settlement, survivorship, and growth of the endangered Caribbean coral, Orbicella faveolata. Chemistry had a minimal effect on the initial settlement of coral larvae, and textured tiles were preferred over smooth tiles. However, substrates that created a more alkaline local environment increased post-settlement survivorship. The increased survivorship of O. faveolata recruits on AE tiles was not due to increased growth on AE tiles, although trends in growth were dependent on chemistry and topography of tiles. Our results indicate that mixing sodium bicarbonate or sodium carbonate into cement used to fabricate artificial reef structures could be an effective means to enhance the development of coral cover. Significance StatementReef-building coral populations in South Florida and the Caribbean are nearing crisis, with low survivorship rates of new recruits being identified as a critical bottleneck limiting their recovery. Since ocean acidification is known to stress corals and hinder their growth, we hypothesized that enhancing substrate alkalinity could help mitigate ocean acidification effects by boosting coral growth and increasing survivorship during their critical early stages. This hypothesis was experimentally tested by adding sodium bicarbonate or sodium carbonate to settlement substrates and studying coral settlement preference, and coral spat growth and survival. Our results showed significantly higher survivorship on alkalinity enhanced substrates, indicating that this could be a promising restoration intervention.

ecology↗

Semaphorin 4A maintains functional diversity of the hematopoietic stem cell pool

Somatic stem cell pools are comprised of diverse, highly specialized subsets whose individual contribution is critical for the overall regenerative function. In the bone marrow, myeloid-biased HSC (myHSC) are indispensable for replenishment of myeloid cells and platelets during inflammatory response but at the same time, become irreversibly damaged during inflammation and aging. Here, we identify an extrinsic factor, Semaphorin 4A (Sema4A), which non cell-autonomously confers myHSC resilience to inflammatory stress. We show that the absence of Sema4A, myHSC inflammatory hyper-responsiveness in young mice drives excessive myHSC expansion, myeloid bias and profound loss of regenerative function with age. Mechanistically, Sema4A is mainly produced by neutrophils, signals via a cell surface receptor Plexin D1 and safeguards myHSC epigenetic state. Our study shows that by selectively protecting a distinct stem cell subset, an extrinsic factor preserves functional diversity of somatic stem cell pool throughout organismal lifespan.

cell biology↗

Identification of targetable epigenetic vulnerabilities in uveal melanoma

Uveal melanoma (UM) is the most common primary intraocular malignancy in adults, with a strong predilection for hepatic metastasis, occurring in approximately 50% of cases. Metastatic UM remains highly resistant to therapy and is almost invariably fatal. The strongest genetic drivers of UM metastasis are loss-of-function mutations in tumor suppressor BAP1, an epigenetic regulator that serves as the ubiquitin hydrolase subunit of the polycomb repressive deubiquitinase (PR-DUB) complex, and a key player in global epigenetic regulation. Inactivation of BRCA Associated Protein 1 (BAP1) has been shown to induce widespread epigenetic alterations across multiple model systems. To identify novel therapeutic strategies, we investigated whether targeting the epigenome could reveal new vulnerabilities in UM. We performed high-throughput compound screening using a curated epigenetic inhibitor library and identified BET (bromodomain and extra-terminal domain) inhibition as a particularly promising approach. Interestingly, we observed significant heterogeneity in the efficacy of different BET inhibitors in UM. While previous clinical trials with two BET inhibitors have failed to show efficacy in UM, our findings highlight substantial differences in the potency of specific BET inhibitors for this malignancy. Notably, the BET inhibitor mivebresib (ABBV-075) significantly improved survival rates by 50% in a metastatic UM xenograft mouse model and completely prevented detectable metastases in the bones, spinal cord, and brain. Unexpectedly, RNA sequencing revealed a strong transcriptional overlap between BET inhibition and histone deacetylase (HDAC) inhibition--an approach currently under clinical evaluation for UM treatment. Both BET and HDAC inhibitors reversed gene expression signatures associated with high metastatic risk and induced a neuronal differentiation-like phenotype in UM cells. Together, our findings demonstrate that UM cells exhibit a distinct vulnerability to BET inhibition and establish BET inhibitors as promising candidates for further clinical evaluation for metastatic UM.

cancer biology↗

Rapid Emergence of Resistance to Broad-Spectrum Direct Antimicrobial Activity of Avibactam

Avibactam (AVI) is a diazabicyclooctane (DBO) {beta}-lactamase inhibitor used clinically in combination with ceftazidime. At concentrations higher than those typically achieved in vivo, it also has broad-spectrum direct antibacterial activity against Enterobacterales strains, including metallo-{beta}-lactamase-producing isolates, mediated by inhibition of penicillin-binding protein 2 (PBP2). This activity is mechanistically similar to that of more potent novel DBOs (zidebactam, nacubactam) in late clinical development. We found that resistance to AVI emerged readily, with a mutation frequency of 2x10-6 to 8x10-5. Whole genome sequencing of resistant isolates revealed a heterogeneous mutational target that permitted bacterial survival and replication despite PBP2 inhibition, in line with prior studies of PBP2-targeting drugs. While such mutations are believed to act by upregulating the bacterial stringent response, we found a similarly high mutation frequency in bacteria deficient in components of the stringent response, although we observed a different set of mutations in these strains. Although avibactam-resistant strains had increased lag time, suggesting a fitness cost that might render them less problematic in clinical infections, there was no statistically significant difference in growth rates between susceptible and resistant strains. The finding of rapid emergence of resistance to avibactam as the result of a large mutational target has important implications for novel DBOs with potent direct antibacterial activity, which are being developed with the goal of expanding cell wall-active treatment options for multidrug-resistant gram-negative infections but may be vulnerable to treatment-emergent resistance.

microbiology↗