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Biology subjects

Ahmed, N.

Publications and source records attributed to Ahmed, N..

At least 19 recordsLinked to original sources

Optimizing copy number profiles for tumor cell phylogeny inference from low-coverage single-cell sequencing data

Motivation: Accurate reconstruction of tumor cell phylogenies is crucial for understanding cancer evolutionary dynamics, inferring intra-tumor heterogeneity, and designing effective and personalized cancer treatment strategies. One of the most promising and cost-effective approaches for reconstructing tumor phylogenies for large numbers of cells is to use copy number alteration (CNA) data obtained from low-coverage single-cell DNA sequencing of sampled tumor cells. However, error and uncertainty in inferred copy number profiles (CNPs), resulting from low sequence coverage, can have a drastic negative effect on the reconstruction accuracy of tumor cell phylogenies. Results: In this work, we show that careful selection of bin size, a critical tuning parameter for inferring CNPs, is necessary to maximize the accuracy of tumor cell phylogeny inference. Specifically, we (i) use three previously published low-coverage single-cell DNA sequencing datasets from real tumors to demonstrate that even small changes in bin size can have a drastic impact on downstream tumor phylogeny reconstruction, (ii) show that this effect persists even when using simulated low-coverage datasets generated under idealized conditions with minimal noise, and (iii) propose a simple computational approach to select bin sizes that maximize tumor phylogeny accuracy. Our experiments span a wide range of bin sizes and account for the impact of various factors including coverage-level, phylogeny reconstruction method, and CNP inference method, and explicitly account for phylogenetic reconstruction uncertainty. Our evaluation of the proposed computational approach for bin size selection shows that it can significantly improve tumor cell phylogeny reconstruction accuracy compared to baseline methods. Availability: Freely available from https://github.com/Tanveer14/BinSelector.

bioinformatics↗

In vivo HSPC gene therapy of hemoglobinopathies without drug selection of corrected cells

In vivo hematopoietic stem/progenitor cell (HSPC) gene therapy remains limited by low gene-editing efficiency and a lack of clinically applicable selection strategies to enrich therapeutically corrected progeny. We used in vivo base and prime editing to introduce a nonpathogenic EPOR variant into HSPCs, conferring erythropoietin hypersensitivity and promoting preferential expansion of gene-corrected erythroid cells. EPOR editing was combined with three therapeutic approaches for the correction of hemoglobinopathies: {gamma}-globin gene addition, {gamma}-globin reactivation, or correction of the sickle cell disease mutation. Tropism-modified helper-dependent adenoviral vectors (HDAd6/3+) targeting HSPCs were used to simultaneously deliver the EPOR-editing machinery and the corresponding therapeutic components. In vitro studies in an erythroid progenitor cell line and primary CD34+ cells demonstrated that the EPORW439* variant conferred a strong proliferative advantage to therapeutically modified erythroid progenitors. Mice humanized with CD34+ cells from a {beta}/{beta}-thalassemia patient were subjected to EPORW439*-mediated EPO hypersensitivity alongside a therapeutic {gamma}-globin transgene which resulted in >70% HbF-positive erythroid cells and substantial reversion of the disease-associated phenotype, including reduced oxidative stress, near-complete elimination of splenic iron deposition, and reduced splenomegaly. Importantly, these effects were achieved after simple intravenous administration of the vectors following HSPC mobilization and cytokine prophylaxis, without subsequent pharmacologic selection. Together, these findings establish a strategy to amplify the therapeutic benefit of otherwise limited in vivo HSPC gene editing for hemoglobinopathies.

bioengineering↗

Enhancer RNA like function of intergenic inherited lncRNAs during maternal to zygotic transition in zebrafish

The maternal-to-zygotic transition (MZT) is a major developmental event during which inherited transcripts are remodeled and zygotic transcription is established. Although parentally inherited long noncoding RNAs (lncRNAs) are present in early embryos, they have been thought to be dispensable. We have identified more than 2000 inherited lncRNAs in zebrafish embryos, but how these RNAs participate in regulatory programs during early development has remained unexplored. Here, the inheritance of selected zebrafish lncRNAs spanning a broad expression range were confirmed at the pre-MZT stage and full-length sequences were captured by Direct RNA nanopore sequencing. We show that 30% inherited intergenic lncRNAs are preferentially associated with active enhancers, annotated as such in DANIO CODE, whereas non-inherited intergenic lncRNAs rarely overlap with enhancers. Perturbation of five inherited intergenic lncRNAs, individually, using antisense oligonucleotides reduced the expression of their respective neighboring genes at 2.5, 4.3, and/or 6 hours post fertilization, indicating that these RNAs act as positive local regulators during MZT. Together, these findings identify inherited intergenic lncRNAs as enhancer-associated regulators with elncRNA-like properties during early embryogenesis.

developmental biology↗

Aberrant expression of stress-related Hsrω-n lncRNA contributes to CGG repeat-mediated toxicity in a Drosophila model of FXTAS

Fragile X-associated tremor/ataxia syndrome (FXTAS) is an adult-onset neurodegenerative disease associated with carriers of premutation (PM) alleles of the fragile X messenger ribonucleoprotein 1 (FMR1) gene. Expanded CGG repeats in the PM alleles are sufficient to cause cellular stress and toxicity in animal and cellular models of FXTAS. Here, we show that in a Drosophila transgenic model of FXTAS, expanded CGG repeats result in the overexpression of heat shock RNA omega-nuclear (Hsr{omega}-n), a stress-related long non-coding RNA (lncRNA) that dominantly enhances expanded CGG-induced neurotoxicity, whereas its reduction suppresses CGG-induced neurotoxicity. Furthermore, overexpression of Hsr{omega}-n lncRNA concomitantly resulted in a significant enhancement in its association with Hrb87F, the fly ortholog of one of the previously identified CGG repeat RNA-binding proteins, hnRNP A2/B1, and with the imitation switch (ISWI) protein, a chromatin remodeling factor. We extended the findings of Hsr{omega}-n to show that the mammalian stress-related lncRNAs SatIII and Neat1 are elevated in cells expressing expanded CGG repeats. Together, these findings highlight the critical roles of stress-related lncRNAs in CGG repeat-mediated toxicity and support a model in which the total steady-state levels of stress-related lncRNAs increase in the presence of expanded CGG repeats as part of the normal regulatory response to combat stress. However, the effects of chronic expanded CGG repeat expression in post-mitotic neurons may result in altered distribution and/or sequestration of specific RNA-binding proteins or chromatin remodeling factors associated with stress-related lncRNAs. This could contribute to the altered cellular homeostasis and genomic instability associated with FXTAS. SignificanceIndividuals carrying the FMR1 allele with expanded CGG repeats (55-200) in the 5' untranslated region are predisposed to fragile X-associated tremor/ataxia syndrome (FXTAS), a late-onset neurodegenerative disorder. FXTAS demonstrates incomplete penetrance, suggesting the possible involvement of genetic modifiers in its pathogenesis. We used a transgenic Drosophila FXTAS model to conduct genetic, molecular, and biochemical analyses. Our findings revealed that the expression of expanded CGG repeats aberrantly activates stress-related Hsr{omega}-n lncRNA in Drosophila melanogaster. Overexpression of this lncRNA exacerbates CGG-mediated neurodegeneration, whereas its repression alleviates this effect. Extending these findings to mammalian systems, we observed the upregulation of SATIII and Neat I, functional homologs of Hsr{omega}-n lncRNA, in cells expressing expanded CGG repeats. Collectively, these findings identify stress-related lncRNAs as key mediators of CGG-mediated toxicity and underscore their potential as therapeutic targets for FXTAS.

genetics↗

Detrimental effects of advanced glycation end-products (AGEs) on a human neuromuscular junction co-culture model

The neuromuscular junction (NMJ) is a specialised synapse between motor neurons and skeletal muscle, and its progressive deterioration contributes to age-related and metabolic disease-associated declines in muscle function. Advanced glycation end-products (AGEs) accumulate in tissues during ageing, diabetes, and chronic metabolic dysfunction and have been implicated in neuromuscular degeneration, yet their effects on the intact NMJ have not previously been examined in a human model system. This study employed a fully human, serum-free, and neural growth factor-free NMJ co-culture system, combining neural progenitor cells with immortalised human myoblasts derived from an 83-year-old donor, to investigate the effects of AGE exposure on neuromuscular integrity across structural, metabolic, functional, and secretory outcomes. AGE exposure induced significant reductions in motor neuron axonal length, myotube remodelling with centralised nuclear positioning, mitochondrial membrane depolarisation, elevated mitochondrial superoxide production, mitochondrial uncoupling, and reductions in spontaneous contraction intensity and frequency. Neurotrophic and myogenic growth factor signalling was significantly downregulated in AGE-treated co-cultures. These findings identify the NMJ as a sensitive target of glycation stress and establish this fully human co-culture platform as a physiologically relevant model for investigating glycation-related neuromuscular pathology and evaluating candidate therapeutic interventions.

cell biology↗

Physiological and Biochemical Responses of Grafted Tomato Plants to Salinity Stress: Evidence from the Syrian Coast

The research was carried out in Tartus (Syria), in a plastic greenhouse during the season 2024-2025. Two tomato hybrids "Levovil and Shannon" were grafted onto two tomato "Spirit and Maxifort" rootstocks. Four levels of salinity (0, 50, 100 and 150 mM NaCl) were applied on the hybrids and grafted plants. Salinty stress induced physiological and biochemical changes in plant. At salinity level of 150 mM, the absolute value of the osmotic pressure increased to -1.18 and -1.12 Mpa, in Levovil and Shannon hybrids, as compared to -0.80 and - 0.74 Mpa in the controls, respectively. While 150 mM salinity level led to an increase of the contents of dry matter, proline, sugars and chlorine, it caused a decrease of K: Na ratio in the two hybrids (1.99 and 1.8) as compared to the controls (5.29 and 4.68, respectively). The salinity stress at 150 mM, reduced the yield of the two hybrids (26.26 and 25.28 kg/m2) as compared to the controls (33.90 and 33.11 kg/m2, respectively). The tomato scions "Levovil and Shannon hybrids" grafted onto the "Spirit and Maxifort" rootstocks enhanced the osmotic adjustment phenomenon, especially when the two hybrids were grafted onto Spirit. While tomato grafting had a significant effect on the accumulation of some osmotic compounds such as proline and sugars, in addition to increasing K: Na ratio, it reduced the absolute value of the osmotic pressure of the plant and its sodium content. Therefore, the yield increased in grafted tomato plants compared with non-grafted plants under saline conditions. The results of PCA analysis showed that the first two principal components (PC1 and PC2) explained 89.08% of the total variation, wherease all the studied parameters were well represented. The dendrogram was obtained based on the same variables as PCA, and it led to classify "Spirit" rootstock as tolerant to salt stress, "Maxifort" rootstock as half tolerant, and the non-grafted hybrids "Levovil and Shannon" as sensitive to salt stress.

physiology↗

ZNF121 recruits YTHDF2 to modulate mRNA stability

N6-methyladenosine (m6A) is the most abundant internal modification in mRNA and has been shown to regulate gene expression through the binding of specific reader proteins, such as YTHDF2, which promotes mRNA decay. Previous studies indicate that YTHDF2 has relatively weak intrinsic RNA-binding affinity, suggesting that additional factors may facilitate its association with target transcripts. Here, we show that the C2H2-Zinc Finger protein, ZNF121, binds mRNA in cells and physically interacts with YTHDF2 in the cytoplasm. We demonstrate that approximately 80% of ZNF121-bound mRNAs are also YTHDF2 targets, and that their binding sites highly correlate with each other. Loss of ZNF121 impairs YTHDF2 binding to shared targets and increases their stability, independent of the presence of m6A modifications. Moreover, co-regulated transcripts are enriched for cell-cycle-related pathways, and ZNF121 depletion leads to elevated expression of the oncogene MDM2, implicating ZNF121 in growth control and the DNA damage response. Our findings identify ZNF121 as a cofactor that enhances YTHDF2-mediated mRNA regulation and reveal a previously unknown layer of control in mRNA decay.

molecular biology↗

BiomarkerKB: FAIR and Integrated Biomarker Knowledge Connecting Biomolecular and Clinical Data Types

Biomarkers are essential tools for disease detection, risk assessment, therapeutic monitoring, and precision medicine. However, biomarker data are dispersed across heterogeneous resources, inconsistently reported in the literature, and rarely standardized for computational use. This fragmentation limits reproducibility, cross-study integration, and the discovery of novel biomarker and disease relationships. We developed BiomarkerKB, a knowledgebase designed to harmonize and integrate biomarker information under a standardized data model. The model follows the FDA-NIH BEST biomarker definition and captures both core fields (biomarker entity, disease/condition, exposure agent) and contextual metadata (specimen, biomarker role, evidence, provenance). Biomarker data and related annotations were either curated from publications or collected from public resources (e.g., OpenTargets, GWAS Catalog, ClinVar, CIViC, OncoMX) and were also contributed by the Common Fund Data Coordinating Centers and the Early Detection Research Network (EDRN). Standardization was achieved using ontologies and reference resources such as Disease Ontology, UBERON, UniProtKB, and HUGO Gene Nomenclature Committee (HGNC) gene symbols. BiomarkerKB data were ingested into a Neo4j-based knowledge graph and integrated with the Common Fund Data Ecosystem (CFDE) Knowledge Graph. The initial release of BiomarkerKB contains over 200,000 biomarker-disease associations spanning genes, proteins, metabolites, glycans, and chemical elements. The knowledge graph comprises more than 300,000 nodes and 1.2 million edges, enabling structured exploration of biomarker relationships within CFDE data as demonstrated through the knowledge graph query-based use cases presented in this study. A publicly accessible web portal (https://biomarkerkb.org) provides keyword search, filtering, data downloads, and access to graph visualization to support both researchers and computational analyses. BiomarkerKB addresses a critical gap in biomarker informatics by providing an integrated, FAIR (Findable, Accessible, Interoperable, and Reusable), and unified framework for biomarker knowledge exploration and discovery.

bioinformatics↗

mRNAfold: Co-optimization of Global Stability, Local Structure, and Codon Choice via Suboptimal Folding

mRNA medicines hold great promise, but designing sequences with high translation efficiency, robust in-solution stability, and manufacturability remains a major challenge due to the vast combinatorial space of synonymous coding sequences. Computational approaches such as mRNA folding algorithms have emerged as powerful tools by co-optimizing for in-solution stability and translation efficiency, yet current methods face important limitations. Here, we present "mRNAfold", an improved mRNA folding algorithm and software package that addresses these gaps by enabling efficient exploration of diverse near-optimal solutions, incorporating untranslated regions (UTRs), parallel execution, and supporting tunable control over local structural features across the mRNA. Thermodynamically optimized mRNAs from mRNAfold were more stable ({approx} 2-fold) in-solution than those generated by simple GC maximization for the same encoded protein. In addition, mRNAs designed to vary local structure near the start codon while maintaining consistent structure and codon optimality elsewhere showed a complex relationship between local structure near the start codon and protein production in cells. We observed no impact of structure in the start codon region for a set of mRNAs with high codon optimality, but it did impact protein production for a set of mRNAs with lower codon optimality. Together, these results underscore the potential of structure-aware, multi-objective design to improve mRNA medicines and offer a framework for exploring how sequence, structure, and expression are interrelated.

bioinformatics↗

Spatially precise neuron formation via hydrogel mediated modulation of the host astrocyte response

The regenerative capacity of the central nervous system after injury or disease is limited. Advancements in genetic and epigenetic reprogramming of non-neuronal cells into induced neurons presents a promising strategy for neuronal replacement and circuit reconstruction yet is hindered by suboptimal reprogramming efficacy. Here, we have achieved high astrocyte-to-neuron reprogramming efficacy through the ectopic expression of a SOX2 transcription factor using adeno-associated viral vectors. We subsequently designed a hybrid composite biomaterial to act as an implantable reprogramming workshop that perform sequential operations. Once injected, this bespoke system forms a tissue mimetic hydrogel encouraging the ingress of astrocytes to act as our raw material. We demonstrate that the system localized and enhanced the spatially confined delivery of reprogramming factors, entrapping astrocytes to then pass through an intermediate neuroblast state to robustly yield mature neurons. Sustained delivery of valproic acid within the system further promoted neuronal maturation. Overall, using design rules informed by requirements of the reprogramming strategy, we have designed a smart delivery system optimized for both the target tissue and reprogramming factors in a minimally invasive manner to enhance neural repair. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=163 HEIGHT=200 SRC="FIGDIR/small/695501v1_ufig1.gif" ALT="Figure 1"> View larger version (65K): org.highwire.dtl.DTLVardef@397e41org.highwire.dtl.DTLVardef@90047forg.highwire.dtl.DTLVardef@1175831org.highwire.dtl.DTLVardef@19f30eb_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

Cold-responsive MtCBF4-MtJMJ13 positive feedback loop negatively regulates anthocyanin biosynthesis in Medicago truncatula

Legumes, being significant food sources for both humans and animals globally, experience several adverse conditions that considerably affect their crop yield. Medicago sp., an important forage crop, thrives under adverse conditions, such as drought, extreme temperatures (heat and cold), and salinity. Several studies have shed light on the regulatory pathways of cold/chilling stress tolerance in Medicago sp. in recent years. In the present study, we found that a cold-repressed transcription factor, MtCBF4, negatively regulates anthocyanin synthesis by directly repressing the expression of MtLAP1, a critical regulatory gene for anthocyanin synthesis in Medicago truncatula. It leads to the downregulation of anthocyanin biosynthesis genes, and reduced anthocyanin accumulation. Apart from the transcriptional cascade, MtCBF4 exerts its effect through histone H3K27 trimethylation. It directly binds to the promoter of MtJMJ13, an H3K27me3 demethylase, to activate its expression, resulting in decreased H3K27 trimethylation. Furthermore, MtJMJ13 demethylates the gene body of MtCBF4 to positively regulate its expression, forming a positive feedback loop that functions in a cold-responsive manner. MtJMJ13 also demethylates the gene body of MtMYB2, MtKFB1, and MtKFB2 genes, which are negative regulators of anthocyanin biosynthesis. Altogether, we have unveiled a new regulatory loop involving a transcriptional regulator and a histone demethylase, operating together to limit anthocyanin production. This regulatory module is repressed under cold stress, leading to increased anthocyanin production and enhanced cold stress tolerance in M. truncatula.

plant biology↗

Modulation of serotonergic, dopaminergic and tubulin-associated pathways by tannin-free natural products in Caenorhabditis elegans reveals effective anthelmintic drug targets

Despite of the Southeast Asias rich medicinal flora, Bangladesh behinds in utilizing the Caenorhabditis elegans model for lead anthelmintic compound discovery. Moreover, target-specific resistance (TSR) and non-target-specific resistance (NTSR) surge have influenced the lead compounds bio-discovery over the past decades. To date, prior researches are limited to basic efficacy tests and conventional extraction protocol. Therefore, the study was designed to assess the anthelmintic potency of 19 tannin-free natural products (NPs) induced alterations of neurotransmission in C. elegans. The anthelmintic activity was assessed by examining the motility (head thrashes and body bends), mortality, egg hatch inhibition (EHI) and expression of cat-1, ser-1, dat-1, and tba-1 genes. Eleven (11) plant extracts, Azadirachta indica A. Juss., Cassia alata L., Portulaca oleracea L., Saraca asoca (Roxb.) W.J.de Wilde, Eleusine indica L. Gaertn., Persicaria hydropiper L. Delarbre, Foeniculum vulgare Mill., Clerodendrum infortunatum L., Linum usitatissimum L., Hibiscus rosa-sinensis L., and Vitex negundo L. revealed a significant neurobehavioral and developmental impairments in C. elegans. Persicaria hydropiper L. Delarbre @ 1{square}mg/mL, caused the lowest body bending (31{square}{+/-}{square}1.7, p{square}<{square}0.01), while Eleusine indica L. Gaertn. reduced head thrashing (66.3{square}{+/-}{square}3.0, p{square}<{square}0.01). L. usitatissimum L. exhibited the highest lethality (87.3{square}{+/-}{square}1.8%, p{square}<{square}0.01) and the LD{square}{square} values of Eleusine indica L. Gaertn. (0.40{square}mg/mL) and Linum usitatissimum L. (0.411{square}mg/mL) were the lowest. Interestingly, C. infortunatum L. exhibited the strongest EHI (97.5{square}{+/-}{square}3.0%, p{square}<{square}0.01). Gene expression analysis revealed a significant down-regulation of the dopaminergic and serotonergic pathway indicating the metabolic and reproductive disruptions. This pioneering study underscores the anthelmintic potency of native plant extracts, highlighting their neuromodulatory and developmental impact on C. elegans. Altogether, these findings suggest that the selected genes may serve as potential drug targets, paving the way for lead compounds identification and development to the advance anthelmintic drug discovery from the NPs.

biochemistry↗

The CFDE Workbench: Integrating Metadata and Processed Data from Common Fund Programs

The NIH Common Fund Data Ecosystem (CFDE) program was established to facilitate data accessibility and interoperability across multiple Common Fund (CF) programs and promote collaborations and new discoveries by combining data from different CF programs. The CFDE Data Resource Center (DRC) was tasked with developing two web-based portals: an Information Portal to serve information about the CFDE, and a Data Portal to host harmonized metadata and processed data contributed by participating CF Data Coordination Centers (DCCs) and other sources. To achieve these goals, the CFDE DRC developed the CFDE Workbench, a web-based platform that hosts highly processed data, metadata, tools, use cases, and analyses developed by the CFDE. The Crosscut Metadata Model (C2M2) and several other processed data formats are hosted by the CFDE Workbench, including set libraries (XMTs), Knowledge Graph (KG) assertions, and attribute tables. These processed data formats make information derived from CF programs more findable, accessible, interoperable, and reusable (FAIR) and artificial intelligence (AI)-ready for cross-DCC knowledge discovery. In addition to these processed data formats, several other assets are served by the Workbench, including Extract Transform Load (ETL) pipelines, OpenAPI-documented and SmartAPI-registered Representational State Transfer (REST) Application Programming Interfaces (APIs), entity pages such as gene, drug, disease, or tissue landing pages, Playbook Workflow Builder (PWB) metanodes which are codified workflow steps involving DCC APIs, and chatbot specifications which are prompts helping chatbots to use DCC APIs to answer a wide range of user queries. Besides serving data, metadata, and code assets, the CFDE Workbench also has several tools that use these data, metadata, and code assets to enable cross-CF-program knowledge discovery use cases. Overall, the CFDE Workbench is a platform that consolidates efforts toward making CF programs funded resources harmonized, FAIR, and AI-ready. The CFDE Workbench website is available from: https://cfde.info/.

bioinformatics↗

Description of two novel Corynebacterium species isolated from human nasal passages and skin

Strains of two novel Corynebacterium species were cultured from samples of human nostrils and skin collected in the United States and Botswana. These strains demonstrated growth on Columbia Colistin-Nalidixic Acid agar with 5% sheep blood and in liquid media (brain heart infusion and tryptic soy broth) supplemented with Tween 80, a source of the fatty acid oleic acid. Cells were Gram-positive, non-spore-forming, non-motile bacilli that showed catalase but not oxidase activity. Major fatty acids in both of these species were 18:1 {omega}9c (oleic acid), 16:0 (palmitic acid), and 18:0 (stearic acid). Analysis of the 16S ribosomal RNA gene sequences identified these strains as belonging to the genus Corynebacterium (family Corynebacteriaceae). Whole-genome sequencing revealed that these strains formed distinct branches on a phylogenomic tree, with C. tuberculostearicum being the closest relative but with average nucleotide identities of < 95% relative to all previously described species. These results indicate that these strains represent novel species of Corynebacterium, for which we propose the names Corynebacterium hallux sp. nov., with the type strain CTNIH22T (=ATCC TSD-435T=DSM 117774T), and Corynebacterium nasorum sp. nov., with the type strain KPL3804T (=ATCC TSD-439T=DSM 117767T). We also describe the characteristics of two strains isolated from human nasal passages that are members of the recently named species Corynebacterium yonathiae. RepositoriesThe sequencing files supporting the conclusions of this study are available in the Sequence Read Archive (PRJNA804245, PRJNA854648, PRJNA842433). The partial 16S ribosomal RNA gene sequences from PCR amplification and Sanger sequencing are available in GenBank for Corynebacterium hallux sp. nov. CTNIH22T (accession number: PQ252679) and Corynebacterium nasorum sp. nov. KPL3804T (accession number: PQ149068). The annotated genomic sequences for the strains characterized in this study have been deposited in GenBank with the following accession numbers: C. hallux sp. nov. CTNIH22T (GCF_032821755.1), C. nasorum sp. nov. KPL3804T (GCF_037908315.1), C. nasorum sp. nov. MSK185 (GCF_030229765.1), C. yonathiae KPL2619 (GCF_037908465.1), and C. yonathiae MSK136 (GCF_022288805.2).

microbiology↗

ACSS2 regulates ferroptosis in an E2F1-dependent manner in breast cancer brain metastatic cells

Brain metastasis diagnosis in breast cancer patients is considered an end-stage event. The median survival after diagnosis is measured in months, thus there is an urgent need to develop novel treatment strategies. Breast cancers that metastasize to the brain must adapt to the unique brain environment and are highly dependent on acetate metabolism for growth and survival. However, the signaling pathways that regulate survival in breast cancer brain metastatic (BCBM) tumors are not known. Primary brain tumor cells can convert acetate to acetyl-CoA via phosphorylation of acetyl-CoA synthetase 2 (ACSS2) by the cyclin-dependent kinase-5 (CDK5) regulated by the nutrient sensor O-GlcNAc transferase (OGT). Here, we show that breast cancer cells selected to metastasize to the brain contain increased levels of O-GlcNAc, OGT and ACSS2-Ser267 phosphorylation compared to parental breast cancer cells. Moreover, OGT and CDK5 are required for breast cancer cell growth in the brain parenchyma in vivo. Importantly, ACSS2 and ACSS2-S267D phospho-mimetic mutant are critical for in vivo breast cancer growth in the brain but not in the mammary fat pad. Mechanistically, we show that ACSS2 regulates BCBM cell survival by suppressing ferroptosis via regulation of E2F1-mediated expression of anti-ferroptotic proteins SLC7A11 and GPX4. Lastly, we show treatment with a novel brain-permeable small molecule ACSS2 inhibitor induced ferroptosis and reduced BCBM growth ex vivo and in vivo. These results suggest a crucial role for ACSS2 in protecting from ferroptosis in breast cancer brain metastatic cells and suggests that breast cancer brain metastatic cells may be susceptible to ferroptotic inducers.

cancer biology↗

Typhi Mykrobe: fast and accurate lineage identification and antimicrobial resistance genotyping directly from sequence reads for the typhoid fever agent Salmonella Typhi

BackgroundTyphoid fever results from systemic infection with Salmonella enterica serovar Typhi (Typhi) and causes 10 million illnesses annually. Disease control relies on prevention (water, sanitation, and hygiene interventions or vaccination) and effective antimicrobial treatment. Antimicrobial resistant (AMR) Typhi lineages have emerged and become established in many parts of the world. Knowledge of local pathogen populations informed by genomic surveillance, including of lineages (defined by the GenoTyphi scheme) and AMR determinants, is increasingly used to inform local treatment guidelines and to inform vaccination strategy. Current tools for genotyping Typhi require multiple read alignment or assembly steps and have not been validated for analysis of data generated with Oxford Nanopore Technologies (ONT) long-read sequencing devices. Here, we introduce Typhi Mykrobe, a command line software tool for rapid genotyping of Typhi lineages, AMR determinants, and plasmid replicons direct from sequencing reads. ResultsWe validated Typhi Mykrobe lineage genotyping by comparison with the current standard read mapping-based approach and demonstrated 99.8% concordance across nearly 13,000 genomes sequenced with Illumina platforms. For the few isolates with discordant calls, we show that Typhi Mykrobe results are better supported by the evidence from raw sequence read data than the results generated using the mapping-based approach. We also demonstrate 99.9% concordance for detection of AMR determinants compared with the current standard assembly-based approach, with similar results for plasmid marker detection. Typhi Mykrobe predicts clinical resistance categorisation (S/I/R) for eight drug classes, and we show strong agreement with phenotypic categorisations generated from reference laboratory minimum inhibitory concentration (MIC) data for n=1,572 Illumina-sequenced isolates (>99% agreement within one doubling dilution). We show strong concordance (>96% for genotype and >98% for AMR and plasmid) between calls made from ONT reads and those made from Illumina reads for isolates sequenced on both platforms (n =93 genomes). Typhi Mykrobe takes less than a minute per sample and is available at https://github.com/typhoidgenomics/genotyphi. ConclusionsTyphi Mykrobe provides rapid and sensitive genotyping of Typhi genomes direct from Illumina and ONT reads, although lower accuracy was observed for R9 ONT data. It demonstrated accurate assignment of GenoTyphi lineage, detection of AMR determinants and prediction of corresponding AMR phenotypes, and identification of plasmid replicons.

microbiology↗

Exploring Effector Protein Dynamics and Natural Fungicidal Potential in Rice Blast Pathogen Magnaporthe oryzae

Rice blast, caused by Magnaporthe oryzae, is a severe agricultural disease leading to significant global economic losses. Genetic and genomic investigations have identified crucial genes and pathways involved in its pathogenesis, particularly highlighting effector proteins like AvrPik variants and MAX proteins. These proteins interact with specific Pik alleles on rice chromosome 11, influencing host immune responses. This study focused on 35 plant-derived metabolites known for their antifungal properties, evaluating their potential as fungicidal agents against M. oryzae. Molecular docking analyses identified Hecogenin and Cucurbitacin E as highly effective binders to MAX40 and APIKL2A proteins, respectively, which are pivotal for fungal virulence and immune evasion. Molecular dynamics simulations further validated strong and stable interactions, affirming the therapeutic potential of these compounds. Additional assessments including Lipinskis rule of five criteria and toxicity predictions indicated their suitability for agricultural use. These findings underscore the promise of Hecogenin and Cucurbitacin E as lead candidates in developing novel fungicidal strategies against rice blast, offering prospects for enhanced crop protection and agricultural sustainability.

bioinformatics↗

Playbook Workflow Builder: Interactive Construction of Bioinformatics Workflows from a Network of Microservices

Many biomedical research projects produce large-scale datasets that may serve as resources for the research community for hypothesis generation, facilitating diverse use cases. Towards the goal of developing infrastructure to support the findability, accessibility, interoperability, and reusability (FAIR) of biomedical digital objects and maximally extracting knowledge from data, complex queries that span across data and tools from multiple resources are currently not easily possible. By utilizing existing FAIR application programming interfaces (APIs) that serve knowledge from many repositories and bioinformatics tools, different types of complex queries and workflows can be created by using these APIs together. The Playbook Workflow Builder (PWB) is a web-based platform that facilitates interactive construction of workflows by enabling users to utilize an ever-growing network of input datasets, semantically annotated API endpoints, and data visualization tools contributed by an ecosystem. Via a user-friendly web-based user interface (UI), workflows can be constructed from contributed building-blocks without technical expertise. The output of each step of the workflows are provided in reports containing textual descriptions, as well as interactive and downloadable figures and tables. To demonstrate the ability of the PWB to generate meaningful hypotheses that draw knowledge from across multiple resources, we present several use cases. For example, one of these use cases sieves novel targets for individual cancer patients using data from the GTEx, LINCS, Metabolomics, GlyGen, and the ExRNA Communication Consortium (ERCC) Common Fund (CF) Data Coordination Centers (DCCs). The workflows created with the PWB can be published and repurposed to tackle similar use cases using different inputs. The PWB platform is available from: https://playbook-workflow-builder.cloud/.

bioinformatics↗