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Ali, A.

Publications and source records attributed to Ali, A..

At least 37 records · Page 2Linked to original sources

Cellquant: a vibecoder's guide to image analysis

Quantitative fluorescence microscopy is central to modern cell biology, yet extracting reproducible measurements from images remains a bottleneck for biologists without programming experience. Here we present cellquant, a single-script command line pipeline for multi-channel fluorescence images that performs cell segmentation, puncta quantification, colocalization analysis, and spatial proximity measurements. Because the interface is entirely text based, the exact command used to generate any result can be recorded and re-executed. We validate cellquant on two biological systems. In human HCT116 cells, the pipeline quantified arsenite-induced stress granule formation. In budding yeast, simultaneous measurement of nucleolar morphology, colocalization, and spatial proximity across a temperature gradient revealed a coordinated sequence of nucleolar reorganization. Applying PCA and UMAP to the multi-parameter output of cellquant resolved a continuous cell state transition across the temperature gradient, with condensate redistribution and nucleolar morphology defining orthogonal axes. The pipeline produces publication-ready quantification with visual quality control and statistically rigorous replicate analysis. All code, documentation, and example datasets are freely available.

cell biology↗

Discrete Transcriptional States Define Biphasic Immune Response and Dynamic CMS Transitions in Colorectal Cancer

BackgroundSequential alterations in APC, KRAS, TP53, and SMAD4 have been proposed as a framework for colorectal cancer progression. Human colorectal cancer datasets have not revealed the biological transitions associated with these mutations. When examining a cohort of TCGA-colorectal tumors grouped as AK (APC/KRAS), AKP (APC/KRAS/TP53), and AKPS (APC/KRAS/TP53/SMAD4), we observed no significant differences in immune-cell composition, four previously defined Consensus Molecular Subtypes (CMS1/2/3/4), or transcriptomic clustering between these genomic groups. Therefore, these canonical alterations do not sufficiently characterize the known properties of metastatic progression in human colorectal cancer. MethodsTo overcome these limitations, we developed a genetically defined, organoid-based, orthotopic mouse model whereby mouse colon organoids modeling sequential APC, KRAS, TP53, and SMAD4 alterations were orthotopically injected into the colon. This was followed by RNA-sequence processing, normalization with DESeq2, differential expression, pathway enrichment, and immune/stromal inference. Gene co-expression modules were identified from variance-stabilized mouse expression data, mapped to 1:1 human orthologs, and summarized as eigengenes. A multinomial logistic regression model trained on mouse eigengenes was applied to TCGA-COAD human tumors to assign them to mouse-informed transcriptomic states (AK-like, AKP-like, AKPS-like), which were then used for downstream visualization and comparative analyses. ResultsWhole-transcriptome analysis revealed discrete transcriptional states and immune-cell differences between the organoid AK/AKP/AKPS groups. Early TP53 loss led to strong activation of immune pathways, accompanied by increased infiltration of NK and T cells. As tumors progressed with SMAD4 loss and metastasis, this immune activity collapsed, giving rise to broad immune suppression. CMS classifications also shifted, with AK tumors resembling epithelial CMS2, AKP tumors displaying immune-rich CMS1 features, and AKPS and metastatic lesions adopting mesenchymal CMS4 characteristics. We then applied a progression-based transcriptomic classifier to 460 human colorectal tumors. This reclassification revealed conserved immune remodeling, CMS transitions, pathway-level differences, and significant differences in patient survival. ConclusionWe show that organoid-derived progression profiles reveal hidden evolutionary structure in human colorectal cancer and provide a transcriptional framework for interpreting metastatic potential and clinical outcomes.

cancer biology↗

An ex vivo gut mucosal explant assay to compare HIV tissue susceptibility shows increased HIV susceptibility with methamphetamine exposure

BackgroundExisting ex vivo mucosal explant models for HIV assess viral replication but do not fully capture the ability of tissue to support productive infection and transfer to target cells, an important indicator of tissue susceptibility. Additional tools to investigate relative mucosal susceptibility to HIV are necessary to better understand risk factors, such as substance use, or to evaluate therapeutics. ResultsWe developed an ex vivo gut mucosal explant assay incorporating a CCR5/CXCR4-expressing GFP-reporter cell co-culture to quantify both viral replication and transfer. Susceptibility was quantified using a composite metric based on time to infection endpoint across viral doses. Modulators of susceptibility, including anti-CD3 stimulation, tenofovir, and methamphetamine, were evaluated. The assay reliably detected differences in tissue susceptibility under various experimental conditions. We detected enhanced susceptibility following anti-CD3 stimulation, while tenofovir pretreatment reduced susceptibility in a dose-dependent manner. Methamphetamine exposure resulted in a modest but significant increase in gut tissue susceptibility to HIV. ConclusionsThis novel ex vivo explant susceptibility assay advances HIV mucosal transmission research by measuring both viral production and transfer. It can detect subtle changes in susceptibility, providing a versatile platform for evaluating prevention therapeutics, host and microbial factors, and substance use effects.

microbiology↗

Platelet-derived CXCL7 induces neutrophil extracellular traps via CXCL7/CXCR2 axis, exacerbating the pathogenesis of diabetic retinopathy

BackgroundProlonged hyperglycemia in diabetes activates platelets and immune cells, forming platelet-immune complexes that damage blood vessels in the retina. However, the role of platelet-neutrophil interactions and neutrophil extracellular traps(NETs) in the development of diabetic retinopathy (DR) was not well studied. In this study, we investigated the mechanisms underlying platelet-mediated NET formation in DR. MethodologyPlatelet activation markers, platelet-neutrophil aggregates (PNA), and NETs markers were assessed by flowcytometry, and the circulatory level of inflammatory markers was measured by Luminex assays in healthy control(HC), type 2 diabetes mellitus(T2DM), non-proliferative DR(NPDR) and proliferative DR(PDR) subjects. In vitro studies investigated platelet-neutrophil interaction in NETs formation using an immunofluorescence assay. Proteomics analysis identified the mechanistic regulators of platelet-induced NETs in DR. Platelet pellet and plasma CXCL7 were quantified using western blot and ELISA, respectively. The role of the CXCL7/CXCR2 axis in inducing NETs formation was examined using CXCL7 recombinant protein, anti-CXCL7 antibody and CXCR2 antagonist (SB225002). ResultsPlatelet activation markers (p-selectin & PF4), PNA, and NETs markers (%NETs, proteinase-3 (PR3), neutrophil elastase (NE)) were significantly increased in the DR group. In vitro studies confirmed that DR-platelets aggregate with healthy neutrophils and form NETs compared to T2DM and HC-platelets. Furthermore, platelet activation and NETs markers were positively correlated with pro-angiogenic (ANGPT2, VEGFA) and inflammatory markers (IL18, ICAM1). In vitro studies reveal that NETs induce inflammation, endothelial dysfunction, disrupt the endothelial monolayer and exacerbate angiogenesis in RF/6A endothelial cell spheroids. Proteomics analysis of platelet-induced NETs in DR revealed dysregulation of proteins involved in platelet activation and NET formation, including CXCL7. Furthermore, increased CXCL-7 levels were observed in platelet pellet and plasma samples from the DR group. Additionally, CXCL7-treated neutrophils formed NETs via the CXCR2 receptor, and inhibition of NETosis was observed in neutrophils exposed to an anti-CXCL-7 antibody and a CXCR2 antagonist. ConclusionOur findings revealed that platelets released CXCL-7 induce NETs formation via the CXCL7/CXCR2 axis and blockade of CXCL7/CXCR2 axis inhibits the NETosis in DR, thereby inhibiting the pathogenesis of DR. Circulating CXCL7 serves as a potential prognostic marker, and the CXCL7/CXCR2 axis may be a therapeutic target for the treatment of DR. What Are the Clinical Implications?Platelets have emerged as immune cells, and platelet-neutrophil interactions are reported to play a significant role in the pathogenesis of various metabolic diseases. The role of platelet-neutrophil interactions and neutrophil extracellular traps (NETs) in the development of diabetic retinopathy (DR) remains poorly understood. Investigating the mechanistic regulators of platelet-mediated NETs in DR is crucial for identifying new therapeutic approaches. Our study observed increased platelet activation, platelet-neutrophil aggregates and NETs among DR subjects. Further, DR-platelet induces NET in healthy neutrophils, and NETs induce inflammation, angiogenesis, and disrupt endothelial barrier function in RF/6A cells in vitro. These findings strengthen the evidence that platelet-neutrophil interactions play a major role in DR pathogenesis. Proteomic analysis identified CXCL7 as a mechanistic regulator of platelet-induced NETs formation in DR. Inhibitors that target the platelet-derived CXCL7/CXCR2 axis for NETosis can be used for the prevention of retinal injury in DR. Overall, our work emphasises the mechanistic understanding of platelet-neutrophil interactions and CXCL7/CXCR2 axis as a therapeutic target for inhibiting the pathogenesis of DR. Graphical abstractScheme for the platelet-derived CXCL7 regulation of NETs in DR. Activated platelets release CXCL7 and platelet aggregates with neutrophils to form NETs via the CXCL7:CXCR2 axis. Blockade of the CXCL7:CXCR2 axis by anti-CXCL7 antibody and CXCR2 inhibitor prevents NETs formation in DR.

immunology↗

A Course-Undergraduate Research Experience (CURE) to explore the effect of structural variants on gene expression in C. elegans balancers

Bioinformatics, a discipline at the crossroads of Biology and Computational Sciences, also referred to as Computational Biology, is nowadays widely spread in research programs. However, implementing any Bioinformatics projects requires the ability to comprehend biological concepts and apply computational approaches, and rare are the undergraduate programs offering such multi-disciplinary training. In addition, understanding the dynamic between Biology research projects and Bioinformatics analyses is challenging with no real-life experience. Course-based undergraduate research experience (CURE) courses are innovative programs that allow more students to acquire research experience and provide the perfect setting to introduce students to applied bioinformatics. As a part of the Bachelor of Health Sciences of the Cumming School of Medicine at the University of Calgary (Canada), a CURE applied bioinformatics was implemented in the Winter of 2023 to 2025. Students investigated the effect of structural variants (SVs, genetic variants larger than 50 bp) on gene expression in the model organism Caenorhabditis elegans (a hermaphrodite 1-mm long roundworm). The students detected and characterized SVs by analyzing genome and transcriptome sequencing data of C. elegans strains called balancers, as they are known to carry large genomic variations balancing regions of the genome by limiting recombination and allowing maintenance of lethal mutations. They used Galaxy, a public web-based supercomputing resource, but also a local High-Performance computing system, and R, to report different effects of SVs on gene expression and splicing. Students research explained the molecular mechanism behind the uncoordinated phenotype caused by the reciprocal translocation eT1(III;V) and uncovered unexpected effects on gene expression on an understudied gene. We evaluated the courses impact on student learning journeys and showed that the CURE favored students understanding of the Bioinformatics field and fostered their research interest. We provide here guidelines to facilitate the CURE implementations to improve access for undergraduate students to bioinformatics research experiences.

bioinformatics↗

Spatial transcriptomic analysis of progressing oral epithelial dysplasia reveals unique differentially expressed genes and microenvironmental changes.

Oral squamous cell carcinoma (OSCC) often arises from oral epithelial dysplasia (OED); however, the gene expression changes during OED progression and its microenvironment are not fully understood. This study used spatial transcriptomics to identify differentially expressed genes and microenvironmental alterations associated with OEDs malignant transformation of OED. A ten-year retrospective analysis of paired OSCC and prior OED samples was conducted at the University of Toronto Oral Pathology Laboratory. A total of 24 paired progressing OED cases and 23 matched non-progressing OED cases were examined using spatial transcriptomics in PanCK+ (dysplastic epithelium or OSCC) and PanCK- (stroma) regions. The analysis included differential gene expression, pathway analysis and spatial deconvolution. Three genes (STOM, KIF26A, and CDKN2A) showed increased expression in the epithelial component of progressing OED compared with non-progressing OED, whereas 41 genes were differentially expressed in OSCC versus the precursor samples. Ubiquitination-related pathways were enriched during OED progression. Functional validation identified TNFRSF12A (Fn14) as a potential regulator of OED progression to OSCC. The OSCC microenvironment displayed increased numbers of fibroblasts, neutrophils, monocytes, and mast cells compared with that of the precursor samples. Our findings suggest that spatial profiling of OED can help identify unique gene signatures and microenvironmental changes that occur before the malignant transformation.

cancer biology↗

Comparative metabolomics of released pollen during dispersal reveals metabolic adaptations to cold and heat stress

Heat and cold stress can severely impair released pollen, reducing pollen viability and ultimately limiting fertilization and crop productivity. Following release, pollen is directly exposed to fluctuating environmental temperatures, necessitating adaptive metabolic mechanisms to sustain viability during dispersal. In response to temperature stress, pollen undergoes metabolic reprogramming that supports biochemical and physiological adaptation. However, the metabolic basis of pollen tolerance to extreme temperatures remains incompletely understood. In this study, biochemical assays combined with LC-MS-based untargeted metabolomic profiling were employed to investigate metabolic changes in released pollen exposed to low (15 {degrees}C) and high (35 {degrees}C) temperature stress. A total of 484 metabolites were detected, of which 147 were significantly altered between cold- and heat-stressed pollen, including 61 upregulated and 86 downregulated metabolites. Differentially regulated metabolites spanned multiple classes, including amino acids, flavonoids, long-chain fatty acids, sugars, and polyamines. Pathway enrichment analysis highlighted biologically relevant perturbations in amino acid and nucleic acid metabolism, including purine metabolism, arginine biosynthesis, and glutathione metabolism. Collectively, these findings demonstrate temperature-dependent metabolic adjustments in released pollen and provide new insights into the biochemical strategies underlying pollen tolerance to heat and cold stress. This work advances our understanding of pollen metabolic adaptation during dispersal and provides a foundation for identifying metabolic indicators relevant to crop fertility under changing climatic conditions. These findings provide a metabolomic framework for understanding pollen thermotolerance during dispersal and offer a resource for future studies on reproductive resilience under climate change.

plant biology↗

Assessing Drought Resilience and Identification of High Yielding Upland Rice Varieties through Phenology, Growth and Yield Traits

Drought stress is the major yield limiting factor in upland rice production where the moisture availability is highly variable. Understanding and evaluating how upland rice responds to drought stress is critical to improving resilience and yield stability. In this study performance of sixteen upland rice varieties were evaluated under non-stressed, moderately stressed and highly stressed conditions. Drought stress was introduced by irrigating upland rice at 70% and 50% field capacity (FC) whereas non-stress treatment was irrigated at 100% FC. Irrigation in moderately stressed and highly stressed conditions was also withheld for six days at lateral crop stages to observe temporary wilting by inducing a stress interval. Data on agronomic traits of upland rice was collected in three experimental replications. Results indicated that performance of upland rice varieties was significantly altered under stress conditions and highest performance was observed under non-stressed conditions. Yield losses for short duration and long duration varieties ranged 35-60% and 24-62% under moderate stress whereas it ranged 43-78% and 52-73% under highly stressed conditions, respectively. Overall varieties Dawk Kha, Khao/ Sai and Dawk Pa-yawm, indicated higher stability under stressed conditions therefore, these long duration varieties could be used for obtaining better yields under diverse agroclimatic conditions and under unpredicted weather patterns. Short duration Ma-led-nai-fai and long duration Goo Meung Lung and Bow Leb Nahag could be used for acquiring traits for higher tillering and panicle bearing capacity. Short heighted varieties such as Jao Daeng, Sahm Deuan and Ma-led-nai-fai could be used in breeding for short heighted new varieties to overcome lodging concerns. Strong significant association of GMP, STI, MPRO, MHAR with grain yield under non-stressed, moderately stressed and highly stressed conditions indicated that these indices were appropriate for their use as selection criteria for drought resilience.

plant biology↗

Organelles harbour pH gradients

Organelle pH is critical to organelle identity and function. Resident proteins that define each organelle modify transiting cargo proteins, with both retention and trafficking between organelles governed by pH-dependent mechanisms. For example, lysosomal enzymes bind mannose-6-phosphate receptors at the higher pH ([~]6.5) of the Golgi and dissociate at the lower pH ([~]5.5) of late endosomes1. Proteins that stray from the endoplasmic reticulum (ER) are captured by KDEL receptors in the acidic Golgi and returned into the neutral ER2,3. This pH-tuned trafficking system compartmentalizes organelle function and prevents mis-localization of critical enzymes4. Dysregulated organelle pH disrupts their function and leads to various diseases. Because protons move rapidly in water, the pH within a single organelle is currently assumed to be spatially uniform5. Here, using a reporter sensitive from pH 5.5 - 10.5 to map a spectrum of organelles at high resolution, we discovered that pH gradients exist within single, large or long organelles such as the ER and mitochondria, and in membrane-less organelles without ion-transporting proteins such as the nucleolus. These new findings upend our understanding of organellar pH, prompting new questions about proton diffusion within the cell, and its potential consequences on organelle function.

cell biology↗

Sleep disturbance is associated with increased repetitive sensorimotor behaviour in autism but not in learning/intellectual disabilities or ADHD: A comparative study

The interplay of sleep and sensorimotor processing is a fundamental neurobiological process crucial for well-being. Individuals with neurodevelopmental conditions like autism, ADHD, and learning disabilities frequently experience both significant sleep disturbance and repetitive sensorimotor behaviours. Although a link between these domains is established, it is unclear whether this relationship and its functional impact are consistent across different neurodevelopmental conditions. In a cross-sectional dataset of individuals aged 5-21 (n = 212), we investigated whether the interplay between sleep disturbance and repetitive sensorimotor behaviours differs between individuals diagnosed with autism (IQ>70), ADHD, a learning disability (IQ<70), as well as typically developed controls. We also examined whether interactions between sleep and repetitive sensorimotor behaviours predicts distinct functional outcomes between neurodevelopmental groups. We found that individuals with autism (IQ>70) were most strongly affected by both sleep disturbance and repetitive sensorimotor behaviours. Crucially, we show that while a robust bidirectional link between sleep and repetitive sensorimotor behaviours exists across the entire sample, sleep problems amplify repetitive sensorimotor behaviours more strongly only in autistic people than in typical controls. Contrary to previous work, we found that learning disabled children and adolescents engaged in more repetitive sensorimotor behaviour but did not experience more sleep disturbance. Although the interaction between sleep problems and repetitive sensorimotor behaviours did not determine functional outcomes in autistic people, the interplay of sleep problems and repetitive sensorimotor behaviour impaired social awareness in people with ADHD. These findings challenge a transdiagnostic view that treats the reinforcement between sleep and sensorimotor problems as a general marker of neurodevelopmental conditions. Understanding the distinct pathways between sleep and waking behaviours is a critical step toward developing personalized, mechanistic interventions targeting specific sleep and sensory impairment profiles in different neurodevelopmental conditions.

neuroscience↗

Integrated proteogenomics uncovers ancestry-specific and shared molecular drivers in localized prostate cancer

Our integrative proteogenomics (genome, proteome and phosphoproteome) of localized prostate cancer (PCa) in an equal-access Military Health System patient cohort (57 Black and 55 White) revealed significant ancestry-associated differences. Somatic and germline regulatory differences converged on androgen, metabolic, PI3K/AKT/mTOR, and DNA damage response (DDR) pathways, with ancestry-specific immune- and stromal-associated signals. Black patients displayed greater genomic variability, enhanced androgen response, fatty-acid metabolism, and epithelial-mesenchymal transition, while White patients showed prevalent DDRG alterations, activated oncogenic signaling (MYC, E2F, mTORC1), and cell cycle regulation. Phosphoproteomics highlighted distinct kinase activities and candidate druggable dependencies. Multiomics integration revealed three exploratory tumor subtypes whose distinct biological programs were reproducibly validated. Ancestry-associated eQTLs supported inherited regulation of the proteome independent of CNAs. Ancestry-specific CNA and protein panels improved progression risk prediction beyond PSA and pathology models. These findings provide a framework for ancestry-informed prognostic models and generate testable hypotheses for precision therapies to reduce outcome disparities.

cancer biology↗

Morphological Identification of Ticks in district Swabi, Mardan, and Charsadda

Ticks are ectoparasitic arthropods that play a significant role in transmitting pathogens to humans and animals. This study aimed to address the research gap in understanding the morphological diversity and distribution of tick species in Swabi, Mardan, and Charsadda districts of Pakistan, where tick-borne diseases are prevalent but poorly documented. The primary objectives were to identify tick species, document their morphological characteristics, and map their distribution patterns across different habitats. Field surveys were conducted over nine months, collecting tick specimens from forests, grasslands, and residential areas. Specimens were preserved and analyzed using stereo-zoom microscopy and taxonomic keys for accurate classification. Key findings revealed a diverse tick fauna, including hard ticks (Ixodidae), with variations in body size, colour patterns, capitulum shape, and scutum features. Some species were widely distributed across all three districts, while others exhibited localized patterns. The study provides the first comprehensive morphological identification of tick species in these regions, highlighting their potential role in disease transmission. The implications of this research are significant for public health and disease control. The data can inform targeted interventions, enhance disease surveillance programs, and support the development of preventive measures to mitigate tick-borne diseases. By contributing to tick taxonomy and epidemiology, this study lays the groundwork for protecting public health in the study regions and beyond. This study addresses the lack of data on tick diversity in high-prevalence regions of Pakistan, providing critical insights for disease control and public health strategies. Limited information exists on the morphological diversity and distribution of tick species in Swabi, Mardan, and Charsadda districts. To identify tick species, document morphological characteristics, and map their distribution patterns in the study areas. Field surveys, specimen collection, stereo-zoom microscopy, and taxonomic classification were employed over nine months. Diverse tick fauna was identified, including hard ticks, with variations in morphology and distribution. Some species were widespread, while others were localized, highlighting potential disease transmission risks. The findings support targeted interventions, disease surveillance, and preventive measures, contributing to public health protection and tick-borne disease control in the region.

zoology↗

Myeloid cell protein tyrosine phosphatase 1B (PTP1B) drives retinal neurodegeneration in diabetic mice

Diabetic retinopathy (DR) is the leading cause of vision loss in the working age population with public health economic implications worldwide. Systemic inflammation and leukocyte activation are early events in diabetes, while microglial activation, neuroinflammation, and retinal neurodegeneration are early events in DR. Protein tyrosine phosphatase 1B (PTP1B) plays a complex role in monocyte / macrophage activation which may impact DR. We therefore investigated the role of myeloid cell-specific PTP1B using LysMcre-PTP1B fl/fl (LysM-PTP1B) mice, as well as a PTP1B inhibitor, MSI-1436, in the early stages of DR. Mice were rendered diabetic for six weeks using anomer-equilibrated streptozotocin (STZ). Retinal changes were evaluated by histology and immunohistochemistry, and systemic leukocyte activation by flow cytometry. Mitochondrial function in high glucose-challenged, cultured bone marrow-derived macrophages (BMDMs) from LysM-PTP1B and MSI-I436-treated mice was determined in vitro. Both myeloid cell-specific depletion and pharmacological inhibition of PTP1B prevent STZ-induced retinal neurodegeneration, development of acellular retinal capillaries, as well as microglial and systemic leukocyte activation without effect on development of diabetes. In vitro, inhibition of PTP1B prevented high glucose-induced mitochondrial dysfunction in BMDMs. We conclude that inhibition of PTP1B prevents DR by decreasing myeloid cell-driven inflammation and PTP1B represents a therapeutic target for prevention DR. HighlightsO_LIMyeloid cell PTP1B is required to induce retinal neurodegeneration in diabetes C_LIO_LIBoth local (microglia) and systemic (bone-marrow derived) myeloid cells are implicated C_LIO_LIInhibition of myeloid cell PTP1B prevents development of acellular retinal capillaries in diabetic mice C_LIO_LIPTP1B mediates superoxide production, decreases mitochondrial membrane potential and increases macrophages cell death in chronic conditions associated with abnormally high glucose. C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=196 SRC="FIGDIR/small/682409v1_ufig1.gif" ALT="Figure 1"> View larger version (43K): org.highwire.dtl.DTLVardef@d8f71dorg.highwire.dtl.DTLVardef@47c8bforg.highwire.dtl.DTLVardef@cc800dorg.highwire.dtl.DTLVardef@1eade18_HPS_FORMAT_FIGEXP M_FIG C_FIG

molecular biology↗

Genetic Determinants of Heart Failure Susceptibility and Response in the Collaborative Cross Mouse Population

Genetic variation and lived experiences shape how our hearts respond to chronic stress. The specific genetic mechanisms which underly cardiac remodeling, however, are still unclear, due in part to the challenge of accounting for environmental effects in human population studies. To overcome this challenge, we used the Collaborative Cross (CC) mouse population to investigate heritable susceptibility to cardiovascular stress by chronic {beta}-adrenergic receptor stimulation. Across 8 founder and 63 CC lines, we measured cardiac structure and function, organ weights, cell and tissue morphology, and left ventricular gene expression. Genome-wide scans detected 49 genome-wide significant loci, collapsing to 20 unique intervals (nine significant for multiple traits and eleven trait-specific), averaging 12.83 Mb in size. To identify high-confidence candidate genes from these loci, we augmented our trait mapping with associations between loci and gene expression, isoproterenol-dependent transcriptional changes, coding variants drawn from sequencing data, tractability in our in vitro rat cardiomyocyte model, and previously reported protein functions and mouse or human phenotypes. This approach recovered both known regulators, such as Hey2, and new candidates. Functional tests in in vitro models highlight three candidate genes that modulate hypertrophic growth: Abcb10, Mrps5 and Lmod3. Abcb10 knockdown increased cell size at baseline and further with isoproterenol, consistent with loss of a mitochondrial stress-buffering role. Mrps5 knockdown blunted stress-induced hypertrophy. Paradoxical upregulation of Lmod3 after siRNA transfection (validated at the protein level) also attenuated hypertrophy, consistent with reinforcement of actin-assembly control under catecholamine stress. Together, these results reveal heritable pathways of {beta}-adrenergic remodeling in mice and provide an interpretable, translational, and stepwise framework to prioritize candidate genes within broad loci for mechanistic studies of heart failure.

genomics↗

The two sides of resistance: aggressiveness and mitotic instability as the Achilles heel of Osimertinib-resistant NSCLC

Non-small cell lung cancer (NSCLC) represents majority of lung cancer cases and remains a leading cause of cancer mortality worldwide. Tumors carrying activating mutations in the epidermal growth factor receptor (EGFR) are highly sensitive to EGFR tyrosine kinase inhibitors (TKIs), with third-generation inhibitors such as Osimertinib now established as standard of care. However, acquired resistance to Osimertinib inevitably develops, involving both genetic and non-genetic mechanisms, the latter playing a major role in sustaining cellular plasticity and promoting tumor aggressiveness. Among regulators of adaptive programs, the Polycomb protein BMI1 has emerged as a key factor driving stemness, epithelial-to-mesenchymal transition (EMT), and therapy resistance in multiple cancers, yet its role in Osimertinib resistance remains poorly defined. Here, we show that Osimertinib-resistant H1975 cells, which display greater aggressiveness than their parental counterparts, are enriched in BMI1 target genes and mitotic cell-cycle pathways, establishing a dependency on microtubule dynamics and mitotic control. Functionally, BMI1 drives migration, invasiveness, and tumor progression in resistant cells. This mitotic dependency creates a therapeutic vulnerability that can be exploited with Unesbulin (PTC596), a BMI1 inhibitor that destabilizes microtubules and induces mitotic catastrophe, thereby effectively suppressing tumor growth in vitro and in vivo. Our findings establish BMI1 as a central mediator of Osimertinib resistance and provide a mechanistic and therapeutic rationale for targeting BMI1 and mitotic weaknesses in refractory EGFR-mutant NSCLC.

cancer biology↗

Metabolic Signatures of Pulmonary Embolism in COVID-19: Insights from Longitudinal Intensive Care Unit Profiles

ABSTRACTO_ST_ABSBackgroundC_ST_ABSPulmonary embolism is a severe complication of COVID-19 infection, associated with a hypercoagulable state and heightened risk of blood clots. As SARS-CoV-2 has become endemic, understanding pulmonary embolisms metabolic effects in COVID-19 patients is warranted. This study investigated the longitudinal metabolic profiles of 66 Intensive Care Unit-admitted COVID-19 patients at Erasmus Medical Center to identify metabolites and mechanisms associated with pulmonary embolism. MethodA total of 1209 metabolic species were measured, including amines and lipids. Metabolic changes were analysed across four timeframes: i) general analysis of pulmonary embolism, ii) 72 hours prior to pulmonary embolism, iii) 48 hours prior and the day of pulmonary embolism, and iv) the day of and 48 hours post-pulmonary embolism. ResultsThe general analysis revealed significant upregulation of amines, triglycerides, phosphatidylethanolamines, ether-linked phosphatidylethanolamines, and eicosanoids in patients who developed a pulmonary embolism. Phosphatidylethanolamines containing the 20:3 fatty acid side chain were notably elevated. Minimal metabolic dysregulation was observed 72 hours before pulmonary embolism, with subtle increases in lysophosphatidylcholines and lysophosphatidylethanolamines. In contrast, there was a strong metabolic response during and post-pulmonary embolism, phosphatidylethanolamines (47%), ether-linked phosphatidylethanolamines(96%) and sphingosines(40%). ConclusionThese findings underscore the critical role of lipid metabolism in pulmonary embolism, particularly triglycerides and specific lipid species. The limited metabolic perturbations before pulmonary embolism suggest early prediction challenges, emphasising the need for further research into temporal metabolic changes and their clinical applications.

molecular biology↗

DDIAS is a single-stranded DNA-binding effector of the TOPBP1-CIP2A complex in mitosis

DNA double-strand breaks and unresolved DNA replication intermediates are particularly dangerous during mitosis. Paradoxically, cells inactivate canonical DNA repair mechanisms during chromosome segregation in favor of alternative pathways that depend on TOPBP1 and CIP2A, but how they function is still poorly defined. Here, we describe the identification of DDIAS as a mitosis-specific DNA damage response protein. We establish DDIAS as a phosphorylation-dependent component and effector of the TOPBP1-CIP2A complex with single-stranded DNA (ssDNA)-binding activity, and thereby delineate a ssDNA protection mechanism that safeguards chromosome integrity during mitosis, particularly in BRCA-deficient cells. We also identify biallelic inactivating mutations in DDIAS in a patient with a severe neurodevelopmental disorder. These findings highlight for the first time a potential physiological role for the DNA damage response in mitosis.

molecular biology↗

Integrated Genomic Profiling Reveals Mechanisms of Broad Drug Resistance and Opportunities for Phenotypic Reprogramming in Cancer Cells

Broad drug resistance is a major barrier to effective cancer therapy, driven by diverse genetic, transcriptional, and metabolic adaptations across tumor types. Here, we developed an integrative computational framework that leverages PRISM drug sensitivity profiles from DepMap, multi-omic datasets, and perturbagen libraries to systematically characterize and identify strategies to reverse broad resistance in cancer cell lines. We found that resistant lines exhibit transcriptional programs enriched for extracellular matrix remodeling, stress adaptation, and survival signaling, with NFE2L2 emerging as a central regulatory hub linked to upstream mutations and downstream oxidative stress pathways. Integrated metabolomics and transcriptomics highlighted metabolic reprogramming as a hallmark of resistance, while mutation analyses revealed convergence on growth factor and ECM-related pathways. These features were also reflected in patient cohorts, where resistance-associated mutations correlated with reduced progression-free survival across diverse cancer types. Computational perturbagen screening identified candidate compounds predicted to reverse resistance-associated gene expression profiles, converging on actionable targets including NFE2L2, ABCB1, and CYP3A4, with compounds such as brefeldin A and nocodazole predicted to have strong activity in resistant lines. This study establishes a scalable, mechanism-informed framework for rationally identifying and prioritizing compounds to overcome broad drug resistance in cancer, providing a roadmap for targeted re-sensitization strategies.

cancer biology↗