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Sridhar, S.

Publications and source records attributed to Sridhar, S..

At least 19 recordsLinked to original sources

A Multi-Kinase Inhibitor Screen Identifies mTOR Signaling as a Vulnerability in Advanced Prostate Cancer.

The receptor tyrosine kinase AXL has context-dependent roles in prostate cancer (PCa). Although AXL signaling has been implicated in PCa progression, it can also promote disseminated tumor cell dormancy. Paradoxically, AXL expression has been reported to be lost in metastatic PCa. This raises two critical questions: Is AXL loss associated with aggressive disease, and does it create therapeutically exploitable vulnerabilities? In this study, we comprehensively analyzed AXL expression across pan-cancer and PCa cohorts and found that AXL downregulation was significantly associated with treatment resistance, aggressive PCa phenotypes, and shorter overall survival. To investigate the functional consequences of AXL loss, we used CRISPR-Cas9 based gene editing to generate isogenic AXLKO cells and characterized their proliferative and migratory properties. To identify therapeutically exploitable vulnerabilities associated with AXL loss, we screened these cells against a library of 1,814 kinase inhibitors using the isogenic cell lines. We identified Torin-2 as a candidate therapeutic agent and validated its activity using dose-response assays, which demonstrated an ~ 18-fold lower IC50 in AXLKO cells than in AXLWT cells. In parallel, PRISM/DepMap analysis provided orthogonal evidence of drug dependencies associated with low AXL expression and revealed a selective vulnerability of AXLKO cells to PI3K/AKT/mTOR pathway inhibition. Collectively, our findings link AXL downregulation to aggressive PCa phenotypes and identify mTOR inhibition as a potential therapeutic strategy for AXL-deficient PCa.

cancer biology↗

Brain pericytes exhibit spatially organized and dynamically regulated molecular heterogeneity

Brain capillary pericytes are essential components of the neurovascular unit, yet the extent of their molecular heterogeneity within intact vascular networks remains poorly understood. Here, we combined spatial imaging with reanalysis of independent single-cell transcriptomic datasets to investigate the molecular organization of adult mouse brain pericytes. We identified spatial organization of pericyte molecular heterogeneity associated with anatomical region and position within the vascular network, including recurrent differences in ACE2, CASQ2, Igf2, and OPN expression. Moreover, pericyte molecular phenotypes varied with aging, acute ischemia, and circadian phase. Notably, light-dark phase emerged as a major axis of transcriptional variation, with pericytes exhibiting distinct circadian phase-associated molecular states. Together, these data demonstrate that adult brain pericytes exhibit spatially organized and dynamically regulated molecular heterogeneity associated with vascular and physiological context.

neuroscience↗

An Integrated Preclinical Platform for Lethal Neuroendocrine Prostate Cancer from Rapid Autopsy Bone and Liver Metastases.

Treatment-emergent neuroendocrine prostate cancer (NEPC) is an aggressive, therapy-resistant disease arising in up to 20% of castration resistant prostate cancers, yet robust biologically relevant preclinical models remain scarce. Here, we describe a technical blueprint for establishing an integrated platform of patient-derived models from visceral and bone metastases collected through a prostate cancer rapid autopsy program (PC RAP). We report the establishment and characterization of patient-derived xenograft (PDX) models from liver metastasis tissue, liver and bone metastasis-derived organoid lines (PDOs), and corresponding patient-derived organoid xenograft (PDOX) models. In addition, we established, to our knowledge, the first mesenchymal stem cell (MSC) cultures derived from neuroendocrine prostate cancer (NEPC) bone metastases. The PDOs preserved intratumoral heterogeneity, displaying both CRPC-NE and CRPC-adenocarcinoma features. These organoids retained neuroendocrine identity across multiple passages, with transcriptomic profiles concordant with the original patient tissue and matched PDX models generated at our institution and at the National Cancer Institute (NCI Patient-Derived Models Repository). To model the bone metastatic microenvironment, we generated novel organoid-based New Approach Methodologies (NAMs) by co-culturing PDOs with iPSC-derived bone marrow organoids, establishing a physiologically relevant vascularized organotypic model of PC bone metastasis. To extend our studies in vivo, we established preclinical models using the liver and bone metastasis-derived organoid models. The PDOX models were tumorigenic and developed spontaneous lymph node metastases, providing clinically relevant models for investigating lethal NEPC biology. Together, these complementary patient-derived models provide a robust and versatile platform for investigating NEPC biology, metastatic progression, and evaluating new therapeutic strategies. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=184 SRC="FIGDIR/small/740121v1_ufig1.gif" ALT="Figure 1"> View larger version (57K): org.highwire.dtl.DTLVardef@1c641f1org.highwire.dtl.DTLVardef@113696aorg.highwire.dtl.DTLVardef@16dcc72org.highwire.dtl.DTLVardef@1898d9f_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LINovel preclinical models of visceral and bone metastases established from a prostate cancer rapid autopsy program. C_LIO_LIThis study is the first to establish mesenchymal stem cell cultures from NEPC bone metastases. C_LIO_LIPDOs preserve heterogeneity, showing both CRPC-NE and CRPC-Adeno features, with transcriptomic profiles concordant with originator tissue and PDX models. C_LIO_LIPC RAP-derived organoids are tumorigenic in vivo and generate spontaneous lymph node metastases. C_LI

cancer biology↗

CXCL10-driven STAT3 signaling programs pathogenic CD4+ T cell responses and limits antiviral immunity during arthritogenic alphavirus infection

Arthritogenic alphaviruses, including onyong nyong (ONNV), chikungunya virus (CHIKV), and Mayaro (MAYV) viruses, cause acute and chronic inflammatory joint disease, with disease severity and resolution influenced by host age. The immunological mechanisms governing persistent inflammation remain incompletely defined. In an age-stratified murine model we show that ONNV infection induced elevated Cxcl10 expression and viral persistence in infected tissues, accompanied by preferential accumulation of CD4+ T cells and limited recruitment of CD8+ T cells. This was associated with skewed T cell differentiation, characterized by increased STAT3 phosphorylation and ROR{gamma}t expression. Infection with CHIKV and MAYV recapitulated these features, supporting a conserved CXCL10-driven pathogenic program across arthritogenic alphaviruses. Perturbation of this axis reduced CD4+ T cell accumulation, altered T cell differentiation states, and decreased tissue viral burden. Thus, we show that a CXCL10-biased immune response is highly proinflammatory but poorly effective in mediating viral clearance, thus setting the stage for chronic disease.

immunology↗

Organoid-T cell co-cultures functionally stratify tumor-reactive T cells and their responses to immune checkpoint inhibitors

Tumor-reactive T cells (TRTs) are critical for anti-tumor immunity but are incompletely captured by current assays, which fail to reproduce tumor-specific antigen diversity. Here, we show that multiplex functional profiling of patient-derived tumor organoid-T cell co-cultures (PDOTs) enables robust identification of TRTs across CD8, CD4, and double-negative (DN) T cell populations. Single activation markers underestimated TRT responses, whereas integrated analysis revealed broader functional repertoire. MHCI blockade abrogated CD8 and DN TRT responses while preserving CD4 reactivity, supporting antigen-dependent recognition across T cell lineages. Tumor PDO expressed MHCI and MHCII, and PDOTs enabled generation and detection of TRTs from peripheral blood. PD1 blockade induced heterogeneous responses, enhancing CD8 and DN activity and unexpectedly augmenting CD4 reactivity. PDOTs further identified additional inhibitory pathways whose therapeutic targeting in combination with PD1 blockade increased TRT responses. These findings establish PDOTs as a platform to identify TRTs and functionally stratify patient-specific tumor-T cell responses to checkpoint immunotherapy.

immunology↗

Mutant KRAS dosage contributes to heterogeneity in lung cancer therapeutic response

Oncogenic KRAS mutations promote tumorigenesis by constitutive activation of multiple, well-characterised signalling pathways. However, there is significant heterogeneity across mutant KRAS tumours in terms of mutation present, mutant allele abundance and downstream signalling strength. It is unclear whether these variations can impact responses to specific therapies. Here, we demonstrate that [~]20% of lung adenocarcinomas (LUAD) show an increase in mutant KRAS dosage (KRASmutant allele fraction > KRASwild-type). Furthermore, we show that KRAS mutant dosage can directly influence clinical outcome and therapeutic susceptibilities in lung cancer. Our findings show that mutant KRAS copy gains specifically affect platinum lung cancer response, promoting resistance to this standard-of-care therapy. Importantly, increases in KRAS mutant dosage are also associated with an increased vulnerability to pS6K inhibition, due to the unique metabolic rewiring of these cells. Together, we show that mutant KRAS dosage contributes to the phenotypic heterogeneity of mutant KRAS NSCLC and that assessment of mutant KRAS content or signalling strength can help optimise treatments strategies for these patients.

cancer biology↗

Generalist method to reconstruct metabolic networks from multi-omics data at large-scale

Metabolic network databases often suffer from inconsistent pathways with blocked reactions. Furthermore, current reconstruction methods lack scalable ways to integrate multi-omics data and cannot flexibly combine diverse modeling pipelines. We introduce SPECTRA, a unified platform that reconstructs metabolic networks across multiple biological scales. SPECTRA uses distinct optimization formulations and achieves a 56-fold speed improvement over existing algorithms for flux consistency based model reconstruction. We demonstrated SPECTRA across three applications. First, we extracted models for 1,479 cancer cell lines, capturing more cancer hallmark reactions than current methods. Second, we gap-filled 7,302 microbial reconstructions from the AGORA2 database, reducing blocked reactions from an average of 482 to 169 per model. Third, we modeled synthetic gut microbiota at the community scale with superior performance in reaction inclusion. This framework provides a scalable and flexible approach for modeling metabolic networks, and generates testable hypotheses for machine learning applications in biology.

systems biology↗

Spatial multi-omics identify an immunosuppressive lipid-laden macrophage niche in primary CNS lymphoma

Primary central nervous system lymphoma (PCNSL) is histologically a subtype of diffuse large B-cell lymphoma (DLBCL), sharing genetic and transcriptomic similarity, but with distinct clinical features, particularly its confinement to the CNS and higher relapse risk. The microenvironmental basis for its divergence from systemic DLBCL remains unclear. Using spatial transcriptomic approaches (Xenium/GeoMx digital spatial profiling) in a comparative study of PCNSL (n=17) and DLBCL (n=76), we found that PCNSL, unlike systemic DLBCL, is dominated by an immunosuppressive macrophage compartment enriched for cholesterol-metabolism programs. In independent cohorts of PCNSL profiled by single-cell RNA sequencing, we validated the presence of a recurrent population of lipid-laden macrophages (LLMs): TREM2/GPNMB-expressing, lipid-remodeled cells transcriptionally distinct from resident microglia and consistent with an infiltrating monocyte origin, not previously characterized in CNS lymphoma. LLMs formed immunosuppressive niches with regulatory T cells, and using Cellscape hyperplex proteomic imaging we demonstrate that LLM-Treg spatial interactions are associated with chemotherapy response. To test whether LLMs are lymphoma-driven and functionally important, we developed an immunocompetent syngeneic PCNSL mouse model, driven by Myd88L252P and Cd79b mutations with Bcl2 overexpression. Monocyte-derived macrophages in lymphoma-bearing brain regions acquired an LLM-like state, not seen in lymphoma-free brain regions or in splenic tumors driven by the same oncogenic lesions. TREM2-SYK signaling sustained this state, and SYK inhibition reversed its tumor-supportive activity ex vivo. These findings identify the LLM program as a targetable immunosuppressive myeloid state in CNS lymphoma.

cancer biology↗

Designing AI-programmable therapeutics with the EDEN family of foundation models

The ability to interpret, modify, and design DNA has driven many of the most significant advances in modern medicine, from diagnostics, biologics, and vaccines to cell and gene therapies. However, the inherent complexity of biological systems means that most modern medicines are still engineered using bespoke, labor-intensive processes. To address the need for a generalisable and programmable approach to therapeutic design, we introduce the EDEN (environmentally-derived evolutionary network) family of metagenomic foundation models, including a 28 billion parameter model trained on 9.7 trillion nucleotide tokens from BaseData1. This dataset, at the time of training, contained more than 10 billion novel genes from over 1 million new species, and is intentionally enriched for environmental and host-associated metagenomes, phage sequences, and mobile genetic elements, enabling the model to learn from diverse and novel cross-species evolutionary mechanisms and apply them to key challenges in human health. EDEN achieves state-of-the-art performance across a series of predictive and generative genomic and protein benchmarks. To demonstrate the models broad applicability across biology, we evaluate EDENs capacity for programmable therapeutic design by challenging a single architecture to design biological novelty across three distinct therapeutic modalities, disease areas and biological scales: (i) large gene insertion, (ii) antibiotic peptide design, and (iii) microbiome design. First, we demonstrate AI-programmable Gene Insertion (aiPGI), in which EDEN designs de novo large serine recombinases (LSRs) capable of inserting large pieces of DNA at desired target sites in the human genome when prompted only on 30 nucleotides of DNA sequence from the desired target site. In low-N experimental validation, EDEN generated multiple active recombinases for all tested disease-associated genomic loci (ATM, DMD, F9, FANCC, GALC, IDS, P4HA1, PHEX, RYR2, USH2A) and 4 potential safe harbor sites in the human genome. EDEN achieves an overall functional hit rate of 63.2% across diverse DNA prompts when prompted on only 30bp of DNA from outside the training data. 50% of EDEN-generated LSRs were active in human cells, achieving therapeutically relevant levels of CAR insertion in primary human T cells. We also show that EDEN can generate active bridge recombinases when prompted on the associated guide RNA alone, with sequence identities to training and public data as low as 65%. These results pave the way for a new generation of cell and gene therapies by opening the door to rapid, programmable and site-specific integration of large genetic payloads without double-strand breaks. This offers an alternative to the safety, efficiency and payload limitations inherent in viral or nuclease-based editing at thousands of currently intractable human therapeutic targets. Second, we use the same model to generate a focused low-N library of novel antimicrobial peptides where 97% showed activity, with top candidates achieving single-digit micromolar potency against critical-priority multidrug-resistant pathogens. Third, to demonstrate that EDEN captures inter-genomic features, we design a gigabase-scale microbiome with over 94,000 synthetic metagenomic assemblies, including prophage genomes and correct cross-species metabolic pathway completions. The EDEN-generated synthetic microbiome covers 9,067 species with a biome-specific taxonomic accuracy of 99%. Over 1,500 of the generated species were outside the fine-tuning dataset while retaining the correct microecological properties and biome association, thus significantly expanding genetic and taxonomic diversity. Together, these results establish a new strategic direction for AI-programmable therapeutics, in which a single foundation model architecture designs candidate therapeutics across diverse modalities and disease areas. This suggests that the combination of billions of years of evolutionary data with specific therapeutic records offers a clear, scaling-driven path to making therapeutic design a predictable engineering discipline. O_FIG O_LINKSMALLFIG WIDTH=141 HEIGHT=200 SRC="FIGDIR/small/699009v2_ufig1.gif" ALT="Figure 1"> View larger version (59K): org.highwire.dtl.DTLVardef@68c20borg.highwire.dtl.DTLVardef@19b8e8corg.highwire.dtl.DTLVardef@1ab9362org.highwire.dtl.DTLVardef@1592cb7_HPS_FORMAT_FIGEXP M_FIG C_FIG

genomics↗

Kinetochore Homeostasis is Maintained by Coordinated Chromatin Stabilization and Soluble Buffering

Faithful chromosome segregation requires the coordinated assembly and maintenance of kinetochore complexes. However, the homeostatic mechanisms that maintain these multi-subunit assemblies remain unclear. CENP-T is a chromatin-bound linker that recruits the outer kinetochore modules via the CENP-T-W-S-X complex to form a functional kinetochore. Although CENP-T must be continuously replenished, how its steady-state levels are maintained is unknown. Here we demonstrate that CENP-T homeostasis is actively sustained by two spatially distinct pathways. Using covalent pulse-labeling, we find that CENP-T undergoes cell-cycle-coupled turnover rather than being stably inherited. In situ proximity labeling of chromatin-bound and soluble pools identifies regulators of CENP-T stability, including an unexpected role for the CENP-O complex in stabilizing soluble CENP-T. Mechanistically, CENP-S-X stabilizes chromatin-bound CENP-T through DNA-binding, whereas the CENP-O complex buffers a replenishment-competent CENP-T pool. Together, these conserved, spatially compartmentalized pathways establish an actively maintained homeostatic system that ensures robust kinetochore assembly and faithful chromosome segregation.

molecular biology↗

Mechanisms of soil carbon preservation illuminated by model mineral-associated organic matter

A central control on atmospheric CO2 is the stabilization of organic carbon in soils. While there are well-described properties associated with stable soil carbon, such as interactions with minerals and conserved chemical features, determining the mechanisms underlying these properties is challenged by soil complexity and heterogeneity. Here we synthesize a naturally complex, tunable model form of mineral-associated organic matter (LabMAOM) to explore mechanisms of stable soil carbon formation and persistence. We show that mineral association and ambient organic matter synergistically stabilize LabMAOM against microbial decomposition. Moreover, different starting compositions of organic matter yield a conserved final LabMAOM chemical composition consistent with natural soils. Our findings open new mechanistic understanding of stable soil carbon and will be of utility to climate modeling and soil health management.

synthetic biology↗

Comparative Analysis of Pathology Foundation Models for Automated Detection of Tertiary Lymphoid Structures in H&E-Stained Digital Pathology Images

Tertiary lymphoid structures (TLS) have been observed in solid tumors and have been associated with better outcomes in patients treated with immunotherapy, but their dynamic nature makes identifying TLS in clinical samples challenging. Recently, pathology foundation models have emerged as powerful tools in computational pathology. In this study, we aimed to develop a computational tool capable of identifying TLS across different cancer types. To this end, we utilized multiple pathology foundation models, along with the ImageNet-pretrained ResNet50 as a baseline, to identify TLS in pancreatic ductal adenocarcinoma (PDAC) and head and neck squamous cell carcinoma (HNSCC) using hematoxylin and eosin (H&E) stained images from The Cancer Genome Atlas (TCGA) and a licensed Real-World Evidence (RWE) cohort. Both pathologist-annotated and transcriptomic signature-based TLS outcomes were employed for performance assessment. Pathologist-identified TLS-positive tumors showed higher expression of TLS signatures in both diseases. Among the models tested, PLIP and CTransPath exhibited strong performance in identifying TLS within PDAC samples (AUC = 0.94 and 0.89, respectively). However, all models struggled in the analysis of HNSCC, likely due to the increased heterogeneity of the tumor microenvironment (TME). Despite their overall utility in detecting TLS in PDAC, all foundation models demonstrated poor performance in predicting transcriptomic signature-based outcomes in both PDAC and HNSCC. This suggests that TLS signatures may reflect broader or more transient aspects of TLS biology, whereas pathology-based assessment anchors on visible morphological features, more closely aligned with the focus of foundation model training. These findings highlight the potential of advanced pathology foundation models for TLS detection and broader tumor immune profiling tasks. These models can also be utilized on routinely collected patient biosamples with nominal costs. However, further refinement is needed to enhance their utility in tumors with more complex TME. Additionally, the identification of transcriptomics-based biomarkers from H&E images remains a significant challenge, despite advancements in digital pathology.

bioinformatics↗

Aging diminishes interlaminar functional connectivity in the mouse cortical V1 and CA1 hippocampal regions

Aging disrupts brain network integration and is a significant risk factor for cognitive decline and neurological diseases, yet the circuit-level mechanisms underlying these changes remain unclear. Most previous studies have utilized cross-sectional or acute approaches, limiting insights into the longitudinal dynamics of the neural network. In this study, we chronically recorded laminar electrophysiological activity in both the primary visual cortex (V1) and hippocampal CA1 region of young (2-month-old) and aged (13-month-old) mice over 16 weeks. This approach allowed us to directly assess how aging modulates functional connectivity within hierarchically connected cortical and hippocampal circuits. We found that single-unit spiking activity and the signal-to-noise ratio were largely preserved in aged versus young mice, suggesting intact neuronal firing properties. However, aged mice showed global reductions in local field potential (LFP) power and a selective decrease in coherence across delta, alpha-beta, and gamma frequency bands within and between cortical layers and V1-CA1 pathways, while phase amplitude coupling remained unaffected. Interestingly, population level excitatory activity in CA1 was increased in aged animals. These findings indicate that aging selectively impairs network-level synchrony and temporal coordination in specific frequency bands and regions, with minimal loss of single-neuron function. Our results highlight the necessity of longitudinal, multi-region measurements to uncover the multi-scale vulnerabilities of the aging brain. Understanding the depth- and region-dependent circuit changes will guide strategies to preserve cortical-hippocampal communication and cognitive function in aging, as well as enhance neural interface technologies for older populations. NEW & NOTEWORTHYAging non-uniformly degrades cortico-hippocampal networks, leading to region- and frequency-specific breakdowns in coordinated activity. While single-neuron activity remains preserved, disruptions in frequency-selective synchrony (delta and alpha-beta) were observed in aged mice, indicating impaired V1-CA1 communication as a marker of aging rather than neuronal degeneration. By separating single-neuron activity and large-scale dynamics, we reveal that aging alters communication between sensory and memory systems, underscoring the need for longitudinal approaches to capture age-related impairments in laminar connectivity.

neuroscience↗

Interpreting convolutional neural networks to study wide-field amacrine cell inhibition in the retina

Wide-field amacrine cells (ACs) play a unique role in retinal processing by integrating visual information across a large spatial area. Their inhibitory influence has been implicated in multiple retinal functions such as differential motion detection and the suppression of retinal activity during eye movements. However, a coherent understanding of their general function is lacking due to difficulties in directly recording from these cells and identifying effective visual stimuli to activate them. In this study, we used convolutional neural networks (CNNs) to investigate widefield inhibition mediated by wide-field ACs in the marmoset retina. We trained CNNs to mimic the function of the retina by predicting retinal ganglion cell (RGC) population responses to naturalistic movie stimuli and optimising the most exciting inputs (MEIs) to visualise RGCs receptive field (RF) structures. We then optimized suppressive surrounds beyond classical RGC RF boundaries, intended to capture the inhibitory effect of wide-field ACs on RGC activity. These optimized surrounds reduced MEI-elicited activity by 10% to 30%, demonstrating that CNNs not only mimic retinal responses but can also reveal hidden computational aspects of wide-field inhibition. However, suppression strength and generalization varied across architectures and datasets, indicating potential model-specific effects, high-lighting the importance of cautious interpretation. Overall, our approach illustrates how interpretability methods applied to artificial neural networks can offer new hypotheses regarding biological retinal computation, paving the way for targeted experimental validation. The code is available here.

neuroscience↗

Integrative immune subtyping of HNSCC reveals clinically relevant phenotypes and treatment-associated transitions

Head and neck squamous cell carcinoma (HNSCC) exhibits profound heterogeneity in clinical presentation, treatment response, and immune landscape. While prior classification systems have identified molecular and immune subtypes in this disease, their applicability to real-world clinical settings remains restricted to small, homogeneous cohorts and limited by lack of multimodal data integration and interpretation. We performed integrated multi-omics analysis including transcriptomic, genomic (copy number, single-nucleotide variants) on 1,149 tumors from 1,102 HNSCC patients across treatment settings. Using the Similarity Network Fusion (SNF) algorithm, we defined immune subtype clusters (ISCs) based on the full immune gene landscape. These clusters were characterized using mutational, transcriptional, and immune cell enrichment analyses, and mapped to hypoxia and traditional subtypes. Associations with clinical outcomes, including progression-free survival, were evaluated across first-line and post-metastatic treatment settings. Four distinct immune subtype clusters (ISC1-ISC4) were identified: ISC1: immune-cold and EMT-enriched; ISC2: immune activated; ISC3: mixed immune-regulatory and stromal-enriched phenotype; and ISC4: immunosuppressed. Distinct treatment response patterns were observed across subtypes in subjects treated with checkpoint inhibitors, chemotherapy, and combination regimens. 44 Patients with matched pre/post treatment tumors revealed treatment-associated transitions between immune subtypes: checkpoint inhibitor treatment enriched for immune activation, while chemotherapy treatment enriched for immunosuppressive signaling pathways. This study provides a clinically relevant immune subtyping framework for HNSCC based on real-world, multi-omics data. These subtypes reflect dynamic tumor-immune states and associated with treatment response and survival, supporting their use in guiding immune-based therapy in HNSCC.

immunology↗

Hypermyelination Improves Strength and Detection of Neuronal Activity in the CA1 Hippocampus and Facilitates Neuroprotection in FusOLcKO Mice

1.0.Loss of oligodendrocytes (OLs) and myelin impairs cortical neuronal firing and network stability, whereas enhancement of oligodendrogenesis improves electrophysiological stability in cortex and, to a lesser extent, hippocampus. OLs exhibit regional heterogeneity, especially in their ability to synthesize cholesterol, a critical driver of myelin wrapping and ensheathment of axons. Conditional depletion of the Fused in sarcoma (Fus) gene in OLs, referred to as FusOLcKO, increases cholesterol biosynthesis, myelin thickness, and tissue cholesterol content. We examine whether this hypermyelination alters extracellular recordings across the layers of visual cortex and the underlying hippocampal CA1 over 16 weeks. In FusOLcKO mice, visually-evoked single-unit detectability and firing rate in CA1 increased relative to wild-type littermates, whereas cortical recordings showed no improvement. At the population level, FusOLcKO cortex exhibited reduced firing rates and lower functional connectivity, indicating altered network dynamics. Post-mortem histology revealed higher neuron density in recorded cortex and greater excitatory synapse density in CA1 of FusOLcKO mice suggesting region-specific neuroprotection and synaptic strengthening. These results demonstrate that cholesterol-driven hypermyelination enhances chronic hippocampal recordings while disrupting cortical network communication. Our study highlights myelins region-dependent roles in supporting single-cell reliability, tuning population dynamics, and maintaining circuit integrity under chronic perturbation. 2.0. SIGNIFICANCE STATEMENTMyelin critically regulates neural circuit function via conduction and metabolic support. Here, we show that cholesterol-driven hypermyelination in FusOLcKO mice augments single-unit detection and firing in hippocampal CA1 but reduces population firing and interlaminar connectivity within the cortex. These findings reveal a dual role for myelin: it can both safeguard specific circuit activity and perturb large-scale cortical communication. Understanding these dynamics is essential for designing myelin-targeted therapies in neurodegenerative disorders.

neuroscience↗

PatternExtract: A facile, scalable pipeline for point pattern generation from spatial imaging data

We present PatternExtract, an open-source pipeline that generates accurate spatial point patterns from RGB pathology images and cell coordinate data without relying on composite channels or proprietary software. Using a novel two-kernel tissue segmentation method combined with automated pixel classification in QuPath, PatternExtract precisely excludes tissue artifacts such as necrosis and blood vessels to define spatial windows. Optimized on 568 diffuse large B-cell lymphoma images and validated on an independent cohort, the pipeline enables robust spatial analyses.

bioinformatics↗

Mapping the Transcriptional Landscape of Drug Responses in Primary Human Cells Using High-Throughput DRUG-seq

To advance our understanding of drug action in physiologically-relevant systems, we developed a high-throughput transcriptomic atlas of compound responses in primary human cell types. Leveraging the scalable and cost-effective Digital RNA with the pertUrbation of Genes (DRUG-seq) assay, we profiled gene expression responses to 89 pharmacologically-active compounds across six concentrations in four distinct primary cell types: aortic smooth muscle cells (AoSMCs), skeletal muscle myoblasts (SkMMs), dermal fibroblasts, and melanocytes. Through rigorous quality control and normalization, we generated reproducible and cell type-resolved transcriptomic signatures, enabling the discovery of both shared and divergent regulatory programs. This dataset revealed core cellular responses, such as brefeldin A-mediated ER stress across all cell types, as well as lineage-specific effects, including dexamethasone-induced hypoxia signaling in AoSMCs, complex inflammatory responses linked to epithelial-to-mesenchymal transition pathways in SkMMs, TGF-{beta}-modulated states in fibroblasts, and dabrafenib-driven transcriptional shifts towards quiescence in melanocytes. By integrating systematic perturbations with primary models, this dataset serves as a resource for building systems-level models of drug response and mechanism. Ultimately, we aim to accelerate predictive pharmacology by enabling high-throughput data generation grounded in human biology and readily usable by artificial intelligence models.

systems biology↗