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Kim, D.

Publications and source records attributed to Kim, D..

At least 37 records · Page 2Linked to original sources

Antibody-dependent priming of spontaneous germinal centers by autoreactive B cells

Autoreactive germinal centers (GCs) are central to autoimmune pathogenesis, yet the mechanisms by which single autoreactive B cell clones prime systemic autoimmunity remain unclear. Using the 564Igi mixed chimera model, we demonstrate that autoreactive 564Igi B cells break tolerance in wild-type B cells through an unexpected mechanism independent of cognate T cell interactions. While B cell-intrinsic TLR7 signaling was essential for spontaneous GC formation, deletion of MHC class II, CD40, or CD80/86 on GC-priming 564Igi B cells failed to prevent GCs. Instead, CRISPR-mediated deletion of Prdm1 (encoding BLIMP-1) in 564Igi B cells ablated spontaneous GCs, implicating autoantibody production as the primary driver. These findings reveal that autoantibodies can initiate feed-forward mechanisms that propagate systemic autoimmunity, independent of B cell-intrinsic antigen presentation.

immunology↗

Structural basis of endogenous lipid recognition and G protein selectivity in GPR119

G protein-coupled receptors (GPCRs) often engage multiple intracellular transducers, yet the structural basis by which endogenous ligands influence G protein selectivity remains poorly understood. GPR119 is a lipid-activated receptor expressed in pancreatic {beta}-cells and enteroendocrine L-cells, where it regulates glucose-dependent insulin and incretin secretion, making it a promising therapeutic target for metabolic disease. Here, we present cryo-electron microscopy structures of GPR119 bound to its endogenous lipid agonist oleoylethanolamide (OEA) in complex with Gs and Gq proteins. These structures reveal that OEA occupies a deeply buried orthosteric pocket but adopts distinct conformations in the two signaling states. Structural and functional analyses further identify an extended TM5 helix that stabilizes the receptor-Gs interface and acts as a key determinant of G-protein subtype selectivity. Together, these findings provide mechanistic insights into endogenous lipid-driven multi-transducer signaling and establish a structural framework for the development of pathway-selective GPR119 therapeutics.

molecular biology↗

CytoGate-Bench: an LLM benchmark for cross-panel cell gating in cytometry

In cytometry, the workhorse single-cell technology of clinical immunology, every study defines its own antibody panel and cell-type vocabulary, so a classifier trained on one cannot annotate the next. Immunologists instead annotate by manual gating, splitting one parent population at a time on a two-marker plot, down an expert-defined hierarchy. We introduce CytoGate-Bench, a benchmark that reformulates this per-step procedure as a zero-shot, panel-agnostic task for large language models. It comprises 23,646 expert-annotated instances re-curated from 11 public flow- and mass-cytometry cohorts spanning eight marker panels. Across six open- and closed-weight backbones, the strongest formulation draws one rectangular gate per candidate and falls within the range of trained, panel-specialized baselines. It degrades less under distribution shift. Walking the hierarchy stepwise outperforms predicting every cell type at once. Ablations trace the signal to the data distribution shape and curated marker priors. However, adding vision or a self-verification loop systematically tightens gates.

bioinformatics↗

Thal-Kak: unifying biomolecular structure predictors reveals a sampling-selection gap

Complementary all-atom structure predictors sample different solutions, but how to allocate a fixed sampling budget across them and select the best output remains unclear. Thal-Kak unifies five released predictors under shared upstream inputs and a common schema. Across FoldBench and CASP16, model mixing improves oracle sampling over single-model runs, but selection remains a bottleneck because confidence scores do not transfer across models and existing quality-assessment methods cannot resolve this gap.

bioinformatics↗

Phenotype-associated spatial biomarker discovery in spatial transcriptomics with spHOT

Spatial transcriptomics now profiles patient cohorts at single-cell resolution, enabling analysis of disease-associated cell organization in situ. However, discovering such spatial biomarkers remains challenging because relevant structures occur at unknown scales and cell-or niche-level annotations are rarely available. We present spHOT, a deep learning framework that localizes phenotype-associated spatial biomarkers from sample-level labels. spHOT combines spatial foundation model embeddings, a hierarchical domain tree for multi-resolution tissue representation, and a teacher-student multiple instance learning architecture that converts sample labels into cell-level biomarker scores. In controlled simulations and real-tissue benchmarks, spHOT outperformed existing spatial and single-cell methods in localizing ground-truth biomarkers. Across fibrotic, metabolic, and autoimmune disease datasets, spHOT recovered disease-relevant niches and tissue states reported by supervised analyses in the original studies. Cross-disease application of spHOT transferred biomarkers across chronic lung diseases without retraining. spHOT enables scalable, annotation-efficient spatial biomarker discovery in cohort-scale spatial transcriptomics.

bioinformatics↗

LEN-Seek: Fast and scalable ligand binding-site similarity search in the latent space of an SE(3)-invariant graph VAE

MotivationLigand binding-site similarity search is a crucial step in drug discovery that reduces the conformational search space for docking and other downstream tasks by comparing a target protein against experimentally identified binding sites. Existing methods rely on either direct structural alignment or lossy compression of structural information, producing a trade-off between scalability and precision. ResultsWe propose LEN-Seek, a ligand binding-site search method based on a graph neural network (GNN)-driven variational autoencoder (VAE) that encodes the 3D structural and physicochemical context of a binding site into a probabilistic latent space, enabling similarity search within a low-dimensional vector space. A binding site is modeled as a graph of amino acid residues, with node features adopted from the protein language model, Ankh, and edges encoded as SE(3)-invariant (roto-translational invariant) geometric relationships, thereby avoiding expensive data augmentation or SE(3)-equivariant models. Compared to ProBiS, the purely geometric graph-clique based method, LEN-Seek successfully retrieves a substantial portion of similar binding sites with a roughly 3,400-fold lower per-comparison cost, demonstrating its potential as a scalable approach to template-based ligand binding-site search in large-scale protein structure databases. Supplementary informationSupplementary data are available at Bioinformatics online.

bioinformatics↗

Green algal symbionts are stably retained and provisioned in the dark despite a clear physiological cost

Photosymbioses, or associations between heterotrophs and photoautotrophs, are widespread and indispensable in today's ecosystems. The chloroplasts of algae and land plants, which are at the heart of most of earth's primary production, stem from ancient photosymbioses. Photosymbioses often combine heterotrophy and autotrophy and must thus efficiently allocate resources between these two costly cellular processes. We currently lack a clear picture of how photosymbioses allocate their valuable cellular resources in response to environmental change. In this study, we combine growth assays, automated fluorescence microscopy, transmission electron microscopy, and mass spectrometry-based proteomics to explore the physiology and cellular resource allocation of the ciliate-green alga photosymbiosis of Paramecium bursaria. In nutrient-rich environments that resemble P. bursaria's natural habitat, the maximum growth rate attained saturates regardless of light intensity. The green algae thus do not provide a benefit in nutrient-replete conditions, and the photosymbiosis primarily functions heterotrophically. The green algae occupy a remarkably similar and constant volume fractions across contrasting light environments despite displaying clear photo-physiological adaptation. This is true regardless of a clear physiological cost of the photosymbionts; aposymbiotic hosts always display higher growth rates in the dark. The host does not decrease 'symbiont load' in environments where green algae are not beneficial. Moreover, in the dark, the green algae are fully dependent on their hosts and take up a larger proteome mass fraction that increases with prey abundance. Differential protein expression analyses suggest that acetate and amino acids are the preferred sources of carbon and nitrogen for the green algae in the dark. The stable persistence and higher resource uptake by the photosymbionts in the dark argue against a view where hosts have full control over and selfishly exploit their symbionts.

physiology↗

AdaGeneBudget: Cell-Adaptive Gene-Token Allocation for Efficient Single-Cell Foundation Models

Single-cell foundation models (scFMs) represent each cell using sequences of gene-associated tokens, making embedding extraction increasingly costly as the number of cells and expressed genes grows. Existing input policies typically rely on fixed input budgets, with retained genes determined by random subsampling, model-native ranking, or a fixed dataset-level highly variable gene (HVG) panel. However, they do not jointly determine, for each cell, which genes to retain and how many tokens to allocate. We introduce AdaGeneBudget, a training-free gene-token selection method that combines each gene's expression with reference-derived inverse detection frequency and retains the shortest ranked prefix that captures a target fraction of the cell's expression-specificity score mass. The resulting cell-specific budget is bounded by predefined minimum and maximum lengths, requires no cell-type labels, and leaves the pretrained backbone unchanged. We evaluated AdaGeneBudget in a frozen-backbone inference setting using pretrained scGPT and Geneformer models on Kang and PBMC reference-mapping tasks, with an additional scPRINT comparison against its official HVG policy and an expressed-only HVG control. Across four scGPT and Geneformer backbone-dataset pairs, AdaGeneBudget substantially reduced mean gene-token counts and peak GPU memory while increasing embedding-extraction throughput by up to 4.63x. Despite this compression, it preserved native-level aggregate annotation utility and consistently outperformed token-matched random selection. AdaGeneBudget also preserved fine-grained and low-support cell identities and retained lineage-marker programs and stimulation-associated pathway genes under compression. In scPRINT, both HVG controls achieved higher annotation macro-F1, whereas AdaGeneBudget more faithfully preserved the stimulation-induced embedding direction. These results establish biologically informed, cell-adaptive gene-token allocation as a practical complement to architectural and systems-level efficiency methods for applying existing scFMs to new datasets. They also suggest a cell-adaptive input-allocation principle for future models operating under finite token budgets.

bioinformatics↗

A modular cranial window enabling maintainable widefield optical access in non-human primates

Longitudinal optical imaging of the primate cortex requires stable yet maintainable cortical access, but existing cranial window approaches remain limited by mechanical instability and tissue responses that progressively degrade optical clarity. These challenges are amplified in primates, where large craniotomies and physiological variability complicate chronic experiments. Moving beyond conventional fixed cranial windows, we present the PRIME cranial window (Primate Reconfigurable Interchangeable Modular Enclosure), a modular platform enabling maintainable, long-term, and widefield optical access in awake macaques. Built on a modular architecture, PRIME supports non-invasive adjustment, repeated cortical access, and on-demand maintenance, including tissue removal, without surgical re-entry, while a dual-ring sealing mechanism prevents cerebrospinal fluid leakage and contamination. Combined with optimized viral delivery, implantation, and maintenance strategies, PRIME preserves cortical integrity and optical clarity for long-term optical imaging. PRIME establishes a versatile, re-accessible experimental framework for chronic, large-area optical studies of the primate brain.

neuroscience↗

Cryptic disease-prone states in human mesocortical assembloids revealed by multimodal profiling

Neurodegenerative diseases are characterized by synaptic failure, aberrant protein accumulation, and neuroglial dysfunction that emerge long before clinical onset. Although brain assembloids, which recapitulate interregional circuit connectivity beyond the scope of single organoids, significantly advance the modeling of circuit pathophysiology, they are predominantly evaluated through single-modality approaches that cannot resolve the functional and molecular heterogeneity underlying differential disease susceptibility. Here we show that morphologically identical human iPSC-derived dorso forebrain-midbrain mesocortical assembloids (MCAs) spontaneously bifurcate into disease-prone and non-prone states under identical culture conditions, revealing that MCA heterogeneity reflects intrinsic neurodegeneration susceptibility rather than stochastic culture variability. Using integrated electrophysiological, molecular, and spatial profiling, we find that disease-prone MCAs exhibit a temporally ordered molecular cascade in which neurofilament light chain elevation precedes tau dysregulation, mirroring the sequential biomarker trajectories observed in pre-symptomatic human neurodegeneration. Disease-prone MCAs further display selective cortical hyperexcitability and aberrant brainwave-like oscillatory dynamics that remain undetectable by any single modality. Spatially, a discrete junction-like neuronal population at the midbrain-forebrain interface shows transcriptional priming for synaptic overactivation alongside impaired astrocytic glutamate clearance, defining a spatially confined neuron-glial uncoupling as a candidate early origin of the disease-prone state. These findings reframe MCA heterogeneity as a biological window into pre-symptomatic neurodegeneration, with broad implications for disease modeling, risk stratification, and therapeutic discovery.

neuroscience↗

Quantitative Modeling of TLR Signaling Reveals Missing Negative Feedback Guiding Identification of TANK-IKKε Checkpoint

Toll-like receptor (TLR) signaling must be activated rapidly and then terminated to support host defense without sustained inflammation. We developed a rule-based model of mouse macrophage TLR4 signaling at the molecular-interaction level using measured protein copy numbers, RNA-seq-based abundance estimates, literature- and structure-informed reaction rates, and 979 dynamic experimental constraints. The trained model reproduced much of the TLR4-induced NF-{kappa}B and MAP kinase response but consistently failed to capture deactivation of MyD88, TRAF6-associated species, and IKK/{beta}. The recurrent model failure conveyed important biological information, localizing missing regulation to the proximal MyD88-IRAK-TRAF6 module and guiding experimental evaluation of IKK{varepsilon} and its scaffold TANK. Loss of IKK{varepsilon} enhanced transcriptional, cytokine, MAP kinase, and NF-{kappa}B responses to MyD88-specific TLR ligands. TANK deficiency produced a similar cellular phenotype and abolished stimulus-induced IKK{varepsilon} phosphorylation. Deficiency of either protein increased IRAK1 and TRAF6 ubiquitination without increasing MyD88 ubiquitination, placing the inhibitory checkpoint at or immediately downstream of the IRAK1-TRAF6 ubiquitin-signaling node. Overlapping but non-identical in vivo phenotypes further supported a shared regulatory axis with additional protein-specific functions. Our study presents a model-experiment discovery cycle where quantitative pathway discordance identifies missing biology and reveals a TANK-dependent IKK{varepsilon} checkpoint that restrains MyD88-driven inflammation.

systems biology↗

Ribonucleotide Reductase Inhibition Overcomes FLT3 Inhibitor Resistance in Acute Myeloid Leukemia

Internal tandem duplication mutations in FLT3 (FLT3ITD) occur in approximately 30% of patients with acute myeloid leukemia (AML) and are among the most common genetic alterations in this disease. FLT3ITD is a major driver of AML and is associated with poor clinical outcomes. Although FLT3 inhibitors (FLT3is) have significantly improved outcomes for patients with FLT3ITD+ AML, acquired resistance remains a major barrier to durable clinical benefit. Reactivation of RAS/MAPK signaling, often driven by activating NRAS mutations, is a major mechanism of FLT3i resistance in AML; however, effective strategies to overcome this resistance remain lacking. Here, we identify ribonucleotide reductase (RNR) as a critical therapeutic vulnerability in NRAS-driven FLT3i-resistant FLT3ITD+ AML. Activation of RAS signaling through SPRY3 loss or oncogenic NRAS mutations confers robust resistance to FLT3is, whereas pharmacologic inhibition of RNR with multiple inhibitors, as well as siRNA-mediated RNR suppression, reverses FLT3i resistance and restores FLT3i sensitivity across multiple FLT3ITD+ AML models in vitro. In vivo, clofarabine, an FDA-approved RNR inhibitor (RNRi), significantly overcomes NRAS mutation-driven FLT3i resistance. In combination with FLT3 inhibition, clofarabine markedly suppresses the progression of FLT3i-resistant AML and significantly prolongs survival in cell line-derived xenograft (CDX) models. Importantly, the therapeutic efficacy of the gilteritinib/clofarabine combination was independently validated in two genetically distinct patient-derived xenograft (PDX) models harboring different NRAS mutations, demonstrating robust reduction of leukemia burden and confirming the generalizability of RNR inhibition in primary FLT3i-resistant AML. Together, these findings identify a previously unrecognized therapeutic vulnerability in FLT3i-resistant FLT3mut+ AML and establish RNR inhibition as an effective strategy to overcome FLT3i resistance, providing a strong rationale for the clinical evaluation of RNRis in combination with FLT3is in patients with resistant AML. SignificanceAlthough FLT3 inhibitors (FLT3i) are an important therapeutic advance in FLT3ITD+ AML, resistance commonly develops. We identified ribonucleotide reductase (RNR) as a new key vulnerability in NRAS-driven FLT3i-resistant AML and demonstrated that multiple RNRis, including the FDA-approved agent clofarabine, restore FLT3i sensitivity and enhance antileukemic activity, supporting a clinically actionable combination strategy.

cancer biology↗

Deep learning representations of human Immune Health for precision immunology

The human immune system is composed of [~]30-50 distinct cell types, each of which can exist in different states of activation or differentiation. Indeed, the mammalian immune system has evolved to sense and respond to infections, cancers, injuries, and changes in tissue or host homeostasis (1). Moreover, an increasingly large fraction of approved drugs target the immune system directly, and/or cause immune changes (2-4). A key feature of the immune system is to store some of this information, for example as innate or adaptive immune memory (5). In addition, rewiring of immune network architecture induced by disease, environmental exposures, drug treatments, and/or chronological age allows the immune system to store information in the pattern of connections and activity across populations of immune cells. This ensemble information storage, in addition to changes to individual cells, functions as a major way the immune system encodes aspects of immune history and future potential. Genetic information can identify inherited risk alleles, but cannot capture the continual remodeling of the immune system shaped by exposures, infection, inflammation, therapy, and aging (6, 7). To define and use such ensemble immunotypes, we developed a self-supervised deep learning framework that transforms high-dimensional immune profiles into representations of immune health. MAESTRO (MAsked Encoding Set TRansformer with self-distillatiOn) encodes a set of cells from an individual into an embedding that captures immune cell population-level organization. Pretrained on 1,792 peripheral blood samples comprising over 418 million immune cells across 13 clinical diagnoses, MAESTRO learns immune fingerprints that are stable within individuals yet diverse across populations, states of health, disease, and treatment, providing a quantitative basis for comparing immune states across individuals and over time. These fingerprints capture immune architecture beyond coarse cell type proportions, enabling patient-efficient clinical prediction using simple task specific models. MAESTRO model embeddings retain a temporal dimension of immune history and potential, reflecting signatures of past exposures and baseline features that predict future immune responses. Finally, we demonstrate a translational precision immunotherapy application by testing this approach in metastatic Pancreatic Ductal Adenocarcinoma (PDAC), where pretreatment immune landscape circuitry maps enable patient stratification and therapeutic response prediction. Overall, we developed a large, attention-based model that captures deep network architecture of immune states through self- supervised representations of immune cytometry data as a reusable foundation for precision immunology, converting immune complexity into clinically actionable embeddings for diagnosis, monitoring, and therapy selection.

immunology↗

Pathway selection for arabinose utilization in Pseudomonas putida reveals a rate-yield tradeoff in muconic acid production from lignocellulosic sugars

Engineering heterologous utilization of substrates requires selection of catabolic pathways that balance strain performance and product biosynthesis. Here, we compare the oxidative and isomerase arabinose utilization pathways in Pseudomonas putida strains engineered for cis,cis-muconic acid production from glucose and xylose. Based on the point of entry into central carbon metabolism, we hypothesized that the oxidative arabinose pathway would enable higher productivity while the arabinose isomerase pathway would enable higher muconate yield. In both strains, additional modifications were engineered to improve muconic acid production including sugar transporter tuning, catechol 1,2-dioxygenase overexpression, a feedback-resistant DAHP synthase, and a flux-stabilizing gltA variant. Consistent with our hypothesis, the oxidative arabinose pathway supported faster growth and higher productivity (0.58 g/L/h), whereas the arabinose isomerase pathway improved carbon efficiency, achieving muconate yields of up to 50 C-mol% in fed-batch bioreactors. Process modeling indicates that these performance metrics can reduce the minimum selling price of muconate-derived adipic acid to $2.74/kg and greenhouse gas emissions to 1.31 kg CO2e/kg, approaching cost parity and reducing emissions by 86% relative to fossil carbon-derived adipic acid. Overall, this study presents a systematic comparison of sugar catabolic pathways that enabled development of strains suited for the tradeoffs between rate and yield.

synthetic biology↗

Cumulative Transfer Function for Assessment of MRI-Induced RF Heating Risk in Pediatric Patients Implanted with Bifurcated Leads

PurposeRF-induced heating remains a major barrier to MRI access for patients with epicardial cardiac implantable electronic devices (CIEDs). Although ISO/TS 10974 Tier-3 transfer function (TF) methods are established for unbranched leads, no analogous framework exists for bifurcated leads, in which branch asymmetry and inter-branch coupling may substantially alter heating. We developed and validated a cumulative transfer function (cTF) framework to address this gap. MethodsFollowing ISO/TS 10974 Tier-3 formalism, we measured, calibrated, and validated cTFs for a commercial 35 cm bipolar epicardial lead at 1.5 T. The framework explicitly accounts for branch-specific response and cross-branch coupling. Validation was performed with 24 canonical lead configurations in a homogeneous phantom and, without recalibration, in a heterogeneous anthropomorphic pediatric phantom with clinically derived trajectories. A single-branch TF approximation served as a comparator. The validated cTF was applied to predict RF heating across adult and pediatric human models at multiple imaging landmarks. ResultsCompared with the single-branch TF approximation, the cTF reduced prediction error by nearly 70% in the primary validation dataset. In secondary validation, the cTF maintained low error across clinically relevant trajectories and imaging landmarks. In human models, the framework revealed marked anatomy- and landmark-dependent variation in predicted heating for the tested 35 cm lead, with low predicted heating in pediatric models and substantially higher heating in selected adult chest and upper abdominal imaging scenarios. ConclusionThe cTF provides a validated framework for RF-heating assessment of bifurcated leads and substantially improves prediction accuracy over single-branch TF approximations that neglect branch coupling.

bioengineering↗

Hepatic stearoyl-CoA desaturase deficiency ameliorates hyperglycemia through bile acid signaling in an insulin-independent manner

Hyperglycemia in Type 1 Diabetes (T1D) is managed almost exclusively via exogenous insulin therapy, an approach restricted by significant glycemic fluctuations, long-term side effects such as weight gain, and high economic burden. Identifying physiological pathways capable of clearing blood glucose independent of insulin is therefore of paramount clinical importance. Here, we demonstrate that liver-specific stearoyl-CoA desaturase-1 (SCD1) deficiency protects against diabetic hyperglycemia and hepatic steatosis in an insulin-independent manner. SCD1 ablation decreases cellular oleate availability, altering lipid flux and redirecting excess cholesterol into alternative biosynthetic pathways. This redirection drives a 2-fold elevation in hepatic bile acids and a striking 10-fold increase in plasma bile acids, predominantly characterized by the accumulation of taurocholic acid. This shifted bile acid pool stimulates the expression of glucose transporter 1 (Glut1) in the liver via activation of the nuclear hormone receptor FXR, facilitating basal glucose clearance in the absence of insulin. Genetic deletion models show that while the hepatokine FGF21 serves as a partial mediator of this phenotype, the local bile acid-FXR axis remains a sufficient driver of systemic glucose clearance. Finally, we show that dietary oleate supplementation completely reverses this protective phenotype, turning down Glut1 expression and restoring overt diabetes. Together, our findings uncover a novel bile acid-FXR-Glut1 signaling axis triggered by SCD1 inhibition, offering a framework for insulin-independent glycemic control. Article HighlightsHere, we demonstrate that liver-specific stearoyl-CoA desaturase-1 (Scd1) deficiency protects against diabetic hyperglycemia and hepatic steatosis in an insulin-independent manner. Mechanistically, Scd1 ablation redirects excess cholesterol into bile synthesis, predominantly characterized by an increase in liver and plasma taurocholic acid. These shifted bile acids stimulate hepatic glucose transporter 1 (Glut1) expression via the farnesoid X receptor (FXR) activation to facilitate basal glucose clearance. While the hepatokine FGF21 acts as a partial systemic mediator, the local bile acid-Fxr axis remains a sufficient driver of clearance, a protective phenotype completely reversed by dietary oleate supplementation.

biochemistry↗

Pre-analytical delay as a dominant confounder in blood RNA-seq: Rapid ex vivo gene expression changes in EDTA blood

Although the impact of delayed processing on gene expression in EDTA blood has been well documented using targeted assays and microarray platforms, the emergence of next-generation RNA sequencing (RNA-seq) has not yet been leveraged to systematically compare these effects against stabilized whole blood collection systems. Notably, no study has performed a time course RNA-seq analysis with human bulk RNA of matched EDTA and PAXgene blood RNA samples drawn from the same subjects. EDTA is still widely used for gene expression analysis studies. Yet, the genome-wide dynamics by which EDTA blood transcriptomes deviate from a stabilized reference over time remain poorly defined. This represents an important methodological gap, given the increasing reliance on RNA-seq for biomarker discovery, clinical transcriptomics and diagnostics.

Molecular Biology↗

Targeting Cellular Pseudo-Senescence to Overcome PARP inhibitors Resistance in BRCA1 -Mutated Breast Cancer

Poly (ADP-ribose) polymerase (PARP) inhibitors (PARPi) are a mainstay therapy for homologous recombination (HR)-deficient cancers; however, resistance remains a major clinical challenge. Previously, through a genome wide CRISPR screen, we identified ZNF251 haploinsufficiency as a novel driver of PARPi resistance. In BRCA1-mutant (BRCA1mut) cells, ZNF251 deficiency led to HR hyperactivation, conferring PARPi resistance that could be reversed by RAD51 inhibition. In this study, we further show that ZNF251 deficiency induces replication stress and a pseudo-senescence state, in which cells exhibit molecular and phenotypic markers of senescence while retaining proliferative capacity. Because senomorphic and senolytic therapies can target senescent cells, we tested whether these approaches could overcome PARPi resistance in ZNF251-deficient breast cancer cells. Critically, targeting this senescence-like state with either senomorphic agents, such as cytokine inhibitors, or senolytic agents, such as BCL-2 and BCL-XL inhibitors, overcame PARPi resistance ex vivo and in vivo. Importantly, pseudo-senescence was also observed in other PARPi-resistant contexts driven by HR hyperactivation, including 53BP1- and Shieldin-mutant cells, suggesting that it may represent a broader mechanism underlying PARPi resistance in breast cancer. Furthermore, in two olaparib-resistant, BRCA-mutant triple-negative breast cancer organoid models, treatment with DT2216, a BCL-XL-targeting PROTAC, sensitized both models to olaparib. Together, our work defines a novel pathway linking HR hyperactivation, replication stress, and pseudo-senescence, and positions both senomorphic and senolytic therapies as promising strategies to overcome PARPi resistance in BRCA1mut breast cancer. HighlightsO_LIZNF251 deficiency drives PARP inhibitor resistance through HR hyperactivation, replication stress, and pseudo-senescence in BRCA1-mutant breast cancer. C_LIO_LISenomorphic and senolytic therapies overcome PARP inhibitor resistance in vitro, in vivo, and in patient-derived organoid models. C_LIO_LIPseudo-senescence represents a shared vulnerability of HR-hyperactivated PARPi-resistant cancers and can be therapeutically targeted. C_LI

Cancer Biology↗