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Biology subjects

Liu, K.

Publications and source records attributed to Liu, K..

At least 19 recordsLinked to original sources

An Injectable Antisense Oligonucleotide (ASO) Approach to Treat Obesity in Preclinical Models

Obesity leads to metabolic dysregulation and thus drives complications including type 2 diabetes, cardiovascular disease, steatotic liver disease and some cancers. Obesity is primarily driven by lifestyle factors, which impacts individuals differently depending on their genetic susceptibility. Studies on this genetic risk have provided major insights into gene networks that modulate obesity development, including incretins and their regulatory circuits. Indeed, incretin-based therapeutics (i.e. GLP1 RA) have proven to be enormously successful in promoting weight loss, but do so with several ongoing challenges including significant weight rebound upon cessation, loss of lean mass, and side effects that reduce therapeutic longevity. Thus, ongoing research is necessary to identify additional interventions that overcome or minimise these concerns. To this end, in our previous work we have identified a novel lipid regulatory protein with potential to influence lipid storage. In the current study, we use antisense oligonucleotides (ASO) to silence this protein in mouse models of diet induced obesity, and demonstrate that once weekly IP injections of ASO prevents the development of obesity, and defends against the progression of existing obesity. This is shown to be effective in multiple mouse strains fed different diets over various time frames, with gene targeting ASOs leading to reductions in fat mass, no change in lean mass, along with improvements in whole body glucose tolerance and steatotic liver disease. Additional studies demonstrate that this phenotype is likely driven by effects in adipose tissue, but not liver, and is not linked to activation of the Type 1 Interferon response, a previously implicated off target effect of ASO administration. Thus, these studies demonstrate that our injectable ASO approach in mice can successfully treat obesity in the absence of major change in lean mass, thus positioning this intervention as a therapeutic program that works alternatively to, or in conjunction with current weight loss medicines.

physiology↗

Venetoclax Resistance Creates Canonical Pyroptosis Vulnerability in AML

Venetoclax has transformed acute myeloid leukemia (AML) therapy, yet resistance is almost inevitable. Its diverse, often difficult-to-target drivers frequently converge on monocytic differentiation, a state that survives on MCL1 rather than BCL2. Here, we show that the very cells that escape BCL2 inhibition retain the canonical pyroptosis machinery of normal monocytes. Across 15 bulk and 4 single-cell cohorts, the canonical pyroptosis program was enriched in monocytic AML and monocytic leukemia stem cells, correlated with venetoclax resistance, and associated with inferior survival specifically after venetoclax-based therapy. In 24 cell lines and 142 patient specimens, both DPP8/9 inhibition by Val-boroPro and NLRP3 activation by lipopolysaccharide (LPS) plus nigericin preferentially killed monocytic AML cells through GSDMD-mediated pyroptosis, largely sparing non-monocytic AML and lymphoid leukemias. Venetoclax preferentially depleted primitive AML blasts but enriched a monocytic residuum vulnerable to canonical pyroptosis; combining venetoclax with Val-boroPro achieved deeper ex vivo clearance than either agent alone. In venetoclax-resistant xenografts, Val-boroPro suppressed leukemia progression, and adding venetoclax further reduced leukemia burden and prolonged survival beyond either monotherapy. Thus, venetoclax resistance itself exposes an alternative, druggable cell-death program that can be therapeutically exploited to overcome this resistance.

cancer biology↗

Simultaneous multi-area recordings in macaque frontoparietal cortex during a manual object-interaction task

Dexterous hand control is essential for daily activities and a top restoration priority for people with tetraplegia, yet neurophysiological datasets suitable for studying and decoding naturalistic hand behavior remain scarce. Most available recordings constrain movements to a small number of degrees of freedom or cover only a single stereotypical grasp-and-hold structure. Moreover, previous brain-computer interface (BCI) studies have largely relied on signals from a single cortical area, even though dexterous hand control is distributed across an interconnected frontoparietal network. To address these limitations, we designed a paradigm that elicits rich hand movements across 40 distinct hand-object interaction conditions. We recorded simultaneous frontoparietal activity with hand kinematics from a rhesus macaque performing them. Neural activity was recorded from the primary motor cortex, anterior intraparietal area, and ventral premotor cortex using microelectrode arrays spanning 1,920 channels, while a multi-camera markerless motion-capture system acquired video from which kinematics were extracted offline. The dataset comprises data from 20 sessions, including processed neural features, synchronized 3D coordinates of hand and elbow keypoints, and trial-level metadata including task conditions and event timing. We further provide technical validation, including a neural decoding benchmark with multiple algorithms for continuous keypoint coordination reconstruction. We found that cross-day and cross-task decoding performance varied among algorithms. Variation in channel availability leads to different performance degradation curves in different algorithms. This task-rich dataset offers a valuable resource for investigating the neural mechanisms of dexterous hand control and for advancing the performance of motor BCIs.

neuroscience↗

Strategies to improve neurite outgrowth from primary neurons ingelatin methacrylate hydrogels polymerized with visible lightexposure inside a microscale 3D model

Gelatin methacrylate (GelMA) hydrogels offer many advantageous properties such as excellent biocompatibility, tunable stiffness, and rapid fabrication, making them attractive materials for neural systems-on-a-chip devices used to study axonal outgrowth in three-dimensional (3D) environments. In this study, we evaluate properties of GelMA hydrogels crosslinked with visible blue light and application of these hydrogels for encapsulation of primary dorsal root ganglion (DRG) explants inside a custom microscale device. We selected low GelMA polymer concentrations of 3 % and 6 % w/v for hydrogel preparation to create soft materials and demonstrate the effect of degree of functionalization (DoF) on stiffness of GelMA in a blue light-induced crosslinking system. Additionally, a decrease in crosslinking efficiency and stiffness of GelMA was observed upon reconstitution of the polymer in DMEM cell culture media compared to PBS. To evaluate the axonal outgrowth, DRG explants were encapsulated in various GelMA formulations and cultured for 7 days. Following incubation, the explants were fixed and immunochemically labelled with anti-beta-III-tubulin antibody. We further describe an application of custom-developed semi-automated image analysis tool together with linear mixed effects modeling to evaluate axonal outgrowth across different GelMA formulations. Among the formulations tested in our study, 3 % GelMA 95 % DoF hydrogels supported the longest neurite outgrowth suggesting that matrix properties influenced neurite extension.

bioengineering↗

AnewDDE: An Agentic Drug Discovery Engine for Biomolecular Interaction Modelling and Closed-Loop Design

Accurate modelling of biomolecular interactions is fundamental to drug discovery, yet current artificial intelligence (AI) workflows remain fragmented across structure prediction, affinity estimation, molecular design, and experimental decision-making. We introduce AnewDDE, an agentic Drug Discovery Engine that connects these capabilities into a closed-loop system for biomolecular interaction modelling and design. We demonstrate several components of this system: AnewFold delivers superior performance on challenging targets, including antibody-antigen complexes and molecular glues, providing more reliable structural hypotheses together with pocket identification and conformational analysis; AnewDesign achieves a 10.7% success rate in identifying binders with single-digit-nanomolar affinities measured by surface plasmon resonance (SPR) in a representative nanobody discovery campaign, substantially reducing the experimental search burden; AnewAffinity combines accuracy approaching that of free-energy perturbation (FEP) methods with the speed required for high-throughput screening and prioritizes promising transformations; and AnewMind, a large language model (LLM) at the hundred-billion-parameter scale that underwent in-house full-parameter post-training, provides scientific reasoning and absorption, distribution, metabolism, excretion, and toxicity (ADMET) prediction for pharmaceutical research and development (R&D). To systematically evaluate AnewMind, we introduce PharmBench, an internal benchmark designed by pharmaceutical R&D experts to evaluate long-horizon decision-making in drug discovery. Evaluations on ADMET tasks and PharmBench show that AnewMind performs competitively with leading frontier models. Together, these capabilities establish AnewDDE as an integrated engine for addressing challenging therapeutic targets and translating molecular insights into experimentally validated candidates at scale.

bioinformatics↗

Long-term voluntary exercise reveals limited translation of hippocampal molecular responses into neuroprotection in 5xFAD mice

Physical exercise promotes systemic and neural adaptations that support healthy brain aging and may mitigate Alzheimers disease (AD) progression. However, the capacity of the AD-afflicted brain to mount and translate exercise-responsive molecular adaptations into neuroprotection remains unclear. Here, we examined the effects of long-term voluntary wheel running (VWR) on molecular, neuropathological, and behavioral outcomes in independently studied male and female 5xFAD mice. VWR elicited expected metabolic and transcriptional remodeling of inguinal white adipose tissue, confirming engagement of exercise-responsive peripheral biology. In contrast, hippocampal transcriptional responses were modest, with few differentially expressed genes and coordinated changes emerging primarily at the pathway level. These responses involved synaptic, neuroimmune, mitochondrial, neurotrophic, and monoaminergic processes and differed qualitatively between the two groups. Several components of the canonical hippocampal exercise response also failed to converge into coordinated cellular adaptations: synaptic protein abundance changed without altering synapse density, while neurotrophic, neurogenic, and vascular responses showed little correspondence across molecular and cellular measures. VWR also produced little change in hippocampal amyloid pathology or behavioral function despite sustained exercise engagement. Together, these findings demonstrate that the 5xFAD brain retains modest molecular responsiveness to prolonged voluntary exercise but may be unable to mount a sufficiently robust or coordinated response to produce broad neuroprotective effects. These findings highlight disease context as an important determinant of the efficacy of exercise-based interventions in neurodegenerative disease.

neuroscience↗

Single-cell multiomic QTL mapping reveals state-dependent genetic regulation and associated gene during cellular senescence

How cellular senescence reshapes inherited regulatory effects across chromatin and transcription, and eventually contribute to non-coding risk loci of complex diseases, remains elusive. We generated single-cell multiome ATAC-RNA profiles from proliferating and replicatively senescent primary HUVECs derived from 100 genotyped donors. Alongside state-resolved eQTL and caQTL mapping, we implemented covariance-aware multivariate QTL mapping to identify chromatin accessibility-expression QTLs (caeQTLs), modeling accessibility and expression as a joint phenotype. Joint analysis recovered moderate, asymmetric and modality-distributed associations missed by single-modality scans, revealed senescence-associated regulatory programs, and prioritized candidate causal variants and effector genes at cardiovascular loci. Fine-mapping, chromatin-state annotation and heritability enrichment, together with independent SNP-to-gene algorithms, provided convergent orthogonal support. Mechanistic dissection revealed that rs2019090 modulates PDGFD expression and endothelial phenotypes through senescence-amplified, allele-specific, HMGA1-associated enhancer-promoter regulation. This study establishes senescence-resolved joint multiomic QTL mapping as a framework for narrowing the missing regulation gap in complex disease genetics.

genetics↗

Adaptive introgression in Tibetans spans focal and distributed architectures

The Denisovan-derived EPAS1 haplotype is a paradigmatic example of adaptive introgression, but whether archaic ancestry contributed beyond such large-effect loci remains unclear. We analyzed genomes from 1,039 Tibetans, Han Chinese, archaic hominins and ancient Tibetan Plateau individuals. Denisovan-derived adaptive variation was highly localized around EPAS1, whereas Neanderthal-derived variation was distributed across the genome; temporal and elevational patterns reinforced this contrast. Despite individual-level heterogeneity, Neanderthal-introgressed gene repertoires repeatedly converged on immune, metabolic and signal-transduction pathways. A Tibetan-enriched Neanderthal PPARGC1A haplotype was associated with lower fasting glucose and showed concordant support from an independent East Asian type 2 diabetes GWAS. These findings reveal that adaptive introgression can contribute to complex human adaptation through functionally convergent combinations of archaic variants, rather than only through shared large-effect haplotypes.

evolutionary biology↗

Glutamine-regulated VCP acetylation creates a proteostasis vulnerability in RB1-deficient tumor

RB1 deficiency defines an aggressive tumor state with limited therapeutic options. Osteosarcoma represents a clinically relevant model of RB1 deficient malignancy, as RB1 alterations are among the most frequent genomic events in this tumor type. However, the non cell cycle vulnerabilities created by RB1 loss remain incompletely understood. We therefore used osteosarcoma to identify therapeutic dependencies associated with RB1 deficiency. Through high-throughput compound screening, we identified a selective vulnerability of RB1-deficient tumor cells to VCP inhibition. NMS-873 and other VCP-targeting compounds preferentially suppressed RB1 deficient cells across two-dimensional cultures, three-dimensional spheroids, patient-derived organoids, and in vivo tumor models. Mechanistically, VCP inhibition exacerbated proteostasis stress and activated endoplasmic reticulum stress responses in RB1-deficient cells. We further found that VCP function was regulated by glutamine-dependent acetylation, with lysine 615 serving as a dominant acetylation site that modulated sensitivity to VCP inhibition. Glutamine restriction phenocopied, whereas glutamine supplementation partially rescued, the effects of NMS-873 on VCP acetylation and tumor cell growth. Clinical and transcriptomic analyses further linked VCP expression and protein-folding stress programs to aggressive disease features. Together, these findings identify glutamine regulated VCP acetylation as a metabolic proteostasis dependency in RB1-deficient tumors and nominate VCP inhibition as a therapeutic strategy for this difficult-to-target tumor state.

cell biology↗

Ensemble tests mask missing dynamics in protein conformational generators

Protein generators are increasingly used to produce conformational ensembles and trajectories as faster alternatives to molecular dynamics (MD). Yet whether agreement with an equilibrium ensemble demonstrates that a generator has learned dynamics remains unresolved. Here we show that this evidence survives the destruction of dynamics. Randomly reordering authentic MD frames leaves ensemble fidelity unchanged but reduces the geometrically gated kinetic pass rate from 0.963 to 0.004; an independent MD replicate reaches 0.95 under the same test. Dynbench separates recovery of conformational states, time-dependent behaviour and trajectory admissibility. Public generators that appear similar under ensemble tests separate sharply, and in a prospective 36-protein lockbox the reference and floor remain stable while two models reverse order. Ensemble tests can therefore systematically overstate evidence of learned dynamics, establishing time-resolved validation as a distinct requirement for claims about protein dynamics generators.

bioinformatics↗

Molecular dynamics descriptors for 1,079 post-translationally modified protein systems

Databases of post-translational modifications (PTMs) catalogue modified sites and increasingly add static structural context, but trajectory-derived descriptors remain scattered across specialised tools and general molecular dynamics archives. Dyna-MO PTM brings together 1,079 AlphaFold 3-seeded systems covering lysine acetylation, lysine and arginine monomethylation, and serine, threonine and tyrosine phosphorylation. Each system is linked to three completed 10 ns replicas generated with CHARMM36m and TIP3P, for 32.37 s of aggregate sampling. A 118-column table joins simulation and quality-control provenance with global relaxation measures, site solvent exposure, rotamers, secondary structure and ionic-contact proxies. Versioned identifiers connect the records to starting structures, trajectories, manifests and analysis scripts. Researchers can use the resource to filter PTM contexts, reproduce descriptors, prioritise longer simulations and evaluate trajectory-analysis or generative methods. The trajectories describe finite-window relaxation rather than equilibrium free energies, kinetics or matched PTM effects.

biophysics↗

An open multimodal spatial resource integrating same-tissue transcriptomics, proteomics, and histology

Spatial transcriptomic and proteomic technologies provide complementary insights into tissue organisation, cellular phenotype and function, yet integrating these modalities on the same tissue section remains technically challenging. Sequential workflows must preserve RNA integrity, antigenicity and tissue morphology while maintaining accurate spatial registration. At present, publicly available multimodal datasets suitable for computational method development remain limited. Here, we present a workflow for sequential 10x Genomics Xenium spatial transcriptomics, COMET cyclic immunofluorescence, and haematoxylin and eosin (H&E) histological staining on the same formalin-fixed paraffin-embedded tissue section. We demonstrate this approach across multiple biologically distinct human tissues, including tonsil, hepatocellular adenoma, and matched tumour and non-tumour hepatocellular carcinoma, illustrating the widespread applicability of the workflow beyond a single tissue type. Following image registration, Xenium-derived cell segmentations were applied to protein images to generate integrated single-cell transcriptomic and proteomic measurements for downstream analyses. To facilitate community reuse, we publicly release four representative aligned tissue cores together with transcript coordinates, multiplex protein images, H&E images, cell segmentations, and integrated single-cell datasets. We additionally introduce UnumLocalia, an open-source visualisation and data extraction tool that enables interactive exploration of aligned multimodal images, supports user-defined cell segmentation, and allows export of integrated single-cell data for downstream analyses. Together, this technical protocol, workflow, software, and openly available dataset provide a reusable resource for multimodal spatial biology, supporting advances in biological discovery, computational method development, multimodal data integration, and validation of emerging analytical approaches across complementary spatial technologies.

immunology↗

PASTRI: Resolving Stage-Specific Cell-State Dynamics from Annotated Cell Lineage Trees

Cellular transitions between phenotypic states are fundamental to development and disease, yet quantitative analysis of their dynamics remains challenging. Here we present PASTRI (Phylogenetic Adjacency-based State Transition Rate Inference), a computational framework that infers transition rates from cell lineages/phylogenies annotated with terminal phenotypic states, such as single-cell transcriptomes. We validate PASTRI using simulated lineages and the Caenorhabditis elegans embryonic lineage. Importantly, by leveraging cell pairs at varying phylogenetic distances, PASTRI accurately resolves stage-specific transition rates, circumventing the issue of developmental changes in dynamics. Applied to three cell phylogeny datasets from our lineage-tracing experiments spanning diverse developmental/disease models and tracing systems, PASTRI uncovers rate-limiting steps in the activation of hepatic stellate cells and the differentiation of primordial lung progenitors, as well as attractor states that support cancer cell proliferation. PASTRI thus opens up a venue for dissecting cell state transition dynamics from annotated cell lineage/phylogeny.

bioinformatics↗

The Maine Coon Cat Harboring the MYBPC3-A31P Mutation: A Genotype-Stratified Phenotypic Characterization of Hypertrophic Cardiomyopathy

Abstract Background: Most animal models of HCM are mouse-based, but the thin interventricular septum in mice makes it difficult to clearly distinguish pathological hypertrophy, which introduces substantial errors and constrains basic HCM research. Cats develop HCM spontaneously, and the common MYBPC3-A31P variant in cats is homologous to human mutations in both genetics and pathology, with a larger body size that makes them suitable as large-animal models. This study examines how heterozygosity or homozygosity for the p.A31P mutation (c.91G>C) in the MYBPC3 gene affects the phenotype and severity of HCM in affected cats, with the aim of establishing an ideal large-animal model for clinical risk stratification and precision diagnosis and treatment of human HCM. Methods: Forty-nine Maine Coon cats were enrolled and stratified into homozygous mutant (HOM, n=8), heterozygous mutant (HET, n=26), and wild-type (WT, n=15) groups. All cats underwent echocardiography, blood pressure measurement, physiological assessment, hematological and biochemical analyses, and cross-species sequence conservation analysis. Results: No significant differences in baseline characteristics including age and body weight were observed among groups (P>0.05). HOM cats exhibited significantly higher left ventricular outflow tract pressure gradients and greater basal septal thickness compared to WT cats (P<0.05), with HET cats showing intermediate values. Analysis of hematological and serum biochemical parameters revealed no evidence of systemic inflammation or hepatic injury. Sequence conservation analysis confirmed that the A31 residue is highly conserved across mammalian species. Conclusions: This study provides a phenotypic characterization of Maine Coon cats carrying the MYBPC3-A31P mutation, revealing marked gene-dose effects on cardiac structure and function, with homozygous individuals exhibiting more severe phenotypic features. This model serves as a large-animal translational platform that not only clarifies genotype-phenotype correlations but also supports risk stratification and precision therapeutic strategies in human HCM. Its spontaneous nature and genetic homology to human disease make it particularly valuable for bridging preclinical findings to clinical application.

genetics↗

Pressure-induced membrane tension mechanically opens the germinant receptor GerA ion channel to trigger bacterial spore germination

High pressure (HP) can trigger bacterial spore germination, acting either through germinant receptors (GRs) or the SpoVA channel. However, the mechanism by which HP activates these membrane-embedded proteins remains elusive. Here, using Bacillus subtilis, we demonstrate that the GerA germinant receptor (GR) is the primary target of moderate HP (50-300 MPa). Mutagenesis reveals that pore-lining residues within the GerA ion channel are essential for the pressure response, whereas canonical ligand-binding and intramembrane signaling residues are dispensable. We then propose a s tretch- t o- o pen (STO) model, in which HP differentially compresses the more compliant inner membrane (IM) relative to the rigid spore core, generating lateral membrane tension that promotes opening of the GerA channel. In situ membrane tension measurements indicate HP-induced compression of IM phospholipids and elevated membrane tension. This tension-dependent gating is further supported by the pressure-dependent phenotypic rescue of GerA channel mutants. Consistently, HP increases IM permeability to water-soluble and membrane-impermeable agents (propidium iodide and formaldehyde), an effect potentiated by GerA, indicating concomitant opening of GerA by HP. Furthermore, modulating IM fluidity via heat activation or decoating altered membrane physical properties and delayed HP-induced germination, establishing the IM as the critical mechanical transducer. Additionally, computational modeling and calculations support faster compression of the IM than of the core under HP, rationalizing the source of tensile stress. Together, our findings establish a novel mechanism of HP-induced GerA activation via the STO model: HP compresses the IM, generates lateral tension, and promotes opening of the GerA ion channel to trigger bacterial spore germination.

microbiology↗

A single-cell and multi-platform spatial atlas of the human pancreas resolves a transformation-associated epithelial axis and a recurrent boundary-organized tumour microenvironment

Most pancreatic ductal adenocarcinoma (PDAC) single-cell and spatial studies analyze one cohort or platform, obscuring recurrent biology. We assembled a human pancreas single-cell and single-nucleus reference of 1,186,130 cells from 19 studies and interpreted 176 Visium sections comprising 458,877 spots across non-diseased pancreas, chronic pancreatitis, PanIN, IPMN, primary PDAC and metastasis. Marker-supported labels were used after two RNA-based copy- number callers failed known-diploid controls. BANKSY domains, two reference-mapping methods and sample-level analyses resolved a cross-sectional epithelial axis extending from acinar-rich to malignant tissue. Five trajectory algorithms recovered similar ordering on a shared embedding; their consensus was interpreted as transformation-associated, not temporal or clonal. Malignant regions were globally segregated from fibroblast and myeloid compartments. Signed- distance analysis refined this pattern into a malignant core, a CAF/myeloid surround beginning at the tumour boundary and a more distal lymphoid compartment. Candidate extracellular-matrix communication, led by COLLAGEN, LAMININ and FN1, concentrated at the interface. Changes were reproduced in six patient-matched Normal-tumour pairs using exact patient-level tests. Visium HD resolved the same organization at single-cell resolution and showed that 8-um bins distorted immune-adjacency estimates. Xenium also revealed recurrent neighbourhoods but sample-specific stromal boundaries. We provide a confound-aware framework for identifying recurrent epithelial and microenvironmental organization in PDAC.

cancer biology↗

Hidden genetic diversity in 320 nearly-complete East Asian genome assemblies

East Asian populations, representing over 20% of the global population, remain critically underrepresented in human genomic studies, limiting our understanding of population-stratified genetic variation and its implications for health and disease. Here we present the first phase of the Asian Pan-Genome project (APG), comprising 320 nearly complete, fully phased haploid genome assemblies from 160 East Asian individuals. These assemblies achieve unprecedented quality, with an average contig N50 of 144.3 megabase pairs and an average quality value of 64.5. Leveraging these superior assemblies, we reveal previously uncharacterized diversity in human repeatome, including population-stratified patterns in centromere satellites and rDNA arrays. Compared to existing global human genome assemblies, the newly generated genomes supplement 152 million base pairs of novel sequences, 355 gene gains, 18,300 structural variation loci and 26 large euchromatic inversions missing from current human pangenomes. We perform population stratification analyses of structural variations, and further resolve the structural haplotypes of complex genomic regions such as Major Histocompatibility Complex and Survival Motor Neuron loci across global pangenomes, exemplifying tandem-duplicate and inversion-rich complex locus architectures in the human genome, respectively. This resource provides a critical foundation for human genetic studies, especially for East Asian populations, promoting more accurate variant discovery, reducing bias, and ultimately advancing the equity and efficacy of genomic medicine.

genomics↗

Multi-compartment immune and tumor cell reprogramming by IFNa2 overcomes colon cancer immunotherapy resistance

Approximately 85-90% of colorectal cancers (CRC) are microsatellite stable and resist immune checkpoint inhibitors (ICI). The recent success of intismeran, a lipid nanoparticle (LNP)-delivered mRNA encoding patient-specific tumor antigens, enhances pembrolizumab responses, showing that LNP-delivered nucleic acids can render ICI-resistant tumors responsive by supplying what the tumor microenvironment lacks. Because an IFN-responsive phenotype predicts CRC response to checkpoint blockade and tumor cell PD-L1 represses IFN-I, we asked whether delivering the missing cytokine, rather than antigen, achieves the same conversion. Here we show that PD-1 blockade failed even when initiated before tumor seeding, yet deleting tumor cell PD-L1 abolished colon cancer lung metastasis, implicating a tumor-intrinsic PD-L1 function that antibody blockade does not neutralize. An LNP-encapsulated IFN2-encoding nanoplasmid (LNP-IFN2) transfected tumor cells within lung metastases, converting them into a self-sustaining and tumor-restricted IFN2 source. LNP-IFN2 suppressed colon cancer lung metastasis in syngeneic and humanized mouse models and sensitized these ICI-resistant tumors to PD-1 blockade. Efficacy improved further with neutralization of co-induced IL6. scRNA-Seq revealed multi-compartment reprogramming: SPP1+ macrophages acquired apoptotic signatures as IFN-responsive monocytes replaced them, progenitor-exhausted T cells exited quiescence and expanded, and tumor cells exited a high-cycling state, gained antigen presentation, and shifted away from cuproptosis resistance. The treated microenvironment transcriptionally recapitulated pembrolizumab-responder signatures, positioning LNP-IFN2 as an off-the-shelf nanomedicine for ICI-resistant CRC.

immunology↗