Biology subjects

Lee, C.-H.

Publications and source records attributed to Lee, C.-H..

At least 19 recordsLinked to original sources

bioRxiv2026

Dual-targeted nanoparticles enable the ex vivo generation of cytotoxic and phagocytic CAR effector cells

Chimeric antigen receptor (CAR)-T cells and CAR-macrophages (or CAR-monocytes) offer complementary antitumor functions. CAR-T cells exert potent cytotoxicity against cancer cells, whereas CAR-macrophages can infiltrate solid tumors, phagocytose cancer cells and promote antigen spreading. Antibody-conjugated lipid nanoparticles (LNPs) have emerged as non-viral mRNA delivery platforms for generating CAR-immune cells. However, these systems have generally been optimized to target a single immune-cell type. Here, to enable the ex vivo generation of CAR-T cells and CAR-monocytes, we developed CLCM08, an anti-CD222 antibody, and co-displayed it with an anti-CD5 antibody on LNPs. CD222 is predominantly expressed by monocytes, macrophages and T cells, and facilitates the internalization of CD222-binding molecules. CD5 is primarily expressed by T cells and has previously been used to enable LNP-mediated mRNA delivery to these cells. Delivery of CAR mRNA by dual CD222/CD5 antibody-conjugated LNPs to primary monocytes and T cells substantially enhanced the ex vivo generation of CAR-monocytes and CAR-T cells compared with non-conjugated LNPs. Moreover, CD19 CAR-T cells and CAR-monocytes generated using dual CD222/CD5 antibody-conjugated LNPs showed enhanced cytotoxic and phagocytic activity, respectively, relative to cells generated using non-conjugated LNPs, with anti-CD222 LNPs producing the largest phagocytic gain in monocytes. These findings establish an adaptable antibody-conjugated LNP-mRNA platform for the ex vivo generation of CAR-T cells and CAR-monocytes.

biochemistry↗

bioRxiv2026

Antibodies with Engineered Fc Domains Having Absolute Binding Selectivity to either FcγRIIa or FcγRI Delineate the Respective Effector Phenotypes by Human Monocytes and Macrophages

IgG1 immune complexes bind to all the Fc{gamma} receptors (Fc{gamma}R) expressed on myeloid cells, making it challenging to determine the precise role of each Fc{gamma}R on Fc effector phenotypes. Here we report the engineering of Fc2KG, an aglycosylated human IgG1 Fc domain that binds with near physiological affinity to Fc{gamma}RIIa/b with no detectable binding to any other Fc{gamma}Rs. Crystallographic analysis elucidated the structural basis of how mutations in the Fc domain compensate for the absence of the N297 glycan and enable selective binding. Using single-cell phagocytosis assays, we show that particles opsonized with Fc-engineered antibodies formatted with Fc2KG or with an Fc domain that binds only Fc{gamma}RI (Fc5), are ingested by THP-1 cells with near identical kinetics. We find that with CD16+ primary human monocytes, selective Fc{gamma}RII engagement is a major contributor to the ADCP mediated by wild-type IgG1. Additionally, we showed that Fc2KG formatted antibodies induce high levels of GM-CSF. With M1-like monocyte-derived human macrophages, trastuzumab formatted with wild-type IgG1 Fc, with Fc2KG or Fc5 were all equally proficient in the trogocytotic killing of opsonized SK-BR-3 HER2+ cells but only Fc{gamma}RI engagement led to secretion of proinflammatory cytokines. Collectively, our results highlight how precisely tuned, Fc-engineered antibodies can be deployed to answer long-standing questions regarding the precise effector functions mediated by human Fc{gamma}Rs, information which is key for the optimization of therapeutic antibodies.

immunology↗

bioRxiv2026

Focused framework sampling recovers binding-positive humanized anti-amyloid-β antibodies in a single sorting round

Therapeutic antibody development requires workflows that integrate antigen-reactive clone discovery with efficient humanization and early developability assessment. Here, we combined immune yeast fragment antigen-binding (Fab) display with single-round focused humanization and applied the workflow to antibodies against amyloid-{beta} (A{beta})-derived preparations. Immunization with A{beta}1-42 aggregate preparations generated a Fab-display library with a diversity of approximately 3.5 x 108. Magnetic enrichment followed by fluorescence-activated cell sorting (FACS) identified three sequence-distinct immunoglobulin G (IgG)-format candidates, of which CLAB17 and CLAB45 were advanced to humanization. Structure-guided libraries sampled framework positions predicted to support complementarity-determining regions (CDRs) or heavy-and light-chain variable-domain packing, and a single FACS round recovered binding-positive variants CLAB17-h2 and CLAB45-h8. Both retained the parental CDRs and showed increased predicted humanness, favorable computational developability triage profiles, and high purity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). By enzyme-linked immunosorbent assay (ELISA), CLAB17-h2 showed a lower apparent half-maximal effective concentration (EC50) for A{beta}1-42AggreSure, whereas CLAB45-h8 showed a lower apparent EC50 for pyroglutamate-modified A{beta}3-42 (A{beta}pE3-42). Because the preparations were not resolved into defined assembly states, these antibodies are considered A{beta}-preparation-binding rather than aggregate-state-selective candidates. This workflow provides a practical route from immune-repertoire discovery to binding-positive humanized antibodies.

bioengineering↗

bioRxiv2026

Intrauterine growth restriction is associated with adaptive hematopoietic reprogramming and selective immune rewiring in monozygotic twins

BackgroundPrenatal growth restriction has been associated with adverse neonatal and long- term health outcomes, yet the epigenetic mechanisms by which an adverse intrauterine environment shapes fetal immune development remain incompletely understood. Monozygotic dichorionic-diamniotic twins with selective fetal growth restriction (sFGR) provide a unique human model for investigating environmentally driven developmental programming independent of genetic variation and inter-twin placental vascular anastomoses. MethodsUmbilical cord blood buffy coat samples were collected from three sFGR and two gestational age-matched concordant control twin pairs. Bulk RNA sequencing and genome-wide DNA methylation analysis were performed, followed by differential expression, pathway enrichment, hematopoietic and immune module analyses, differential methylation, and integrative transcriptomic-epigenomic analyses. ResultsCompared with concordant control twin pairs, discordant twins exhibited broad attenuation of immune and inflammatory transcriptional programs alongside enrichment of erythroid- and hypoxia-related pathways, consistent with adaptive hematopoietic responses to intrauterine stress. Within discordant twin pairs, the growth-restricted co-twins displayed marked transcriptional asymmetry characterized by selective enrichment of cytotoxic lymphoid signatures despite global suppression of myeloid and antigen-presenting cell-associated programs. Integrated transcriptomic and epigenomic analyses further revealed coordinated epigenetic remodeling, with hypomethylated regions in growth-restricted twins enriched for immune regulatory pathways, including T cell differentiation and leukocyte activation. At selected loci, concordant hypomethylation and increased gene expression suggested a potential epigenetic basis for the observed immune remodeling. ConclusionsThese findings suggest that intrauterine growth restriction is associated with coordinated hematopoietic and immune reprogramming at birth, consistent with both compositional and cell-intrinsic alterations. In genetically identical twins, relative growth divergence was associated with polarized transcriptional states, highlighting how intrauterine environmental differences may shape early immune development independent of genetic background.

genomics↗

bioRxiv2026

Mild and Reversible Proprioception Perturbation Suggests Causal Biomechanics for Memory-Dependent Spatial Behavior in Mice

The neural system at the periphery is a frontline for embodied cognition, yet an acute, mild perturbation to dissect functional causality is hard to achieve. Peripheral neural processes and the central nervous system may work in concert to generate sophisticated representations regarding self and environments in the brain. This hypothesis, together with the underlying mechanisms, is particularly difficult to test for certain sensory inputs due to the lack of reversible manipulation techniques. Long postulated as a component for path integration, proprioception is one of such modalities. In this study, we developed a murine experimental system to manipulate proprioceptive inputs during memory-dependent localization task (which required precise operant-conditioned licks) in spatial virtual reality (VR). Through bioluminescent optogenetics (luminopsin) selectively expressed in the parvalbumin-positive neurons of the dorsal root ganglia in mice, proprioceptive processing was compromised directly from the periphery to bypass the bottleneck of specific central targeting, which results from the lack of anatomically or genetically dedicated proprioceptive circuits in the brain. In-vivo IVIS imaging and behavior suggested the effects of luminopsin last for roughly 20 minutes. While mice exhibited normal performance in tasks relying on gross motor skills, they showed subtle deficits in challenging spatial tasks that required integration of past movements. These observations support a task-specific role for proprioception, and demonstrate a potential of chemogenetics-like, rapidly reversible strategies for characterizing peripherally defined sensory contribution to spatial cognition. Future work will optimize this approach; for instance, to activate opsins by light with millisecond precision. To our knowledge, this is a first causal demonstration for acute participation of proprioception in path-integration biomechanics, enabling the first temporally defined method for mild perturbation of path-integration mechanisms.

neuroscience↗

bioRxiv2026

Coexisting but dissociable place and spatial view codes in the primate hippocampus

In rodents, hippocampal activity is classically described in terms of position-anchored place cells, whereas nonhuman primate work has emphasized spatial view cells, in which activity depends on the region of space being viewed. This contrast has fueled a longstanding debate over whether primate and rodent spatial codes are biologically distinct. We recorded single units in the hippocampus of macaques as they navigated linear tracks in two visually distinct contexts, intermittently making context-dependent object choices. Using a generalized linear model to dissociate position coding from spatial view coding, we identified three coexisting populations in the macaque hippocampus: Position cells, View cells, and Conjunctive cells encoding both variables. Position cells were prevalent and showed rodent-like properties--localized firing, directional tuning, and contextual remapping--even after accounting for visual sampling, whereas View cells stably encoded allocentric visual space. Population decoding confirmed this dissociation at the ensemble level. Robust position codes thus coexist with, yet remain dissociable from, view codes in the primate hippocampus, suggesting that the apparent scarcity of place-like responses in earlier primate work reflects methodological and task-related differences rather than a fundamental species-level divergence.

neuroscience↗

bioRxiv2026

Combinatorial docking and molecular generation to navigate over 100-billion molecules for prospective ligand discovery

Commercially available make-on-demand libraries now exceed 100 billion compounds, requiring over 50 years to screen on 2,000 CPU cores using conventional docking. We present two complementary approaches to address this challenge. CombiDOCK, a combinatorial docking framework, enables exhaustive screening at the 100-billion scale within 40 days. MINT-Dock, a generative framework, accelerates navigation of this space by integrating CombiDOCK with Monte Carlo Tree Search. Benchmarked on 46 diverse targets, CombiDOCK matched full-molecule docking accuracy, and MINT-Dock achieved a 4,800-fold enrichment over random selection. Compared with prior billion-scale brute-force campaigns against {sigma}2, VMAT2, and VAChT, prospective CombiDOCK screens of the 100-billion-molecule library yielded higher hit rates and more potent ligands, while MINT-Dock achieved comparable outcomes across single- and multi-target objectives with >20-fold computational cost reductions. Docking-predicted poses of the best VAChT-binding compounds were confirmed by cryo-EM structures. These methods provide exhaustive and generative paths for navigating the trillion-molecule frontier of drug discovery.

bioinformatics↗

bioRxiv2026

A Lineage-Specific Peptide Suppresses Juvenile Hormone to Drive Reproductive and Longevity Reprogramming in Ants

In ants and other eusocial insects, reproductive division of labor is tightly regulated by juvenile hormone (JH), which suppresses reproduction in most individuals to maintain the worker caste. However, how social cues trigger systemic JH suppression to permit reproductive activation remains unclear. In the ant Harpegnathos saltator, workers respond to queen loss by engaging in sustained ritualistic dueling and transitioning into reproductive, long-lived pseudoqueens (gamergates). This provides a powerful model for investigating the molecular basis of socially induced plasticity. We examined the hemolymph proteome of transitioning ants and identified HCRG1, a lineage-restricted peptide upregulated during dueling, as a circulating factor that physically interacts with Hex70c, a broadly conserved JH-binding Hexamerin. HCRG1 promotes ovarian development by antagonizing JH signaling. Expression of ant HCRG1 in the heterologous solitary model Drosophila also extends lifespan. These findings identify an evolutionarily derived circulating factor that links social sensory perception to systemic JH suppression, enabling coordinated reproductive and longevity transitions in a social insect.

evolutionary biology↗

bioRxiv2026

Layered social competition coordinates reproductive hierarchy formation in ants

Social interactions establish reproductive hierarchies and regulate division of labor in eusocial insect colonies. In the ant Harpegnathos saltator, queen loss induces a subset of workers to transition into reproductive pseudo-queens, known as gamergates, through a ritualized antennal-dueling tournament that can last for more than a month. Gamergate transition is associated with sustained dueling engagement, yet the interaction networks and social dynamics that shape reproductive hierarchy formation remain unclear. Here, we combine long-term automated tracking with behavioral classifiers to systematically quantify social interactions, including antennal dueling and two distinct biting behaviors: aggressive grooming and mandible locking. We find that antennal dueling is organized into two activity states, an engaged state and a disengaged state, and that persistent maintenance of the engaged state, rather than transient entry into it, predicts successful transition to gamergate fate. Early access to abundant food shapes dueling activity and biases ants toward entry into the engaged state. Moreover, engaged-state individuals exhibit a strong assortative preference for dueling with one another and cluster spatially within the colony. We further show that mandible locking precedes transitions out of the engaged state, whereas aggressive grooming reinforces the disengaged state, together consistent with a role in stabilizing worker fate. Our findings reveal how structured social interactions organize reproductive hierarchy formation by regulating stable behavioral states associated with caste fate.

neuroscience↗

bioRxiv2026

Staged heavy-chain filtering enables Fab discovery from combinatorially intractable library spaces

BackgroundCombinatorial fragment antigen-binding (Fab) libraries encode an immense heavy-light chain pairing space, often exceeding 10{superscript 1} possible combinations, which far surpasses the diversity that can be experimentally constructed and screened in display systems. As a result, direct Fab screening samples only a small fraction of the theoretical search space, creating a practical bottleneck for functional binder discovery. ResultsHere, we frame Fab discovery as a staged search problem by decoupling heavy-chain (HC) and light-chain (LC) exploration. We implemented a sequential HC preselection-remating workflow in yeast surface display, in which antigen-reactive HC variants are first enriched and subsequently recombined with a diverse LC repertoire to reconstruct a focused Fab library. In a SARS-CoV-2 spike-targeted campaign, HC and LC libraries of 2.05 x 10 and 2.33 x 10 members corresponded to a theoretical pairing space of approximately 4.8 x 10{superscript 1} combinations. Sequential HC enrichment followed by LC remating allowed recovery of multiple functional Fab clones from a tractable library scale of approximately 10, including clones that shared a common HC scaffold but carried distinct LC partners. A representative recombinant IgG output showed broad but heterogeneous spike/RBD binding, measurable pseudovirus neutralization activity (EC = 11.1 nM), and compatibility with standard early biophysical characterization after full-length IgG reformatting. ConclusionsThese results provide proof of principle that combinatorial Fab discovery can be approached as a staged exploration problem under realistic library-size constraints. By focusing downstream Fab reconstruction on an antigen-compatible HC subspace, sequential HC preselection followed by LC remating offers a practical strategy for exploring otherwise intractable antibody pairing landscapes in eukaryotic display systems.

bioengineering↗

bioRxiv2026

A sterol-binding pocket in iRhom1 underlies paralog-specific regulation of the sheddase ADAM17

ADAM17, the major sheddase in mammalian cells, releases membrane-tethered EGFR ligands and inflammatory cytokines, and is a central regulator of cell signalling. The rhomboid pseudoproteases, iRhom1 and iRhom2, function as essential cofactors of ADAM17, controlling its maturation and activation. In contrast to the well-characterized iRhom2, the mechanism and regulation of its ubiquitously expressed paralog iRhom1 remain undefined. Here, we present a 2.5 [A] cryo-EM structure of the full-length human iRhom1/ADAM17 complex, revealing a previously unrecognized sterol-binding pocket located between TMD2 and TMD5. Structure-guided mutagenesis and pharmacological perturbation of sterol binding demonstrate that sterol binding is required to stabilize the iRhom1/ADAM17 complex and sustain its shedding activity. Strikingly, this regulation is paralog-specific: iRhom2 precludes sterol binding and instead stabilizes ADAM17 through direct intramolecular interactions. Furthermore, two human iRhom1 variants associated with cardiac disease localize adjacent to the sterol-binding pocket and disrupt ADAM17 maturation and activity. Together, these findings uncover mechanistic divergence between iRhom paralogs and establish a sterol-binding pocket in iRhom1 as a critical determinant of ADAM17 stability, revealing a potential avenue for paralog-selective therapeutic targeting.

cell biology↗

bioRxiv2026

Structure-Function Analysis of the FCRL5-IgG1 Fc Complex Reveals an Unappreciated Effect of Fc-Silent Antibodies on B cells

Human Fc receptor-like 5 (FCRL5) is a low-affinity IgG Fc receptor expressed on various B cell subsets and a potential therapeutic target. We discovered that commonly used Fc-silencing mutations, designed to prevent interactions between the Fc{gamma} receptors on immune cells and the Fc domain of therapeutic IgG, do not prevent binding to FCRL5. As a result, unintended interactions between Fc-silent therapeutic IgG and human B cells may occur. We isolated a well-expressed variant of the Fc-binding portion of human FCRL5 by directed evolution and used structural modeling to guide the engineering of a human IgG1 Fc variant with approximately 100-fold higher affinity for FCRL5, enabling us to produce FCRL5:Fc complexes in solution. Native mass spectrometry, size exclusion chromatography, and the crystal structure of the FCRL5- IgG1 Fc complex solved at 3.4 [A] indicate that the two proteins bind in a 1:1 stoichiometry. Furthermore, the structure revealed that FCRL5 binds to IgG1 Fc in a manner completely distinct from that of previously characterized Fc-binding proteins, such as Fc{gamma} receptors, explaining why most Fc-silencing mutations do not disrupt FCRL5 binding. We demonstrate that selective cross-linking of FCRL5 with the B cell receptor (BCR) in cis, using Fc-engineered antibodies with either physiological or enhanced FCRL5 affinity, inhibits Ca2+ flux in FCRL5-expressing B cells. We compare this effect with the selective co-ligation of Fc{gamma}RIIb with the BCR. Our work demonstrates that FCRL5 interacts with human IgG Fc in a distinctive manner and that engagement of FCRL5 by Fc-silent therapeutic IgG could influence B cell function.

bioengineering↗

bioRxiv2026

Structure and mechanism of human sphingosine-1-phosphate transporter MFSD2B

Sphingosine-1-phosphate (S1P) is an essential signaling lipid that maintains vascular integrity and regulates immune cell trafficking. The major facilitator superfamily domain-containing protein 2B (MFSD2B) serves as the main S1P exporter in red blood cells and platelets; however, its structure and transport mechanism are unclear. Here, we report the 3.0 [A] cryo-EM structure of human MFSD2B bound to S1P. S1P is captured in a distinctive binding state, deeply buried within the C-domain, with its sphingoid tail accommodated by a hydrophobic pocket and its phosphate group coordinated by a cluster of polar residues within the transporters cavity. Mutagenesis and molecular dynamics simulations identify the TM2/TM11 lateral opening as the primary pathway for S1P translocation, with key charged residues acting as sequential anchors during transport. Furthermore, we demonstrate that MFSD2B functions as a uniporter, and that subtle rewiring of local charge networks can alter its coupling mechanism. Our work provides a molecular framework for understanding S1P transport mediated by MFSD2B in hematopoietic cells.

biochemistry↗

bioRxiv2025

Tissue-guided multi-omics profiling identifies extracellular vesicle biomarkers indicative of lung pathology in acute respiratory distress syndrome

BackgroundAcute respiratory distress syndrome (ARDS) remains a lethal inflammatory lung condition lacking reliable biomarkers that reflect lung-specific pathology. Extracellular vesicles (EVs) circulate systemically and may carry molecular signals from injured organs, but the correspondence between EV cargo and lung tissue alterations remains unclear. MethodsWe established aspiration-, lipopolysaccharide (LPS)-, and COVID-19-induced murine ARDS models and applied a tissue-guided multiomics framework integrating proteomic and metabolomic analyses of lung tissue and plasma-derived EVs to identify lung-originating circulating biomarkers. ResultsFour proteins--haptoglobin (HP), inter-alpha-trypsin inhibitor heavy chains 3 and 4 (ITIH3, ITIH4), and clusterin (CLU)--were consistently upregulated in both lung tissue and plasma EVs across all ARDS etiologies. Metabolomic integration revealed dysregulation of arachidonic acid metabolism as a unifying inflammatory axis. Multiomics network analysis further distinguished etiology-specific molecular programs, including glycolytic activation in aspiration-induced, platelet aggregation in LPS-induced, and vascular smooth muscle dysregulation in COVID-19-induced ARDS. ConclusionsThis study establishes a tissue-informed EV profiling framework that links local lung pathology to systemic molecular signatures, revealing HP, ITIH3, ITIH4, CLU, and arachidonic-acid-related metabolites as potential diagnostic markers for ARDS. These findings provide a foundation for developing clinically translatable, EV-based biomarker assays for early detection and molecular subtyping of lung injury.

biochemistry↗

bioRxiv2025

Distinct regulation of AEBP2 isoforms on PRC2 activity

Polycomb Repressive Complex 2 (PRC2) represses gene expression through catalyzing H3K27me3, a histone modification essential for gene repression and maintenance of cellular identity. PRC2 accessory proteins modulate its catalytic activity, chromatin localization, and propagation along the chromatin. AEBP2, one of the PRC2 accessory proteins required for mouse embryogenesis, exists in distinct isoforms. The short isoform enhances PRC2 catalytic activity and promotes H3K27me3 spreading, facilitating robust gene repression while the function of the long isoform has remained unclear. Here, we found that the N-terminal region of AEBP2 long isoform contains a conserved DE-rich motif that inhibits PRC2 activity, both EZH2 automethylation and H3K27 methylation. In addition, re-expression of the AEBP2 long isoform in Mtf2/Jarid2/Aebp2 triple-knockout mESCs failed to restore H3K27me3 and caused defective differentiation, unlike the short isoform. Together, our findings uncover an isoform-specific regulatory mechanism by which AEBP2 controls PRC2 activity and contribute to a broader understanding of how PRC2 is dynamically regulated during development.

molecular biology↗

bioRxiv2025

The Fallacy and Bias of Averages on Vegetation Indices based Plant Phenotyping

BackgroundVegetation indices (VIs) from remote sensing are widely used for non-destructive plant phenotyping, often averaged across plots or image regions to represent each plot. However, according to Jensens inequality, which is known as the "fallacy of the average", it can bias estimates when nonlinear relationships exist between VIs and target traits. To examine this issue, we systematically assessed the severity of this bias and tested a correction method. VI values were simulated using six beta distributions with varying shapes and skewness, and with normalized difference vegetation index (NDVI) images from a paddy rice experiment to evaluate bias under real conditions. Nonlinear link functions (concave, convex, logistic) with different noise levels were applied to model VI-trait relationships. ResultThe results showed that averaging under nonlinear relationships reduced predictive performance, lowering the coefficient of determination (R2) between true and predicted traits by up to 82%. In the rice NDVI simulation, R2 was reduced by up to 58% around the tillering stage. Our correction method, which predicts traits from VI before averaging, substantially mitigated bias, improving R2 by up to 0.68 depending on noise level, VI distribution, and link function. To facilitate application, we established an interactive R Shiny website enabling users to quantify potential biases and the efficacy of corrections within this workflow based on their own research conditions ConclusionIn summary, averaging VIs without accounting for nonlinear relationships can introduce substantial bias and degrade phenotyping accuracy. This bias should be explicitly considered in phenotyping analyses, and correction methods applied when appropriate to improve reliability.

plant biology↗

bioRxiv2025

Rapid CAR Screening and circRNA-driven CAR-NK Cells for Persistent Shed-resistant Immunotherapy

Chimeric antigen receptor (CAR)-based immunotherapies against solid tumors face two major hurdles, the "decoy effect" of shedding antigens that sequester CARs, and the limited persistence of immune effectors within the immunosuppressive tumor microenvironment. Here, we present a mechanistic approach to overcome these barriers by integrating a physiologically relevant screening platform with circular RNA (circRNA) engineering. Unlike conventional screens using immortalized cell lines, we performed rapid functional screening directly in human primary natural killer (NK) cells to identify a novel scFv, CLMS10. Structural modeling revealed that CLMS10 targets a membrane-proximal epitope that overlaps the proteolytic cleavage site, thereby evading inhibition by soluble mesothelin (solMSLN). Furthermore, we demonstrated that circRNA-mediated CAR expression, when codelivered with interleukin-21 (IL-21), confers sufficient stability to withstand the continuous antigen shedding induced by cancer-associated fibroblasts (CAFs), resulting in reduced CAR downregulation. In an in vivo metastatic pancreatic cancer model, IL-21-augmented circCAR-MS10-NK cells exhibited potency comparable to that of lentivirally engineered CAR-NK cells while offering superior manufacturability. Overall, this study establishes a paradigm for generating shed-resistant CAR therapeutics through the strategic integration of epitope-specific functional selection and enhanced RNA stability.

bioengineering↗

bioRxiv2025

EZH1 Q571R-mediated chromatin compaction and its oncogenic potential in thyroid cancer

Dysregulation of Polycomb Repressive Complex 2 (PRC2) is established to contribute to cancer. Of its catalytic subunits, EZH1 and EZH2, EZH2 mutations in cancer have been extensively identified and studied, but the role of EZH1 in cancer remains largely unexplored. The recurrent presence of the EZH1Q571R mutation in follicular thyroid cancer suggests its involvement in tumor progression. Using EZH1 ChIP-seq, ATAC-seq, and H3K27me3 CUT&Tag, we demonstrated that EZH1Q571R enhances chromatin binding and compaction and stimulates PRC2-EZH1 catalytic activity, leading to increased H3K27me3 levels and repression of tumor suppressor genes. Purified PRC2-EZH1Q571R demonstrated a significant increase in histone methyltransferase activity compared to PRC2-EZH1WT via enhanced nucleosome binding and DNA compaction. Notably, this effect is particularly observed with EZH1Q571R but only to a lesser extent with the corresponding EZH2Q570R mutation, highlighting it as an EZH1-specific mechanism. We additionally demonstrated PRC2-EZH1Q571R to efficiently methylate H3K27 in pre-existing H3K36me2/3 nucleosomes, disrupting chromatin homeostasis. Our findings provide key insights into the molecular pathogenesis of EZH1Q571R-driven follicular thyroid cancer.

molecular biology↗