Biology subjects

Wang, J.

Publications and source records attributed to Wang, J..

At least 307 records · Page 17Linked to original sources

bioRxiv2026

Holomycin from Yersinia ruckeri mediates fish infection at low temperature

Why microbes invest energy in producing structurally complex natural products at extremely low concentrations within hosts remains an important topic of interest in microbial ecology. This is because such compounds with potential "biomolecular activities" can reflect molecule-target interactions, thereby providing conceptual and mechanistic insights that inform the rational design of essential clinical agents. Here, we uncover the in situ ecological function of the dithiolopyrrolone antibiotic holomycin produced by the fish pathogen Yersinia ruckeri. We show that holomycins biosynthesis is strictly gated by low temperature and that trace amounts of holomycin accumulate in vivo during infection in rainbow trout. In addition, the holomycin-deficient mutant leads to markedly reduced virulence. Integrated multiomics, phenotypic analyses, and in vivo bioassay test reveal that holomycin is not a direct lethal factor but instead remodels host-associated microbiota and enhances Y. ruckeri biofilm formation. These coordinated effects promote pathogen colonization and infection progression. Our findings establish holomycin as a host-associated effector and illustrate how its existence and ecological function are mechanistically intertwined in the evolution of pathogenic bacterial chemical arsenals.

microbiology↗

bioRxiv2026

Distinct HPO axis responses and ovarian aging trajectories to chronic unpredictable mild stress in reproductively young versus middle-aged female mice

Psychosocial stressors are key contributors to ovarian functional decline. Chronic unpredictable mild stress (CUMS) is widely used to model stress-induced premature ovarian insufficiency (POI) in mice; however, current animal models do not adequately reflect middle-aged women, who represent a key population exposed to chronic psychosocial stress, nor do they capture the dynamic progression toward POI. Here, female C57BL/6 mice aged 2 or 6 months were subjected to CUMS for 8 or 12 weeks. Estrous cyclicity, endocrine profiles, ovarian histology, and transcriptomic changes in HPO axis-related tissues were systematically analyzed. After 8 weeks of exposure, 2-month-old mice exhibited impaired pituitary responsiveness to estradiol negative feedback, as evidenced by dysregulated FSH secretion, indicating reduced stress tolerance compared with 6-month-old mice. Following 12 weeks of CUMS exposure, both age groups showed significant reductions in ovarian size and follicle numbers across all developmental stages. These findings demonstrate that CUMS induces an age-dependent progression toward POI, with short-term exposure eliciting compensatory phases preceding overt ovarian insufficiency, accompanied by distinct endocrine and reproductive alterations and differential responsiveness of the HPO axis. Transcriptomic analyses revealed age-dependent stress responses: ovaries of 2-month-old mice displayed marked activation of inflammatory and immune-related pathways, whereas 6-month-old mice showed sustained upregulation of protein kinase-related signaling networks. Notably, the 6-month-old CUMS model more closely recapitulates stress-associated reproductive aging in adult women. In briefCUMS has been widely used to establish mouse models of psychosocial stress-induced POI. However, current animal models do not adequately reflect middle-aged women, who represent a key population exposed to chronic psychosocial stress, nor do they capture the dynamic progression toward premature ovarian insufficiency (POI). In this study, we demonstrate that different durations of CUMS exposure induce distinct stages of ovarian dysfunction in both young and middle-aged mice, with short-term exposure driving age-dependent compensatory phases and prolonged exposure leading to overt POI, both accompanied by divergent endocrine and reproductive alterations, alongside age-dependent changes in HPO axis responsiveness to CUMS. Notably, the 6-month-old CUMS model shows greater clinical relevance in recapitulating chronic psychosocial stress and stress-related reproductive aging in adult women.

physiology↗

bioRxiv2026

Integration of spatial single-cell proteomics and spatial metabolomics reveals tumor microenvironment predictive of immunotherapy response in mucosal melanoma

Mucosal melanoma (MuM) is a rare but aggressive malignancy with limited benefit from immune checkpoint inhibition and few predictive biomarkers. We integrated single-cell spatial proteomics (COMET) and spatial metabolomics (MALDI-IMS) to profile 97 tissue cores from 26 patients treated with PD-1/PD-L1 and/or CTLA-4 inhibitors. We profiled 695,444 cells and resolved 25 cell states across eight major cell types. Cellular neighborhood (CN) analysis revealed distinct tumor- and stromal-associated spatial architectures. Responders were enriched for tumor-associated CNs (invasive tumor and tumor boundary) with close spatial proximity among Ki67 tumor cells, CD163 macrophages, and CD11c dendritic cells (DCs), and increased proliferating/cytotoxic CD8+ T-cell subsets. Non-responders showed stromal CN dominance with reduced immune infiltration. Spatial metabolomics identified lower abundance of indole-derived metabolites and reduced indole/tryptophan pathway activity in responders within tumor and TME regions that tracked with DC/macrophage-enriched spatial contexts. This study advances MuM spatial biology and provides a framework for biomarker-driven immunotherapy strategies. Statement of significanceMucosal melanoma (MuM) responds poorly to immune checkpoint blockade, and predictive biomarkers are limited. Integrated spatial proteomics and metabolomics reveal response-associated tumor-immune neighborhood architecture, stromal contexts linked to immune exclusion, and altered indole/tryptophan metabolism in the microenvironment. These spatial features nominate biomarkers and therapeutic hypotheses to improve immunotherapy for MuM.

cancer biology↗

bioRxiv2026

Parkinsonism disrupts the population-level organization of cortical dynamics

Parkinsons disease (PD) is marked by impairments in voluntary movement, including prolonged movement preparation and execution, yet how parkinsonism alters neural processing to produce these deficits remains unresolved. Prior work examining M1 spiking activity in parkinsonian states has largely characterized firing-rate changes and motor representations at the level of individual neurons, with inconsistent results and limited insight into population-level organization. Here we investigated how parkinsonism reshapes the population-level organization of neural activity in M1 during movement. We simultaneously recorded large populations of neurons from M1 in two nonhuman primates performing reaching tasks before and after induction of parkinsonism with the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). In the parkinsonian state, both preparation and reach durations were significantly prolonged. Population-level analyses revealed that parkinsonism increased the dimensionality of M1 activity during both preparation and movement and reduced the orthogonality between preparatory and movement-related subspaces. Moreover, trial-by-trial variability in reach duration was explained by the alignment of the preparatory and reach subspaces, indicating the functional role of subspace orthogonality. Together, these findings suggest that parkinsonism disrupts the population-level organization of cortical dynamics across computations, providing a population-level framework linking altered cortical dynamics to the movement-related dysfunction observed in PD.

neuroscience↗

bioRxiv2026

Biomimetic virus-like mesoporous silica nanoparticles activate NK cells indirectly via monocyte crosstalk

Cancer immunotherapies show clinical promise but often rely on T-cell priming and are limited by tumor heterogeneity and the immunosuppressive tumor microenvironment (TME). Innate immune activation offers a complementary strategy, with specific aim in natural killer (NK) cell activation for antigen-independent response. Biomimetic nanoparticles combining virus-like morphology with cell membrane (CM) coating offer a strategy to engage this innate immune axis. This study investigates virus-like mesoporous silica nanoparticles (VLPSi) with tunable spikes, surface functionalization, and CM coating as innate immunity modulators. Optimization revealed that longer spikes, amine functionalization, and CM coating synergistically enhance NK cell activation within human PBMCs, as indicated by CD69/CD25 upregulation and IFN-{gamma} secretion. CD14+ monocyte depletion attenuated activation, identifying monocyte-dependent crosstalk as a key mechanism. In purified NK cells, engineered CM-coated VLPSi induced early activation and supported feeder-free expansion. These results define topology, surface chemistry, and CM coating as parameters for innate immune modulation. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=69 SRC="FIGDIR/small/720074v1_ufig1.gif" ALT="Figure 1"> View larger version (18K): org.highwire.dtl.DTLVardef@db5cdorg.highwire.dtl.DTLVardef@1ab41eorg.highwire.dtl.DTLVardef@127428dorg.highwire.dtl.DTLVardef@82609a_HPS_FORMAT_FIGEXP M_FIG C_FIG

immunology↗

bioRxiv2026

Origin and evolution of grapevine genomes

Grapevines (Vitis) belonging to grape family (Vitaceae) are symbolic fruit crops pivotal to human civilization. The evolutionary history of grapevines divergent from other Vitaceae plants remains mysterious, requiring a family-wide whole-genome phylogenomic analysis. Here, we conduct chromosome-level phylogenomics to investigate the origin and evolution of grapevines using 29 genome assemblies of five genera Vitis, Parthenocissus, Ampelopsis, Tetrastigma, and Cissus, 27 of which are newly released in this study. Phylogenomic and macrosynteny analysis unanimously support Ampelopsis as a sister lineage to Parthenocissus, placing both closer to Vitis, with introgression and incomplete lineage sorting contributing to these relationships. Ancestral genome reconstruction delineates the major chromosome rearrangement events in Vitaceae karyotype evolution, highlighting the conserved karyotype in Vitis and the extensive karyotypic reorganization in Tetrastigma and Cissus. Pan-3D genome analysis highlights the contributions of structural variants (SVs) to the variation of A/B compartments and topologically associated domains (TADs), revealing a strong purifying selection of SVs at TAD boundaries. We further demonstrate that Helitron transposons drive the expansion and expression regulation of NLR immune-receptor genes in Vitis. Importantly, we discovered an NLR gene VbRpv35 from wild grapevine V. bellula resistant to downy mildew (DM), whose heterologous expression in V. vinifera confers enhanced DM resistance. Taken together, we provide phylogenomic insight into the origin and evolution of grapevines and valuable resources for grapevine improvement and understanding angiosperm evolution.

genomics↗

bioRxiv2026

The ABI5-WRKY45-LSU1 axis confers tolerance of Arabidopsis thaliana to cadmium

Abscisic acid (ABA) is involved in Cd tolerance in Arabidopsis, but the underlying mechanisms are unclear. In this study, we revealed that the ABI5-WRKY45-LSU1 axis confers the tolerance of Arabidopsis to Cd stress. Under Cd stress, the biosynthesis of ABA is increased, and the expression of transcription factor ABI5 is upregulated. Accordingly, the abi5-8 mutants show increased Cd sensitivity. ABI5 directly binds the ABRE element in the WRKY45 promoter to activate its transcription. Overexpression of WRKY45 rescues the Cd-hypersensitive phenotype of the abi5-8 mutant, placing WRKY45 downstream of ABI5. Transcriptome analyses identified LSU1 as a potential WRKY45 target. qRT-PCR, DUAL-LUC and EMSA experiments verified that WRKY45 binds the W-box cis-element in the LSU1 promoter to activate its expression. Overexpression of LSU1 enhances Cd tolerance by promoting the biosynthesis of non-protein thiols (NPT), glutathione (GSH), and phytochelatins (PC). Moreover, overexpression of LSU1 suppresses Cd sensitivity in the wrky45 mutant, confirming LSU1 acts downstream of WRKY45. On the other hand, we found that ATP sulfurylase 1 (APS1) interacts with LSU1 based in vitro and in vivo evidences. LSU1 stabilizes APS1, slows its degradation, and enhances APS1 activity, thus leading to increased NPT, GSH, and PC accumulation and improved Cd detoxification. Notably, overexpressing LSU1 did not rescue the Cd sensitivity of the aps1-1 mutant, indicating that LSU1 acts upstream of and depends on APS1. In short, we demonstrated a novel ABI5-WRKY45-LSU1 axis that regulates Cd tolerance through sulfur assimilation and phytochelatin synthesis. HighlightsO_LICadmium stress triggers ABA biosynthesis and ABI5 expression; ABI5 directly binds to ABRE motifs in the WRKY45 promoter and activates its transcription. C_LIO_LIWRKY45 transcriptionally activates LSU1, and LSU1 interacts with APS1 to stabilize it and elevate ATP sulfurylase activity, acting in an APS1-dependent manner. C_LIO_LIThe ABI5-WRKY45-LSU1 module enhances Arabidopsis Cd tolerance by boosting sulfur assimilation and GSH/PC-mediated Cd detoxification, rather than reducing Cd uptake. C_LI

molecular biology↗

bioRxiv2026

Autoluminescence characteristics of 25 Phalaenopsis orchids following transient transformation with eFBP2

Engineering autoluminescent plants, especially horticultural crops, has recently emerged as a promising research area, with one current approach involving the transgenic introduction of fungal bioluminescence pathway genes. Although autoluminescent plants such as tobacco and petunia have been created, it remains unclear whether all horticultural plants have the potential to be autoluminescent after genetic modification, especially those having flowers with dark colors and thick cuticles. To understand whether, and what if any, floral traits affect autoluminescence potential, we assess the autoluminescence characteristics of 25 representative Phalaenopsis cultivars after transient transformation of fungal bioluminescence pathway genes, alongside several key morphological and biochemical traits. Our results demonstrate that autoluminescence characteristics are correlated with floral color lightness, organ textures and epidermal cell types. In contrast, the content of the substrates of luciferin--caffeic acid and tyrosine, and the infiltration ease of inoculation solution into floral organs after injection, have limited effects on autoluminescence characteristics. Autoluminescence intensity can be reasonably predicted using five floral traits investigated, as 80.4% of variation can be explained by these traits. Our study not only identifies specific Phalaenopsis cultivars with high potential for developing autoluminescent lines but also provides a selection framework applicable to other horticultural crops.

plant biology↗

bioRxiv2026

Generation and long-term expansion of human pancreatic islet organoids in vitro

The scarcity of expandable, functional human islet cells remains a major barrier to diabetes therapy. Here, we identify PROCR+ cells within adult human islets and establish a defined culture system to generate pancreatic islet organoids. These organoids self-organize into -, {beta}-, {delta}-, and PP cells at near-native ratios, exhibit regulated insulin and glucagon secretion, and support exponential in vitro expansion. Single-cell transcriptomics reveals a unique progenitor-like cell population that is transcriptionally primed for endocrine differentiation but shares molecular features with fetal trunk cells and endocrine progenitors. When transplanted, the organoids rapidly ameliorate hyperglycemia in diabetic mice. Importantly, in a non-human primate model, intraportal transplantation of these organoids reduced exogenous insulin requirements, restored glucose-stimulated C-peptide secretion, and achieved sustained glycemic control-representing a critical step toward clinical translation. This study provides a strategy for expanding human islet organoids, offering a scalable platform for diabetes treatment, disease modeling, and regenerative medicine.

cell biology↗

bioRxiv2026

Application of a High-Biomimetic Tumor Organoid-CAF Co-Culture Model for the Efficacy Evaluation of CAR-T Drugs

The tumor microenvironment (TME) is a complex ecosystem composed of tumor cells, cancer-associated fibroblasts (CAFs), immune suppressive cells, and the extracellular matrix (ECM), playing a crucial role in tumor development and CAR-T cell therapy efficacy. CAR-T therapy has shown promise in hematological malignancies but faces challenges in solid tumors due to the TMEs ability to suppress CAR-T cell infiltration, proliferation, and cytotoxicity. Traditional drug evaluation models, such as 2D cell cultures and animal models, have significant limitations due to oversimplification of the in vivo environment or physiological differences between species. Organoid models offer a more biomimetic approach but often fail to fully recapitulate the TMEs complexity and heterogeneity. Our research developed a tumor organoid and CAF co-culture model using the IBAC co-culture chip, demonstrating that CAFs significantly impact CAR-T cell therapy efficacy by forming physical (e.g., fibronectin) and chemical (e.g., IL-10) barriers that prevent CAR-T cell infiltration and cytotoxicity. This model provides a high-biomimetic platform for investigating the TMEs effects on CAR-T therapy and highlights the importance of incorporating a comprehensive stromal component into in vitro models to enhance their predictive power for cancer treatment.

cell biology↗

bioRxiv2026

Cardiac oxidative stress monitoring enabled by hierarchical mechanical adaptation

Soft bioelectronics have advanced cardiac monitoring through electrophysiological tracking, yet this alone cannot resolve the metabolic pathology essential to surgical decision-making. However, real-time molecular sensing on beating hearts remains unresolved due to deformation-induced sensor failure and stress-induced metabolite artifacts. This challenge is exemplified by ischemia-reperfusion injury (IRI), a major cardiac surgery complication characterized by reactive oxidative species (ROS) bursts, where true pathological ROS signals being confounded by mechanotransduction-induced ROS artifacts. Herein, we propose an enzymatic cardiac oxidative stress biosensor (E-cardiac) with hierarchical mechanical adaptation: macro-scale biofluid-mediated contact, micro-scale fiber reorganization, and nano-scale enzymatic confinement within gold nanoarches dissipate interfacial stress. This produces ultrathin ([~]460 nm), soft (0.79 kPa) E-cardiac with robust electrochemical stability (100% strain), low detection limit (380 nM), rapid adhesion (<3 s), stable biosensing on beating heart, as well as minimal invasive deployment capability. Mechanical analysis and cellular studies confirm mitigated stress-induced ROS and absent PIEZO channel activation. Validated across cardiomyocytes, ex vivo tissues, multi-species ischemia models (mouse, rat, rabbit, pig), rat ischemia-reperfusion injury, and Langendorff hearts simulating graded perfusion deficits, E-cardiac quantitatively differentiates IRI severity (sham < ischemia < reperfusion) as well as detecting the "ECG blind window". The E-cardiac platform provides real-time metabolic feedback for surgical guidance during cardiac procedures, enabling timely intervention before irreversible damage.

bioengineering↗

bioRxiv2026

Phosphoregulated SMCR8-FIP200 interaction connects the ALS/FTD-linked C9orf72 complex to autophagy initiation and mitochondrial quality control

Hexanucleotide (GGGGCC) repeat expansions in the non-coding region of C9ORF72 are a major genetic cause of ALS/FTD and reduce C9orf72-SMCR8-WDR41 complex levels, but how this contributes to autophagy-lysosome dysfunction and previously reported mitochondrial quality-control defects in C9ORF72-ALS/FTD remains unclear. Here we identify a direct interaction between SMCR8 and the FIP200 subunit of the ULK1/2 autophagy initiation complex, mediated by two FIP200-interacting region (FIR) motifs in a disordered SMCR8 loop. Phosphorylation of these motifs by ULK1/2 or TBK1 strengthens binding and promotes ULK1/2 complex association in cells. Stabilising the SMCR8-FIP200 interaction suppresses Parkin-dependent mitophagy, whereas both stabilisation and weakening impair deferiprone-induced mitophagy, while leaving bulk autophagy, lysophagy and ivermectin-induced mitophagy largely intact. These findings define a regulated C9orf72-ULK1/2 axis and provide a mechanistic framework by which repeat-expansion-associated reduction in C9orf72 complex abundance may contribute to previously observed mitochondrial quality-control defects in C9ORF72-ALS/FTD.

cell biology↗

bioRxiv2026

Modular Deep Learning for Direct RNA Sequence Design via Self-Contained RNA Units

RNA sequence design is a pivotal challenge in synthetic biology, yet state-of-the-art deep learning methods face a fundamental bottleneck: the scarcity of high-resolution 3D structures. To compensate for limited training data, existing approaches like NA-MPNN and RiboDiffusion employ computationally expensive autoregressive or iterative diffusion sampling, substantially limiting their throughput and scalability. In this work, we propose that this data limitation is largely a problem of accessibility and granularity. We introduce SCRU-DB, a comprehensive database that systematically decomposes complex RNAs into over 61,000 Self-contained RNA Units (SCRUs). This scale far exceeds previous RNA motif libraries, capturing over 8,200 unique structural clusters. Crucially, SCRUs are rigorously defined as structurally autonomous modules identified via tertiary contact clustering, ensuring they act as self-stabilizing, foldable physical units. Leveraging this massive, modular prior, we present SCRU-Seq (a direct, O(1) prediction GNN) and SCRU-Diff (an iterative diffusion model). On our high-fidelity set112 benchmark, SCRU-Seq achieves a native sequence recovery (NSR) of 63.7%, while SCRU-Diff reaches a superior Best NSR of 79.2%. We demonstrate high structural fidelity via 3D backbone superposition using the C4 RMSD (reaching 1.5[A] for complex targets) and validate the structural isomorphism of our modular fragments. This framework provides a scalable, physically grounded solution for generating diverse and structurally accurate RNA sequences.

bioinformatics↗

bioRxiv2026

Differential effects of α-Synuclein monomers and seeds on the material properties of Tau condensates

Tau and -Synuclein (Syn) frequently co-aggregate in various neurodegenerative disorders. Recently, Tau has been shown to form dynamic, liquid-like condensates that can recruit Syn, and potentially serve as a precursor to pathological aggregation. However, the quantitative impact of Syn on the material properties of these condensates remains elusive. Here, we measure the viscosity and interfacial tension of Tau condensates and determine how these properties are modulated by Syn monomers and fibril seeds. We find that while both forms of Syn partition efficiently into Tau condensates, they exert vastly different effects on the condensates material state. The viscosity of Tau condensates remains unchanged in the presence of Syn monomers at concentrations up to 200 {micro}M, accompanied by a moderate reduction in the condensates interfacial tension. In contrast, the addition of only 5 {micro}M Syn fibril seeds triggers rapid solidification of Tau condensates, manifested by a nearly 100-fold increase in condensate viscosity within one hour. These findings provide quantitative insights into condensate mechanics, highlighting the unique capacity of Syn seeds to drive the liquid-to-solid transition of Tau condensates that may underlie the formation of pathological aggregates.

biophysics↗

bioRxiv2026

Biomolecular condensates provide a unique environment for redox-mediated protein crosslinking

Biomolecular condensates, often formed through liquid-liquid phase separation, are dynamic cellular compartments. Here, we demonstrate that a wide range of fluorescently tagged proteins undergo inadvertent, condensate-mediated crosslinking, resulting in rapid solidification of condensates under common fluorescence imaging conditions. The process is driven by excitation-induced, short-lived reactive oxygen species (ROS), whose otherwise limited crosslinking potential becomes uniquely enabled in the dense phase. In live cells, excitation-induced ROS potently trigger stress granule formation, while the ROS-driven solidification of condensates is modulated by compartment-dependent antioxidant buffering. Our findings demonstrate that condensates create a distinct environment that enables ROS chemistry unlikely to occur in the bulk cytosol. Furthermore, the cellular redox level can be a general regulator of condensate rheology. Beyond biological insights, our findings underscore the need for scrutiny when examining fluorophore-labeled condensates.

biophysics↗

bioRxiv2026

Enforced ZFP281 expression delays breast cancer initiation and can provide lifelong protection against breast cancer metastasis

Breast cancer metastatic reactivation and its links to mammary development are largely unknown. Here, using conditional knockout and overexpression in normal and HER2+ mammary glands, we show that the dormancy regulator ZFP281 promotes branching and dissemination while suppressing growth, and its overexpression can even override HER2-driven cancer initiation. Notably, while ZFP281 does not limit HER2-driven early dissemination, it constrains DCC plasticity, confining cells to a dormant mesenchymal/hybrid-like state and effectively suppressing metastasis throughout the mouse lifespan. ZFP281 is induced by estrogen, progesterone, and glucocorticoid signaling, and RNA sequencing of early lesions revealed that it regulates glutathione metabolism and ferroptosis, potentially supporting fitness during dormancy, while repressing angiogenesis, Th17-like inflammation, innate immune genes, and pro-inflammatory programs that might otherwise trigger awakening. Integrating these findings with human data, we show that DCIS and IBC (invasive breast cancer) lesions that later relapse are selectively enriched for ZFP281-regulated M-like and dormancy signatures and, after pregnancy, depleted for a ZFP281-induced stress-autophagy module, indicating that erosion or imbalance of these programs marks lesions that seed DCCs with higher reactivation potential. We propose that ZFP281 acts as a hormone-regulated dormancy gatekeeper that uncouples dissemination from growth, enforcing a metabolically fit, angiogenesis-low, immune-evasive dormant state in breast DCCs, thereby shaping the timing of metastatic relapse and potentially exploitable for durable prevention of metastasis. STATEMENT OF SIGNIFICANCEZFP281, a hormone-regulated dormancy gatekeeper, uncouples dissemination from growth and constrains DCCs into long-term arrest, defining human gene signatures that distinguish dormancy-prone from awakening-prone lesions and predict breast cancer relapse dynamics.

cancer biology↗

bioRxiv2026

A C. elegans model for functional analysis of ADPKD variants in cilia, extracellular vesicles, and sensory signaling

Interpreting the pathogenic significance of missense variants in human disease gene candidates remains a major challenge in precision medicine. Autosomal dominant polycystic kidney disease (ADPKD) is the most common genetic cause of kidney failure and caused by mutations in the PKD1 or PKD2 genes that encode polycystin-1 and polycystin-2. Here, we establish C. elegans as a platform for the functional classification of PC2 variants by characterizing PKD-2C180S, the C. elegans ortholog of the likely pathogenic human variant PC2C331S. Using CRISPR/Cas9 endogenous genome editing combined with dual-color fluorescent reporters and super-resolution imaging, we show that PKD-2C180S severely reduces protein stability, abolishes ciliary and extracellular vesicle (EV) localization, and eliminates sensory function comparable to a pkd-2 null allele. In heterozygous animals, PKD-2C180S is recessive and exerts no dominant-negative effect on wild-type PKD-2 trafficking, protein levels, or function, establishing that PKD-2 is haplosufficient in this model. PKD-2C180S also abolishes ciliary and EV localization of the PC1 homolog LOV-1 and reduces LOV-1 cell body levels comparable to pkd-2 null animals, consistent with PC2 functioning as a molecular chaperone for PC1 stability and trafficking. Genetic epistasis experiments show that PKD-2C180S protein levels are unaffected in lov-1 mutants, indicating that the PKD-2C180S mutation acts prior to complex assembly. Quantitative analysis reveals that LOV-1*PKD-2 complexes are more stable at the ciliary membrane and more efficiently packaged into EVs than PKD-2 lacking LOV-1. Together, this work demonstrates that PC2C331S may act recessively via loss of polycystin complex function and establishes a C. elegans pipeline for the mechanistic classification of ADPKD-associated variants.

genetics↗

bioRxiv2026

Discovery and Development of First-in-Class Cereblon-Recruiting RIPK1 Degraders

Receptor-interacting protein kinase 1 (RIPK1) is a critical regulator of programmed cell death and is implicated in various pathological conditions, particularly in mediating tumor resistance to immune checkpoint inhibitors (ICBs). In this study, we have pioneered the development of a novel cereblon (CRBN)-recruiting RIPK1 degrader, LD5095, through systematic optimization of linker and CRBN ligand portion. LD5095 demonstrates potent and selective RIPK1 degradation across cell lines, with rapid kinetics and sustained degradation over 72h post-washout. Functionally, RIPK1 degradation by LD5095 significantly sensitized Jurkat cells to TNF-induced apoptosis. Furthermore, LD5095 exhibited favorable pharmacokinetics, including metabolic stability and an extended half-life. Strikingly, in vivo, a single dose of LD5095 achieved durable RIPK1 degradation in xenograft tumors over 6 days. These findings underscore the potential of LD5095 as a chemical probe for studying RIPK1 biology and a promising candidate for cancer treatment.

pharmacology and toxicology↗