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Lin, Y.

Publications and source records attributed to Lin, Y..

At least 37 records · Page 2Linked to original sources

A novel AR molecular glue degrader via bivalent engagement of the N-terminal domain and RNF213-UBE2J2 complex

Targeted protein degradation enables the elimination of disease-relevant proteins through induced proximity between E3 ligases and substrates. Current androgen receptor (AR) therapies act via the ligand-binding domain (LBD) but fail against constitutively active LBD mutants or splice variants such as AR-V7. Here, we identify GZL626, a small-molecule degrader that induces proteasome-dependent degradation of both full-length AR and AR-V7. GZL626 suppresses AR transcriptional activity, inhibits proliferation of AR-positive prostate cancer cells, and blocks SARS-CoV-2 infection. Mechanistically, GZL626 binds the intrinsically disordered AR N-terminal domain (NTD) and promotes formation of a noncanonical RNF213-UBE2J2-AR ternary complex that drives AR ubiquitination and degradation. The AR DNA-binding domain (DBD) acts as a structural hub within this assembly. Distinct from PROTACs or classical molecular glues, GZL626 engages both AR and RNF213 bivalently, remodeling protein interactions to stabilize a functional E3-E2-substrate complex. These findings establish GZL626 as a first-in-class NTD degrader with potential to overcome resistance in advanced prostate cancer.

biophysics↗

Assessing Microcirculation Impairment in Ischemic Stroke Mice Using Arteriovenous Co-fluctuation Analysis

Accurate assessment of cerebral hemodynamics impairment traditionally relies on arterial metrics, yet often overlooks venous drainage and arteriovenous dynamics, thereby limiting the evaluation of ischemia-induced microvascular dysfunction. To address this limitation, we implemented a signal-averaging framework, combined with co-fluctuation analysis, to extract predominantly arterial and venous hemodynamic signals and construct a dynamic arteriovenous co-fluctuation index that quantifies frame-by-frame coordination between arterial inflow and venous outflow activity. This time-resolved index enables spatial characterization of large-scale cortical arteriovenous coordination beyond conventional static correlation-based analyses. Comparative analyses between healthy controls and acute ischemic stroke mice demonstrated that the arteriovenous co-fluctuation index sensitively detects disruption of vascular coordination, revealing a slower state transition that occurs alongside distinct temporal abnormalities and regional heterogeneity between ischemic core and penumbral regions. These findings underscore the utility of arteriovenous coordination as a sensitive indicator of microcirculatory dysfunction, offering a practical analytical tool for assessing stroke-induced microvascular impairment.

neuroscience↗

Field-Correcting GRAPPA (FCG): a technique to correctspatiotemporal-varying phase errors in Echo Planar Imaging

PurposeTo develop a Field-Correcting GRAPPA (FCG) technique to correct the spatiotemporal-varying phase errors in EPI caused by eddy currents. MethodsThe fast-changing gradient in EPI causes strong eddy current effects and associated spatiotemporal-varying phase errors, producing significant image artifacts. The use of higher gradient amplitude, slew rate, and ramp sampling factor for faster imaging exacerbates this problem. In this work, FCG was developed to address this challenge by using a multi-layer perceptron (MLP) to provide a compact representation of a family of GRAPPA-like kernels that correct the spatiotemporal-varying phase errors in the data. A dedicated calibration pipeline was designed to acquire high-quality source and target data for MLP training in both slice-by-slice and simultaneous multi-slice (SMS) acquisitions. To validate FCGs assumptions and performance, a field camera was used to provide ground-truth measurement of phase patterns. The performance of FCG was further validated on phantom and in vivo experiments using demanding EPI trajectories across multiple 3T and 7T systems. ResultsField camera measurements revealed strong spatiotemporal phase variations along the kx direction that repeat along ky during EPI readouts. The experiments on high-performance systems across 3T and 7T demonstrate that FCG can provide superior correction for the artifacts induced by spatiotemporal-varying phase errors compared with existing approaches. ConclusionFCG is an effective and robust method for correcting spatiotemporal phase errors in EPI, enabling improved image quality on high-performance systems.

bioengineering↗

Visual uncertainty and task demands shape active sensing strategies in mice

In natural environments, animals actively sample visual information to guide behavior. Sensory feedback is dynamic and often requires active movements, whether saccading across the lines of this page or walking through a park. From high-acuity vision in hawks to low-acuity mice, many animals actively navigate to seek information, which can be called infotaxis. Although mice have relatively low-acuity vision, they still rely on sight for critical behaviors including navigation and prey capture. Yet, how sensitive they are to visual information and performing infotaxis has not been established. Here, we develop a virtual reality object discrimination task to investigate visual decision-making under naturalistic conditions. We show that mice perform infotaxis by actively seeking out informative views to guide their choices. Stimulus manipulations confirm that this strategy is modulated by the amount of available visual information. These results reveal that mice use principled active strategies to resolve visual uncertainty, highlighting a key role for information-seeking in natural vision. HighlightsO_LIMice perform active visual sensing in a free-range VR object discrimination task. C_LIO_LIThey perform infotaxis and select the correct object under variable visibility. C_LIO_LIDecisions decoded from head-body movements and speed reflect occlusion sensitivity. C_LI

neuroscience↗

Scout-based Multi-Echo NAvigator (SMENA) for high temporal resolution motion and B0 estimation and correction: applications to multi-echo GRE and EPTI

PurposeTo develop a data-driven technique, Scout-based Multi-Echo NAvigator (SMENA), for joint estimation of motion and B0 inhomogeneity ({delta}B0) at a temporal resolution of [~]200 ms with minimal additional scan time for gradient-echo acquisition. MethodsSMENA consists of two key acquisition components: SMENA-scout and SMENA-nav. SMENA-scout is a rapid 3D 4-mm multi-echo acquisition completed in less than 8 seconds, providing images with matched contrast and phase at multiple echo times. SMENA-nav captures signal variations induced by motion and{delta} B0 during the scan using compact multi-echo navigator trajectories (3.5 ms) embedded within each TR. Motion and{delta} B0 maps were jointly estimated every [~]200 ms through a model-based optimization framework relating SMENA-scout to SMENA-nav. The estimation accuracy and correction performance of SMENA were evaluated in simulations and in vivo using multi-echo GRE and GRE-EPTI acquisitions. Multiple prospective motion experiments, including large continuous movement and deep breathing, were investigated. ResultsIn both simulations and in vivo experiments, accurate motion and{delta} B0 estimation were achieved. Compared with motion-only estimation, joint estimation reduced rotation and translation errors. Joint motion and{delta} B0 correction resulted in substantial improvements in image quality, particularly at longer echo times, producing an NRMSE of 10.4% compared to 31.6% with motion-only correction. High-temporal-resolution tracking of motion and{delta} B0 enabled improved reconstruction quality in scenarios involving continuous motion and deep breathing. ConclusionSMENA enables high-temporal-resolution joint estimation of motion and{delta} B0 with minimal additional acquisition cost, providing a practical solution for motion- and{delta} B0-robust MRI.

neuroscience↗

Type I interferon primes the alveolar epithelium to receive reparative signals from tissue-resident macrophages

Lung repair in response to viral infection requires integrated communication between epithelial and immune compartments, yet the impact of antiviral mediators on epithelial regenerative capacity remains poorly defined. Here, we demonstrate that type I interferon (IFN-I) signaling primes the lung for alveolar renewal following viral challenge. IFN-I contributes to the induction of an interferon-stimulated gene-high (ISG)hi Sca-1Pos population of alveolar type II (ATII) epithelial cells. Sca-1Pos ATIIs exhibit enhanced proliferative capacity and increased organoid-forming efficiency compared with their Sca-1Neg counterparts. Viral challenge concurrently drives phenotypic reprogramming of tissueresident alveolar macrophages (trAMs). Sca-1Pos ATIIs display heightened responsiveness to oncostatin M (OSM) and, following viral challenge, require trAM-derived OSM for their proliferation. Together, these findings reveal that viral stimuli induce coordinated IFN-I-dependent epithelial and macrophage states that poise the lung for regeneration, positioning IFN-I not only as a central antiviral defense mechanism but as a priming signal that prepares lung tissue for renewal.

immunology↗

A Brugada-related KCNT1 mutation unveils its conductance-independent activation of store-operated Ca 2+ entry

BackgroundPotassium sodium-activated channel subfamily T member 1 (KCNT1) is a Na+-activated K+ channel, associated with epilepsy and cardiac diseases. Besides K+ conductance, KCNT1 is also involved in Ca2+ handling. KCNT1:R1106Q was identified in a Brugada Syndrome (BrS) patient; however, its pathological characteristics regarding either K+ or Ca2+ homeostasis remain to be fully elucidated. MethodsFura-2-loaded HEK293T cells were used for intracellular Ca2+ investigations, and whole-cell patch-clamp technique were used as complementary tests. Stromal interaction molecule 1 (STIM1) aggregation and endoplasmic reticulum (ER)-plasma membrane (PM) junctions in HeLa cells were visualized using spinning disc confocal microscope. Multielectrode array (MEA) assays were utilized to assess the field potential duration (FPD) and spontaneous beating rate of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). ResultsKCNT1 overexpression increased SOCE, which was further upregulated by the KCNT1R1106Q mutant. Furthermore, KCNT1 upregulated ER Ca2+ release and was found to localize at ER-PM junctions. The ER Ca2+ dynamics in KCNT1R1106-overexpressing cells was disrupted, while its SOCE remained intact. The KCNT1 cytoplasmic tail alone sufficiently modulated Ca2+ homeostasis. Given that KCNT1 possesses prominent charge-concentrated segments within its cytoplasmic domain, we substituted charged amino acids in either the 740-DDE-742 or 1114-RRLSR-1118 motifs with alanine; these substitutions abolished the KCNT1-mediated increases in ER Ca2+ release and ER-PM contact formation. MEA recordings of hiPSC-CMs revealed that the overexpression of KCNT1 shortened the FPD and accelerated the cardiomyocyte beating rate compared to baseline. However, the magnitude of beating rate acceleration induced by KCNT1R1106Q was significantly attenuated compared to that of KCNT1WT. ConclusionsOur findings indicate that, independent of KCNT1-mediated ion conductance, the charged motifs located at both ends of the KCNT1 cytoplasmic tail serve as structural anchors to facilitate ER-PM contact formation. This non-conducting role endows KCNT1 with a K+ current-independent mechanism to modulate intracellular Ca2+ homeostasis and cardiomyocyte physiology. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=133 SRC="FIGDIR/small/731813v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@104e234org.highwire.dtl.DTLVardef@19eceb3org.highwire.dtl.DTLVardef@1148512org.highwire.dtl.DTLVardef@2c8118_HPS_FORMAT_FIGEXP M_FIG C_FIG Novelty and SignificanceO_ST_ABSWhat is known?C_ST_ABSO_LIKCNT1 is a Na+-activated K+ channel, and mutations within this gene are often associated with epilepsy and epilepsy-related cardiac diseases. C_LIO_LIBesides functioning as a rectifier, KCNT1 is implicated in cellular Ca2+ handling, such as regulating Ca2+ oscillations in rat cortical neurons. C_LIO_LIKCNT1:R1106Q missense mutation has been identified in a patient with Brugada Syndrome (BrS); however, its pathophysiological characteristics and underlying mechanisms remain uncharacterized. C_LI What New Information Does This Article Contribute?O_LIKCNT1 overexpression upregulates both ER Ca2+ release and store-operated Ca2+ entry (SOCE). KCNT1R1106Q further increased SOCE intensity, however, its increase in ER Ca2+ release and [Ca2+]ER were attenuated. C_LIO_LIThe cytoplasmic tail of KCNT1 alone is suNicient to modulate Ca2+ closely mimicking the eNects of full-length KCNT1. C_LIO_LIThe charged clusters distributed along the KCNT1 cytoplasmic tail facilitate ER-PM contact. Alanine substitutions abolish the enhancements in ER Ca2+ release and ER-PM contact formation, while SOCE modulation remains intact. C_LIO_LIThe KCNT1 cytoplasmic tail alone shortens the field potential duration (FPD) and accelerates the spontaneous beating rate in hiPSC-CMs. However, the magnitude of beating rate acceleration induced by KCNT1R1106Q is significantly attenuated compared to KCNT1WT. C_LI This study uncovers a novel, conductance-independent role for the KCNT1 channel in structural organization, demonstrating that the charged motifs on its cytoplasmic tail serve as physical anchors facilitating ER-PM junctions. This structural mechanism regulates intracellular Ca2+ homeostasis and subsequently alters cardiomyocyte electrophysiology. By decoupling KCNT1s structural impact from its ion-conducting function, these findings provide new mechanistic insights into how specific KCNT1 variants, such as the BrS-associated KCNT1:R1106Q mutation, may contribute to arrhythmogenesis through imbalanced Ca2+ handling rather than classical K+ current alterations.

Molecular Biology↗

Cell-Type-Selective Cortical Pathology and Functional Deficits in Synucleinopathy

Aggregates of -synuclein (-syn), a hallmark of synucleinopathies, accumulate in the cerebral cortex accompanied by the emergence of motor symptoms, which are associated with altered cortical neuronal activity. However, the mechanism by which -syn pathology drives cortical network dysfunction, and how these alterations contribute to impaired motor execution and learning, remain unknown. Here, we adopted a multi-disciplinary approach to elucidate the pathophysiological characteristics in transgenic mice that express mutant human -syn, with minimal nigrostriatal degeneration. In vivo two-photon imaging revealed distinct alteration patterns in excitatory and parvalbumin (PV)-expressing inhibitory cortical neurons accompanying fine motor deficits during learning. Cell type specific ex vivo whole-cell recording further revealed selectively altered intrinsic properties in excitatory but not PV neurons, consistent with the preferential accumulation of -syn inclusions in excitatory rather than PV neurons within the same cortical region. These results indicate cell-type selective vulnerability in motor cortex of early stage synucleinopathy, leading to disrupted excitatory/inhibitory balance and dysregulated cortical plasticity, driving early-stage motor symptoms. This study provides evidence for selective vulnerability of excitatory neurons in cortical synucleinopathy.

neuroscience↗

Preserved self-other integration during social decision making among individuals with elevated autistic traits

Autistic people can find social interactions difficult to navigate, traditionally attributed to difficulties in taking others perspectives. However, we have a limited understanding of how autistic people integrate self and other information efficiently during social decision-making. We conducted four highly powered experiments (total N = 1,621) to determine whether autistic traits affect two aspects of self-other integration during social decision making: self-bias and social basis function use. Using Bayesian analyses, we found strong support for the absence of a relationship between autistic traits and either aspect of social decision making, even after controlling for potential confounds (BF01 = 32.13 for self-bias, BF01 = 7.04 for social basis function use). Our results indicate that variations along autistic traits do not impact how people prioritise self-relevant information (self-bias) or utilize compressed social patterns of interaction (social basis function use) to guide their decisions about oneself and other people. These findings nuance the conceptualisation of social-cognitive processes across autistic traits while highlighting the need for large samples to validate null effects.

neuroscience↗

EZH1/2 inhibition improves immunotherapy response through MHC Class II de-repression and neutrophil reprogramming

Squamous cell carcinoma of the lung is a difficult-to-treat cancer with high prevalence in the US, and particularly in Kentucky. Here, we sought to test if the EZH1/2 inhibitor valemetostat improves anti-PD1 responses in squamous cell lung cancer models, and to develop ex vivo models to test immunotherapy drug combinations. We found that valemetostat augmented anti-tumor responses to anti-PD1 therapy, led to up-regulation of tumor cell specific Major Histocompatibility Complex Class II (MHC Class II), and drove systemic neutrophil maturation. Likewise, Ezh2 knock-out mice produced neutrophils that were more apoptotic, less migratory, and less able to produce extracellular nets, but had similar ability to kill bacteria as Ezh2-WT neutrophils. To test tumor responses to differing neutrophil populations, we engineered three-dimensional air-liquid interface cultures with tumoroids, lung mesenchymal cells, and T cells, with and without bone marrow containing neutrophils and myeloid progenitors from distinct donors. Bone marrow from tumor-naive or mice with actively growing untreated tumors boosted tumoroid growth, while bone marrow from spontaneously tumor-rejected or mice with tumors treated with valemetostat was anti-tumor. MHC Class II blockade lowered the ability of bone marrow to boost tumor growth, and reduced the ability of valemetostat with anti-PD1 to reduce tumoroid growth. Patient samples revealed a strong negative correlation between EZH2 and MHC Class II, suggesting that targeting EZH2 activity could lead to marked increase in MHC Class II and improve treatment responses in lung squamous cell carcinomas.

cancer biology↗

Assessing and Optimizing Low-Frequency Somatic Mutation Detection: A Multi-Platform High-Throughput Sequencing Perspective

The availability of multiple commercial short-read sequencing platforms necessitates systematic cross-platform performance comparisons, particularly for challenging applications such as low-frequency somatic mutation detection. Here, a large-scale targeted sequencing dataset from five Genome in a Bottle (GIAB) human genomic DNA reference standards, HG001 to HG005, alongside Twist Biosciences cfDNA reference standards featuring 1% variant allele frequency (VAF), was generated by six platforms (NovaSeq 6000, NovaSeq X, FASTASeq 300, GenoLab M, SURFSeq 5000, and MGISEQ-T7). To build a realistic benchmark while keeping authentic sequencing backgrounds, we developed PosMix, a simulating tool that generates position-specific VAFs. To overcome the limitations of conventional variant callers (high recall with poor precision for VarScan2, higher precision with lower recall for Strelka2/Mutect2), we developed SomaticXGB, a machine learning-based caller. In this study, SURFSeq 5000 consistently exhibited the lowest error rates and achieved superior accuracy for VAFs as low as 0.5%, outperforming all other sequencing platforms. On the other hand, SomaticXGB attained F1 scores of approximately 0.92 on simulated datasets with VAFs ranging from 0.5% to 1.5% and 0.89 on Twist 1% standards, substantially outperforming conventional methods. This work delivers a valuable rich multi-platform data resource, offering a standardized pipeline for performance benchmarking and a machine learning-based strategy for optimized somatic mutation detection.

bioinformatics↗

Trichome acylsugar defenses are equally effective against leaf-mining and free-living insects

Trichomes are epidermal projections, crucial components of plant defense systems. Whether these epidermal defenses are effective against leafminers, which infest the internal mesophyll layers, remains unclear. Using the devasting invasive tomato pest, Phthorimaea absoluta (tomato leafminer), and the trichome-rich wild tomato, Solanum pennellii, we demonstrate that trichome produced acylsugars effectively inhibit the growth of endophagous P. absoluta larvae feeding inside leaves. High spatial resolution leaf- and frass-metabolite analysis by MALDI-MSI and Stimulated Raman Scattering microscopy revealed that trichome-synthesized acylsugars are translocated into the mesophyll tissues when larvae infest leaves. We identified an ATP-binding cassette (ABC) transporter, SpABCB5, as the mediator of this translocation and as an essential component of defense against leafminers. These data reveal that epidermal trichomes not only defend against free-living (ectophagous) herbivores through direct contact or rapid deposition of chemical defenses onto leaf surfaces, but also defend against endophagous leafminers by rapidly depositing chemical defenses into internal tissues. By delivering mines against leaf miners, trichomes offer new avenues for breeding crops with resistance to leafminers.

plant biology↗

Inhibition of hexokinase 2 undermines cartilage health and accelerates osteoarthritis

ObjectiveEnhanced glycolysis is a metabolic hallmark of chondrocytes in osteoarthritis (OA); however, the roles of the glycolytic rate-limiting enzyme hexokinase 2 (HK2) in cartilage remain poorly understood. MethodsPharmacological approach (3-bromopyruvate (3-BrPA) treatment) and mice model involving HK2 knockout in Col2a1-expressing chondrocytes are utilized to access the impact of HK2 blockage on cartilage ex vivo and in vivo. The in vivo effects of HK2 inhibition on OA progression were evaluated using a destabilization of the medial meniscus (DMM)-induced OA mouse model, through both intra-articular 3-BrPA administration and chondrocyte HK2 deletion. Additionally, we analyzed published single-cell RNA sequencing (scRNA-seq) datasets from human articular cartilage and integrated these with bulk RNA-seq data from HK2-deficient chondrocytes to characterize HK2 expression features across conditions. ResultsBoth pharmacological inhibition and genetic deletion of HK2 impair cartilage formation ex vivo. Bulk RNA-seq analysis and ex vivo studies demonstrated a promoted ossification-like process due to HK2 ablation in chondrocytes. Through pseudotime analysis of published single-cell RNA sequencing (scRNA-seq) datasets from human articular cartilages, we further identified that HK2 is differentially expressed across conditions, with a feature of a relatively high expression level at terminal stages of chondrocyte differentiation in the context of OA. We next confirmed HK2 deficiency in chondrocytes significantly exacerbated OA progression but having no impact on skeletal development in mice. ConclusionsHK2 plays a critical role in maintaining cartilage health, likely through the regulation of calcification, thereby highlighting the potential risks associated with targeting glycolytic enzymes as a therapeutic strategy for OA.

pathology↗

High-Dimensional Sensitivity Analysis for Genomic Studies: An Adversarial Framework for Learning Worst-Case Latent Confounders

High-dimensional genomics studies are frequently confounded by unmeasured biological processes that obscure disease-specific signals. While existing workflows can estimate these latent confounders, they fail to quantify how robust a discovery is to varying levels of hypothetical confounding. We introduce sensGAN, a deep-learning adversarial framework that systematically explores the confounding spectrum by learning "worst-case" latent variables that nullify the most gene associations under novel predictive-gain constraints. By identifying the minimum confounding strength required to explain away an observed effect, our method shifts the paradigm toward a formal, quantitative sensitivity analysis. In diverse simulations, sensGAN accurately recovers latent structures and outper-forms existing methods in identifying confounder-sensitive genes. Applied to human Alzheimers disease microglia, our framework prioritizes robust disease pathways while successfully isolating signals driven by unmeasured co-occurring neurodegenerative pathologies. Our method is publicly available, deposited at the GitHub repository yifanlinz/AD_sensitivity_ICML.

genomics↗

Genos-m: a foundation model for human-associated microbial genomes

Human-associated microbial genomes encode extensive strain-level diversity and niche-specific gene repertoires that are critical to host health. However, these complex sequence features remain difficult to capture using general-purpose DNA foundation models, highlighting the need for dedicated representation learning tailored to the human microbiome. Here, we introduce Genos-m, an open-source foundation model for human-associated microbial genome representation. Genos-m was pretrained on approximately 1.2 trillion nucleotide tokens from a curated microbial genome corpus, including human-associated prokaryotic isolates, high-quality metagenome-assembled genomes (MAGs) and bacteriophages, supplemented with GTDB species-level representative genomes to broaden prokaryotic taxonomic breadth. The model uses a sparsely activated Mixture-of-Experts (MoE) Transformer architecture, with 4.7 billion total parameters, approximately 330 million activated parameters per forward pass and a maximum context length of one million base pairs. We evaluated frozen Genos-m representations across short-sequence and gene-level tasks, biosynthetic gene cluster (BGC)-based regional sequence tasks, whole-genome strain phenotype prediction, and zero-shot transfer on prokaryote-related RNAfitness assays. Across these benchmarks, Genos-m consistently ranked among the leading comparison models, with the best performance in five of eight gene-fitness regression tasks and in BGC type classification. Using sparse autoencoders, we identified sparse features in Genos-m hidden activations that aligned with annotated ORFs, intergenic regions, and tRNA and rRNA loci. In downstream applications, Genos-m-derived genome-informed species representations in-corporated into a human microbiome self-supervised learning model improved colorectal cancer (CRC)-control classification over conventional species-abundance random forest models. Genos-m also generated stable sample-level embeddings from as few as 10,000 metagenomic reads, retaining gut microbial community structure that distinguished geographic origin and aligned with enterotypes defined from full-depth taxonomic profiles. Together, these results support Genos-m as a reusable representation model for microbial genomes and metagenomes, with conclusions bounded by the reported datasets, task definitions and evaluation protocols. Genos-m model weights, inference code, and usage documentation are publicly available on GitHub (https://github.com/BGI-HangzhouAI/Genos-m) and Hugging-Face (https://huggingface.co/BGI-HangzhouAI/Genos-m).

bioinformatics↗

The HOG MAPK - Transcription Factor CsAtf1 - CsErg5B Regulatory Module Mediates Conidial Germination and Fludioxonil Sensitivity in Colletotrichum siamense

Colletotrichum siamense is a predominant causal agent of anthracnose in rubber tree and numerous economically important crops, causing severe yield losses worldwide. Conidial germination represents a critical early step for successful infection, while the high-osmolarity glycerol (HOG) MAPK pathway and ergosterol biosynthesis individually govern fungal development, stress adaptation and fungicide responses. However, the molecular crosstalk between these two modules remains largely elusive in phytopathogenic fungi. Here, we identified CsErg5B, a sterol C-22 desaturase homolog, as a direct target of the HOG- regulated transcription factor CsAtf1 in C. siamense. CsErg5B was indispensable for ergosterol biosynthesis, conidial germination, appressorium formation, and full virulence. The {Delta}CsErg5B mutant showed increased conidiation but severely impaired germination, and exhibited elevated resistance to fludioxonil while hypersensitivity to azole fungicides. Epistasis analysis using the {Delta}CsErg5B/{Delta}CsCyp51G1 double mutant - where CsCyp51G1 serves as another downstream target of CsAtf1 - revealed that CsErg5B functions as the predominant downstream effector of CsAtf1 in modulating conidial development and fludioxonil sensitivity. Furthermore, overexpression of CsErg5B significantly rescued the defects in conidial germination and fludioxonil sensitivity in both {Delta}CsAtf1 and {Delta}CsPbs2 mutants. Taken together, our findings uncover a HOG MAPK - CsAtf1 - CsErg5B regulatory axis that connects HOG MAPK signaling to ergosterol homeostasis, thereby governing conidial germination and fungicide sensitivity in C. siamense. This study provides novel insights into the regulatory network underlying fungal development and fungicide response, and offers promising molecular targets for the integrated management of plant anthracnose.

microbiology↗

Selective Pyroptosis in NF1-Deficient Cells through PKCδ Agonism

Pyroptosis, a lytic and immunogenic form of cell death, holds broad therapeutic potential, yet its selective induction in specific cell populations remains a fundamental challenge. Loss of the NF1 tumor suppressor, one of the most frequent events across pediatric and adult cancers, elevates RAS-GTP and drives tumorigenesis through hyperactivated RAS signaling. Here we demonstrate that protein kinase C{delta} (PKC{delta}) agonism selectively triggers pyroptosis in NF1-deficient cells by exploiting their dependency on KRAS. PKC{delta} directly phosphorylates KRAS at S39 and S181, inducing KRAS-GDP accumulation and driving endoplasmic reticulum translocation. The dually phosphorylated KRAS-GDP interacts with caspase-8 and competitively displaces inhibitory BCL2, promoting caspase-8/caspase-3/gasdermin-E-mediated pyroptosis. This vulnerability is conserved across multiple NF1-deficient tumor types, and PKC agonism suppresses NF1-deficient neurofibroma and malignant peripheral nerve sheath tumor growth in vivo. These findings establish the inactive KRAS-GDP as a functionally active signaling molecule and PKC{delta} agonism as a selective therapeutic strategy for NF1-deficient cancers.

cancer biology↗

Widespread use of invalid statistical tests in biomedical machine learning

Cross-validation is routinely used to compare performance in biomedical artificial intelligence. Standard tests ignore correlation across cross-validation folds, inflating false-positive rates. In a PRISMA-guided meta-analysis of 184 studies (impact factor [≥] 15) across 30 biomedical fields, 97% use invalid tests. Among studies with abstract-level claims supported by invalid tests, 59% rely on a spurious comparison: one that loses significance after correcting for this correlation. On average, regaining significance requires a 43% larger effect. Extending these findings to the broader literature, we estimate that spurious comparisons occur in nearly two in three studies and support abstract-level claims in nearly one in three studies. Simulations confirm false-positive rates paradoxically approach 100% when cross-validation is repeated to improve stability. We introduce SHARP, a redesign of cross-validation, which best balances false-positive control and power among 13 benchmarked tests. These results reveal widespread fragility in biomedical artificial intelligence and offer a practical route to valid comparisons.

bioinformatics↗