Biology subjects

Wang, J.

Publications and source records attributed to Wang, J..

At least 541 records · Page 30Linked to original sources

bioRxiv2025

Ultra-High-Throughput LC-MS Method for Targeted Protein Degradation Compound Screening Using the Orbitrap Astral Mass Spectrometer

High-throughput proteomics analysis is crucial for targeted protein degradation (TPD) compound screening. Here, we developed a 300 sample per day (SPD) LC-MS method using the Orbitrap Astral mass spectrometer for ultra-high-throughput TPD compound screening. We identified close to 8,000 protein groups from a single cell line with a coefficient of variance (CV) of less than 10%, highlighting the deep proteome coverage and method reproducibility even at 300 SPD. This high degree of precision provides the statistical confidence to detect subtle, yet significant, changes in protein abundance that were previously challenging to quantify in high-throughput workflows. To evaluate the quantitation accuracy of this method, we further mixed the digests from two or three species at different ratios. Our three-proteome mixture results demonstrated highly accurate quantitation for proteins with both small and large fold changes. Moreover, our two-proteome mixture experiment, where 20 to 160 ng of yeast digest was spiked into 200 ng of HeLa digest, showed an R2 of 0.999 for the yeast proteome, underscoring the quantitation accuracy of the method. Utilizing this workflow, we studied dose-dependent protein degradation patterns induced by pomalidomide, iberdomide, and mezigdomide. Our results indicate that mezigdomide may possess enhanced efficacy in T cells by degrading additional proteins such as IKZF2, thereby boosting anti-cancer immunity. Together, we developed an ultra-high-throughput LC-MS method with excellent proteome coverage and quantitation accuracy that is highly suitable for chemoproteomics screening of drug libraries.

biochemistry↗

bioRxiv2025

Targeted α-Synuclein mRNA Degradation by PMO-Based RNA-Degrading Chimeras

-Synucleinopathies are devastating neurodegenerative diseases characterized by pathological accumulation of a neuronal protein, -synuclein (Syn). Lowering soluble Syn levels is a promising therapeutic strategy to limit aggregation and neurotoxicity, but directly targeting this protein is hindered by its intrinsically disordered structure and other factors, such as its conformational heterogeneity and intracellular drug delivery barriers. Consequently, increasing attention has been directed toward targeting the SNCA transcript, which encodes Syn. Here, we developed phosphorodiamidate morpholino oligonucleotide (PMO)-based RNA-degrading chimeras (RDCs) that selectively bind the 5' untranslated region of SNCA mRNA and recruit RNase L for targeted RNA degradation. Through the systematic evaluation of 10 RDCs, we identified and optimized 4-D1, which effectively reduced SNCA mRNA and Syn protein expression in HEK293T cells in an RNase L-dependent manner. 4-D1 lowered SNCA transcript and Syn protein levels in both primary cortical neurons from humanized SNCA mice and in human induced pluripotent stem cell-derived cortical neurons. This reduction prevented prion-like seeding induced by patient-derived Syn fibrils and protected neurons from fibril-induced cytotoxicity. Finally, in vivo studies confirmed the efficacy of 4-D1 in reducing Syn mRNA expression in humanized SNCA mice. These findings indicate that PMO-based RDCs may represent a promising therapeutic modality for -synucleinopathies. Significance StatementAbnormal aggregation of the neuronal protein -synuclein is central to Parkinsons disease and related disorders, yet therapeutic candidates that directly target this protein have yet to demonstrate efficacy. in clinical trials. We developed a new strategy that lowers -synuclein production at the RNA level using phosphorodiamidate morpholino oligonucleotide (PMO)-based RNA-degrading chimeras (RDCs). These molecules recruit a natural RNA-degrading enzyme to selectively destroy the RNA transcript encoding -synuclein. Our lead RDC reduced -synuclein levels in cultured cells, humanized mouse and human neurons, blocked -synuclein pathological aggregation, and protected neurons from toxicity. This study establishes RDCs as a promising therapeutic platform for Parkinsons disease and other neurodegenerative diseases driven by -synuclein.

biochemistry↗

bioRxiv2025

Noncanonical amino acid incorporation enables minimally disruptive labeling of stress granule and TDP-43 proteinopathy

We report a minimally disruptive labeling strategy for stress granule protein, G3BP Stress Granule Assembly Factor 1 (G3BP1), and ALS-linked protein, TAR DNA-binding protein 43 (TDP-43), using the fluorescent noncanonical amino acid Anap. By integrating genetic code expansion (GCE) with rational site selection, we achieved precise incorporation of Anap that preserves protein structure and function. In live cells and neurons, Anap labeling faithfully recapitulated localization, stress-induced dynamics, and recovery behavior, outperforming conventional fluorescent tags and enabling physiologically relevant visualization of protein pathobiology.

cell biology↗

bioRxiv2025

A newly identified Duck orbivirus as the etiological agent of egg-production decline in Chinese breeder ducks

The genus Orbivirus comprises double-stranded RNA viruses, many of which are transmitted by arthropods and cause febrile, hemorrhagic, or reproductive diseases in animals. In recent years, unexplained outbreaks characterized by marked egg-production decline have occurred in breeder duck flocks in China. To identify the causative agent, samples were collected from affected farms in multiple regions across China between 2022 and 2025. Representative samples were analyzed by high-throughput sequencing, followed by phylogenetic analysis, transmission electron microscopy, host cell infection assays, and animal challenge experiments. A previously unreported Orbivirus, designated Duck orbivirus (DORV), was identified. Epidemiological investigation indicated that outbreaks mainly affected breeder ducks during the peak laying period (21-55 weeks of age), with higher incidence in summer and autumn. The DORV genome contains ten double-stranded RNA segments encoding seven structural and five nonstructural proteins. Phylogenetic analysis showed that DORV is closely related to Parrys Lagoon virus (PLV) and Corriparta virus (CORV). Electron microscopy revealed icosahedral particles 30-40 nm in diameter. DORV replicated efficiently in DEF, DEL and C6/36 cells but not in MDCK and LMH cells, suggesting a restricted host range with potential arthropod-borne features. Infection of breeder ducks with DORV-SD01 reduced egg production by 30-40% and caused lesions in the ovary, liver, and spleen. This study describes a novel duck-origin Orbivirus linked to egg-production decline, offering new etiological insights and extending the host spectrum of Orbivirus. The results provide a foundation for future surveillance and prevention of emerging waterfowl diseases.

microbiology↗

bioRxiv2025

Cancer cell-selective ecotopic expression of CD20 as an antigen enables rituximab repurposing for solid tumor immunotherapy

Despite the clinical success of cancer immunotherapies, their efficacy is often compromised by antigen-related problems, including heterogeneity, downregulation, loss, and off-tumor toxicity. To overcome these limitations that challenge the current immunotherapies dependent on native antigens, we here describe a new cancer immunotherapy strategy, which artificially and specifically expresses a clinical validated antigen on variant tumors and thus repurposes clinical antibody drugs to treat cancers not belonging to their indications. To authenticate the strategy, we delivered a CD20 gene under a control of NF-{kappa}B-specific promoter to tumors by adeno-associated virus and then treated them with a CD20 antibody, rituximab. We found that CD20 was selectively expressed in tumors and the followed rituximab treatment activated natural killer (NK) cell to kill cancer cells by antibody-dependent cellular cytotoxicity. We demonstrated that this strategy is effective not only in variant cultivated cancer cells, HCT116 spheroids, and patient-derived organoids of human colorectal cancer, but also in humanized mouse with HCT116 xenograft and immunocompetent mouse with CT26 transplant. The strategy showed high cancer cell specificity in both in vitro and in vivo treatments, leading to high security in animal treatments. This strategy thus creates a new modality of cancer immune-redirection therapy by repurposing the clinical validated both antigen and antibody.

cancer biology↗

bioRxiv2025

Highly Adaptive Conductive Polymer Electronics Enhance Neural Data and Learning Accuracy

Human skin, the bodys largest organ, plays a vital role in sensing and transmitting neuronal, mechanical, and biochemical signals, making it an essential non-invasive interface for health monitoring, rehabilitation, and human-machine interaction. However, aging-related changes, including thinning, increased wrinkling, dryness, and altered collagen structure, significantly impact electrical impedance, conductance, and contact stability, challenging the fidelity and consistency of bioelectronic signal acquisition. Here, we address this gap by developing "AdapSkin," an age-adaptive, skin-mimicking, bio-adhesive, and stretchable polymeric electronic skin interface that seamlessly conforms to diverse skin properties, enabling high-fidelity and high-density electrophysiological recording. The soft electrodes of AdapSkin are composed of an aqueously processed, homogeneously mixed organic nanocomposite with a conductive polymer percolation network, forming a gel-like interface that reduces modulus and enhances skin-electrode contact. The materials platform achieves extraordinary softness and electrical stretchability of up to 1200% through a double-network composite structure. AdapSkin significantly minimizes age-induced variations in interfacial impedance and signal-to-noise ratio (SNR), improving signal consistency for neuromuscular assessment, prosthetic control, and rehabilitation applications. Scalable fabrication enables the creation of large-area electrode arrays, which reduces motion artifacts, improves sEMG mapping reliability, and ensures long-term signal stability across various age groups. Machine learning analysis further demonstrates AdapSkins superior accuracy in gesture classification for elderly users, highlighting its potential to enhance prosthetic control, assistive robotics, and rehabilitation for individuals with sarcopenia and neuromuscular decline. By improving signal quality and adaptability in aging populations, AdapSkin advances fair bioelectronic interfaces, fostering more equitable and effective healthcare technologies for age-related conditions.

bioengineering↗

bioRxiv2025

Single-cell exploration of ovarian aging across vertebrate models

Mammalian female reproductive span is thought to be limited by a fixed "ovarian reserve" determined at birth. With age, a dwindling ovarian reserve leads to infertility, culminating in menopause in humans. In addition to infertility, accumulating evidence has shown that age-related ovarian functional decline contributes to multisystem aging and frailty, making post-menopausal women most susceptible to an array of chronic diseases. However, due to limited tissue accessibility and lack of reliable research models, molecular drivers of ovarian aging remain poorly understood. A key barrier in the field has been the limited establishment and benchmarking of preclinical models faithfully recapitulating human ovarian biology. To address this, we curated publicly available single-cell/nucleus ovarian RNA-seq datasets from human, macaque, mouse, and goat, and processed them using a consistent and stringent pipeline. Datasets were then annotated in a harmonized fashion across studies in order to conduct a robust, integrative, cross-species analysis of ovarian aging with single cell resolution. We systematically evaluated cell-type composition, global transcriptional perturbations, gene-level changes, pathway and network features, and drug-response alignments. Across analyses, granulosa and theca cells emerged as the cell-types most affected by aging. We observed limited but promising consistencies across species, including granulosa-specific signature genes (FSHR and OSGIN2) and cell type-linked pathways, with extracellular matrix/adhesion programs in granulosa and ribosomal/mitochondrial programs in theca cells. These convergences suggest that cross-species modeling likely capture core aspects of ovarian aging. Together, our meta-analysis approach may help refine model selection, generate testable hypotheses, and cautiously inform preclinical and translational work in ovarian aging.

genomics↗

bioRxiv2025

An ultra-long acting insulin enables glucose-synchronised release

Delayed and weak glucose-responsive kinetics, coupled with short plasma exposure and complicated formulation, remain major obstacles to the clinical translation of glucose-responsive insulin formulations. Here we report a rapidly absorbable, ultra-long acting, and glucose-responsive insulin analogue via engineering recombinant human insulin with two phenylboronic acids. The insulin analogue has a long circulating half-life (t1/2{beta} [~]150 h) due to its unique, unprecedented, glucose-responsive binding to blood circulation-associated proteins. This interaction drives the formation of a unique systemic reservoir that allows synchronised dynamic response in insulin levels to glucose fluctuations. In type 1 diabetic mice and minipigs, a single subcutaneous dose can maintain normoglycaemia for over one week (47.5-fold compared to insulin glargine). After the last injection of four consecutive weekly administrations in mice, the glucose-lowering effect can last 800 hours. Administering this glucose-responsive insulin analogue with a fully automated insulin delivery system can leverage dual closed loops and achieve a glucose time in range of over 85% and a coefficient of variation of approximately 30%, better than the recommended targets for human therapy. Importantly, no toxicity was identified in the study.

bioengineering↗

bioRxiv2025

Primary metabolism determines the outcome of salicylic acid-mediated immune induction

Controlling the deleterious effects of immune responses is as vital as fighting infection. In plants, this is achieved, in part, by circadian clock-mediated regulation, such as the synthesis of and response to the immune hormone salicylic acid (SA)1,2. Application of SA at the same concentration under light/dark cycles induces immunity with minimal impact on growth, however, prolonged darkness leads to plant death2. To uncover what determines this life-or-death outcome, we identified twenty survival of SA-induced death (ssd) mutants through genetic screening. These mutants are defective in starch, glucose, and nitrate metabolism, and circadian regulation, and accumulate excessive starch and/or glucose. Likewise, glucose application rescues SA-treated plants in prolonged darkness. Surprisingly, SA treatment does not deplete glucose, but instead, induces amino acid and fatty acid catabolism. Through transcriptomic analyses of glucose-rescued WT plants and ssd mutants for shared pathways, we found that SA triggers plant death in darkness by inducing oxidative stress, and water loss, while glucose antagonizes these processes, boosts ER protein processing and re-establishes the anabolism-catabolism balance. Interestingly, the programmed cell death induced by effector-triggered immunity shares common transcriptomic patterns with those observed during SA-induced cell death in darkness and could also be attenuated by glucose treatment. Therefore, coordination with the cellular metabolic context plays a central role in determining immune outcomes and optimizing plant health.

plant biology↗

bioRxiv2025

scRetinaDB: A Comprehensive Database of Single-Cell and Spatial Omics from Cross-Species Retinas

The retina is essential for encoding visual signals, and its dysregulation can lead to retinal diseases. Recent advances in single-cell and spatial sequencing technologies have yielded extensive omics data from retinal tissues across species and biological conditions. However, existing retinal omics data are dispersed across various repositories without uniform processing, which limits integrative analysis. To address this we developed scRetinaDB (https://casapp.dnayun.com/scretina/), a comprehensive resource that aggregates single-cell RNA sequencing (scRNA-seq), single-cell assay for transposase-accessible chromatin using sequencing (scATAC-seq), and spatial RNA sequencing (spRNA-seq) data from retinas across species and diverse biological conditions. The database comprises over 2.79 million retinal cells collected from 453 scRNA-seq datasets spanning 34 studies, 17 species and 27 biological conditions. For each species, these scRNA-seq datasets were integrated to construct a retinal cell atlas. In addition, scRetinaDB also contains 107 scATAC-seq and 18 spRNA-seq datasets from human and mouse retinas. The scRetinaDB website provides four major modules separately for browsing species-specific omics data, searching cross-species omics profiles, performing analyses of cell type annotation and cell similarity analysis, and downloading preprocessed multi-omics datasets. Overall, scRetinaDB is a valuable resource for retinal single-cell and spatial omics, advancing cross-species studies particularly in vision research.

bioinformatics↗

bioRxiv2025

Integrating Mutation and Stop Signals for Improved RNA Structure Analysis and Insight Discovery

BackgroundUnderstanding RNA structures is essential for exploring its diverse cellular roles. Chemical modification-based RNA structure probing remains a key approach to revealing RNA structures in complex in vivo conditions. Practically, chemical modifications generate both mutation and stop signals during reverse transcription within a single experiment. However, traditional analysis pipelines often rely on only one of the two signals without fully leveraging both. ResultsTo address this, we developed a machine learning-based approach, STONE, that integrates both signals from a single experiment. STONE improved RNA structure analysis across multiple independent validation regions, notably 28S rRNA, viral RNA and regulatory RNAs. In genome-wide datasets, especially single-cell data, STONE significantly increased nucleotide coverage per transcript and improved gene detection reliability. In genome-wide datasets, especially single-cell data, STONE substantially enhanced structural information coverage at both transcript and nucleotide levels. This maximized signal utilization, yielding RNA structures in single-cell data comparable to those from bulk datasets. Furthermore, STONE-derived structural scores allow direct identification of RNA-binding protein binding sites directly from a single probing experiment, with results on RNAs such as U1 snRNA, RNase P RNA, and XIST lncRNA closely matching established techniques like CLIP-seq and RNP-MaP. ConclusionsBy integrating mutation and stop signals from one single experiment, STONE advances RNA structure analysis accuracy, extends nucleotide coverage, and facilitates complex RNA-protein interaction studies, broadening the methods applications in RNA-based research.

bioinformatics↗

bioRxiv2025

The role of medicinal Cu2+ salts in expanding the multi-pathway application of tigecycline against drug-resistant bacteria

Tetracyclines are highly susceptible to photolysis, causing pollution in water, land and other ecosystems, as well as posing threats to human health. In this study, we found that only tigecycline and minocycline reduce the antibacterial activity in light, and this inferiority has severely hindered their scope of clinical use. By strategically regulating the photolysis pathway, we identified chalcanthite, which primarily contains Cu2+. By conducting antibiotic susceptibility test and electron microscopy analysis, Cu2+ salts (including copper sulfate, copper chloride, and copper gluconate) were confirmed to restore the antibacterial activity of tigecycline under light exposure against all tested bacterial strains. Spectroscopic characterization combined with quantum chemical calculations elucidated the molecular mechanism by which Cu2+ selectively modulates the photolytic degradation pathway of tigecycline and structural determinants (dimethylamino groups on the D ring). Multi-omics technologies revealed the mechanism underlying the Cu2+-mediated (non-antibacterial) inhibition of bacterial ferric citrate transporter. Animal infection models were developed using diverse drug-resistant pathogens to validate the in vivo therapeutic efficacy of copper gluconate-tigecycline combination therapy under photic conditions. These findings revealed effective adjuvants that facilitate the development of topical tigecycline formulations, and the establishment of a scientific foundation for synthesizing photostable tetracycline-based antibiotics through structural optimization.

biochemistry↗

bioRxiv2025

Multi-platform framework for mapping somatic retrotransposition in human tissues

Mobile element insertions (MEI) shape the human genome in both germline and somatic tissues. While inherited MEIs are well characterized, mapping somatic MEIs (sMEI) in non-cancer tissues remains challenging due to their low allelic fraction and repetitive nature. We established an integrative framework for sMEI analysis leveraging modern sequencing technologies and analytical innovations. We first benchmarked sMEI detection and demonstrated advantages of long-read and MEI-targeted sequencing for ultra-low-frequency events using a mixture of well-established cell lines. We then showed that haplotype phasing and donor-specific assemblies refine sMEI detection, effectively distinguishing from germline and false signals in in-silico tumor-normal mixtures. We further developed a source-tracing strategy based on internal sequence variation, expanding the catalogue of active source elements beyond traditional transduction-based methods. Applying this framework to donor tissues, we identified 18 rare somatic L1 insertions, revealing structural and source diversity. Our work provides a foundational framework and biological insight into sMEIs.

genomics↗

bioRxiv2025

An Empirical Study on Quantifying Physical Energy Expenditure during High-intensity Exercise

In the process of high-intensity exercise (HIE), the accurate evaluation of Physical Energy Expenditure (PEE) is of great significance to many occupational fields. However, due to the limitation of current technology, the quantitative ability of existing PEE evaluation methods in HIE situations is still insufficient. Based on the existing physiological coupling mechanism between cardiovascular parameters and energy metabolism, this study analyzed the dynamic changes of high-frequency blood pressure (BP) and heart rate (HR) during incremental load exercise and high-intensity interval training. The results show that although HR has a nonlinear response to the increase of exercise intensity, its time pattern-when analyzed synergistically with BP fluctuation-can provide reliable metabolic indicators. We used Doubly Labeled Water (DLW) as the gold standard for PEE determination and measured the total energy expenditure (TEE) in three consecutive days. During the monitoring period, high frequency invasive BP and HR monitoring was performed simultaneously, and the time integral value of SBP x HR and its ratio to TEE (i.e. Quotient value) were calculated. By dividing the corresponding SBP x HR integral value during HIE by the above quotient value, the PEE of this exercise can be estimated. Specifically, TEE can be accurately estimated by time integration of the product of SBP and HR. The short-term PEE rate can be solved by the differential equation constructed by the systolic blood pressure(SBP) trend function in this period, and can also be obtained by dividing the PEE in this period by the duration of this period. In this study, based on the results of invasive measurements, a new method for real-time monitoring of HIE using non-invasive cardiovascular parameters during PEE was proposed, providing new ideas and methodological support for applications in related fields.

physiology↗

bioRxiv2025

ceQTL: A co-expression QTL model to detect a variant that affects transcription factor binding and its target regulation

Expression quantitative trait locus (eQTL) mapping is used to identify a functional link between a genomic variant, such as single nucleotide polymorphism (SNP), and gene expression (often close-by pair for cis-eQTL) by linear regression, commonly done when matching genotype and expression data from same individuals are available. Millions of significant eQTLs have been reported by both individual studies and coordinated large consortiums such as Genotype-Tissue Expression project (GTEx). A significant eQTL association does not establish a causal relationship or provide any underlying mechanism so further investigation is needed to understand how a SNP impacts gene expression. One of the plausible explanations for eQTL is that a genomic variant affects transcription factor (TF) binding and thus impacts its regulation on its target genes (TGs). However, data-driven or formal statistical methods to prove that hypothesis are still lacking. To address the gap, we propose a new method called differential co-expression QTL (ceQTL) among different alleles using Chow statistics to specifically detect eQTLs that are modulated by a particular TF. We start with building a trio of TF, its TG, and related SNP and then test the significant coefficient difference among different levels of SNP in terms of TF and TG correlation. We applied this ceQTL model to simulated data and the lung tissue datasets from the GTEx project. The simulated data results showed that the model was robust to detect true ceQTLs at variable sample sizes and different minor allele frequencies as measured by Area Under the Curve (AUC). In normal lung tissue, a small fraction of eQTLs were found to have strong ceQTLs, i.e., eQTLs where SNP affects gene expression though TF binding. Some ceQTLs may not be detected by traditional eQTL analysis. Our tool also performed a TF binding affinity analysis to add another layer of evidence for functional interpretation. Comparisons with other similar tools were also presented. In summary, ceQTL analysis provides a more interpretable and biological insight into the mechanism of eQTL, which would help us better understand how genomic variants affect phenotypes and diseases.

bioinformatics↗

bioRxiv2025

Integrated analysis reveals neuro-immune pathway in the central nervous system that supports SGLT2i`s protective effects in treatment of cardiac remodeling

BackgroundDespite growing evidence of sodium-dependent glucose transporters 2 inhibitors (SGLT2i) improving heart failure, their mechanisms remain unclear. This study explored the effects of empagliflozin (EM) on sympathetic nerve activity and potential neuroimmune pathways. MethodsSingle-nucleus RNA sequencing (snRNA-seq) was performed in hypothalamic tissue of a DOCA-salt mouse model to screen for cellular changes. Validation of relevant mechanisms in DOCA-salt and high-salt diet (HSD) mouse models. Additionally, clinical data from 1,189 hospitalized heart failure patients were analyzed to evaluate the translational relevance of the experimental findings. ResultsEM treatment was associated with protective effects against cardiac remodeling in both DOCA-salt and HSD mice. Both models exhibited sympatho-excitation and neuronal hyperactivity in pre-autonomic brain regions, which appeared to be blunted by EM co-treatment. snRNA-seq analysis suggested that cellular interplay among vascular cells, microglia, and inhibitory neurons might be involved in these processes. Specifically, high-salt intake was linked to peripheral immune activation, including lymphocytosis and elevated plasma interferon-{gamma}, which potentially mediated microglial state transitions. EM might counteract the disease-associated upregulation of protein ubiquitination. Clinically, SGLT2i use was significantly associated with reduced lymphocyte counts in specific patient subgroups. ConclusionsEmpagliflozin may attenuate cardiac remodeling caused by high sodium intake through neuroimmune effects by restoring microglial homeostasis and inhibiting sympathetic nerve activity.

molecular biology↗

bioRxiv2025

Absolute hand determination of glycofibrils from natural sources in cryo-EM

Glycans are one of the four fundamental macromolecules that constitute life. However, compared with proteins and nucleic acids, our understanding of the structures and functional mechanisms of glycans remains limited. Moreover, it is generally challenging to obtain high-resolution three-dimensional structures of glycans. Recent studies have demonstrated the potential of the CryoSeek strategy in enabling high-throughput structural determination of glycans, thereby presenting novel avenues for their structural investigation. Meanwhhile, unlike proteins, near atomic resolution density maps of glycofibrils do not inherently facilitate the determination of absolute hand, which is a prerequisite for building atomic models of glycofibrils. Existing absolute hand determination methods have severe limitations in the cases of glycofibrils from natural sources. In this study, we introduce Ahaha, a straightforward and efficient method for determining such absolute hand in cryo-EM. With their absolute hand measured by Ahaha, we built atomic models of four glycofibrils derived from a natural water sample, facilitating the study of glycans. The online service of Ahaha is available at https://cryoseek.org/ahaha.

biophysics↗

bioRxiv2025

Oncogenic PTPN11/SHP2 drives immune escape in juvenile myelomonocytic leukemia (JMML) through activation of ectonucleotidase/adenosine signaling

Juvenile myelomonocytic leukemia (JMML) is a myelodysplastic/myeloproliferative neoplasm of early childhood driven by RAS pathway mutations. Allogeneic hematopoietic stem cell transplantation (HSCT) is the therapy of choice for most patients. However, relapse rate is high, in patients with adverse features, frequently noted in PTPN11-mutated JMML, or in patients without evidence of graft-versus-host disease (GvHD). Here we set out to understand the mechanisms associated with oncogenic PTPN11 immune escape. Analyzing primary PTPN11-mutated JMML samples and MxCre;Ptpn11D61Y/+ mice, we observed elevated expression of immune checkpoint molecules, including ectonucleotidases CD39 and CD73 - key mediators of the adenosine pathway - on monocytic and granulocytic leukemic cells. Stimulation with GM-CSF, a central mediator of JMML pathogenesis, induced ectonucleotidases expression on granulocytes and monocytes. In contrast, MEK inhibition downstream of Ptpn11D61Y/+ reduced ectonucleotidases expression. Functionally, Ptpn11D61Y/+-mutated myeloid cells suppressed activation and proliferation of wild-type (WT) T lymphocytes, an effect recapitulated by adenosine and reversed by pharmacological CD39 inhibition with POM-1. In vivo, POM-1 treatment of MxCre;Ptpn11D61Y/+mice presenting with myeloproliferation reduced spleen size and partially restored immune responsiveness. Moreover, POM-1 induced apoptosis in murine Ptpn11D61Y/+ myeloid cells, highlighting a dual therapeutic benefit of CD39 inhibition in JMML. Together, these findings suggest that targeting the adenosine pathway may represent an immunomodulatory approach to enhance T cell-mediated control of JMML, particularly in the context of HSCT and relapse prevention.

cancer biology↗