Biology subjects

Du, J.

Publications and source records attributed to Du, J..

At least 19 recordsLinked to original sources

bioRxiv2026

Spatially resolved host-fungal transcriptomics profiling by fungal-DBiT

Hosts and their associated fungi coexist within a shared tissue environment, engaging in dynamic interactions that influence immune regulation, tissue homeostasis, and disease progression. Dissecting these interactions requires spatially resolved and unbiased approaches capable of capturing gene expression from both host and fungal cells within intact tissue architecture. Here, we present fungal-DBiT, a spatial transcriptomic platform that enables simultaneous, high-resolution mapping of host and fungal RNAs, directly in tissue sections. Using fungal-DBiT, we profiled the spatial distribution of host and fungal transcripts in a fungal infection mouse model, and review pronounced spatial heterogeneity in fungal localization and activity, accompanied by region-specific host transcriptional responses. We further applied fungal-DBiT to intestinal tissues under physiological conditions, demonstrating its sensitivity in detecting low-abundance fungal species in the absence of overt inflammation. Together, these results demonstrate the ability of fungal-DBiT to resolve host-fungal interactions in situ and highlight its utility for studying mycobiome associated immune responses and disease processes with spatial precision.

microbiology↗

bioRxiv2026

Shotgun metagenomic and untargeted metabolomic analysis of gut microbiome associated with peripheral artery disease

Background: While the human gut microbiome is a potentially important and modifiable risk factor for cardiovascular diseases, the links between the gut microbiome and peripheral artery disease (PAD) and its clinical subgroups (claudication and chronic limb-threatening ischemia; CLTI) are not well understood. Methods: We used shotgun metagenomics and untargeted metabolomics to analyze the fecal samples of people with PAD and age- and sex-matched non-PAD controls collected in a cross-sectional manner. We used functional prediction pathways, a network analysis, and weighted gene co-expression network analysis of metagenomic, metabolomics, and clinical data to better understand PAD-associated dysbiosis. Results: Alpha and beta diversity were similar between the PAD (n=78) and non-PAD (n=20) groups. Differentially abundant species were dominated by those belonging to the Lachnospiraceae, Ruminococcaceae and Enterobacteriaceae families. Fecal butyrate levels were reduced between the non-PAD and PAD groups, but differences were not statistically significant. There was significantly reduced alpha diversity in the samples of the CLTI compared to claudication subgroup. Abundance of multiple butyrate producers were greater in the claudication group compared to CLTI. Network analysis revealed 5 meta-modules, each with specific associations to PAD presentation and metabolomic profiles, and species that are both top module hubs and differentially abundant between groups. Community interconnectedness appears to increase in claudication, followed by community disintegration in CLTI. Conclusions: This metagenomic-metabolomic analysis of the gut microbiome of individuals with PAD, highlights the importance of studying functional capacity, diversity, and connectivity between species. Microbial modules may allow an integrative view of the microbial ecology relevant to presence and progression of PAD.

microbiology↗

bioRxiv2026

Aging Differentially Remodels the Retina and RPE/Choroid Proteome and Metabolism

Purpose: The neural retina and retinal pigment epithelium/choroid (RPE/Cho) are metabolically coupled. Metabolic dysfunction of the RPE/Cho contributes to photoreceptor death in age-related macular degeneration (AMD). This study aimed to determine how aging, a major risk factor for AMD, alters the proteome and metabolic pathways in the retina and RPE/Cho. Methods: We performed quantitative proteomics on neural retina and RPE/Cho from young (8-week) and old (73-week) C57BL/6J mice. The differentially expressed proteins between old and young, as well as between the retina and RPE/Cho, were visualized using volcano plots and heatmaps, and functional analyses were assessed by Gene Ontology enrichment tools. Results: Aging altered the expression of 85 retinal proteins and 272 RPE/Cho, with 16 shared between tissues. In the aged retina, proteins associated with NADP(H) metabolism, neurodegeneration, and immunity were mostly upregulated, while proteins in lipid metabolism, transcription, and epigenetic regulation decreased. In the aged RPE/Cho, all 21 significantly altered lysosomal enzymes and transport proteins increased, together with complement components, and photoreceptor outer segment proteins. These changes accompanied increases in proteins associated with lipid metabolism and protection against lipid peroxidation, while several glutathione metabolism enzymes decreased. Seven shared proteins involved in lipid and NADP(H) metabolism, changed in opposite directions between retina and RPE/Cho. Comparison of the retinal and RPE/Cho proteomes identified 3073 differentially abundant proteins, of which 2452 maintained tissue-specific differences across both age groups, while approximately 17% showed age-dependent shifts in tissue enrichment. These shifts involved proteins associated with metabolism, extracellular matrix, visual function, lysosomal pathways, and innate immunity. Within the metabolic proteome, several glycolytic and fatty acid oxidation proteins showed greater relative enrichment in RPE/Cho with age, whereas glucose transporters and mitochondrial complexes shifted toward the retina. Conclusions: Overall, these findings show that aging differentially remodels the retina and RPE/Cho proteome, accompanied by altered metabolic specialization between the two tissues.

biochemistry↗

bioRxiv2026

CryoFAST: A Fab-Aided Cryo-EM Workflow for High-Resolution Structure Determination of Sub-50-kDa Protein Targets

Single-particle cryo-electron microscopy (cryo-EM) plays an increasingly important role in structure-based drug discovery (SBDD) by revealing target protein-ligand interactions at the molecular level, but its application has largely focused on large- and medium-sized protein complexes. Many therapeutically important targets are smaller than 50 kDa and remain highly challenging for cryo-EM analysis because of low signal-to-noise ratios and difficulties in particle alignment. Here, we describe cryoFAST, an integrated workflow that couples ligand-compatible Fab discovery and structure-guided computational maturation with biochemical triage, fiducial-complex assembly, and single-particle cryo-EM. Through phage-display screening and/or AI-guided design, we developed high-affinity Fab binders of approximately 50 kDa that stabilize the target without disrupting ligand-target interactions and then assembled Fab-target complexes for cryo-EM analysis. Using the therapeutically relevant extracellular domain (~20 kDa) of interleukin-4 receptor (IL-4R) as a proof of concept, we determined its cryo-EM structure at 2.74 Angstrom resolution and established a framework for elucidating ligand-binding modes. This workflow provides a practical, efficient, and potentially broadly applicable route to the structural characterization of small protein targets that are otherwise challenging to study. By extending the applicability of cryo-EM to high-value but technically challenging small targets, cryoFAST may open new opportunities for structure-based drug discovery.

biophysics↗

bioRxiv2026

Causal event structure shapes convergent reactivation signatures in human cortex and language model

Humans adapt to changing environments by understanding the latent causal structure of events, and modern large language models (LLMs) increasingly exhibit analogous abilities. However, elucidating how causal knowledge is represented and integrated in the human brain and in LLMs poses a substantial challenge for cognitive science. To address this, we combine topological analysis of representational manifolds with neuroscientific analyses of reactivation. We find that event-level causal relations modulate temporally structured neural reactivation in the human brain, particularly in the angular gyrus, precuneus and prefrontal cortex. By contrast, LLMs express these relations through positionally structured representational reactivation. Despite these differences, the resulting reactivation patterns exhibit similar intrinsic manifold structure across the two systems. Together, these findings suggest that the human brain and LLM share a common principle for organizing causal knowledge.

neuroscience↗

bioRxiv2026

A reversible GNAS-PKA-RUNX2 checkpoint governs skeletal stem cell fate in fibrous dysplasia

Fibrous dysplasia (FD) arises from activating GNAS mutations that increase cAMP signaling in skeletal stem/progenitor cells (SSPCs), but the underlying mechanisms and whether the resulting fate imbalance can be reversed remain unclear. Using controlled GNAS induction with transcriptomic, pharmacologic, and cell-fate analyses, we identify a PKA-dependent checkpoint governing SSPC lineage commitment. GNASR201C redirects SSPCs toward osteoclastogenesis while suppressing osteoblast maturation; genetic or pharmacologic PKA inhibition reverses both effects, revealing this fate decision is dynamically reversible. Mechanistically, PKA phosphorylates RUNX2, the master regulator of osteogenesis, enforcing its cytoplasmic retention and blocking osteogenic transcription; a non-phosphorylatable RUNX2 mutant restores nuclear localization and osteoblast differentiation, defining RUNX2 phosphorylation as a disease-initiating molecular switch. Critically, PKA inhibition in vivo reduces fibrosis and osteoclast burden in established bone lesions, demonstrating reversibility even in advanced FD. These findings define a druggable GNAS-PKA-RUNX2 checkpoint and identify PKA inhibition as a strategy to reverse existing disease in FD.

cell biology↗

bioRxiv2026

Spatiotemporal atlas resolves the mesenchymal-epithelial stem cell axis governing human hair follicle aging and pathology

How adult stem cell populations coordinate tissue regeneration across physiological and pathological states remains unclear. Hair follicles contain epithelial and mesenchymal stem cells that codetermine hair regeneration. Here, we integrate a multimodal spatiotemporal atlas with functional validation to define the mesenchymal-epithelial stem cell dynamics governing human hair follicle homeostasis, aging, and androgenetic alopecia. Aging is characterized by passive depletion of hDSC-derived regenerative cues, whereas AGA involves a shift of hDSC toward an inhibitory signaling hub that promotes HFSC dysfunction. We further pinpoint hDSC-secreted PTN as a key regenerative factor that promotes hair growth and counteracts androgen-induced growth suppression. This study provides a holistic framework of adult stem cell crosstalk across physiological and pathological states, offering a mechanistic roadmap to reinstate tissue regeneration.

cell biology↗

bioRxiv2026

Risk factors and immunological characteristics of SARS-CoV-2 RNA re-positivity in convalescent COVID-19 Patients: A retrospective study

Purpose: To investigate the incidence, clinical features, and risk factors for SARS-CoV-2 nucleic acid re-positivity in recovered COVID-19 patients. Methods: We enrolled 441 convalescent COVID-19 patients from the rehabilitation ward of the Fifth Hospital of Shijiazhuang between January and April 2021. Based on nucleic acid test results, patients were categorized into repositive (n = 69) and non-repositive (n = 372) groups. Demographic, clinical, laboratory, and imaging data were compared between groups. Logistic regression was used to identify factors associated with re-positivity. Results: The overall re-positivity rate was 15.65%. Compared with the non-repositive group, the repositive group presented with a significantly higher proportion of moderate disease severity (72.46% vs. 50.54%) and a lower proportion of patients under 14 years old (7.25% vs. 23.66%). Additionally, multi-lobar involvement ([≥] 3 lobes) during the acute phase was more common in the repositive group (43.48% vs. 26.61%), with all differences being statistically significant (P < 0.001). Laboratory analysis revealed that repositive patients had significantly lower lymphocyte and eosinophil counts, as well as reduced albumin (ALB), aspartate aminotransferase/alanine aminotransferase (AST/ALT) ratio, and albumin/globulin (A/G) ratio. Furthermore, circulating CD3+, CD4+, and CD8+ T-cell counts were markedly decreased. In contrast, the neutrophil-to-lymphocyte ratio (NLR), immunoglobulin M (IgM), immunoglobulin A (IgA), and neutralizing antibody (NAb) levels were significantly elevated in re-positive patients (all P < 0.05). Multivariate logistic regression analysis identified the CD8+ as independent risk factors for re-positivity. Conclusion: In convalescent patients, SARS-CoV-2 RNA re-positivity correlates with distinct clinical and immunological characteristics. The CD8+ T-cell level is an independent predictor, highlighting that dysregulated immune and inflammatory responses contribute to delayed viral clearance.

immunology↗

bioRxiv2026

Multimodal substrate recruitment enables CTLH-MKLN1 E3 ligase to target N-, C-, and internal degrons

The GID/CTLH family of E3 ubiquitin ligases employs several substrate receptor subunits that recruit distinct degrons, but the substrate recognition mechanism of the CTLHMKLN1 assembly has largely remained elusive. Here, we reconstitute CTLHMKLN1-dependent ubiquitylation of three biochemically distinct substrates - MKLN1 itself, ZMYND19, and FAM72A-recruited UNG2, and determine cryo-EM structures of each substrate bound to MKLN1. The structures reveal how MKLN1's discoidin and Kelch {beta}-propeller domains engage substrates through multivalent contacts. MKLN1 recruits itself through discoidin - Kelch interactions between MKLN1 dimers, forming an assembly that competes with other substrates. ZMYND19 and FAM72A each bind to MKLN1 through loops engaging the discoidin trench domain while their C-terminal Arg residues engage the MKLN1 Kelch central channel, identifying MKLN1 as an Arg/C-degron receptor. The acetylated N-terminus of UNG2 is positioned within a Y-shaped tunnel of FAM72A, defining an adaptor-mediated Ac/N-degron recognition mechanism. Thus, our data reveal how combinatorial deployment of MKLN1 discoidin and Kelch domains enable a single receptor subunit to recognize a diverse substrate repertoire, adding to a complex picture of degron recognition across the GID/CTLH family.

biochemistry↗

bioRxiv2026

A bispecific antibody targeting a membrane-proximal ASGPR1 region promotes TNF-α recruitment and uptake

Abstract Recruiting an endocytic receptor through an antibody offers a route to redirect extracellular cytokines into cells. Here, we developed BiAb J31, an IgG like bispecific antibody linking TNF recognition to a membrane-proximal region of asialoglycoprotein receptor 1 (ASGPR1). J31 was isolated by immunization with an engineered ASGPR1 extracellular domain followed by screening against wild-type ASGPR1 and HepG2 cells. Its variable domains were combined with unmodified adalimumab variable regions using knobs into holes and CrossMab engineering, with L234A/L235A/P329G Fc substitutions. BiAb J31 retained HepG2 binding and engaged both antigens in bridging assays. Sequential binding experiments demonstrated recruitment of TNF to BiAb J31 treated cells. Live cell imaging showed uptake of fluorescent antibody-containing complexes and overlap with LysoTracker-positive compartments. In HepG2 cultures containing BiAb J31, TNF and a PE conjugated detection antibody, supernatant PE fluorescence fell by approximately 60% over 90 minutes. Truncation and synthetic-peptide assays localized J31 recognition to ASGPR1 residues 62~100, outside the carbohydrate-recognition domain, and AlphaFold 3 modeling proposed an Fv peptide interface. These findings identify a membrane-proximal ASGPR1 binding antibody that is compatible with bispecific engineering and supports cellular recruitment and uptake of TNF containing complexes. BiAb J31 provides a molecular starting point for antibody based cytokine redirecting approaches.

immunology↗

bioRxiv2026

H3K4me3 recruits the chromatin remodeling DILL-PICKLE-PEPPERCORN complexes to promoters in plants

The plant CHD chromatin remodeler PICKLE (PKL) is a master regulator of cellular identity and differentiation, controlling developmental, hormonal and stress-response processes. Yet the molecular mechanisms underlying its function remain unclear. Here, we show that PKL forms three complexes, each composed of a protein of previously unknown function and one of three mutually exclusive novel DNAJ proteins that recruit HSP70-1. Simultaneous loss of all three DNAJs phenocopies the pkl mutant, indicating functional redundancy among PKL complexes. In vitro activity assays and cryo-electron microscopy reveal that PKL clamps nucleosomal DNA via its ATPase motor domain and recognizes H3K4me3 through its double chromodomain. Genome-wide profiling shows that H3K4me3 recognition positions PKL complexes at genic promoters to bidirectionally fine-tune gene expression. Together, these results provide structure-function insight into PKL recruitment and its control of developmental gene expression, and reveal a chaperone-coupled complex assembly that offers a broader perspective on how chaperone networks may support chromatin remodeling complexes across eukaryotes.

plant biology↗

bioRxiv2026

Metabolic signatures of cone-dominant and cone-degenerating retinas

Cone photoreceptors are essential for daylight vision, and their degeneration has more profound visual consequences than rod loss in retinal degenerative diseases. Metabolic dysfunction is closely associated with cone degeneration, however the relatively small population of cones in mice and humans have limited our understanding of cone-specific metabolism. Here, we leveraged cone-dominant and cone-degeneration mouse models including Nrl-/-, Cnga3-/-, and high-dose Triiodothyronine (T3) treatment to investigate cone-specific metabolism and their metabolic impacts on the retinal pigment epithelium (RPE). Across models, cone-dominant retinas consistently showed lower pyruvate abundance alongside increases in glutathione, purines, pentose phosphate pathway intermediates, and one-carbon metabolites. Increases in several key amino acids were also associated with higher cone abundance, such as proline, arginine, alanine, valine, leucine, and hypotaurine. Strikingly, aminoadipate, an intermediate in lysine catabolism, was the most robustly changed metabolite in the retina, showing highly consistent increases across models. Relative cone increases were also associated with metabolic changes in the RPE/choroid. Like the retina, RPE/choroids showed consistent increases in aminoadipate, proline, and hypotaurine, as well as xanthosine and betaine, alongside decreases in uracil. Moreover, proteomic analysis of Nrl-/- mice showed decreases in many key metabolite transporters in the RPE/choroid, including carriers for glucose, lactate, aspartate, glutamate, serine, lysine, taurine, and proline. Collectively, these findings further our understanding of cone-specific metabolism and highlight potential cone-specific metabolic vulnerabilities in retinal degeneration.

biochemistry↗

bioRxiv2026

Regional and tissue-specific metabolic differences in human neural retina and RPE/choroid

It is clear that the human retina and its underlying retinal pigment epithelium and choroid (RPE/choroid) form an interdependent metabolic ecosystem, but how metabolism differs between the cone-rich macula and rod-rich periphery remains unclear. Using targeted metabolomics, we quantified 133 metabolites in paired macular and peripheral neural retina and RPE/choroid explants from human donor eyes following short-term culture to restore metabolic activity. Distinct metabolic differences were identified between retinal regions and between tissues. Compared with the peripheral retina, the macula showed metabolic features consistent with greater glycolytic activity, increased NADH availability and higher levels of the neurotransmitter-associated metabolites N-acetyl-aspartate (NAA) and N-acetyl-aspartyl-glutamate (NAAG), consistent with increased energetic and neuronal activity. Compared with peripheral RPE/choroid, the macular RPE/choroid had higher levels of the flavin cofactor FAD together with NAD-related metabolites, including NAD, NADP and NAAD. Comparisons between the neural retina and RPE/choroid further showed that the neural retina was primarily associated with energy production and neurotransmission, whereas the RPE/choroid was associated with cofactor metabolism, nucleotide salvage and lipid metabolism. These findings are consistent with metabolic coupling between the neural retina and RPE/choroid. The macula has metabolic features consistent with high energetic demand, providing a potential metabolic basis for its selective vulnerability in macular disease.

neuroscience↗

bioRxiv2026

Multiple Nested Distributed Language Networks in the Human Brain

Brain regions specialized for language have been extensively described, yet their arrangement into one or multiple networks remains debated. Using precision functional mapping across three independent cohorts of intensively scanned individuals (22 individuals scanned over 216 separate MRI sessions), we dissociated two nested left-lateralized perisylvian networks: an intermediate language network (intLANG) and an anatomically distinct association language network (aLANG). intLANG is anchored to precentral speech areas and the Sylvian parietal-temporal area (Spt), whereas aLANG surrounds intLANG and extends into higher-order prefrontal and temporal association cortices. The two networks can be fully recapitulated by functional connectivity from adjacent cerebellar regions, indicating that they are segregated, brain-wide networks. Task-based analyses further reveal that intLANG and aLANG are functionally distinct: intLANG responds robustly during rhyme judgments and nonword reading that emphasize phonology, whereas aLANG is preferentially recruited during meaning-based sentence processing. These findings indicate that human language engages nested distributed networks each specialized for distinct components of language processing: a lower-order network biased toward phonology, and a surrounding association network that subserves higher-order syntax and semantics. This nested organization is similar to other brain systems suggesting a shared hierarchical motif that may give rise to specialized cognitive functions across the human brain.

neuroscience↗

bioRxiv2026

Peripheral sensory terminal degeneration is linked to sensory fiber hyperexcitability in paclitaxel-induced peripheral neuropathy

Paclitaxel-induced peripheral neuropathy (PIPN) is the most common dose-limiting side effect of paclitaxel chemotherapy, yet the relationship between structural damage to peripheral nerve terminals and functional impairment of nociceptors has not been directly examined. Here we combined ex vivo skin-nerve electrophysiology with three-dimensional (3D) imaging of fDISCO-cleared glabrous skin to characterize both morphological and functional changes in peripheral sensory terminals following paclitaxel treatment in rats. Five weeks after paclitaxel administration, skin-nerve recordings revealed a marked increase in the proportion of C- and A{delta}-fibers exhibiting spontaneous discharge (approximately 2.4- and 2.6-fold increases, respectively) compared to vehicle-treated controls. Paclitaxel also selectively reduced the mechanical activation threshold of C-fibers without affecting A{delta} fibers. Three-dimensional reconstruction of PGP9.5-immunolabeled nerve terminals showed that sensory fibers in glabrous skin form a vertically oriented, tree-like architecture. Paclitaxel treatment severely reduced both the terminal branch length and the density of free nerve endings in the epidermis. Strikingly, co-administration of the Kv7 channel activator retigabine prevented both the electrophysiological and morphological alterations induced by paclitaxel. These findings provide direct evidence that peripheral nerve terminal degeneration and hyperexcitability co-occur in PIPN and that Kv7 channel activation can protect against both structural and functional damage. SignificanceThis study provides the first direct evidence linking intraepidermal sensory terminal degeneration with peripheral sensory fiber hyperexcitability in paclitaxel-induced peripheral neuropathy by combining skin-nerve electrophysiology with three-dimensional imaging of sensory terminals. Paclitaxel induces distal degeneration of intraepidermal sensory terminals, increases spontaneous discharge, and lowers the mechanical activation threshold of C-fibers, demonstrating that structural degeneration and functional abnormalities occur concurrently in the peripheral terminal. Retigabine prevents both structural and functional alterations, supporting peripheral sensory terminals as a therapeutic target for preventing chemotherapy-induced neuropathy.

neuroscience↗

bioRxiv2026

Bone Mechanosensing Dictates Hematopoietic Stem Cell Fate and Immune Homeostasis

The vertebrate water-to-land transition was accompanied by a six-fold increase in gravitational force, followed by the migration of hematopoietic stem cells (HSCs) from kidney or liver to the bone marrow, and the acquisition of enhanced immune functions to cope with novel environmental pressures. Bone senses mechanical loading and provides a microenvironment for HSC development, yet whether bone mechanosensation affects immune cell development and immune homeostasis remains unclear. Here, we unveil bone as a mechanosensory organ that translates mechanical force into hematopoietic instructions. Mechanical loading of bone directs HSCs toward lymphoid lineages in mice and non-human primates, whereas unloading favors myeloid commitment. This process requires osteocyte mechanosensor Piezo1, which induces loading-responsive bone-derived factors such as IL1R2, SERPINC1 and INMT, thereby restraining inflammatory signaling and guiding HSC differentiation. Osteocyte Piezo1 deficiency recapitulates the hematopoietic and immune alterations observed during unloading. Functionally, this pathway enhances acute infection resistance and suppresses immunosenescence in mice and in aged long-tailed macaques. Notably, skeletal mechanoregulation of immune homeostasis is conserved across vertebrate species. Our research defines the mechano-bone-immune axis (mechano-osteoimmunology), offering a novel evolutionary perspective on the interconnected development of the skeletal and immune systems and presenting a promising non-pharmacological target to address immune dysfunction.

immunology↗

bioRxiv2026

Generation and MHC class II loading of an endogenous influenza epitope revealed by a T cell receptor-like antibody

CD4+ T cells coordinate immune responses to infections and cancers and initiate many autoimmune diseases, yet the intracellular pathways that generate the peptide antigens they recognize remain incompletely understood. Progress has been limited in part by the scarcity of reagents that directly detect defined peptide:MHC class II (pMHC-II) complexes. Here, we established an mRNA vaccine-based pipeline to generate and characterize T cell receptor (TCR)-like antibodies. One result was 1D6, a monoclonal antibody specific for NA25:Ab, an influenza A/PR8 neuraminidase-derived epitope presented by MHC-II. 1D6 bound NA25:Ab with high affinity and specificity. 1D6 recognizes NA25:Ab through binding determinants partially distinct from those used by the cognate TCR. Using this reagent, we confirmed the proteasome dependence of NA25 presentation and showed that NA25:Ab accumulates in intracellular MHC-II loading compartments without a major requirement for canonical macroautophagy, pointing to an unconventional mode of antigen presentation. These findings establish TCR-like antibodies as powerful tools for dissecting noncanonical MHC-II antigen processing pathways. One Sentence SummaryAn mRNA immunization strategy generated a highly specific antibody to study how a noncanonical natural epitope from influenza virus is produced and loaded onto MHC-II.

immunology↗

bioRxiv2026

A Selective Multivalent NET-Associated Chromatin Neutralizer Resolves Infection-Associated Inflammation in Severe Sepsis

Targeting the overwhelming inflammation driven by neutrophil extracellular traps (NETs) during infection provides an opportunity to manage severe sepsis. This potential needs to be realized by exploring selective NET-neutralization materials, which remains a challenge. Herein, we report a multivalent macromolecular strategy that targets NET-associated DNA-histone chromatin complexes while preserving antibacterial activity of aminoglycoside. We identify 8-arm PEG-conjugated netilmicin (8-arm Netil) as a lead NETs-neutralizer from a library of multivalent aminoglycoside-displayed materials. When compared with 2- and 4-arm counterparts, 8-arm Netil exhibits potent antibacterial activity and high-affinity binding to DNA-histone chromatin complexes through stable multivalent noncovalent interactions, thereby suppressing NET-induced TLR4/TLR9 activation and macrophage inflammatory responses. In severe septic mice, intravenously administered 8-arm Netil preferentially accumulates in inflamed tissues, leading to improved survival protection, owing to the reduction of bacterial dissemination, NET accumulation, systemic cytokine production, and multiple-organ injury. These findings establish NET-associated DNA-histone chromatin complexes as actionable extracellular targets and demonstrate multivalent chromatin targeting as a rational material design strategy for selective NET neutralization and inflammation control in severe sepsis.

bioengineering↗