Biology subjects

Huang, W.

Publications and source records attributed to Huang, W..

At least 109 records · Page 6Linked to original sources

bioRxiv2025

Metabolic modulation of intratumoral cholesterol with gut microbiota for the treatment of colorectal cancer

Excess cholesterol is positively correlated with colorectal cancer (CRC). Current therapeutic strategies for modulating cholesterol levels in CRC are limited and often come with complications. Here, we demonstrated that microbiome shunting of intestinal cholesterol to anticancer metabolites is a safe and applicable therapeutic approach for CRC. By screening major microbial metabolic products of cholesterol, we found that 4-cholesten-3-one (4-C-3) was selectively depleted in fecal samples and tumor tissues of patients with CRC. 4-C-3 exhibits strong antitumor effects on human CRC cell lines, patient-derived organoids, and patient-derived xenograft (PDX) models. Mechanistically, 4-C-3 suppresses CRC tumorigenesis by dually targeting the epidermal growth factor receptor (EGFR) and Kirsten rat sarcoma viral oncogene homologue (KRAS). 4-C-3 directly binds to EGFR, blocking its signal transduction by inhibiting the binding of EGFR ligands. Additionally, 4-C-3 inhibits oncogenic KRAS variants, such as KRASG12D, by suppressing nucleotide exchange activity and effector engagement. By targeting both EGFR and KRAS, 4-C-3 reduces primary resistance to anti-EGFR therapies caused by KRAS mutations in CRC. As a proof-of-concept study, we showed that delivery of 4-C-3 by Oscillibacter ruminantium, a 4-C-3- producing commensal bacterium that is reduced in CRC patients, or a nonpathogenic Escherichia coli strain engineered to specifically convert intestinal cholesterol into 4-C-3, removed intratumoral cholesterol and led to rapid tumor regression in multiple models of CRC. These results suggest that microbial therapies for restoring intestinal cholesterol homeostasis represent a new therapeutic avenue for CRC.

cancer biology↗

bioRxiv2025

NanoLoop: A deep learning framework leveraging Nanopore sequencing for chromatin loop prediction

Chromatin loops play a crucial role in gene regulation and cellular function, providing key insights into understanding the three-dimensional structure of the genome and its impact on cellular homeostasis. Nanopore sequencing technology, with its advantages in simultaneously detecting sequences and methylation patterns, brings new opportunities for studying three-dimensional genome structures. We introduce NanoLoop, the first algorithmic framework attempting to predict genome-wide chromatin interactions using Nanopore data. In experiments across four human lymphoblastoid cell lines, NanoLoop demonstrated excellent predictive performance and cross-cell line generalization capabilities. We also discovered four distinct methylation patterns at loop anchors that influence histone modification levels and determine various loop types. NanoLoop further predicted previously uncharacterized long-range chromatin loops, highlighting DNA methylations role in three-dimensional genome regulation and providing new insights into the complex regulatory relationships between epigenetic modifications and three-dimensional genome organization.

bioinformatics↗

bioRxiv2025

Endothelial AGO1 Drives Vascular Inflammation and Atherosclerosis via a Non-Canonical Nuclear Mechanism

BACKGROUNDEndothelial cell (EC) dysfunction is a cause and consequence of vascular inflammation and lipid dysregulation in atherosclerosis, yet the molecular drivers linking EC dysfunction to systemic metabolic derangements remain incompletely understood. Moreover, whether inhibiting an endogenous gene in ECs can impact liver function, lipid profile, and the vascular inflammation in the context of atherosclerosis has not been demonstrated. We previously identified Argonaute 1 (AGO1), a component of the RNA-induced silencing complex, as a regulator of EC function in angiogenesis and obesity. However, the role of endothelial AGO1 in vascular inflammation and liver function in the context of hyperlipidemia and atherosclerosis is unknown. METHODSEC-conditional AGO1 knockout (EC-AGO1-KO) and wildtype mice were subjected to pro-atherosclerotic models induced by AAV9-PCSK9 coupled with a Western diet or partial carotid ligation. Metabolic and vascular phenotype and gene expression were analyzed. In human liver sinusoidal and aortic ECs, AGO1 was knocked down using antisense oligonucleotides (ASO), followed by assessment of inflammatory responses (qPCR, RNA-seq, ELISA, and monocyte adhesion assays). To identify the molecular mechanisms linking AGO1 and EC inflammation, Cut&Tag sequencing, chromatin immunoprecipitation, immunofluorescence, proximal ligation assay, and co-immunoprecipitation were performed. The therapeutic effect of AGO1 inhibition was assessed using ASO-delivered via lipid nanoparticle (LNP) for systemic distribution and monocyte membrane-coated nanoparticles (MoNP) to target the inflamed endothelium. RESULTSEC-AGO1-KO mice exhibited improved plasma lipid profiles, reduced hepatic steatosis, inflammation, and fibrosis, and decreased aortic atherosclerotic burden. AGO1 knockdown in ECs attenuated inflammatory responses. Mechanistically, AGO1 interacted with NF-{kappa}B p65 and promoted p65 nuclear translocation and the transcriptional activation of pro-inflammatory genes, including ICAM1 and THBS1. AGO1-ASO delivered through LNP or MoNP achieved the anti-inflammatory, anti-hyperlipidemic, and anti-atherosclerotic effects, recapitulating the phenotypes observed with EC-AGO1-KO. CONCLUSIONSEndothelial AGO1 promotes vascular inflammation and liver dysfunction in the context of hyperlipidemia and atherosclerosis, in part through a non-canonical nuclear action of AGO1 as an NF-{kappa}B coactivator. Inhibition of endothelial AGO1 provides the dual benefits of ameliorating lipid dysregulation and suppressing vascular inflammation. These results highlight EC-AGO1 as a possible therapeutic target for atherosclerosis and cardiometabolic diseases.

pathology↗

bioRxiv2025

Gene Editing and Aging: A Bibliometric Analysis of Global Trends and Frontier Themes (2015-2024)

ObjectiveThe accumulation of DNA damage and mutations is a key contributor to aging. Recent studies have shown that disrupting the Beclin 1-BCL2 autophagy regulatory complex through gene editing can extend lifespan in mice. The precise application of gene editing technologies offers a promising strategy for aging. This study conducted a bibliometric analysis to map the knowledge landscape of gene editing and aging. MethodsWe retrieved publications related to genome editing and aging from the Web of Science Core Collection, covering the period from 2015 to 2024. The data were analyzed using VOSviewer and R package Bibliometrix. These tools enabled us to identify the most productive researchers, journals, institutions, countries and visualized current trends, emerging research hotspots. ResultsA total of 982 publications on genome editing and aging were identified. The United States (n=285) and China (n=214) form a dual-core structure leading global output. Harvard University (n=116) emerged as the most prolific institution. Scientific Reports was the top-publishing journal, with 23 articles in 2024. ZHANG Y (n=12, citations=102, H-index=6) was identified as the most productive author. KIM Es 2017 publication in Nature Communications (TC=494, TC/year=54.9, NTC=9.33) has had a significant and ongoing impact. The analysis indicates that future directions will include CRISPR optimization and AI-assisted genomic analysis. ConclusionThis study presents the first comprehensive bibliometric analysis and visualization of the knowledge structure in gene editing and aging research up to 2024. It offers researchers a detailed overview of current developments, trends, and emerging frontiers in this rapidly evolving domain.

scientific communication and education↗

bioRxiv2025

Oncogene Silencing via ecDNA Micronucleation

Extrachromosomal DNA (ecDNA) contributes to cancer genome instability by enabling high-copy oncogene amplification, intratumoural heterogeneity and rapid genetic change. Micronuclei (MN) are frequently observed in chromosomally unstable cancers, yet their origins and relevance in ecDNA-driven tumours remain incompletely understood. Here, we investigate the relationship between ecDNA segregation errors and MN formation. We find that ecDNA frequently localizes to MN and represents a prominent source of MN content in ecDNA-positive cancer cells. Mitotic clustering of oncogene-bearing ecDNAs is associated with asymmetric inheritance and mis-segregation into MN. Transfer of oncogenes from ecDNA into MN is accompanied by reduced transcriptional output. Single-MN sequencing shows that individual MN are enriched for ecDNA to a degree that exceeds expectations from stochastic mis-segregation and that in MN oncogenes originating from multiple distinct ecDNAs coalesce. Using live-cell imaging, we observe that cells inheriting ecDNA-positive MN show limited proliferative capacity and an increased likelihood of cell death. In neuroblastoma patients with MYCN-amplified ecDNA, higher frequencies of ecDNA-positive MN at diagnosis are associated with improved event-free and overall survival. Together, these findings link ecDNA mis-segregation to MN formation and reduced cellular fitness, suggesting that ecDNA-positive MN may reflect a state of impaired oncogenic ecDNA function with potential relevance for clinical outcome.

cancer biology↗

bioRxiv2025

Shared genomic architecture of the brain white matter structural connectome and intelligence

White matter (WM) connections, which facilitate communication between brain regions, substantially impact intellectual performance. However, the shared genetic underpinnings of WM connections and intelligence remain unclear. In the present study, we first conducted genome-wide association studies on the global and regional topological properties of WM connectomes constructed from diffusion-weighted and T1-weighted magnetic resonance images of 26,655 participants from the UK Biobank. Forty-one independent significant single-nucleotide polymorphisms (SNPs) associated with global WM connectome efficiency and 45 SNPs linked to nodal efficiency for 246 brain regions were identified. Then, regional heterogeneity was demonstrated for the genetic correlations between WM connectome efficiency and intelligence. The nodal efficiency of 128 regions with nominally significant genetic correlations with intelligence was determined. Among these, 44 regions and 3 regions shared SNPs with intelligence within the chromosome 6q21 and 3p21.1 locus respectively, and significant causal effects on intelligence were found for 63 regions, mainly in the orbital gyrus and superior frontal gyrus. Finally, we found that the integration of polygenetic score and WM connectome efficiency data was more accurate for predicting individual intelligence than the use of a single data type. These findings improve our understanding of how genes shape the individual WM connectome and intelligence and the relationship between WM connectomes and intelligence.

genomics↗

bioRxiv2025

Super-Resolution Diffusivity Mapping Reveals Spatial Correlations Between Lateral Mobility and Structural Heterogeneities on Cellular Membranes

Cellular membranes orchestrate critical processes such as molecular transport and signal transduction, both regulated by the lateral mobility of lipids and proteins. However, resolving nanoscale diffusional heterogeneities and elucidating their underlying mechanisms remains a formidable challenge due to the membranes intricate architecture and compositional diversity. Here, we present point-cloud single-molecule diffusivity mapping (pc-SMdM), a cutting-edge super-resolution technique that offers a point-cloud data format with enhanced spatial resolution for diffusivity mapping. Using pc-SMdM, we visualize nanoscale diffusion slowdown clusters with [~]50 nm in diameter on plasma membranes. These clusters are predominantly governed by cholesterol content, alongside contributions from membrane protein assemblies and topographical features. Leveraging two-color pc-SMdM, we concurrently imaged multiple lipids and membrane probes, revealing distinct "fingerprint" diffusivity maps shaped by their interactions within lipid bilayers. Our findings position pc-SMdM as a transformative tool for spatially resolving molecular interactions and membrane dynamics in live cells, offering new insights into the underlying mechanisms that govern membrane mobility at the nanoscale.

biophysics↗

bioRxiv2025

Boosting Antitumour Efficacy and Immunity by Boron Neutron Capture Therapy with Size-Controlled Nanoparticles

Boron neutron capture therapy (BNCT) is emerging cancer radiotherapy requiring 10B sensitizer. Although boronophenylalanine (BPA)-BNCT is approved clinically in Japan, the low tumour selectivity and retentivity result in the long-time infusion of high doses. Based on our finding of the size-controllable mechanochemical synthesis of boron-10 carbide nanoparticles (10B4C NPs), the 50 nm size NPs grafted with poly(glycerol), {superscript 1}BC(50)-PG, is found to show superior tumour selectivity and retentivity to enhance the eradication efficacy at much lower dosage (5 mg [10B] / kg (mouse)). The dosage is further reduced by twice neutron irradiation or combination with an immune checkpoint inhibitor (ICI). Antitumour immunity is found to be boosted by {superscript 1}BC(50)-PG-BNCT to induce abscopal effect to treat remote or metastatic tumours and long-term memory to prevent cancer recurrence. Additionally, minimal side effects and gradual NP excretion are observed for one year. The 10B4C(50)-PG is concluded to be a promising 10B carrier for clinical application of BNCT due to the prominent antitumour efficacy and immune-activation with minimal toxicity.

cancer biology↗

bioRxiv2025

Diverse microbial exposure exacerbates the development of allergic airway inflammation in adult mice

BackgroundExposure to a diversity of microbes has been implicated in playing a major role in susceptibility to the development of allergic lung type diseases. The hygiene hypothesis suggests that those exposed to a broad diversity of microbes are more likely to be protected against developing allergic type diseases. However, changes in exposure to microbial diversity can occur in both younger individuals, as well as in adults, and the effects are not always understood. ObjectiveWe investigated the effect of exposure to broad microbial diversity on the airway T cell response in house dust mite (HDM) induced allergic airway disease (AAD, a model of allergic asthma). MethodsWe increased exposure to broad microbial diversity by co-housing specific pathogen free (SPF) adult or newborn mice with pet store mice (PSE or BiPSE, respectively). Mice were then exposed to HDM to induce AAD. ResultsWe found that the effect of increased microbial exposure on the development of allergic airway inflammation differs by age. Increasing exposure to diverse microbes as adults exacerbates the development of allergic airway inflammation, whereas this was not observed when exposure occurred at birth. ConclusionWe suggest that experimental evaluation of the hygiene hypothesis in inflammation, particularly those using mouse models, may need to consider age of the host and time of microbial exposure. Capsule SummaryMouse models of increased exposure to diverse microbial environment shown to differentially affect the development of allergic airway inflammation, depending on the age of microbial exposure.

immunology↗

bioRxiv2025

Spatial single-cell interactome and niche-specific molecular signatures in alcohol-related liver disease

Alcohol-related liver disease (ALD) remains a major global health burden with limited therapeutic options due to an incomplete understanding of its underlying molecular mechanisms and cellular crosstalk. Here, we applied ultra-high resolution (on 2 {micro}m spots) spatial transcriptomics to a cirrhotic liver tissue obtained from an end-stage ALD patient, analyzing >265,000 spatially resolved cells with further validation on single-cell and single-nuclei datasets from patients with ALD cirrhosis. Our analysis delineated distinct cellular sub-populations and molecular landscapes across fibrotic, vascular, and parenchymal niches of ALD cirrhosis. We identified robust zonation of hepatocytes, hepatic stellate cells, and diverse immune subpopulations, including enrichment of pro-inflammatory T cells and dendritic cells in the fibrotic niche and MARCO+ tissue-resident macrophages localizing mostly in parenchymal areas. Analysis of spatial metrics assigned expression of WNT4, RCAN3, PPIAL4G, PLA2G5, and SLC6A9 to the fibrotic environment in ALD. Differential expression and ligand-receptor interactome analyses revealed niche-specific signaling, with marked CCL19-CCR7 activity in fibrotic regions and DLL4-NOTCH3 crosstalk in vascular compartments. Notably, WNT4+ fibroblasts emerged as key mediators of extracellular matrix remodeling and chemoattraction, particularly via CCL19-mediated signaling towards CD8+ T cells, which was validated on single-cell resolution within the ALD cirrhotic liver in external datasets. These spatial and single-cell findings highlight novel potential therapeutic targets for patients with ALD cirrhosis.

systems biology↗

bioRxiv2025

Aedes aegypti aminopeptidase N3 is a functional binding receptor of Bacillus thuringiensis subsp. israelensis Cry4Ba toxin

Bacillus thuringiensis is widely employed for biological control. It can effectively suppress populations of various mosquito species, including Aedes aegypti. However, the precise mechanism underlying the action of cry toxin secreted by Bacillus thuringiensis on Ae. aegypti remains elusive. In this study, we investigated one of the binding receptors of cry toxin, aminopeptidase N. Through comprehensive bioinformatics analysis involving whole-genome screening, genetic mapping, structural characterization, phylogenetic analysis, and spatiotemporal expression profiling, we identified twenty-nine homologs of Ae. aegypti aminopeptidase N. Further, we successfully expressed GST-APN3 protein in E. coli and demonstrated through ligand blot and ELISA assays that APN3 exhibits high affinity binding to Cry4Ba toxin (Kd = 20.53 nM). To elucidate the functional role of APN3 as a receptor mediating Cry4Ba activity in Ae. aegypti midgut cells (some of which express this gene at high levels), CRISPR/Cas9 technology was employed to knock out APN3. Our bioassay results revealed that APN3 knockout mosquito larvae had 2.9 to 4.1-fold higher resistance against Cry4Ba, indicating its crucial involvement as an active receptor mediating Cry4Ba activity. Overall, this study provides a foundation for elucidating the specific larvicidal mechanisms of Bt against mosquito populations.

molecular biology↗

bioRxiv2025

Accurate Somatic SV detection via sequence graph model-based local pan-genome optimization

Somatic structural variations (SVs) are critical genomic alterations in cancer genomes. Long-read sequencing (LRS) is theoretically optimal for detecting somatic SVs. However, influenced by reads-to-reference alignment errors, particularly in low-complexity or highly repetitive genomic intervals, current LRS-based somatic SV callers fail to accurately detect SVs. Moreover, the lack of comprehensive ground-truth datasets hinders accurate evaluation. Here, we introduce SVscope, a novel algorithm that fundamentally addresses these challenges by leveraging full-length sequence information from span-reads and integrating local graph-genome optimization with a random forest strategy. SVscope outperforms state-of-the-art methods on six paired long-read whole-genome sequencing (WGS) benchmark cell lines, achieving a maximum F1-score improvement of 16.7%. In simulated datasets, SVscope demonstrates superior performance in both somatic SV detection and read phasing tasks. Based on the findings from SVscope, we validated 47 somatic SVs, thereby significantly expanding the existing experimentally validated ground-truth somatic SV dataset by 69.1%.

bioinformatics↗

bioRxiv2025

Early-life infection with Cryptosporidium parvum induces inflammatory responses to dietary antigens

To examine the effect of early-life infection with Cryptosporidium parvum on the development of oral tolerance, we developed a low-dose infection model in neonatal mice. C. parvum infection in neonatal mice results in immunopathology in the colon. IL-1{beta} released during C. parvum infection blocked the formation of colonic goblet cell associated antigen passages, which normally serve as a conduit for antigen uptake and development of peripheral regulatory T cells (pTregs), responsible for long-term oral tolerance. Following infection with C. parvum, adoptively transferred OT-II cells, which respond to ovalbumin (ova), developed reduced frequency of Foxp3+Ror{gamma}t+ cells in mesenteric lymph nodes with an expansion of TH1-like Tregs in the colon. The altered pTreg profile was accompanied by a strong TH1 immune response and robust IgG2c antibody responses to orally administered ova. Our findings suggest that intestinal inflammation and altered pTreg development leads to loss of oral tolerance during early life infection with C. parvum.

microbiology↗

bioRxiv2025

Cryo-EM structures reveal the PP2A-B55α and Eya3 interaction that can be disrupted by a peptide inhibitor

We have previously shown that Eya3 recruits PP2A-B55 to dephosphorylate pT58 on Myc, increasing Myc stability and enhancing primary tumor growth of triple-negative breast cancer (TNBC). However, the molecular details of how Eya3 recruits PP2A-B55 remain unclear. Here we determined the cryo-EM structures of PP2A-B55 bound with Eya3, with an inhibitory peptide B55i, and in its unbound state. These studies demonstrate that Eya3 binds B55 through an extended peptide in the NTD of Eya3. The Eya3 peptide and other PP2A-B55 substrates and protein/peptide inhibitors including B55i bind to a similar area on the B55 surface but the molecular details of the binding differ. We further demonstrated that the B55i peptide inhibits the B55 and Eya3 interaction in vitro. B55i peptide expressed on a plasmid increases pT58 and decreases Myc protein level in TNBC cells, suggesting the potential of B55i or similar peptides as therapies for TNBC.

molecular biology↗

bioRxiv2025

Complex exchanges among plasmids and clonal expansion of lineages shape the population structure and virulence of Borrelia burgdorferi

The many plasmids (commonly >20 per strain) of Borrelia burgdorferi (Bb) pose challenges for studies of Lyme disease. The genetic content of most plasmids cannot be resolved using short-read sequencing. We generated long-read assemblies (LRA) of 183 isolates of human-derived Bb and analyzed patterns of genome variation in the Lyme spirochete. LRAs confirm that populations consist of strongly-structured genotypes with nearly-clonal structure and tight-knit blocks of accessory genome elements. Notably, these patterns of linkage were statistical but not physical, with linkage blocks distributed across multiple plasmids. A consequence of this structure is that plasmid name and/or plasmid subtype does not capture strain-specific genetic content. We used network methods to characterize patterns of genetic linkage. We demonstrate that co-occurring gene networks, here termed genetic modules, are the fundamental unit of genome variation in Bb and designate genetic modules consisting of co-occurring genes. We linked genetic modules to Bb phenotype by identifying modules that influence dissemination in humans. This modular decomposition clarifies previously observed associations between strain and virulence. For example, virulent RST1/OspC type A strains are distinguished by the presence of virulence-associated modules containing gene content from lp28-1, lp56, and the chromosome along with the absence of gene content on lp28-1 and lp28-4 associated with localized disease. LRAs also demonstrate that the well-established statistical linkage between physical unlinked genetic markers (e.g. RST and OspC) is a general pattern among accessory genome elements. In summary, genetic modules containing genes linked across multiple replicons, rather than strain-defining plasmids, organize the Bb accessory genome and the strain-specific variation responsible for differences in human virulence.

microbiology↗

bioRxiv2025

Anterior cingulate cortex in complex associative learning: monitoring action state and action content

Environmental changes necessitate adaptive responses, and thus the ability to monitor ones actions and their connection to specific cues and outcomes is crucial for survival. The anterior cingulate cortex (ACC) is implicated in these processes, yet its precise role in action monitoring versus outcome tracking remains unclear. To investigate this, we developed a novel discrimination-avoidance task for mice, designed with clear temporal separation between actions and outcomes. Our findings show that ACC neurons primarily encode post-action variables over extended periods, reflecting the animals preceding actions rather than the outcomes or values of those actions. Specifically, we identified two distinct subpopulations of ACC neurons: one encoding the action state (whether an action was taken) and the other encoding the action content (which action was taken). Importantly, increased post-action ACC activity was associated with better performance in subsequent trials. These findings suggest that the ACC supports complex associative learning through extended signaling of rich action-relevant information, thereby bridging cue, action, and outcome associations.

neuroscience↗

bioRxiv2025

Identification and Engineering of UDP-rhamnosyltransferase from Trillium tschonoskii for Heterologous Biosynthesis of Polyphyllin II in Engineered Yeast

AbstractPolyphyllins, a class of isospirostan-type steroidal saponins, exhibit potent cytotoxicity against a variety range of cancer cells. Although extensive research efforts have been made, the complete biosynthetic pathway of these compounds remains unclear. In order to elucidate these pathways, different tissues from Trillium tschonoskii were collected for sequencing that yielded 173,382 high-quality unigene sequences, among which 353 were annotated as glycosyltransferases. Then, a novel rhamnosyltransferase gene, UGT738A3, was characterized, which catalyzes the conversion of triglycoside polyphyllin III and pennogenin 3-O-beta-chacotrioside into tetraglycoside polyphyllin II and polyphyllin VII. The key residues that affect the catalytic activity of UGT738A3 were identified through site-directed mutation that the mutant A158T/P101L exhibited improved catalytic activity towards polyphyllin III and pennogenin 3-O-beta-chacotrioside by 2.5-fold and 6.5-fold respectively. We thus reconstructed the biosynthesis pathway of polyphyllin II in yeast by introducing the UGT93M3(catalyzing the formation of polyphyllin VI) and UGT738A3A158T/P101L3. This study not only elucidates the pivotal role of UGT738A3 in catalyzing the formation of tetraglycoside, but also provides highly efficient enzymatic components essential for the heterologous biosynthesis of polyphyllin saponins.

synthetic biology↗

bioRxiv2025

Peptide Design through Binding Interface Mimicry

Peptides offer distinct advantages for targeted therapy, including oral bioavailability, cellular permeability, and high specificity, which set them apart from conventional small molecules and biologics. In this work, we developed an AI algorithm, named PepMimic, to transform a known protein receptor or an existing antibody of a target into a short peptide drug by mimicking the binding interfaces between targets and known binders. The structural root mean square deviation and interface DockQ with reference binders were 61% and 75% better than the best existing methods on the PepBench datasets. We then applied this novel peptide-design methodology to five drug targets: PD-L1, CD38, BCMA, HER2, and CD4. SPRi results show that 8% of the peptides exhibited dissociation constant (KD) values at the 10-8M level, and 26 peptides achieving KD values as low as 10-9M. This success rate was 20,000 times higher than that observed in a random library screening conducted under identical conditions. PepMimic was applied to target proteins lacking available binders by first utilizing AI algorithms to design protein binders, followed by the generation of peptides through simulation of these artificial interfaces. The top-ranked peptides underwent extensive cellular validation and in vivo testing through tail vein injections in breast, myeloma, and lung tumor mouse models. Experimental results demonstrated effective membrane binding and highlighted the strong potential of these peptides for clinical diagnostic imaging and targeted therapeutic applications.

biochemistry↗