Biology subjects

Liang, Z.

Publications and source records attributed to Liang, Z..

At least 91 records · Page 5Linked to original sources

bioRxiv2024

Cytotoxicity of Activator Expression in CRISPR-based Transcriptional Activation Systems

CRISPR-based transcriptional activation (CRISPRa) has extensive research and clinical potential. Here, we show that commonly used CRISPRa systems can exhibit pronounced cytotoxicity. We demonstrate the toxicity of published and new CRISPRa vectors expressing the activation domains (ADs) of the transcription factors p65 and HSF1, components of the synergistic activation mediator (SAM) CRISPRa system. Based on our findings for the SAM system, we extended our studies to additional ADs and the p300 acetyltransferase core domain. We show that the expression of potent transcriptional activators in lentiviral producer cells leads to low lentiviral titers, while their expression in the transduced target cells leads to cell death. Using inducible lentiviral vectors, we could not identify an activator expression window for effective SAM-based CRISPRa without measurable toxicity. The toxicity of current SAM-based CRISPRa systems hinders their wide adoption in biomedical research and introduces selection bottlenecks that may confound genetic screens. Our results suggest that the further development of CRISPRa technology should consider both the efficiency of gene activation and activator toxicity.

molecular biology↗

bioRxiv2024

Chain Termination Effects of Chemically Modified Nucleoside Analogs on SARS-CoV-2 RNA Polymerase

Nucleoside analogues (NuAs) targeting the SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) represent a key antiviral strategy. However, their efficacy is fundamentally limited by the viral proofreading exoribonuclease nsp14, which excises misincorporated NuAs from nascent viral RNA. To identify NuAs capable of achieving chain termination while resisting nsp14-nsp10-mediated excision, we sequentially assessed a panel of nucleotide analogues encompassing ribose 2', 3', and nucleobase modifications for RdRp chain termination followed by nsp14-nsp10 ExoN resistance. Among the candidates, 5-methyl-3'-dUTP emerges as a standout inhibitor. It functions as an efficient immediate chain terminator, and the 5-methyl modification confers substantial resistance to nsp14-nsp10-mediated proofreading excision, notably outperforming its unmodified counterpart 3'-dUTP. RdRp also exhibited improved incorporation efficiency for this analogue. RNA chains terminated with 5-methyl-3'-dUTP exhibit barely detectable primer extension even following nsp14-nsp10-mediated exonucleolytic cleavage. The superior chain termination and ExoN resistance of 5-methyl-3'-dUTP were independently corroborated using an smFRET assay. Mechanistically, molecular dynamics simulations reveal that this enhanced ExoN resistance arises from destabilization of F146 loop stacking in the nsp14-nsp10 active site. Collectively, these findings establish 5-methyl-3'-dUTP as a promising antiviral lead compound and provide a structural framework for designing next-generation nucleoside analogues capable of evading coronavirus proofreading surveillance.

microbiology↗

bioRxiv2024

Modular Nano-Scaffold Biocatalysis for Superior PET Depolymerization and Valorization

The global crisis of polyethylene terephthalate (PET) waste demands new strategies for sustainable management. Biocatalytic recycling offers a promising route, but direct depolymerization under moderate conditions is often limited by kinetic constrains of individual enzyme. Here, we engineer and optimize modular protein scaffolds that utilize orthogonal interaction domains, including the cohesin-dockerin pairs from bacterial cellulosomes and the SH3 ligand/domain pair from mouse Crk, to assemble complementary hydrolases (PETase, MHETase, and ICCG) into a synergistic multi-enzyme cascade. This architecture overcomes intermediate inhibition, efficiently converting PET hydrolysis products into the final monomers, terephthalic acid (TPA) and ethylene glycol (EG), outperforming previous multi-enzyme systems. The systems application is further evaluated by stability enhancement via immobilization on metal-organic frameworks (MOFs), coupling depolymerization to the valorization of EG into glycolic acid (GA), and adapting the system for scalable whole-cell biocatalysis. This scaffold-based multi-enzyme approach provides a new strategy for developing integrated biocatalytic systems to advance a circular PET economy.

synthetic biology↗

bioRxiv2024

Validity of a New Stress Induction Protocol using Speech Improvisation (IMPRO)

This paper proposes a novel stress induction protocol utilizing speech improvisation -- Interactive Multitask Performance Response Observation (IMPRO) -- in front of an audience. This approach aims to induce stress through a combination of public speaking, cognitive effort, and challenge to maintain a spontaneous narrative. We investigate the validity of this novel approach by examining physiological responses of 35 healthy participants aged 18 to 38. Saliva cortisol was measured as the ground truth for mental stress assessment. In addition, electrodermal activity (EDA) was used as a non-invasive measure of sympathetic activation, offering real-time data with high resolution. Functional near-infrared spectroscopy (fNIRS) measurement was performed to assess the cortical hemodynamic responses induced by the IMPRO protocol. We focused on hemodynamics in the prefrontal cortex (PFC), a region associated with stress processing. We found that cortisol levels and EDA significantly increased in response to the stress task compared to the baseline. We also observed a significant change in PFC hemodynamic levels in a set of channels compared to the baseline, with a higher overall increase in the right frontopolar area compared to the left. In conclusion, our findings validated IMPRO as an effective and easy to perform method for mental stress induction.

neuroscience↗

bioRxiv2024

Leveraging large language models for metabolic engineering design

Establishing efficient cell factories involves a continuous process of trial and error due to the intricate nature of metabolism. This complexity makes predicting effective engineering targets a challenging task. Therefore, it is vital to learn from the accumulated successes of previous designs for advancing future cell factory development. In this study, we developed a method based on large language models (LLMs) to extract metabolic engineering strategies from research articles on a large scale. We created a database containing over 29006 metabolic engineering entries, 1210 products and 751 organisms. Using this extracted data, we trained a hybrid model combining deep learning and mechanistic approaches to predict engineering targets. Our model outperformed traditional metabolic engineering target prediction algorithms, excelled in predicting the effects of gene modifications, and generalized well to out-of-distribution products and multiple gene combinations. Our study provides a valuable dataset, a chatbot, and an engineering target prediction model for the metabolic engineering field and exemplifies an efficient method for leveraging existing knowledge for future predictions.

systems biology↗

bioRxiv2024

Sapphire-Based Optrode for Low Noise Neural Recording and Optogenetic Manipulation

Electrophysiological recordings of neurons in deep brain regions using optogenetic stimulation are essential to understanding and regulating the role of complex neural activity in biological behavior and cognitive function. Optogenetic techniques have significantly advanced neuroscience research by enabling the optical manipulation of neural activities. Because of the significance of the technique, constant advancements in implantable optrodes that integrate optical stimulation with low-noise, large-scale electrophysiological recording are in demand to improve the spatiotemporal resolution for various experimental designs and future clinical applications. However, robust and easy-to-use neural optrodes that integrate neural recording arrays with high-intensity light emitting diodes (LEDs) are still lacking. Here, we propose a neural optrode based on Gallium Nitride (GaN) on sapphire technology, which integrates a high-intensity blue LED with a 5x2 recording array monolithically for simultaneous neural recording and optogenetic manipulation. To reduce the noise interference between the recording electrodes and the LED, which is in close physical proximity, three metal grounding interlayers were incorporated within the optrode, and their ability to reduce LED-induced artifacts during neural recording was confirmed through both electromagnetic simulations and experimental demonstrations. The capability of the sapphire optrode to record action potentials has been demonstrated by recording the firing of mitral/tuft cells in the olfactory bulbs of mice in vivo. Additionally, the elevation of action potential firing due to optogenetic stimulation observed using the sapphire probe in medial superior olive (MSO) neurons of the gerbil auditory brainstem confirms the capability of this sapphire optrode to precisely access neural activities in deep brain regions under complex experimental designs.

neuroscience↗

bioRxiv2024

A causative SNP in the promoter of myogenin is essential for myogenic differentiation

Single nucleotide polymorphisms (SNPs) widely existing in different breeds genome represent population-specific. Under the influence of long-term evolution and artificial selection, there are a large number of SNPs between western lean-type pig breeds and Chinese indigenous pig breeds, but until now, little is known about their roles in inter-breed differences. Our study revealed SNP rs3471653254 C>T generated from the two types of pigs mentioned above, located in the promoter shared by MyoG and Myoparr, played an important role in the differentiation of myoblast by influencing the enrichment of HOXA5 to regulate the transcription of MyoG and Myoparr. Meanwhile, Myoparr could be used as the sponge of mir-30b-3p which repressed myogenic differentiation and muscle regeneration through targeting MyoD. Our results indicated that SNP rs3471653254 C>T is essential for myogenic differentiation and regeneration and could be used as an ideal site for increasing lean meat production in pigs.

genetics↗

bioRxiv2024

Genomic and transcriptomic profiles influence on brain morphology and their interactions with pain sensitivity

Pain sensitivity varies widely among individuals and is influenced by a complex interplay of multi-omics factors, including genetic variations, gene expression, and brain morphology. While previous studies have identified associations between pain sensitivity and brain morphology, the exact mechanisms by which genetic profiles interact with brain structure to influence individual pain sensitivity remain unclear. In this study, we used aggregated datasets, including magnetic resonance imaging (MRI) and single nucleotide polymorphism (SNP) genotypes from 432 healthy participants, along with gene expression data from the Allen Human Brain Atlas (AHBA), to explore this multi-omics interplay. We first measured individual pain thresholds using laser stimuli and discovered structural brain differences between high and low pain sensitivity groups. We then identified two key gene sets with polarized expression patterns linked to brain morphology variations, enriched in functions related to ion channels and transmembrane transporter activities. Further statistical and mediation analyses revealed specific SNPs from ECM1, SLC24A2, and SCN9A genes that influence pain sensitivity, mediated through brain morphological changes in multiple basal ganglia regions. Our findings suggested that these SNPs not only affect brain structure but also modulate how individuals pain perception. Finally, we proposed an interpretation model integrating genomic, transcriptomic, and neuroimaging data, providing a detailed framework that illustrates the multi-omics contributions to individual difference in pain sensitivity. This study advances our understanding of how genetic and brain structural factors combine to shape pain perception, offering potential targets for personalized pain management strategies.

bioinformatics↗

bioRxiv2024

SMYD5 is a ribosomal methyltransferase that catalyzes RPL40 lysine methylation to enhance translation output and promote hepatocellular carcinoma

While lysine methylation is well-known for regulating gene expression transcriptionally, its implications in translation have been largely uncharted. Trimethylation at lysine 22 (K22me3) on RPL40, a core ribosomal protein located in the GTPase activation center (GAC), was first reported 27 years ago. Yet, its methyltransferase and role in translation remain unexplored. Here, we report that SMYD5 has robust in vitro activity toward RPL40 K22 and primarily catalyzes RPL40 K22me3 in cells. The loss of SMYD5 and RPL40 K22me3 leads to reduced translation output and disturbed elongation as evidenced by increased ribosome collisions. SMYD5 and RPL40 K22me3 are upregulated in hepatocellular carcinoma (HCC) and negatively correlated with patient prognosis. Depleting SMYD5 renders HCC cells hypersensitivity to mTOR inhibition in both 2D and 3D cultures. Additionally, the loss of SMYD5 markedly inhibits HCC development and growth in both genetically engineered mouse (GEM) and patient-derived xenograft (PDX) models, with the inhibitory effect in the PDX model further enhanced by concurrent mTOR suppression. Our findings reveal a novel role of the SMYD5 and RPL40 K22me3 axis in translation elongation and highlight the therapeutic potential of targeting SMYD5 in HCC, particularly with concurrent mTOR inhibition. This work also broadens the understanding of lysine methylation, extending its significance from transcriptional regulation to translational control.

cancer biology↗

bioRxiv2024

Anoctamins mediate polymodal sensory perception and larval metamorphosis in a non-vertebrate chordate.

An exceptional feature of most animals is their ability to perceive diverse sensory cues from the environment and integrate this information in their brain to yield ethologically relevant behavioral output. For the myriad of marine species, the ocean represents a complex sensory environment, which acts as a crucible of evolution for polymodal sensory perception. The cellular and molecular bases of polymodal sensory perception in a marine environment remain enigmatic. Here we use Ca2+ imaging and quantitative behavioral analysis to show that in the tunicate Ciona intestinalis two members of the evolutionarily conserved Anoctamin family1-4 (Tmem16E/Ano5 and Tmem16F/Ano6), are required for sensing chemosensory and mechanosensory metamorphic cues. We find that they act by modulating neuronal excitability and Ca2+ response kinetics in the primary sensory neurons and axial columnar cells of the papillae, a widely conserved sensory-adhesive organ across ascidians5-9. Finally, we use electrophysiological recordings and a scramblase assay in tissue culture to demonstrate that Tmem16E/Ano5 acts as a channel, while Tmem16F/Ano6 is a bifunctional ion channel and phospholipid scramblase. Our results establish Ano5 and Ano6 as novel players in the zooplanktonic, pre-vertebrate molecular toolkit that controls polymodal sensory perception in aquatic environments.

neuroscience↗

bioRxiv2024

Design nonrepetitive and diverse activity single-guide RNA by deep learning

Multiplex and precise control of the gene expression based on CRISPR/Cas9 is important to metabolic regulation in synthetic biology. However, employing single guide RNAs (sgRNAs) that possess repetitive DNA sequences and exhibit uniform activity could detrimentally affect the editing process, undermining both its stability and regulatory potential. In this study, we developed a deep generative model based on a decoder-only Transformer architecture (sgRNAGen) for the de novo generation of a series of nonrepetitive and diverse sgRNAs with activity. To assess the quality of sgRNAs generated by sgRNAGen, we evaluated their activity by targeting essential genes, with the results indicating that 98% of the generated sgRNAs were active in Bacillus subtilis. The generated sgRNAs were further validated for applications in single-gene editing, large fragment knockouts, and multiplex editing. Notably, the efficiency of knocking out long fragments up to 169.5 kb reached 100%, and targeting multiple sites allowed for the creation of strains with various combinations of mutations in a single editing. Furthermore, we developed a CRISPRi system utilizing the designed sgRNAs to regulate gene expression with desired strength and high precision. SgRNAGen offers a method for devising nonrepetitive and diverse activity sgRNAs, enhancing metabolic control and advancing applications within synthetic biology. TOC O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=112 SRC="FIGDIR/small/596019v1_ufig1.gif" ALT="Figure 1"> View larger version (30K): org.highwire.dtl.DTLVardef@f68faaorg.highwire.dtl.DTLVardef@151f5e2org.highwire.dtl.DTLVardef@1e5cb28org.highwire.dtl.DTLVardef@17cd3f8_HPS_FORMAT_FIGEXP M_FIG C_FIG

synthetic biology↗

bioRxiv2024

Unveiling the Secrets of Extracellular Vesicles in Urban Water Systems: Understanding the Link Between Human and Environmental Health

It is crucial to gain valuable insights into the ecological health of rivers to inform management decisions and protect sustainable living conditions. Understanding biological information is vital to gaining insight into river ecosystem biodiversity, but reliable methods are challenging. Here, We investigate the potential impact of extracellular vesicles (EVs) in urban water systems on human and environmental health to promote urban sustainability. We used EXODUS to detect EVs and perform metaproteomic analysis on samples from an urban water system that contained human feces, wastewater, bacteria, plants, arthropods, and soil. We analyzed EVs collected from urban and green areas, observing taxonomic variations and discovering bacterial contributions to their protein content. According to our research, the abundance and expression levels of proteins in EVs can indicate how human activities affect microbial communities in rivers, potentially impacting public health. Our study offers a fresh perspective on the interconnectedness of urban sustainability, public health, and river ecosystem biodiversity.

ecology↗

bioRxiv2024

A beta-Galactosidase acting on unique galactosides: the structure and function of a beta-1,2-galactosidase from Bacteroides xylanisolvens, an intestinal bacterium

Galactosides are major carbohydrates that are found in plant cell walls and various prebiotic oligosaccharides. Studying the detailed biochemical functions of {beta}-galactosidases in degrading these carbohydrates is important. In particular, identifying {beta}-galactosidases with new substrate specificities could help in the production of potentially beneficial oligosaccharides. In this study, we identified a {beta}-galactosidase with novel substrate specificity from Bacteroides xylanisolvens, an intestinal bacterium. The enzyme did not show hydrolytic activity toward natural {beta}-galactosides during the first screening. However, when -D-galactosyl fluoride (-GalF) as a donor substrate and galactose or D-fucose as an acceptor substrate were incubated with a nucleophile mutant, reaction products were detected. The galactobiose produced from the -GalF and galactose was identified as {beta}-1,2-galactobiose using NMR. Kinetic analysis revealed that this enzyme effectively hydrolyzed {beta}-1,2-galactobiose and {beta}-1,2-galactotriose. In the complex structure with methyl {beta}-galactopyranose as a ligand, the ligand is only located at subsite +1. The 2-hydroxy group and the anomeric methyl group of methyl {beta}-galactopyranose faces in the direction of subsite -1 and the solvent, respectively. This observation is consistent with the substrate specificity of the enzyme regarding linkage position and chain length. Overall, we concluded that the enzyme is a {beta}-galactosidase acting on {beta}-1,2-galactooligosaccharides. SynopsisThe structural and functional analysis of {beta}-galactosidase from an intestinal bacterium led to the discovery of a new {beta}-galactosidase hydrolyzing unique {beta}-1,2-galactooligosaccharides.

biochemistry↗

bioRxiv2024

Nuclear Translocation of SIRT4 Mediates Deacetylation of U2AF2 to Modulate Renal Fibrosis Through Alternative Splicing-mediated Upregulation of CCN2

TGF-{beta} stimulates CCN2 expression which in turn amplifies TGF-{beta} signaling. This process promotes extracellular matrix production and accelerates the pathological progression of fibrotic diseases. Alternative splicing plays an important role in multiple disease development, while U2 small nuclear RNA auxiliary factor 2 (U2AF2) is an essential factor in the early steps of pre-mRNA splicing. However, the molecular mechanism underlying abnormal CCN2 expression upon TGF-{beta} stimulation remains unclear. This study elucidates that SIRT4 acts as a master regulator for CCN2 expression in response to TGF-{beta} by modulating U2AF2-mediated alternative splicing. Analyses of renal biopsy specimens from patients with CKD and mouse fibrotic kidney tissues revealed marked nuclear accumulation of SIRT4. The tubulointerstitial fibrosis was alleviated by global deletion or tubular epithelial cell (TEC)-specific knockout of Sirt4, and aggravated by adeno-associated virus-mediated SIRT4 overexpression in TECs. Furthermore, SIRT4 was found to translocate from the mitochondria to the cytoplasm through the BAX/BAK pore under TGF-{beta} stimulation. In the cytoplasm, TGF-{beta} activated the ERK pathway and induced the phosphorylation of SIRT4 at Ser36, which further promoted its interaction with importin 1 and subsequent nuclear translocation. In the nucleus, SIRT4 was found to deacetylate U2AF2 at K413, facilitating the splicing of CCN2 pre-mRNA to promote CCN2 protein expression. Importantly, exosomes containing anti-SIRT4 antibodies were found to effectively mitigate the UUO-induced kidney fibrosis in mice. Collectively, these findings indicated that SIRT4 plays a role in kidney fibrosis by regulating CCN2 expression via the pre-mRNA splicing.

molecular biology↗

bioRxiv2024

Chemotherapy induces myeloid-driven spatial T-cell exhaustion in ovarian cancer

To uncover the intricate, chemotherapy-induced spatiotemporal remodeling of the tumor microenvironment, we conducted integrative spatial and molecular characterization of 97 high-grade serous ovarian cancer (HGSC) samples collected before and after chemotherapy. Using single-cell and spatial analyses, we identify increasingly versatile immune cell states, which form spatiotemporally dynamic microcommunities at the tumor-stroma interface. We demonstrate that chemotherapy triggers spatial redistribution and exhaustion of CD8+ T cells due to prolonged antigen presentation by macrophages, both within interconnected myeloid networks termed "Myelonets" and at the tumor stroma interface. Single-cell and spatial transcriptomics identifies prominent TIGIT-NECTIN2 ligand-receptor interactions induced by chemotherapy. Using a functional patient-derived immuno-oncology platform, we show that CD8+T-cell activity can be boosted by combining immune checkpoint blockade with chemotherapy. Our discovery of chemotherapy-induced myeloid-driven spatial T-cell exhaustion paves the way for novel immunotherapeutic strategies to unleash CD8+ T-cell-mediated anti-tumor immunity in HGSC.

cancer biology↗

bioRxiv2024

A hemagglutinin and neuraminidase biased immunological memory shapes the dynamics of antibody responses to Influenza A virus

Influenza A virus (IAV) infection establishes a more diverse immunological memory to different viral proteins compared to vaccination. We hypothesized that the relative abundance of pre-existing immune memory to different viral antigens could skew post-infection antibody responses. To explore this, we generated mouse models with either an IAV hemagglutinin (HA)- or neuraminidase (NA)-biased immunological memory. We inoculated groups of mice with cocktails of isogenic viruses bearing antigenically-distinct HA (H3v) or NA (N2v) chosen to span the IAV H3N2 human circulation history. We challenged the mice with two H3N2 strains of opposing virulence and antigenic distance (AD) and examined the post-infection antibody landscapes. In both challenges, immune-naive mice seroconverted to both HA and NA whereas in primed mice, antibody response was detected to the antigen for which there is no pre-existing memory. In cases where the homologous antibody response was blunted, there was diversification on the breadth of response to antigenically-related strains with low baseline titers. Our findings clarifies the concept of "original antigenic sin" and demonstrate a mechanism by which the dynamics of antibody responses to HA and NA after infection can be altered by pre-existing immunity.

immunology↗

bioRxiv2024

A multi-modal image fusion workflow incorporating MALDI imaging mass spectrometry and microscopy for the study of small pharmaceutical compounds

Multi-modal imaging analyses of dosed tissue samples can provide more comprehensive insight into the effects of a therapeutically active compound on a target tissue compared to single-modal imaging. For example, simultaneous spatial mapping of pharmaceutical compounds and endogenous macromolecule receptors is difficult to achieve in a single imaging experiment. Herein, we present a multi-modal workflow combining imaging mass spectrometry with immunohistochemistry (IHC) fluorescence imaging and brightfield microscopy imaging. Imaging mass spectrometry enables direct mapping of pharmaceutical compounds and metabolites, IHC fluorescence imaging can visualize large proteins, and brightfield microscopy imaging provides tissue morphology information. Single-cell resolution images are generally difficult to acquire using imaging mass spectrometry, but are readily acquired with IHC fluorescence and brightfield microscopy imaging. Spatial sharpening of mass spectrometry images would thus allow for higher fidelity co-registration with higher resolution microscopy images. Imaging mass spectrometry spatial resolution can be predicted to a finer value via a computational image fusion workflow, which models the relationship between the intensity values in the mass spectrometry image and the features of a high spatial resolution microscopy image. As a proof of concept, our multi-modal workflow was applied to brain tissue extracted from a Sprague Dawley rat dosed with a kratom alkaloid, corynantheidine. Four candidate mathematical models including linear regression, partial least squares regression (PLS), random forest regression, and two-dimensional convolutional neural network (2-D CNN), were tested. The random forest and 2-D CNN models most accurately predicted the intensity values at each pixel as well as the overall patterns of the mass spectrometry images, while also providing the best spatial resolution enhancements. Herein, image fusion enabled predicted mass spectrometry images of corynantheidine, GABA, and glutamine to approximately 2.5 m spatial resolutions, a significant improvement compared to the original images acquired at 25 m spatial resolution. The predicted mass spectrometry images were then co-registered with an H&E image and IHC fluorescence image of the - opioid receptor to assess co-localization of corynantheidine with brain cells. Our study also provides insight into the different evaluation parameters to consider when utilizing image fusion for biological applications.

biochemistry↗

bioRxiv2024

Antibody-drug conjugates targeting EGFR ligand Epiregulin inhibit colorectal tumor growth irrespective of RAS mutational status

As colorectal cancer (CRC) remains a leading cause of cancer-related death, identifying therapeutic targets and approaches is essential to improve patient outcomes. The EGFR ligand epiregulin (EREG) is highly expressed in RAS wildtype and mutant CRC with minimal expression in normal tissues, making it an attractive target for antibody-drug conjugate (ADC) development. In this study, we produced and purified an EREG monoclonal antibody (mAb), H231, that had high specificity and affinity for human and mouse EREG. H231 also internalized to lysosomes, which is important for ADC payload release. ImmunoPET and ex vivo biodistribution studies showed significant tumor uptake of 89Zr-labeled H231 with minimal uptake in normal tissues. H231 was conjugated to either cleavable dipeptide or tripeptide chemical linkers attached to the DNA-alkylating payload duocarmycin DM, and cytotoxicity of EREG ADCs was assessed in a panel of CRC cell lines. EREG ADCs incorporating tripeptide linkers demonstrated the highest potency in EREG-expressing CRC cells irrespective of RAS mutations. Preclinical safety and efficacy studies showed EREG ADCs were well-tolerated, neutralized EGFR pathway activity, caused significant tumor growth inhibition or regression, and increased survival in CRC cell line and patient-derived xenograft models. These data suggest EREG is a promising target for the development of ADCs for treating CRC and other cancer types that express high levels of EREG. While the efficacy of clinically approved anti-EGFR mAbs are largely limited by RAS mutational status, EREG ADCs may show promise for both RAS mutant and wildtype patients, thus improving existing treatment options. Significance: EREG-targeting antibody-drug conjugates demonstrate acceptable safety and robust therapeutic efficacy in RAS mutant and wildtype colorectal cancer, suggesting their potential as an alternative to EGFR-targeted therapy to benefit a broader patient population.

cancer biology↗