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Li, V.

Publications and source records attributed to Li, V..

At least 19 recordsLinked to original sources

A global signature of microbiome resilience to climate stress

As climate change accelerates, the microbial communities sustaining organisms and ecosystems worldwide face an uncertain future, with some proving resilient to stress while others collapse, yet no generalizable framework exists to distinguish the two beforehand. Using amplicon sequencing data from ~29,000 microbiomes, we constructed over 440 matched network pairs spanning three of the most pressing stressors: drought, warming, and salinity. We show that keystone microbes with high connectivity, functional redundancy, and metabolic flexibility, are disproportionately likely to survive stress, and that baseline network and functional properties predict community-level structural and functional stability even as taxonomic composition shifts stochastically. These relationships held across the diverse hosts, ecosystems, and regions we tested. Because these properties can be derived from standard, low-cost sequencing collected before disturbance occurs, our findings turn baseline microbiome data into an early-warning system for identifying which communities are most vulnerable to climate stress.

systems biology↗

Do microbial effects on hosts vary across life stage and vitalrate?

Interactions with microbial symbionts are widespread and can influence host fitness through effects on survival, growth, and reproduction. Demographic and evolutionary theory suggests that symbiont effects on components of host fitness should often vary in strength and even in direction as host and symbiont allocate limited resources. Through a meta-analysis of 1,736 host-symbiont effect size measurements, we evaluated how commonly the direction of symbiont effects varies across host life history stages and vital rates, and whether these effects across stages and vital rates differ across taxa. Contrary to our prediction, we found that, compared to a null expectation, published effects on host vital rates more often occurred consistently in the same direction, rather than in opposition. This finding suggests that symbiosis often provides benefits to host performance that manifest across vital rates. While the direction of symbiotic effect was often consistent, the magnitude of effects differed across fitness components and across host taxonomic groups, highlighting diverse demographic mechanisms by which symbiosis can influence host fitness. For hosts in the Poaceae family, we also found that effects of symbiosis varied across symbiont type: vertically-transmitted Epichloe fungi most strongly benefited seed germination, while the benefits of mycorrhizal fungi and bacterial endophytes were stronger for adult growth and reproduction. Importantly, this literature survey also highlights gaps in the current literature on host-microbe symbiosis. Most studies measure growth during juvenile stages for taxa from iconic, well-studied symbioses. Meta-analytic estimates of symbiotic effects on host life history provide a path towards quantitatively modeling host-symbiont demography across the tree of life.

ecology↗

NC4touch: An open-source networked touchscreen apparatus for rodent behavioral testing

In recent years, rodent touchscreen-based paradigms have gained popularity for their flexible task design, automated data collection and improved standardization. These devices minimize experimenter intervention and contribute to more replicable and reliable research. However, the high costs and proprietary hardware / software associated with commercially available systems present a financial barrier for smaller labs or researchers with limited resources. To address this issue, we developed "NC4Touch", an open-source scalable, modular rodent testing apparatus. It features three independent touchscreens and a feeding port for easy use in a wide array of tests comparable to those available with standard operant chambers, with the added benefit that visual stimuli can be highly customized. The system includes a user-friendly graphical interface that offers real-time control, task customization, video recording, and data management. We demonstrate its effectiveness in a visual discrimination task using both rats and mice. Several devices can be operated in parallel from the same computer and user interface, allowing high- throughput data collection. We provide detailed assembly instructions for the hardware and well- documented and easily-configurable software. We hope that the affordable open-source nature of NC4Touch will expand the scope of behavioral testing, allowing researchers to overcome traditional financial barriers and create a more collaborative community. Significance StatementTouchscreen-based testing is a powerful tool for assessing cognition in rodent models, however commercial systems remain cost-prohibitive for many researchers. NC4Touch is a rodent touchscreen apparatus that provides an open-source, customizable, and affordable alternative that enables high-throughput, cognitive testing in both mice and rats. By lowering financial barriers and promoting hackability, this system enables greater accessibility and collaboration in research.

animal behavior and cognition↗

Ultramutagenesis through combined Msh2 depletion and proofreading-deficient DNA polymerase ε expression enhances small molecule target identification in forward genetic screens

DNA mismatch repair (MMR) deficiency has been widely utilized in forward genetic screens to identify drug resistance mutations and elucidate the mechanisms of action of cytotoxic small molecules. However, MMR deficiency generates a characteristic mutational signature enriched for C>T transitions that precludes saturation of mutagenesis. We hypothesized that expression of proofreading deficient DNA polymerase epsilon, as has been observed in patients with biallelic MMR deficiency, would enhance mutagenesis and enable identification of new resistance mutations and cytotoxin mechanisms. Here, we combine auxin-inducible degradation of Msh2 with doxycycline-inducible expression of the proofreading-deficient Pol{varepsilon}P286R mutant in murine small cell lung cancer (SCLC) cells. Simultaneous MMR deficiency and mutant Pol{varepsilon} expression markedly increased the emergence of bortezomib-resistant clones relative to either perturbation alone and targeted sequencing of Psmb5 in bortezomib-resistant clones identified recurrent mutations previously reported in chemical mutagenesis-based resistance screens. We next applied this platform to investigate resistance to lurbinectedin, a clinically relevant therapy for SCLC. Whole-exome sequencing revealed recurrent loss-of-function alterations in nucleotide excision repair (NER) genes, most notably Ercc5 and Ercc4, implicating NER deficiency as a major mechanism of lurbinectedin resistance and supporting a central role for NER in mediating lurbinectedin-induced cytotoxicity. Collectively, these findings establish inducible ultramutagenesis as a powerful and versatile platform for the unbiased discovery of drug resistance mechanisms and therapeutic vulnerabilities.

Cancer Biology↗

Maternal Influenza A Virus Infection Induces Antiviral and Immune Dysregulation in the Placenta and Fetus Without Vertical Transmission

Influenza A virus (IAV) infection during pregnancy is associated with stillbirth and preterm birth, but the degree to which IAV alters placental and fetal immunity is poorly understood. The objective of our study was to determine the immunologic impact of maternal IAV infection on the placenta and fetus in a pigtail macaque (Macaca nemestrina) model. Pregnant pigtail macaques were inoculated with 107 plaque forming units (PFU) of IAV [A/California/07/2009 (H1N1)] and underwent necropsy 5 days post-infection (N=11). Results were compared to uninfected historical controls (N=16). IAV inoculation induced maternal pneumonia in all cases. Stillbirth occurred in 18% (2/11) of IAV-infected pregnancies, but not in controls. While vertical transmission was not observed, low-level IAV viral RNA was detected in two placentas. In the placenta, maternal IAV infection was associated with increased IL-1{beta}, IL-18, and IFN-{beta} levels, and an upregulated type I interferon (IFN) transcriptional response. IAV infection was also associated with significantly higher frequencies of intermediate and non-classical monocytes, plasmacytoid dendritic cells, CD4 T cells, and NKT cells in the fetus (lung, lymph node, blood). Although placental immune and transcriptional perturbations were rarely correlated with maternal IAV disease indicators (e.g., maternal lung viral load/IFN-/IFN-{beta}/IL-6), there were consistent and significant correlations between these metrics and perturbed immune cell populations in the fetus (CD4+ and CD8+ T cells, plasmacytoid dendritic cells, monocyte sub-populations). Maternal IAV infection disrupted both placental and fetal immune environments, but only fetal immune alterations correlated with maternal lung disease severity. One Sentence SummaryMaternal influenza A virus infection in pregnant pigtail macaques dysregulates placental and fetal immunity, with disease severity correlating strongly with fetal, but not placental, immune perturbations.

systems biology↗

Ambient temperature storage in DESS supports molecular studies of benzimidazole resistance from canine hookworm eggs

Emerging benzimidazole resistance in canine hookworms poses a growing concern for veterinary and public health. Molecular diagnostics targeting {beta}-tubulin gene mutations are essential for resistance surveillance but traditionally rely on refrigerated faecal samples. This study evaluates dimethyl sulfoxide, EDTA, and saturated NaCl (DESS) as a room-temperature preservation medium for canine faeces. Using ITS-2 rDNA and isotype-1 {beta}-tubulin gene (tubb-1) amplicon metabarcoding, we compared DNA integrity and diagnostic performance of DESS-preserved samples (4, 28, and 106 days) to refrigerated controls. No significant differences in PCR amplification or sequencing outcomes were observed. DESS enabled reliable detection of hookworm species and resistance-associated SNPs, including F167Y in tubb-1, with mutation frequencies consistent across treatments. Therefore, DESS can preserve samples from remote settings without cold chain logistics. Our findings validate DESS as a robust alternative for sample collection for molecular parasitology, facilitating expanded surveillance of anthelmintic resistance in field conditions. HighlightsO_LIDESS preserves canine faeces for DNA analysis without refrigeration. C_LIO_LIHookworm species and resistance SNPs detected reliably from DESS samples. C_LIO_LIPCR and NGS results from DESS samples match refrigerated controls. C_LIO_LIF167Y mutation found in A. caninum and U. stenocephala samples. C_LIO_LIDESS enables remote hookworm surveillance without cold chain logistics. C_LI

microbiology↗

Integral Synthesis and Clearance Analysis via DIA (ISDia) Reveals Coordinated Protein Dynamics Regulation during Endoplasmic Reticulum Stress

Endoplasmic Reticulum (ER) stress disrupts protein homeostasis, driving cellular responses critical for survival, development and disease. However, systematic proteome-wide analyses of protein synthesis and clearance during ER stress remain limited. In this study, we developed Integral Synthesis and clearance analysis via DIA (ISDia), a robust mass spectrometry (MS)-based workflow that integrates multiple pulsed-SILAC labeling timepoints under one condition into a single sample and utilize data-independent acquisition (DIA) to quantify heavy and light peptide changes across conditions. It enables proteome-wide tracking of protein synthesis and clearance with high coverage by directly comparing light and heavy peptides in pSILAC-DIA experiments across multiple conditions. Using ISDia, we uncover diverse regulatory mechanisms by which protein synthesis and clearance are modulated to regulate protein abundances during ER stress, revealing PERK dependent and independent regulatory mechanisms across subcellular compartments, complexes and isoforms. These findings highlight the potential of ISDia as a powerful and widely applicable workflow for revealing protein abundance regulatory mechanisms.

systems biology↗

Fatty Acid Transport Protein-2 (FATP2) Inhibition Enhances Glucose Tolerance through α-Cell-mediated GLP-1 Secretion

Type 2 diabetes affects more than 30 million people in the US, and a major complication is kidney disease. During the analysis of lipotoxicity in diabetic kidney disease, global fatty acid transport protein-2 (FATP2) gene deletion was noted to markedly reduce plasma glucose in db/db mice due to sustained insulin secretion. To identify the mechanism, we observed that islet FATP2 expression was restricted to -cells, and -cell FATP2 was functional. Direct evidence of FATP2KO-induced -cell-mediated GLP-1 secretion included increased GLP-1-positive -cell mass in FATP2KO db/db mice, small molecule FATP2 inhibitor enhancement of GLP-1 secretion in TC1-6 cells and human islets, and exendin[9-39]-inhibitable insulin secretion in FATP2 inhibitor-treated human islets. FATP2-dependent enteroendocrine GLP-1 secretion was excluded by demonstration of similar glucose tolerance and plasma GLP-1 concentrations in db/db FATP2KO mice following oral versus intraperitoneal glucose loading, non-overlapping FATP2 and preproglucagon mRNA expression, and lack of FATP2/GLP-1 co-immunolocalization in intestine. We conclude that FATP2 deletion or inhibition exerts glucose-lowering effects through -cell-mediated GLP-1 secretion and paracrine {beta}-cell insulin release. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=136 SRC="FIGDIR/small/635976v1_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@190c799org.highwire.dtl.DTLVardef@203076org.highwire.dtl.DTLVardef@14dbf26org.highwire.dtl.DTLVardef@140869e_HPS_FORMAT_FIGEXP M_FIG C_FIG

physiology↗

Structure-Based Computational Analysis of Interactions between Insulin Receptor and Insulin Inhibitory Receptor

The recently discovered insulin inhibitory receptor (inceptor) plays a crucial role in insulin resistance and diabetes by reducing the insulin receptor count on cell membranes, resulting in higher blood glucose levels and decreased insulin sensitivity. Therefore, understanding the mechanism of how the inceptor insulin receptor complex interacts is exceedingly important. This study uses computational drug discovery to inhibit this interaction. Initially, we employed AlphaFold-Multimer to model the inceptor-insulin receptor protein complex and subsequently identified specific inceptor residues likely involved in binding to the insulin receptor. Through virtual screening, thousands of potential small molecules were found to bind to the inceptor, and 10 with the highest probability were chosen for docking. Beta-L-fucose, beta-D-fucose, and alpha-L-fucose showed the most promising binding energies, meaning these three small molecules can effectively interrupt the binding between the complex. We also computationally mutated the binding site of the insulin receptor and calculated the change in binding energy of the inceptor insulin receptor complex, the most dramatic being a 0.4 kcal mol^-1 change when Arginine mutated to Tryptophan at residue 926. Our study suggests that the mutations led to disease primarily due to the change in interactions of the inceptor insulin receptor complex.

bioinformatics↗

Persistent cross-species transmission systems dominate Shiga toxin-producing Escherichia coli O157:H7 epidemiology in a high incidence region: a genomic epidemiology study

BackgroundSeveral areas of the world suffer notably high incidence of Shiga toxin-producing Escherichia coli, among them Alberta, Canada. We assessed the impact of persistent cross-species transmission systems on the epidemiology of E. coli O157:H7 in Alberta. MethodsWe sequenced and assembled 229 E. coli O157:H7 isolates originating from collocated cattle (n=108) and human (n=121) populations from 2007-2015 in Alberta. We constructed a timed phylogeny using BEAST2 using a structured coalescent model. We then extended the tree with human isolates through 2019 (n=430) to assess the long-term disease impact of locally persistent lineages. Shiga toxin gene (stx) profile was determined for all isolates. ResultsDuring 2007 to 2015, we estimated 108 (95% HPD 104, 112) human lineages arose from cattle lineages, and 14 (95% HPD 5, 23) from other human lineages; i.e., 88.5% of human lineages arose from cattle lineages. We identified 11 persistent lineages local to Alberta, which were associated with 38.0% (95% CI 29.3%, 47.3%) of human isolates. Of 117 isolates in locally persistent lineages, 6.0% carried only the Shiga toxin gene stx2a and the rest both stx1a and stx2a. During the later period, six locally persistent lineages continued to be associated with human illness, including 74.7% (95% CI 68.3%, 80.3%) of reported cases in 2018 and 2019. The stx profile of isolates in locally persistent lineages shifted from the earlier period, with 51.2% encoding only stx2a. ConclusionsOur study identified multiple locally evolving lineages transmitted between cattle and humans persistently associated with E. coli O157:H7 illnesses for up to 13 years. Of concern, there was a dramatic shift in locally persistent lineages toward strains with the more virulent stx2a-only profile. Locally persistent lineages may be a principal cause of the high incidence of E. coli O157:H7 in locations such as Alberta and offer opportunities for understanding the disease ecology supporting E. coli O157:H7 persistence, as well as for local prevention efforts.

microbiology↗

Learning the language of proteins and predicting the impact of mutations

Mutations in proteins directly impact their structure and function. Understanding the "language" of proteins, or the sequence to function (genotype-phenotype) relationship has many real-world applications. One set of applications includes those in biodefense, such as biological threat detection and biosurveillance, antibody engineering, and medical countermeasure development. In this study, we present a novel language model-based approach that can rapidly analyze vast collections of sequences, and make near real-time functional predictions that compare favorably to those made using conventional bioinformatic and experimental methods. Our findings reveal that tailored protein language models can predict protein mutation phenotypes, such as binding affinity or level of expression, when they are trained with high-throughput functional data. Protein language models applied to viral genomes can also discern the lineage within a family (e.g., sarbecovirus sequences). Coupled with sequenced-based biosurveillance, this type of model may provide early warning signals of potential zoonotic spillovers (i.e. host jumping) or "escape" from existing medical countermeasures posed by novel mutations. This research not only underscores the potential of ML and language models in addressing pressing challenges in understanding the mapping of sequence to function, but further elucidates their potential application in accelerating the response to biological threats as they evolve.

bioinformatics↗

An engineered human cardiac tissue model reveals contributions of systemic lupus erythematosus autoantibodies to myocardial injury

Systemic lupus erythematosus (SLE) is a highly heterogenous autoimmune disease that affects multiple organs, including the heart. The mechanisms by which myocardial injury develops in SLE, however, remain poorly understood. Here we engineered human cardiac tissues and cultured them with IgG fractions containing autoantibodies from SLE patients with and without myocardial involvement. We observed unique binding patterns of IgG from two patient subgroups: (i) patients with severe myocardial inflammation exhibited enhanced binding to apoptotic cells within cardiac tissues subjected to stress, and (ii) patients with systolic dysfunction exhibited enhanced binding to the surfaces of viable cardiomyocytes. Functional assays and RNA sequencing (RNA-seq) revealed that IgGs from patients with systolic dysfunction exerted direct effects on engineered tissues in the absence of immune cells, altering tissue cellular composition, respiration and calcium handling. Autoantibody target characterization by phage immunoprecipitation sequencing (PhIP-seq) confirmed distinctive IgG profiles between patient subgroups. By coupling IgG profiling with cell surface protein analyses, we identified four pathogenic autoantibody candidates that may directly alter the function of cells within the myocardium. Taken together, these observations provide insights into the cellular processes of myocardial injury in SLE that have the potential to improve patient risk stratification and inform the development of novel therapeutic strategies.

bioengineering↗

Taste cells expressing Ionotropic Receptor 94e reciprocally impact feeding and egg laying in Drosophila

Chemosensory cells across the body of Drosophila melanogaster evaluate the environment and play a crucial role in neural circuits that prioritize feeding, mating, or egg laying. Previous mapping of gustatory receptor neurons (GRNs) on the fly labellum identified a set of neurons in L-type sensilla defined by expression of Ionotropic Receptor 94e (IR94e), but the impact of IR94e GRNs on behavior remained unclear. To understand their behavioral output, we used optogenetics and chemogenetics to activate IR94e neurons and found that they drive mild suppression of feeding but enhanced egg laying. In vivo calcium imaging revealed that IR94e GRNs respond strongly to certain amino acids, including glutamate. Furthermore, we found that IR94e is necessary and sufficient for the detection of amino acid ligands, and co-receptors IR25a and IR76b are also required for IR94e GRN activation. Finally, IR94e mutants show behavioral changes to solutions containing amino acids, including increased consumption and decreased egg laying. Overall, our results suggest that IR94e GRNs on the fly labellum discourage feeding and encourage egg laying as part of an important behavioral switch in response to certain chemical cues.

neuroscience↗

Mixed Alkyl/Aryl Phosphonates Identify Metabolic Serine Hydrolases as Antimalarial Targets

Malaria, caused by Plasmodium falciparum, remains a significant health burden. A barrier for developing anti-malarial drugs is the ability of the parasite to rapidly generate resistance. We demonstrated that Salinipostin A (SalA), a natural product, kills parasites by inhibiting multiple lipid metabolizing serine hydrolases, a mechanism with a low propensity for resistance. Given the difficulty of employing natural products as therapeutic agents, we synthesized a library of lipidic mixed alkyl/aryl phosphonates as bioisosteres of SalA. Two constitutional isomers exhibited divergent anti-parasitic potencies which enabled identification of therapeutically relevant targets. We also confirm that this compound kills parasites through a mechanism that is distinct from both SalA and the pan-lipase inhibitor, Orlistat. Like SalA, our compound induces only weak resistance, attributable to mutations in a single protein involved in multidrug resistance. These data suggest that mixed alkyl/aryl phosphonates are a promising, synthetically tractable anti-malarials with a low-propensity to induce resistance.

microbiology↗

Competition-driven eco-evolutionary feedback reshapes bacteriophage lambda's fitness landscape and enables speciation

A major challenge in evolutionary biology is explaining how populations navigate rugged fitness landscapes without getting trapped on local optima. One idea illustrated by adaptive dynamics theory is that as populations adapt, their newly enhanced capacities to exploit resources alter fitness payoffs and restructure the landscape in ways that promote speciation by opening new adaptive pathways. While there have been indirect tests of this theory, none have measured how fitness landscapes deform during adaptation, or test whether these shifts promote diversification. Here, we achieve this by studying bacteriophage , a virus that readily speciates into co-existing receptor specialists under controlled laboratory conditions. We used a high-throughput gene editing-phenotyping technology to measure s fitness landscape in the presence of different evolved- competitors and found that the fitness effects of individual mutations, and their epistatic interactions, depend on the competitor. Using these empirical data, we simulated s evolution on an un-changing landscape and one that recapitulates how the landscape deforms during evolution. heterogeneity only evolved in the shifting landscape regime. This study provides a test of adaptive dynamics, and, more broadly, shows how fitness landscapes dynamically change during adaptation, potentiating phenomena like speciation by opening new adaptive pathways.

evolutionary biology↗

A Cell Cycle-Dependent Ferroptosis Sensitivity Switch Governed by EMP2

Ferroptosis is a non-apoptotic form of cell death characterized by iron-dependent lipid peroxidation. Ferroptosis can be induced by system xc- cystine/glutamate antiporter inhibition or by direct inhibition of the phospholipid hydroperoxidase glutathione peroxidase 4 (GPX4). The regulation of ferroptosis in response to system xc- inhibition versus direct GPX4 inhibition may be distinct. Here, we show that cell cycle arrest enhances sensitivity to ferroptosis triggered by GPX4 inhibition but not system xc- inhibition. Arrested cells have increased levels of oxidizable polyunsaturated fatty acid-containing phospholipids, which drives sensitivity to GPX4 inhibition. Epithelial membrane protein 2 (EMP2) expression is reduced upon cell cycle arrest and is sufficient to enhance ferroptosis in response to direct GPX4 inhibition. An orally bioavailable GPX4 inhibitor increased markers of ferroptotic lipid peroxidation in vivo in combination with a cell cycle arresting agent. Thus, responses to different ferroptosis-inducing stimuli can be regulated by cell cycle state.

cancer biology↗

Overexpressed Malat1 Drives Metastasis through Inflammatory Reprogramming of Lung Adenocarcinoma Microenvironment

Metastasis is the main cause of cancer deaths but the molecular events leading to metastatic dissemination remain incompletely understood. Despite reports linking aberrant expression of long noncoding RNAs (lncRNAs) with increased metastatic incidence, in vivo evidence establishing driver roles for lncRNAs in metastatic progression is lacking. Here, we report that overexpression of the metastasis-associated lncRNA Malat1 (metastasis-associated lung adenocarcinoma transcript 1) in the autochthonous K-ras/p53 mouse model of lung adenocarcinoma (LUAD) is sufficient to drive cancer progression and metastatic dissemination. We show that increased expression of endogenous Malat1 RNA cooperates with p53 loss to promote widespread LUAD progression to a poorly differentiated, invasive, and metastatic disease. Mechanistically, we observe that Malat1 overexpression leads to the inappropriate transcription and paracrine secretion of the inflammatory cytokine, Ccl2, to augment the mobility of tumor and stromal cells in vitro and to trigger inflammatory responses in the tumor microenvironment in vivo. Notably, Ccl2 blockade fully reverses cellular and organismal phenotypes of Malat1 overexpression. We propose that Malat1 overexpression in advanced tumors activates Ccl2 signaling to reprogram the tumor microenvironment to an inflammatory and pro-metastatic state.

cancer biology↗

Stabilisation of β-Catenin-WNT signalling by USP10 in APC-truncated colorectal cancer drives cancer stemness and enables super-competitor signalling

The contribution of deubiquitylating enzymes to {beta}-Catenin stabilisation in intestinal stem cells and colorectal cancer (CRC) is poorly understood. Here, we report the deubiquitylase USP10 as an APC-truncation- specific enhancer of {beta}-Catenin stability, potentiating WNT signalling in CRC and cancer stem cells. Mechanistically, interaction studies in various CRC cell lines and in vitro binding studies, together with computational modelling, revealed that USP10 binding to {beta}-Catenin is mediated via the unstructured N-terminus of USP10 and requires the absence of full-length APC. Notably, loss of USP10 in CRISPR engineered intestinal organoids reduces tumorigenic properties of CRC and blocks the super competitor-signalling of APC-mutated CRC. Furthermore, reduction of USP10 induces the expression of differentiation genes, and opposes the APC-truncated phenotype in an intestinal hyperplasia model of D.melanogaster. Taken together, our findings reveal USP10s role in intestinal tumourigenesis by stabilising {beta}-Catenin, leading to aberrant WNT signalling, enhancing cancer cell stemness and implicate the DUB USP10 as a cancer specific therapeutic vulnerability in Apc truncated CRC.

cancer biology↗