BACKGROUND:Colorectal cancer (CRC) is characterized by aberrant mucin expression, but the full spectrum of mucin profiles and their clinical relevance remain unclear. Here, we developed a novel mucin-based CRC subtyping using multiple datasets to tailor therapeutic strategies. METHODS:Mucin-related signatures were used to stratify CRC into four novel mucin-based subtypes: metabolism initiated, inflammatory, immune activated, and progressing. Highly accurate predictions were achieved, with under the curve (AUC) values of 0.977 for PC1 and 0.997 for PC2. RESULTS:The novel subtypes demonstrated prognostic potential in predicting CRC patient survival outcomes. We identified interactions between mucin-based subtypes and tumor-associated macrophages (TAMs). Mechanistic research revealed that MUC20 overexpression promoted M2 macrophage-dependent tumor glycolysis and hypoxia. Specifically, M2 phenotype TAMs secreted TNF-α, which enhanced tumor glycolysis and upregulated MUC20 expression in cancer cells. This established a MUC20/glycolysis positive feedback loop that exacerbated hypoxia and aerobic glycolysis in CRC. Additionally, chemotherapy responses in patient organoids were highly matched to these subtypes. Patients in cluster 2 were sensitive to 5-Fu or CPT-11 but potentially resistant to routine FOLFOX treatment. CONCLUSION:We identified a novel four mucin-characterized subtypes, which advance precision medicine by refining chemotherapy regimen selection. Our study provides valuable insights for tailoring therapeutic strategies.

ABSTRACT Organoids are innovative three‐dimensional (3D) cellular constructs, offering a unique platform to replicate the architectural and functional complexity of organs and tissues. In oncology, the tumor microenvironment (TME) dictates tumor evolution and therapeutic resistance. Consequently, therapies targeting TME components have emerged as a burgeoning frontier in cancer treatment. However, accurately recapitulating the dynamic, multicellular crosstalk of TME remains a significant hurdle for clinical translation. This review encapsulates the spectrum of current organoid coculture methodologies, ranging from direct coculture and air–liquid interface to advanced microfluidics and 3D bioprinting. These models not only deepen our understanding of the fundamental mechanisms at play in cancer but also evaluate emerging therapeutic modalities, such as antibody–drug conjugates and immunotherapy. By closely mimicking the in vivo tumor milieu, organoid cocultures enhance our ability to predict therapeutic outcomes and pave the way for the development of precision medicine approaches, thereby propelling forward the frontiers of oncology. This review aims to provide a comprehensive overview of organoid coculture models, spanning from construction methodologies to clinical applications. We envision this work serving as a definitive guide for the field, ultimately accelerating the transition from theoretical research to clinical practice.

PURPOSE:We hypothesized that a dynamic surveillance strategy guided by circulating tumor DNA (ctDNA) methylation would increase the rate of curative-intent therapy for recurrence in patients with nonmetastatic colorectal cancer (CRC) after curative resection. METHODS:The FIND trial (ClinicalTrials.gov identifier: NCT05904665) is a prospective, multicenter, randomized, phase III study. Patients with nonmetastatic CRC were randomly assigned to ctDNA-guided surveillance or standard computed tomography (CT)-based monitoring. In the ctDNA-guided group, a positive ctDNA result triggered immediate CT imaging; if negative, bimonthly CT continued alongside quarterly ctDNA testing. After two consecutive ctDNA-negative results, imaging reverted to standard frequency. The primary end point was the proportion of patients with recurrence receiving curative-intent metastasis-directed therapy. RESULTS:Among 584 eligible patients (289 ctDNA-guided, 295 control) in the modified intention-to-treat population, with a median follow-up of 23.3 months, recurrence rates were similar (18.0% v 18.6%, P = .919). The ctDNA-guided group had a significantly higher rate of curative-intent treatment (48.1% v 23.6%, relative risk 2.03, P = .008). The median time to clinical recurrence was significantly shorter in the ctDNA-guided group than in the control group (9.5 v 13.4 months; P < .001), representing a lead time of 3.9 months. Among recurrences confined to the liver and/or lungs, the ctDNA-guided group showed higher curative resection rates (42.3% v 18.2%, P = .002). These patients had more favorable hepatic metastatic features: fewer lesions (≤3: 75.0% v 28.6%, P = .005), smaller tumor size (≤3 cm: 90.0% v 57.1%, P = .033), and more unilobar disease (80.0% v 28.6%, P = .002). CONCLUSION:ctDNA methylation-guided dynamic surveillance improves the rate of curative-intent therapy for recurrence in patients with initially nonmetastatic CRC through earlier detection of resectable metastases, pending validation of long-term survival benefit in future analyses with mature data.

BACKGROUND:Current therapeutic outcomes for advanced colorectal cancer (CRC) remain suboptimal, and chemotherapy-based regimens continue to be the mainstay of treatment. Circular RNAs (circRNAs) can serve as templates for translating short peptides or proteins, and the resulting products actively regulate malignant tumour progression, making them attractive therapeutic targets. METHODS:We identified the novel protein PVT1-104aa translated from circPVT1, which is generated by back-splicing of the non-coding PVT1 gene. Its expression and clinical significance were examined in CRC clinical specimens and cell lines. Proliferation, metastasis, and tumour growth were assessed by CCK-8, colony formation, transwell, wound healing, and xenograft syngeneic tumour models. Mechanistic studies were performed by immunoprecipitation, ubiquitination assays, and protein half-life analysis. The relationship between PVT1-104aa, c-Myc, and PD-L1 was evaluated by promoter reporter assays, ChIP-qPCR, and immunohistochemistry. The therapeutic efficacy of combining PVT1-104aa inhibition with anti-PD-L1 therapy was tested in vivo. RESULTS:PVT1-104aa was significantly overexpressed in CRC and correlated with poor patient prognosis. Functionally, it drove tumour progression by promoting proliferation and metastasis. Mechanistically, PVT1-104aa enhanced c-Myc phosphorylation at Ser62, disrupted the c-Myc-FBW7 interaction, and thereby inhibited ubiquitin-mediated degradation of c-Myc, as shown by accelerated c-Myc turnover upon PVT1-104aa knockdown. In addition, PVT1-104aa regulated PD-L1 expression through c-Myc. Combining anti-PVT1-104aa with anti-PD-L1 therapy suppressed CRC growth and increased CD4+ and CD8+ T cell infiltration in xenograft syngeneic tumour models and CRC tissues. CONCLUSIONS:Our results uncover a pathogenic role of the PVT1-originated molecular species PVT1-104aa and suggest that targeting this pathway represents a promising therapeutic strategy for CRC treatment. KEY POINTS:Circ-PVT1 is upregulated in CRC patients and encoded a novel protein: PVT1-104aa. PVT1-104aa promotes CRC progression and predicts worse prognosis. PVT1-104aa enhance Myc expression through inhibition of Myc ubiquitination. PVT1-104aa regulate PD-L1 expression of CRC cells and modulate T cells infiltration in vivo.

BackgroundThe optimal choice of chemotherapy regimen for resectable colorectal liver metastases (CRLM) is still undetermined. The study aimed to estimate the impact of perioperative or adjuvant chemotherapy on disease-free survival (DFS), and explore the underlying mechanisms which influence chemotherapy efficacy.MethodsEight hundred twenty-two patients undergoing curative resection of CRLM were retrospectively collected from May 2018 to December 2023. Treatment effects between perioperative and adjuvant chemotherapy were compared in full subgroup analyses by Kaplan–Meier and Cox proportional hazards methods. Single-cell RNA sequencing and tumor derived organoids were used to explore the molecular mechanisms.ResultsAfter propensity score matching, 630 patients were enrolled with ratio 1:1 in adjuvant and perioperative chemotherapy group. The median DFS in adjuvant and perioperative group was comparable with 28 and 32.5 months, respectively (HR = 0.99, P = 0.945). Perioperative chemotherapy was associated with improved DFS (25 vs. 13 months, P = 0.039) in patients with a clinical risk score (CRS) of 5. Notably, within the high-risk CRS 4-5 population receiving perioperative chemotherapy, patients with chronic hepatitis B (CHB) had significantly worse DFS (HR = 4.31, 95% CI 1.93–9.64; P < 0.001). Single-cell analyses revealed TSPAN8+ stem-like epithelial cells were enriched in sample with CHB. TSPAN8 knockdown sensitized colorectal cancer cells to 5-fluorouracil, and anti-TSPAN8 antibody showed synergistic efficacy with 5-fluorouracil in patient-derived liver metastasis organoids. In addition, the enhanced SPP1-CD44 signaling between TSPAN8+ epithelial cells and SPP1+ macrophages also contributed to 5-fluorouracil resistance.ConclusionsHigher CRS identify a subgroup of CRLM patients who may derive greater benefit from perioperative chemotherapy. HBV-related serological profiles is a potential predictor of chemotherapy resistance. The exploratory findings warrants further validation.

Mesothelial cells play an important role in colorectal cancer peritoneal metastasis (CRC-PM), where they support tumor growth and invasion. In this study, we investigate the molecular mechanisms by which mesothelial cells contribute to CRC metastasis. Using single-cell RNA sequencing (scRNA-seq) on tissue samples from 12 CRC patients with peritoneal metastasis, we identify PDK4 as a key gene in mesothelial cells during metastasis. The expression of PDK4 is significantly greater in mesothelial cells undergoing mesothelial-to-mesenchymal transition (MMT) compared to normal peritoneal cells, suggesting its involvement in mesothelial cell reprogramming during peritoneal metastasis. In vitro experiments show that coculturing mesothelial cells with CRC cells leads to increased PDK4 expression, which in turn enhances mesothelial cell migration and invasion. Knockdown of PDK4 reduces mesothelial cell invasion, while overexpression of PDK4 increases invasive ability, highlighting its critical role in mesothelial cell invasion. Additionally, PDK4 promotes metabolic changes, specifically increasing fatty acid oxidation (FAO), which is necessary for mesothelial cell invasion. Blocking FAO reduces the invasive ability of PDK4-overexpressing mesothelial cells, while restoring FAO in PDK4-knockdown cells rescues their invasion potential. Further analysis shows that PDK4 enhances the acetylation of β-catenin, a protein involved in cell movement, and that this modification is crucial for mesothelial cell invasion. Our results suggest that PDK4 regulates mesothelial cell invasion through β-catenin acetylation following metabolic reprogramming, offering a potential target for therapies aimed at inhibiting CRC-PM.

BACKGROUND:BRAFV600E-mutant colorectal cancer (CRC) represents an aggressive molecular subtype characterized by poor prognosis and resistance to standard therapies. Predictive models to guide treatment selection and overcome drug resistance are urgently needed. METHODS:We established a biobank of patient-derived organoids (PDOs) and matched xenografts (PDOXs) from BRAFV600E-mutant CRC, capturing inter- and intratumoral heterogeneity. PDOs were characterized by histopathology, whole-exome sequencing, bulk and single-cell RNA sequencing, and drug sensitivity testing to chemotherapies and targeted agents. Immunogenicity was assessed via PDO-T cell co-culture and humanized xenograft models. RESULTS:PDOs faithfully preserved parental tumor histology, mismatch repair status, mutational landscapes, and copy number alterations (91.6% concordance in driver genes). Single-cell transcriptomics confirmed high fidelity between PDOs and original tumors. Drug sensitivity assays revealed marked interpatient heterogeneity and accurately recapitulated clinical responses to FOLFIRI, FOLFOXIRI, and BRAF/EGFR inhibitor combinations. Notably, RNF43-mutant PDOs exhibited significantly enhanced sensitivity to encorafenib plus cetuximab. Integrative transcriptomic analysis (TCGA and PDO cohorts) identified that RNF43 mutations are associated with upregulation of E2F, G2M, interferon-α/γ, and inflammatory pathways, along with elevated MHC-I expression. In co-culture with CD8⁺ T cells, RNF43-mutant PDOs triggered stronger T-cell activation, enhanced cytotoxicity, and increased tumor cell apoptosis. In humanized PDOX models, RNF43-mutant tumors showed suppressed growth and heightened CD8⁺ T-cell infiltration compared to wild-type controls. CONCLUSIONS:This functional precision oncology platform integrates multi-omics and immune co-culture to uncover RNF43 mutation as a dual biomarker for targeted therapy sensitivity and tumor immunogenicity in BRAFV600E-mutant CRC. Our findings provide mechanistic rationale for combining BRAF/EGFR inhibition with immunotherapy to overcome drug resistance and improve outcomes in this aggressive subtype.

Colorectal cancer liver metastasis (CRLM) and chemotherapy resistance remain major clinical challenges, with the underlying molecular mechanisms yet to be fully elucidated. In this study, based on analyses of five initial clinical cohorts from FUSCC, the E3 ubiquitin ligase COP1 was identified as a critical driver of CRLM and resistance to oxaliplatin-based chemotherapy. Using an organoid biobank derived from paired primary colorectal tumors and liver metastases, integrated multi-omics analyses (WES, bulk RNA-seq, scRNA-seq) of patient-derived organoids (PDOs) from CRLM revealed significantly elevated COP1 expression in liver metastases compared to primary tumors. High COP1 levels were associated with poor prognosis, increased liver metastatic burden, and resistance to oxaliplatin-based chemotherapy. In vitro and in vivo functional experiments demonstrated that COP1 facilitates CRLM progression by ubiquitinating and degrading LUZP1, thereby releasing DAPK3 from LUZP1-mediated suppression. This process leads to enhanced MYL9 phosphorylation and activation of epithelial-mesenchymal transition (EMT) as well as the JAK2-STAT3-CCND2 signaling axis—pathways crucial for liver metastasis and resistance to oxaliplatin-based chemotherapy. These findings establish the COP1-LUZP1-MYL9 axis as a therapeutic target for CRLM and oxaliplatin-based chemoresistance. Clinically, COP1 expression profiling in PDOs from postoperative specimens enables a precision strategy for managing oxaliplatin-based chemoresistance, especially in the context of FOLFOX.

Colorectal cancer (CRC) frequently exhibits an immune-excluded tumor microenvironment that limits the efficacy of PD-1 blockade. Through systematic profiling of 106 protein tyrosine phosphatases (PTPs), we identify three PTP-based molecular subtypes, among which the immune-excluded Cluster 2 is highly resistant to immunotherapy. Integrative analyses across patient cohorts highlight PTPRN2 as a prominent PTP enriched in immune-excluded tumors and strongly associated with anti-PD-1 non-response. Functional studies demonstrate that PTPRN2 promotes CRC cell proliferation and invasion, and more importantly, suppresses tumor-intrinsic MHC-II expression. Mechanistically, PTPRN2 may promote STAT1 Y701 dephosphorylation and inhibit STAT1 activation, thereby downregulating CIITA expression and impairing antigen presentation. PTPRN2 knockdown restores MHC-II, enhances CD4⁺ and CD8⁺ T-cell activation in organoid co-culture, and increases T-cell infiltration in vivo. Therapeutically, PTPRN2 knockdown combined with PD-1 blockade achieves more effective tumor suppression than either treatment alone. Furthermore, IL-15 rescues STAT1 phosphorylation and MHC-II expression in PTPRN2-overexpressing cells, revealing a cytokine-sensitive regulatory axis. Collectively, our study identifies PTPRN2 as a potential regulator of immune exclusion and a candidate therapeutic target to improve immunotherapy efficacy in CRC.

Colorectal liver metastasis (CRLM) is one of the leading death causes among colorectal cancer (CRC) patients, yet its underlying molecular events remain poorly understood, particularly at the proteomic and phosphoproteomic levels. A proteogenomic analysis combining genomics, transcriptomics, proteomics, and phosphoproteomics is performed on 102 samples from 34 treatment-naïve CRLM patients, including primary CRC, adjacent normal colorectal, and matched liver metastasis tissues. CRC cell lines, organoids, mouse models, and an independent patient cohort are used to validate the findings. Proteomics and phosphoproteomics show profoundly dysregulated pathways in liver metastasis tissues, notably disruptions in carbon metabolism. Functional validation using CRC organoids and mouse models demonstrates that the one-carbon metabolism enzyme SHMT1 promotes CRC tumorigenesis and metastasis via formate-mediated AMPK inhibition, whereas PIM kinase-dependent NDRG1 Ser330 phosphorylation exacerbates liver metastasis by promoting ubiquitin-dependent degradation of NDRG1. Unsupervised clustering identifies two proteomic subtypes of liver metastasis samples with distinct clinical outcomes: a poor-prognosis C1 (metabolism) subtype and a better-prognosis C2 (RNA function) subtype. Considering expression frequency, specificity, and functional relevance, FTCD, GPD1, SOD2, and EIF4B Ser422 phosphorylation are further identified and validated as subtype prognostic biomarkers. This study provides critical insights into the molecular mechanisms underlying CRLM and offers resources for high-risk metastatic CRC.

In the original publication [...].

Background:Microsatellite stability (MSS) colorectal cancers (CRCs) have a limited response to immune checkpoint inhibitors (ICIs) compared to microsatellite instability-high (MSI-H) CRCs. Nevertheless, previous studies have shown that some MSS CRCs are sensitive to ICIs, although established criteria for treatment justification are still lacking. Objective:This study aimed to test the tumor-infiltrating lymphocyte (TIL) features of MSS and develop a novel computational tool for the similarity prediction between MSS and MSI-H status in patients with CRC based on multiple factors. Methods:We collected and analyzed data from 188 patients with CRC, including MSI status, immune cell distributions, clinical features, and gene mutations, using statistical methods and Cox regression. An ensemble machine learning-based MSI-H score was developed using stacked extreme gradient boosting classifiers to quantify the similarity of patient data to MSI-H data based on immune cell distributions, clinical features, and gene mutations. The model was robust and could address missing input data for immune cell distributions and gene mutations. Results:The scorer performed well (mean Cohen κ of 0.40, SD 0.05, over 10 random seeds) in identifying MSI-H-like MSS samples with TIL distributions similar to genuine MSI-H CRCs. No significant difference was observed between the TIL features of MSI-H-like MSS CRCs and MSI-H CRCs. The disparity between MSI-H-like MSS CRCs and MSS CRCs potentially lies in the T regulatory cells (P=.09) and macrophage (P=.16) populations within the tumor stromal region. Conclusions:Some patients with MSS CRC presented similar immune cell distributions with high immunoactivity compared to patients with MSI-H CRC. The MSI-H score serves as a metric to quantify the similarity of MSS CRCs to MSI-H CRCs and presents a promising avenue for more personalized and effective cancer immunotherapy treatment, offering a clinical reference for potential ICI targets in MSS CRCs.

Background MRI has helped to identify mesorectal fascia (MRF) positivity as a poor prognostic factor in patients with locally advanced rectal cancer. However, outcomes in patients with MRF positivity may vary. Purpose To investigate the prognosis of the four types of MRF-positive classifications and their genetic and protein-level interpretations. Materials and Methods In this multicenter retrospective study (January 2009 to May 2023), patients with locally advanced rectal cancer who underwent rectal MRI were analyzed and categorized as follows: MRF caused by tumor invasion (tumor invasion vs no tumor invasion), extramural vascular invasion (EMVI; EMVI vs no EMVI), lymph node metastasis (LNM; LNM vs no LNM), and tumor deposits (TD; TD vs no TD). Disease-free survival (DFS) was estimated using the Kaplan-Meier method and compared using the log-rank test. Prognostic factors were evaluated using multivariable Cox proportional hazard models. Tumor-associated genes were sequenced, tumor mutational burden (TMB) was calculated, and immunohistochemistry was performed to quantify the expression of Ki-67, as well as the infiltration densities of CD3+ and CD8+ T cells. Results A total of 959 patients from three centers were included (median age, 56 years [IQR, 48-63 years]; 622 men). Kaplan-Meier analysis showed MRF with TD and MRF with EMVI were associated with worse DFS compared with MRF without TD (median DFS, 18 vs 99 months, respectively; P < .001) and MRF without EMVI (median DFS, 39 vs 99 months, respectively; P = .01). At multivariable Cox regression analysis, MRF with TD and MRF with EMVI were independent risk factors for poorer DFS (hazard ratio, EMVI vs TD: 1.95 [P = .006] vs 2.55 [P < .001], respectively). TMB and T cell infiltration (CD3+ and CD8+) were lower in patients with MRF and EMVI compared with MRF without EMVI (log-transformed TMB: -0.49 vs -0.44, respectively [P = .04]; mean counts, CD3+: 103.4 vs 251.4, respectively; mean counts, CD8+: 72.3 vs 183.0, respectively [P < .001 for all]). Ki-67 expression was higher in patients with MRF with TD versus MRF without TD (median, 70% vs 60%, respectively; P = .01). Conclusion MRF caused by TD or EMVI at pretreatment rectal MRI predicted poor prognosis in patients with locally advanced rectal cancer. © The Author(s) 2025. Published by the Radiological Society of North America under a CC BY 4.0 license. Supplemental material is available for this article. See also the editorial by Horvat and Kim in this issue.

BACKGROUND:Radiotherapy (RT) is an essential treatment for colorectal cancer (CRC), yet the factors influencing radiosensitivity remain unclear. In the quest to enhance the therapeutic efficacy in CRC, the interplay between genetic mutations and RT sensitivity has emerged as a pivotal yet enigmatic area. METHODS:We harness the fidelity of patient-derived organoids (PDOs) to dissect the molecular landscape of radiosensitivity, with a particular emphasis on BRAFV600E mutations. To further investigate, a cohort of 9 BRAFV600E-mutant and 10 BRAF wild-type PDOs is constructed to systematically assess the radiobiological traits of BRAFV600E-mutant CRC, including morphology, cell viability, and DNA damage, while also evaluating their responses to chemotherapy and chemoradiotherapy. RESULTS:Our systematic investigation unveils a profound correlation between BRAFV600E mutation status and radioresistance, which is validated by clinical treatment responses. Intriguingly, BRAFV600E-mutant PDOs exhibit reduced sensitivity to conventional chemotherapy, yet demonstrate an enhanced response to combined chemoradiotherapy, characterized by increased apoptosis. The results are validated through in vivo analyses using patient-derived organoid xenograft mouse models and aligned with patient clinical outcomes. CONCLUSIONS:This study outlines the distinct radiobiological profile of BRAFV600E-mutant CRC, underscoring the critical role of radiotherapy in comprehensive treatment strategies. This work not only advances our molecular understanding of CRC but also paves the way for precision medicine, offering valuable insights for therapeutic decision-making in the clinical management of BRAFV600E-mutant CRC.

Simultaneous Fe+H ion irradiation (denoted as Fe+H) and Fe+H ion irradiation with pre-implantation of He (denoted as He/Fe+H) were carried out on Chinese Low-Activation Martensitic (CLAM) steels at 350 ℃-550 ℃ to investigate the synergistic effect between H and He. The peak dose was 15 dpa, the He concentration in the observation area was 11 appm/dpa and the H concentration was 44 appm/dpa. The microstructure and hardening of the irradiated samples were evaluated by TEM and nanoindentation, respectively. Bubbles were not observed in irradiated CLAM steels at all temperatures, which implies that the promotion of bubbles by H may not be as strong as that by He and that H attenuates the promotion of bubbles by He. The hardening induced by He/Fe+H irradiation at 450°C is about 1.6 times that of Fe+H irradiation, the synergistic effect of H and He is most significant at this temperature. Comparison with Fe+He irradiation and single Fe irradiation reveals that Fe+H irradiation at 350°C induces lower interstitial defect damage than Fe+He irradiation, but the level of interstitial defects induced by the synergistic effect of both H and He with displacement damage is comparable at higher temperatures. The average size of dislocation loops induced by Fe+H irradiation is between that of single Fe irradiation and Fe+He irradiation, but the number density of dislocation loops induced by Fe+H irradiation is higher than that of Fe+He irradiation. The synergistic effect of He with displacement damage tends to promote the growth of dislocation loops, while the synergistic effect of H tends to promote the nucleation of dislocation loops.

Background: Markers of aging hold promise in the context of colorectal cancer (CRC) care. Utilizing high -resolution metabolomic profiling, we can unveil distinctive age -related patterns that have the potential to predict early CRC development. Our study aims to unearth a panel of aging markers and delve into the metabolomic alterations associated with aging and CRC. Methods: We assembled a serum cohort comprising 5,649 individuals, consisting of 3,002 healthy volunteers, 715 patients diagnosed with colorectal advanced precancerous lesions (APL), and 1,932 CRC patients, to perform a comprehensive metabolomic analysis. Results: We successfully identified unique age -associated patterns across 42 metabolic pathways. Moreover, we established a metabolic aging clock, comprising 9 key metabolites, using an elastic net regularized regression model that accurately estimates chronological age. Notably, we observed significant chronological disparities among the healthy population, APL patients, and CRC patients. By combining the analysis of circulative carcinoembryonic antigen levels with the categorization of individuals into the "hypo" metabolic aging subgroup, our blood test demonstrates the ability to detect APL and CRC with positive predictive values of 68.4% (64.3%, 72.2%) and 21.4% (17.8%, 25.9%), respectively. Conclusions: This innovative approach utilizing our metabolic aging clock holds significant promise for accurately assessing biological age and enhancing our capacity to detect APL and CRC.

Background: Approximately 60% of patients with colorectal liver metastases (CRLM) experience relapse within 2 years after radical resection, previous studies have proven that repeat local treatment (LT) could prolong survival, however, it is difficult to seize the window for LT due to the lack of a high-sensitive surveillance method. In this study, the authors aim to examine the value of longitudinal circulating tumor DNA (ctDNA) in guiding adjuvant chemotherapy, optimizing clinical surveillance strategy, and thereby improving CRLM outcomes. Materials and methods: The authors conducted a prospective clinical trial using a personalized, tumor-informed ctDNA assay to monitor 60 CRLM patients undergoing resection with curative intent. Formalin-fixed paraffin-embedded tumor samples were collected after surgery. Blood samples were collected before surgery, 30 days after surgery (post-OP), and every third month until relapse or up to 2 years. Results: A total of 394 plasma samples from 60 eligible patients were analyzed, with a median follow-up time of 31.3 months. Landmark analyses revealed that detectable ctDNA at post-OP (HR, 4.8), postadjuvant chemotherapy (HR, 6.0), and end-of-treatment (HR, 5.6) were associated with higher recurrence risk ( P <0.001). Post-OP ctDNA positivity served as the only independent prognostic marker in the multivariant analysis (HR, 5.1; P <0.001). Longitudinal ctDNA analysis identified relapsed patients at both sensitivity and specificity of 100%. Most (75%) patients were found with radiological relapse within 6 months after the first detectable ctDNA with a median lead time of 3.5 months. In relapsed patients, 73.2% had oligometastatic disease and 61% were liver-restricted, of which 72.0% received repeat LTs, and 60.0% achieved a secondary no evidence of disease status. Conclusions: Longitudinal ctDNA monitoring assists in early prediction of relapse, and thereby improves survival of CRLM patients by increased secondary resection rate and secondary no evidence of disease rate.

Three-dimensional cultured organoids recapitulate histopathologic and molecular characteristics of the primary tissues. The rise and development of organoids provide an important platform for clinical medicine research. Patient-derived organoids could be used as models to help clinicians to better understand the pathophysiology of related diseases and formulate corresponding treatment strategies; Organoids derived from tumors could be utilized as a drug screening system for precision medicine, helping clinicians to select the most appropriate regimen for cancer patients and to improve the clinical outcome. This review would summarize the application and research advances of organoids in clinical medicine.

Simultaneous Fe+He ion irradiations of China Low Activation Martensitic (CLAM) steel and oxide-dispersion-strengthened ferritic/martensitic (ODS-F/M) steel were carried out at 350 °C-550 °C to the damage dose of 11 dpa with different He concentrations to investigate the effect of oxide particles on the evolution of He bubbles and dislocation loops. The microstructures of all irradiated samples were observed by transmission electron microscopy (TEM). The average size of oxide particles decreased and the density increased after irradiation. The aggregation of He bubbles was observed at the edge of oxide particles. As the He concentration increased, the aggregation of helium bubbles appeared at the edge of more oxide particles. Furthermore, He bubbles were not observed inside ODS-F/M steel grains or at grain boundaries, which suggests that the oxide particle interface is more capable of capturing helium atoms than other sinks. The average dislocation loop size of ODS-F/M steels is only 58% of that in CLAM steel at 450 °C, showing that ODS-F/M steel has excellent inhibition on the growth of dislocation loops. Nevertheless, with the increase of temperature and He concentration, the number density and size of dislocation loops in ODS-F/M steels gradually approach those in CLAM steels. The ability of ODS-F/M steels to inhibit irradiation hardening relative to CLAM steels increases with increasing temperature and decreases with increasing He concentration. The result of first-principles calculations shows that the formation energy of interstitial Fe atom at the interface increases with the increasing of the number of He atoms, which means that the aggregation of excessive He atoms at the interface will inhibit superior irradiation resistance of ODS-F/M steel.

Background Organoids are three-dimensional structures that closely recapitulate tissue architecture and cellular composition, thereby holding great promise for organoid-based drug screening. Although growing in three-dimensional provides the possibility for organoids to recapitulate main features of corresponding tissues, it makes it incommodious for imaging organoids in two-dimensional and identifying surviving organoids from surrounding dead cells after organoids being treated by irradiation or chemotherapy. Therefore, significant work remains to establish high-quality controls to standardize organoid analyses and make organoid models more reproducible. Methods In this study, the Z-stack imaging technique was used for the imaging of three-dimensional organoids to gather all the organoids’ maximum cross sections in one imaging. The combination of live cell staining fluorescent dye Calcein-AM and ImageJ assessment was used to analyze the survival of organoids treated by irradiation or chemotherapy. Results We have established a novel quantitative high-throughput imaging assay that harnesses the scalability of organoid cultures. Using this assay, we can capture organoid growth over time, measure multiple whole-well organoid readouts, and show the different responses to drug treatments. Conclusions In summary, combining the Z-stack imaging technique and fluorescent labeling methods, we established an assay for the imaging and analysis of three-dimensional organoids. Our data demonstrated the feasibility of using organoid-based platforms for high-throughput drug screening assays. Graphical Abstract