Preoperative exercise induces anti-tumor Kupffer cells to prevent surgical stress-promoted colorectal cancer liver metastasis

Yunwei Zhang; Yiyu Zhang; Chengli Shen; Jiye Li; Han Wang; Ahmad Hamad; Chunyan Cao; Mohamad El Moheb; Yu Wang; Yujia Xia; Kaelyn C Cummins; Joal Beane; Aslam Ejaz; Hai Huang; Allan Tsung; Hongji Zhang

doi:10.1016/j.xcrm.2026.102589

. 2026 Feb 9;7(2):102589. doi: 10.1016/j.xcrm.2026.102589

Preoperative exercise induces anti-tumor Kupffer cells to prevent surgical stress-promoted colorectal cancer liver metastasis

Yunwei Zhang ¹, Yiyu Zhang ¹, Chengli Shen ¹, Jiye Li ¹, Han Wang ², Ahmad Hamad ^2,³, Chunyan Cao ¹, Mohamad El Moheb ¹, Yu Wang ², Yujia Xia ², Kaelyn C Cummins ¹, Joal Beane ², Aslam Ejaz ^2,⁴, Hai Huang ⁵, Allan Tsung ^1,^∗, Hongji Zhang ^1,^6,^∗∗

PMCID: PMC12923984 PMID: 41666921

Summary

Colorectal cancer mortality is primarily driven by hepatic metastasis, with 50–60% of patients relapsing following liver metastasis resection due to micro-metastases or tumor cell dissemination. Surgery-induced immunologic disturbances contribute to liver recurrence. Exercise modulates immune responses, yet its role in surgical stress-promoted liver metastasis remains unclear. We demonstrate that 4 weeks of preoperative exercise (PEx) limits tumor growth in a murine model of surgical stress-promoted liver metastasis by shifting Kupffer cells toward an anti-tumor phenotype. PEx promotes Kupffer cell cytotoxic cytokines release and enhances CD8⁺ T cells recruitment and activation via the CXCL9-CXCR3 axis. Elevated CXCL9 levels are observed in murine and patient sera post exercise, with Kupffer cells identified as the primary source. Furthermore, exercise-induced butyrate accumulation in Kupffer cells inhibits histone deacetylase 3 activity, promoting CXCL9 expression. These findings suggest that PEx may serve as a non-invasive strategy to reduce recurrence and provide potential targets for exercise-mimetic therapies.

Keywords: preoperative exercise, colorectal liver metastasis, surgical stress, hepatic immune microenvironment, Kupffer cells, CD8+ T cells, butyrate, histone deacetylase 3, CXCL9, CXCR3

Graphical abstract

Highlights

•
PEx enhances hepatic anti-tumor immunity, preventing surgical stress-promoted CRLM
•
CXCL9⁺ KCs recruit CXCR3⁺ CD8⁺ T cells and augment their anti-tumor activity post PEx
•
PEx-induced butyrate accumulation in KCs upregulates CXCL9 expression via HDAC3 inhibition
•
Exercise elevates CXCL9 levels in KCs and peripheral blood in CRLM patients

Preoperative exercise (PEx) prevents surgical stress-promoted liver metastasis by enhancing anti-tumor immunity. Zhang et al. demonstrate that PEx-induced gut microbiota-derived butyrate accumulates in Kupffer cells, where it inhibits HDAC3 to upregulate CXCL9. This chemokine then recruits and activates CXCR3⁺ CD8⁺ T cells, thereby suppressing surgical stress-promoted colorectal cancer liver metastasis.

Introduction

Colorectal cancer (CRC) is the third most commonly diagnosed cancer and ranks second in cancer-related deaths worldwide, approximately 50% of CRC patients eventually developing liver metastases.¹ Surgical resection of hepatic metastases, when feasible, offers 15%–20% of patients a chance for improved overall and disease-free survival.²^,³ However, not all patients benefit from curative surgery, as 50–60% of colorectal cancer liver metastasis (CRLM) patients experience a relapse within the liver following resection.⁴^,⁵ This recurrence is often fueled by the growth of undetected micro-metastases or the dissemination of tumor cells during the surgical procedure, both of which are influenced by changes in the perioperative period.⁶^,⁷^,⁸ Surgery-induced immune disturbances can alter the host-tumor immune environment, which may facilitate tumor recurrence.⁹ Both systemic and local inflammation following surgery have been directly correlated with poorer cancer outcomes in CRC.¹⁰^,¹¹ Despite these known correlations, our understanding of how surgery-triggered inflammation enhances tumor recurrence remains limited.

During liver resection in CRLM patients, the Pringle maneuver, a vascular inflow occlusion technique involving hepatic pedicle clamping to control intraoperative hemorrhage, induces liver ischemia/reperfusion (I/R) injury.¹² The direct ischemic insult and excessive inflammation following the restoration of blood flow¹³ significantly increase the risk of liver recurrence after resection of CRLM.¹⁴^,¹⁵^,¹⁶ Inflammation is a potent driver of tumorigenesis, promoting both tumor initiation and metastasis by cultivating a tumor-favorable microenvironment.¹⁷ The injury sites are preferential areas for tumor regrowth, and surgical trauma has been shown to enhance loco-regional metastasis.¹⁸ Therefore, alleviating perioperative surgical stress-induced hepatic inflammation is crucial for reducing the recurrence of CRLM after surgery.

Regular exercise is increasingly recognized as a therapeutic strategy for cancer due to its ability to modulate the immune response by enhancing anti-tumor immune cell mobilization and infiltration.¹⁹^,²⁰^,²¹ Exercise has been associated with a lower risk of CRC-specific mortality both before and after CRC diagnosis.²² Preoperative exercise therapy (PEx) has proven effective in alleviating postoperative complications and improving clinical outcomes in cancer patients undergoing surgery.²³^,²⁴ Exercise-induced tumor suppression has been shown to be immune-related,²⁵ and PEx has been demonstrated to directly modulate the hepatic immune microenvironment²⁶. Our previous study demonstrated that just four weeks of aerobic PEx significantly attenuated liver inflammation and injury from hepatic I/R injury. Specifically, PEx drives Kupffer cells (KCs) toward an anti-inflammatory phenotype with trained immunity through metabolic reprogramming.²⁷ However, whether PEx protects against hepatic I/R injury-promoted CRLM remains unknown. Gaining insights into the mechanisms of PEx could pave the way for developing pharmacological exercise mimetics, offering potential benefits for exercise-intolerant CRLM patients.

KCs, the resident macrophages of the liver, play pivotal roles in maintaining liver homeostasis²⁸ and exhibit dual roles in tumor metastasis.²⁹^,³⁰ While KCs can directly eliminate tumor cells and secrete chemokines to recruit immune cells and enhance anti-tumor immune response,³¹^,³²^,³³^,³⁴ they can also facilitate hepatic metastasis by secreting pro-tumor cytokines that create a favorable environment for tumor growth.³⁵ These contrasting functions of KCs are shaped by the surrounding microenvironment and external stimuli.³⁶ Understanding how to modulate KC activity is essential for developing strategies to prevent hepatic metastasis.

In this study, we utilized a clinically relevant mouse model of surgical stress-promoted CRLM, along with a randomized controlled trial involving CRLM patients undergoing PEx, to investigate the cellular and molecular mechanisms underlying the effects of PEx on surgical stress-promoted CRLM. Comprehensive analyses of the hepatic immune microenvironment revealed that 4 weeks of PEx reprogrammed KCs toward an anti-tumor phenotype. This reprogramming was marked by increase of cytotoxic cytokines release, which directly eliminated tumor cells, as well as enhanced CD8⁺ T cells recruitment and activation via the CXCL9-CXCR3 axis. Furthermore, PEx-induced butyrate accumulation increased CXCL9 expression by inhibiting histone deacetylase 3 (HDAC3) activity in KCs. Notably, PEx also elevated CXCL9 expression in KCs and peripheral blood in CRLM patients. These findings elucidate the mechanisms by which PEx modulates the anti-tumor function of KCs, ultimately mitigating surgical stress-promoted CRLM.

Results

PEx suppresses surgical stress-promoted highly immunogenic CRLM and remodels the liver immune microenvironment

To assess the impact of PEx on surgical stress-promoted CRLM, we employed a moderate-intensity treadmill-running exercise regimen (12.5 meters/min, 60 min/day, and 5 days/week, for 4 weeks) in a surgical stress-promoted liver metastasis murine model.²⁷^,³⁷ We implanted luciferase-expressing MC38 CRC cells into the livers of sedentary (Sed) or PEx mice via portal vein injection followed by liver I/R to mimic surgical stress (Figure 1A). Tumor growth was monitored weekly using in vivo imaging system (IVIS). We found that metastasis progression was significantly reduced in the PEx group compared to the Sed mice after 2 weeks of tumor injection (Figure 1B). Notably, PEx mice exhibited lower tumor burdens, evidenced by the fewer metastatic nodules and the reduced liver-to-body weight ratio (Figures 1C and 1D). In contrast, in the absence of surgical stress (without liver I/R), the PEx and Sed mice exhibited comparable tumor growth rate and tumor burden (Figures S1A–S1D). To further validate these findings, we used CT26 cells, a model of poorly immunogenic CRC cell lines, in our model. However, PEx did not suppress surgical stress-promoted CRLM in the CT26 tumor cells model (Figures S1E–S1H).

PEx suppresses surgical stress-promoted CRLM and remodels the liver immune microenvironment

(A) Schematic diagram showing the surgical stress-promoted colorectal liver metastasis (CRLM) models in sedentary (Sed) and preoperative exercise (PEx) wild-type (WT) mice.

(B) Weekly bioluminescence imaging was performed to monitor tumor progression. Representative images were shown, and bioluminescence was quantified as photons per second at the indicated time points.

(C and D) Representative images of livers with metastatic tumors (C) (n = 10/group), the number of gross tumor nodules, and the liver-to-body weight ratios (D) in Sed and PEx mice (n = 10/group).

(E) Single-cell RNA sequencing (scRNA-seq) analysis was performed on live leukocytes (CD45⁺) isolated from liver tissues of Sed and PEx mice 3 weeks after establishing the surgical stress-promoted CRLM model. Uniform Manifold Approximation and Projection (UMAP) plots illustrated the clustering of distinct immune cell populations, color-coded for different cell types (n = 2 pooled livers per group).

(F) UMAP plots displaying subpopulations of Kupffer cells (KCs) from Sed and PEx groups.

(G) Volcano plot showing the differentially expressed genes in PEx KCs compared with Sed KCs, with 407 genes significantly altered. All experiments: error bars represent mean ± SEM, statistical analysis by Student’s t test. ∗∗p < 0.01, ∗∗∗p < 0.001. See also Figure S1.

To investigate the effects of PEx on the hepatic immune microenvironment in surgical stress-promoted CRLM, we performed unbiased single-cell RNA sequencing (scRNA-seq) of the immune milieu on liver tissues from Sed and PEx mice 3 weeks after model establishment. Using Uniform Manifold Approximation and Projection (UMAP), we identified the distribution and identities of cellular clusters. After qualitative filtering and excluding CD45⁻ cells, we obtained 23,922 cells with high-quality transcriptomes and classified them into nine cell types: KCs, dendritic cells (DCs), monocytes, neutrophils, B cells, CD4⁺ T cells, CD8⁺ T cells, natural killer T (NKT) cells, and natural killer (NK) cells (Figure 1E). PEx significantly altered the transcriptome of KCs, characterized by a decreased abundance of KC cluster in the lower UMAP region and an increased abundance in the upper region. Additionally, PEx increased the abundance of B cell and NKT cell clusters while reducing neutrophil and monocyte clusters compared with the Sed group. Furthermore, differentially expressed genes (DEGs) analysis across hepatic immune cells revealed that the PEx liver exhibited up-regulation of anti-tumor genes, including Cxcl9,³⁸ Cd40Ig,³⁹ Ahnak,⁴⁰ Cxcl10,⁴¹ and Ifngas1⁴² in KCs, CD8⁺ T cells, B cells, DCs, and NKT cells, respectively (Figure S1I). Collectively, these results suggest that PEx may remodel the hepatic immune microenvironment and potentially associate with anti-tumor immunity in the context of surgical stress-promoted CRLM.

Consistent with our previous findings that 4 weeks of PEx shifts KCs phenotype,²⁷ we observed significant transcriptomic changes in KCs following PEx in our tumor model. To further explore the role of KCs, we identified eight distinct KC subpopulations in the liver based on marker expression (Figure 1F). Clusters 2, 6, and 7, enriched in genes related to anti-tumor phenotype (Cxcl9³⁸, H2-Aa,⁴³ H2-Eb1,⁴³ and H2-Ab1⁴³), were predominantly observed in PEx mice, whereas clusters 0, 1, 3, and 4, characterized by genes related to pro-tumor or immunosuppressive phenotypes (Apoc1,⁴⁴ Ccl24,⁴⁵ Fabp7,⁴⁶ Cd5l,⁴⁷ and Vsig4⁴⁸), were primarily presented in Sed mice (Figures 1G and S1J). These findings suggest that PEx promotes the expression of anti-tumor genes while suppressing the expression of pro-tumor genes in KCs under the condition of surgical stress-promoted CRLM.

KCs are essential for alleviating surgical stress-promoted CRLM following PEx

We then hypothesize the PEx-induced anti-tumor transcriptomic profile in KCs is potentially beneficial on surgical stress-promoted CRLM. To evaluate the necessity of KCs in PEx-induced protective effects, we generated KC-DTR mice with diphtheria toxin receptor (DTR) expression specifically on KCs. Compared with the control R26^LSL-DTR mice, administration of diphtheria toxin (DT) in KC-DTR mice resulted in nearly complete depletion of KCs within 24 h, as confirmed by flow cytometry (Figure S2A). DT was administered intraperitoneally to both R26^LSL-DTR control and KC-DTR mice in the Sed and PEx groups 24 h prior to tumor injection²⁹ (Figure 2A). Depletion of KCs abolished the protective effects of PEx, with no significant difference in tumor growth observed between Sed and PEx KC-DTR mice (Figures 2B–2D and S2B).

To further validate the role of KCs in PEx-induced anti-tumor effects, we performed adoptive transfer experiments. KC-depleted Sed or PEx KC-DTR mice were adoptively transferred with wild-type (WT) KCs from Sed or PEx mice 12 h before tumor injection (Figure 2E). Adoptive transfer of KCs from PEx WT mice significantly protected against surgical stress-promoted CRLM, regardless of the recipient mice’s exercise status, whereas adoptive transfer of Sed-derived KCs had no protective effect (Figures 2F–2H and S2C). These results demonstrate that the protective effects of PEx in alleviating surgical stress-promoted CRLM are dependent on KCs.

PEx enhances KC’s anti-tumor immunity by modulating cytokine secretion

To investigate how PEx modulates KCs in the surgical stress-promoted CRLM model, we first assessed its effects on KC abundance and found no difference in the percentage of hepatic KCs between the Sed and PEx groups (Figure S2D). We subsequently explored the molecular alterations and pathways involved in PEx-associated KCs compared to Sed-associated KCs. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that the DEGs were enriched in cytokine-cytokine receptor interaction (Figure S2E); Gene Ontology (GO) analysis identified enrichment in terms related to leukocyte migration and chemotaxis (Figure S2F). To further investigate how PEx influences the anti-tumor activity of KCs, we performed gene set enrichment analysis. The analysis revealed significant enrichment of gene sets associated with “pathways in cancer” (Figure S2G). Additionally, Reactome pathway analysis revealed enrichment of the signal recognition particle-dependent co-translational protein targeting to membrane pathway, which is essential for mediating protein secretion⁴⁹ (Figure S2H). Together, these data connected PEx to cytokine secretion, immune cell recruitment, and anti-tumor functions.

KCs prevent liver metastasis by directly eliminating tumor cells through phagocytosis and releasing cytolytic molecules.³⁰ To further investigate the impact of PEx on KCs cytokine secretion, we isolated KCs from Sed and PEx mice and co-cultured with MC38-GFP cells⁵⁰ to measure levels of anti-tumor and pro-tumor cytokine (Figure 2I). Interestingly, PEx-KCs produced significantly lower levels of pro-tumor cytokines (interleukin [IL]-10 and transforming growth factor [TGF] β)⁵¹ and higher levels of anti-tumor cytokines (interferon [IFN]-γ, tumor necrosis factor alpha [TNF-α], and IL-12)³⁴^,⁵² compared with Sed-KCs (Figures 2J and 2K). Moreover, we did not detect a significant difference in the phagocytic activity of KCs between Sed and PEx groups (Figures 2L and S2I). These results suggest that PEx exerts its protective effect in the surgical stress-promoted CRLM by modulating the cytokine secretion profile of KCs, rather than altering their quantity or phagocytic activity against tumor cells.

PEx increases CXCL9 release from KCs and promotes CXCR3⁺ CD8⁺ T cells recruitment

Since KCs also secrete chemokines and cytokines to recruit and activate immune cells, thereby amplifying the anti-tumor response,³⁰ we used CellPhoneDB analysis to examine differences in cellular communication between KCs and other immune cells. This revealed significantly increased interactions between KCs and CD4⁺ T cells, CD8⁺ T cells, neutrophils, and NK cells following PEx (Figure 3A). scRNA-seq analysis identified Cxcl9, a chemokine that regulates immune cell migration, differentiation, and activation through its receptor CXCR3,⁵³ as the most significantly upregulated DEG in PEx-KCs compared to Sed-KCs (Figure 1G). Notably, hepatic CXCL9-expressing immune cells in PEx mice were predominantly KCs from PEx-specific clusters 2, 6, and 7 (Figures 3B and S3A). Based on these findings, we hypothesized that PEx-KCs recruit CXCR3⁺ immune cells via CXCL9 to exert anti-tumor effects. Flow cytometry confirmed that PEx increased the CXCL9 expression in KCs (Figure 3C), and circulating and hepatic CXCL9 levels were significantly elevated in PEx mice compared with the Sed mice 3 weeks after surgical stress-promoted CRLM model establishment (Figures 3D and 3E). These results suggest that PEx stimulates CXCL9 secretion by KCs.

To validate these preclinical findings in humans, we examined sera from patients with CRLM undergoing surgery. These patients were enrolled in our randomized controlled trial (NCT04602026) evaluating a prehabilitation program aimed at improving the postoperative outcomes for gastrointestinal malignancies. Patients enrolled in the PEx arm completed a tailored 4-week exercise regimen prescribed by a physical therapist based on assessments during their initial clinic visit and subjective evaluation. Baseline characteristics, including tumor burden, stage, RAS and BRAF mutation status, and MSS/MSI status are summarized in Table S1. Intriguingly, CRLM patients in the PEx arm exhibited increased serum CXCL9 levels post exercise compared to their baseline levels and non-PEx controls (Figures 3F and S3B). Furthermore, CXCL9 expression in KCs isolated from peritumoral liver tissues was higher in patients from the PEx arm compared to the non-PEx arm (Figure 3G).

Our scRNA-seq data revealed that Cxcr3, which codes for the receptor for CXCL9, was enriched in CD8⁺ T cells, NKT cells, and NK cells from PEx mice (Figure 3H). Flow cytometry confirmed that PEx increased the percentage of CD8⁺ T cells, NK cells, and NKT cells in the liver (Figures S3C and S3D) and upregulated CXCR3 expression on these cells (Figures 3I, S3E, and S3F). Notably, CD8⁺ T cells exhibited the highest fold increase in CXCR3 expression, rising approximately 6.12-fold, compared to 3.93-fold in NKT cells and 2.63-fold in NK cells. Similarly, CRLM patients in the PEx arm showed elevated CXCR3 levels on peripheral CD8⁺ T cells compared to non-PEx controls (Figure 3J). PEx also increased CD8⁺ T cell infiltration into tumors (Figure S3G) and enhanced their cytotoxicity in both the liver and tumor microenvironments, as indicated by elevated expression of Granzyme B, IFN-γ, and TNF-α in surgical stress-promoted CRLM model (Figures 3K and S3H). These findings demonstrate that PEx promotes the recruitment and activation of CD8⁺ T cells, enhancing their anti-tumor activity.

The CXCL9-CXCR3 axis is crucial for PEx-mediated protection against surgical stress-promoted CRLM

To verify the role of the CXCL9-CXCR3 axis in PEx-mediated protection, we established the surgical stress-promoted CRLM model in Cxcl9 knockout (KO) and Cxcr3 KO mice (Figures 4A and 4D). PEx failed to confer protective effects in Cxcl9 KO mice and Cxcr3 KO mice, which showed similar tumor growth rates (Figures S4A and S5A) and tumor burdens (Figures 4B, 4C, 4E, and 4F) compared to Sed Cxcl9 KO mice and Sed Cxcr3 KO mice, respectively. Moreover, PEx did not enhance CD8⁺ T cell recruitment or their anti-tumor activity in Cxcl9 KO mice and Cxcr3 KO mice (Figures S4B and S5B).

To further evaluate the role of CXCL9-expressing KCs, we performed adoptive transfer experiments in KC-depleted Sed and PEx KC-DTR mice by transferring KCs from 4-week PEx WT and Cxcl9 KO mice (Figure 4G). Mice receiving WT KCs exhibited significantly slower tumor growth (Figure S4C) and reduced tumor burdens (Figures 4H and 4I) compared to those receiving Cxcl9 KO KCs, regardless of the recipient’s exercise status. Additionally, the adoptive transfer of WT KCs increased CD8⁺ T cell infiltration and enhanced their anti-tumor activity, whereas Cxcl9 KO KCs failed to do so (Figure S4D). To assess whether PEx induces the long-term anti-tumor effects in KCs, KCs-depleted Sed and PEx KC-DTR mice were adoptively transferred with KCs from 4-week PEx WT or Cxcl9 KO mice, followed by Sed or PEx regimens for an additional 4 weeks (Figure S4E). Notably, Sed mice receiving WT KCs showed a significant reduction in tumor burdens compared to those receiving Cxcl9 KO KCs. In contrast, this effect was absent in PEx mice receiving Cxcl9 KO KCs (Figures S4F and S4G). Given the significant upregulation of CXCR3 on CD8⁺ T cells following PEx, we next investigated the role of CXCR3⁺ CD8⁺ T cells in our model. We isolated CD8⁺ T cells from Sed WT or Cxcr3 KO mice and transferred into CD8⁺ T cell-depleted Sed or PEx mice (Figure 4J). Transfer of WT CD8⁺ T cells into PEx mice suppressed tumor growth, increased CD8⁺ T cell infiltration, and enhanced their anti-tumor function, whereas transfer of Cxcr3 KO CD8⁺ T cells did not (Figures 4K, 4L, S5C, and S5D). We further confirmed the role of CXCL9 by neutralizing it in vivo (Figure 4M). Administration of anti-CXCL9 antibody abolished PEx-induced protective effects, leading to comparable tumor growth rates (Figure S4H), tumor burdens (Figures 4N and 4O), CD8⁺ T cell recruitment, and anti-tumor function (Figure S4I) between Sed and PEx mice. Similarly, pharmacologic inhibition of CXCR3 with a specific antagonist⁵⁴ administered eliminated the PEx-induced suppression of tumor growth, CD8⁺ T cell recruitment, and anti-tumor activity (Figures 4P–4R, S5E, and S5F).

To determine whether the CXCL9-CXCR3 axis mediates the interaction between KCs and CD8⁺ T cells in vitro, we co-cultured WT and Cxcl9 KO KCs with CD8⁺ T cells using a transwell assay (Figure S6A). The results showed that KCs from PEx WT mice promoted CD8⁺ T cell migration and anti-tumor function compared to Sed KCs. However, these effects were abolished by adding the anti-CXCL9 antibody or using Cxcl9 KO KCs (Figures S6B and S6C). We next sought to determine whether CXCR3 expression on CD8⁺ T cells is equally critical for this protective interaction. We co-cultured WT and Cxcr3 KO CD8⁺ T cells with PEx KCs using a transwell assay (Figure S6D). The results revealed that CD8⁺ T cells lacking CXCR3 exhibited reduced migration and anti-tumor function when exposed to PEx KCs in vitro, a phenotype recapitulated by adding a CXCR3 antagonist to WT CD8⁺ T cells (Figures S6E and S6F). Together, these findings demonstrate that PEx-mediated interaction between KCs and CD8⁺ T cells through the CXCL9-CXCR3 axis is essential for PEx-mediated protection against surgical stress-promoted CRLM, driving enhanced CD8⁺ T cell recruitment and anti-tumor activity.

CXCL9 enhances anti-tumor activity of CXCR3⁺ CD8⁺ T cells

To further investigate whether PEx modulates CD8⁺ T cell activity via CXCL9, we stimulated WT hepatic CD8⁺ T cells with serum from Sed and PEx WT mice 3 weeks after establishing surgical stress-promoted CRLM model (Figure 5A). CD8⁺ T cells incubated with PEx serum for 24 h exhibited increased production of cytotoxic molecules, including Granzyme B, IFN-γ, and TNF-α, compared to cells stimulated with Sed serum (Figure 5B). This effect was abolished when WT hepatic CD8⁺ T cells were stimulated with serum from Sed and PEx Cxcl9 KO mice (Figures 5C and 5D) or when Cxcr3 KO CD8⁺ T cells were cultured with serum from Sed and PEx WT mice (Figures 5E and 5F). Additionally, recombinant mouse CXCL9 significantly enhanced the anti-tumor activity of hepatic CD8⁺ T cells (Figures S7A and S7B).

Given the distinct immune microenvironments of the liver and tumor, we next examined the effect of CXCL9 on tumor-infiltrating CD8⁺ T cells. Consistent with hepatic CD8⁺ T cells, both PEx serum and the recombinant CXCL9 enhanced the anti-tumor activity of tumor-infiltrating CD8⁺ T cells (Figures 5G, 5H, S7C, and S7D). In contrast, serum from PEx Cxcl9 KO mice did not increase CD8⁺ T cell anti-tumor activity compared to serum from Sed Cxcl9 KO mice (Figures 5I and 5J). Similarly, PEx serum did not enhance the anti-tumor function of tumor-infiltrating CD8⁺ T cells from Cxcr3 KO mice compared to Sed serum (Figures 5K and 5L).

To further verify the in vivo role of CXCL9 in promoting CD8⁺ T cell anti-tumor activity, we administered recombinant CXCL9 or control (0.1% bovine serum albumin in PBS) to Sed or PEx mice after tumor injection⁴¹ (Figure S7E). Administration of recombinant CXCL9 in Sed mice enhanced their anti-tumor response, resulting in comparable tumor burdens between Sed and PEx mice (Figures S7F and S7G).

To extend these findings to humans, we isolated peripheral CD8⁺ T cells from healthy volunteers and stimulated them with pre- and post-exercise sera from the same CRLM patient (Figure 5M). Post-exercise serum enhanced CD8⁺ T cell anti-tumor activity compared to pre-exercise serum (Figure 5N). Similarly, recombinant human CXCL9 significantly increased the anti-tumor activity of CD8⁺ T cells (Figures S7H and S7I). Collectively, these results demonstrate that PEx-induced CXCL9 enhanced the anti-tumor activity of CD8⁺ T cells via the CXCL9-CXCR3 axis, with effects conserved across murine and human systems.

PEx-induced remodeling of the gut microbiota contributes to the alleviation of surgical stress-promoted CRLM

Next, we explored the mechanisms by which PEx induces CXCL9 secretion in KCs. As shown in Figure S2E, KEGG pathway enrichment analysis revealed significant enrichment of metabolic pathways in PEx KCs compared to Sed KCs. Metabolic processes in innate immune cells are critical regulators of epigenetic reprogramming, as metabolites can directly or indirectly influence gene expression.⁵⁵ Using liquid chromatography-tandem mass spectrometry, we conducted non-targeted steady-state metabolomics profiling of the Sed and PEx KCs 3 weeks after tumor injection. Sparse Partial Least Squares Discriminant Analysis revealed distinct intracellular metabolite profiles in PEx-KCs compared to Sed-KCs (Figure 6A). Pathway enrichment analyses identified arginine and proline metabolism, butanoate metabolism, and tyrosine metabolism as the top three enriched pathways in PEx-KCs compared to Sed-KCs (Figure 6B), with butanoate metabolism showing the highest enrichment ratio.

Butanoate, also known as butyrate,⁵⁶ is a short-chain fatty acid produced by gut microbiota fermentation of dietary fibers.⁵⁷ It serves as both an energy source for intestinal cells and a modulator of gene expression with anti-inflammatory and anti-tumor properties.⁵⁸ To investigate whether PEx exerts anti-tumor effects in KCs through butyrate, we collected fecal pellets from Sed and PEx mice before and after establishing the surgical stress-promoted CRLM model and performed 16s rRNA sequencing to analyze the abundance of butyrate-producing bacteria. This analysis revealed a significant increase in butyrate-producing bacteria, particularly Clostridia,⁵⁹ in PEx mice compared to Sed mice, regardless of surgical stress-promoted CRLM model establishment (Figure 6C). Elevated butyrate levels were also detected in the nuclei of KCs and serum of PEx mice compared to Sed mice (Figure 6D), as well as in human KCs and peripheral blood samples from the PEx arm compared to the non-PEx arm (Figure 6E).

To assess the role of PEx-induced microbiota changes in alleviating surgical stress-promoted CRLM, we administered antibiotics (ABx) to deplete the gut microbiota prior to model establishment (Figure 6F). ABx treatment abolished the protective effects of PEx, with no difference in tumor progression between PEx and Sed mice (Figures 6G and 6H). Interestingly, co-housing Sed mice with PEx mice reduced tumor burden in Sed mice to PEx mice levels (Figures 6I–6K), suggesting that PEx-induced remodeling of microbiota played a key role in mediating the anti-tumor effects.

PEx promotes CXCL9 expression in KCs via butyrate-mediated HDAC3 inhibition

Given that butyrate-producing bacteria were increased after PEx, we verified whether butyrate could directly inhibit tumor cell growth in vitro. These results revealed that stimulation with sodium butyrate (NaB) significantly suppressed the growth of MC38 and CT26 CRC tumor cells (Figures S8A and S8B). We then administered NaB or PBS daily via intragastric injection to Sed mice after surgery (Figure 7A). NaB-treated mice exhibited suppressed tumor growth (Figure 7B) and reduced tumor burdens (Figures 7C and 7D) compared to PBS-treated controls, accompanied by increased CXCL9 expression in KCs (Figure 7E).

Butyrate is known to act as a histone deacetylase (HDAC) inhibitor, modulating gene transcription.⁶⁰ Previous studies have shown that HDAC can influence the production of chemokines and cytokines, with HDAC3 inhibition upregulating CXCL9 expression in fibroblast-like synoviocytes.⁶¹^,⁶² Our scRNA-seq data identified HDAC3 as the most downregulated HDAC in PEx KCs (Figure 7F). Consistently, we confirmed reduced HDAC3 expression in PEx KCs compared to Sed KCs (Figure 7G). To validate the role of HDAC3 in regulating CXCL9 expression, we administered an HDAC3 inhibitor (RGFP966) to Sed mice after surgery (Figure 7H). RGFP966 reduced tumor growth (Figure 7I) and tumor burdens (Figures 7J and 7K) while significantly increasing CXCL9 expression in KCs (Figure 7L). In addition, RGFP966 directly inhibited the growth of CRC tumor cells in vitro (Figures S8C and S8D). Together, these results reveal that PEx-induced butyrate enhances CXCL9 expression in KCs by inhibiting HDAC3, contributing to the anti-tumor effects of PEx in surgical stress-promoted CRLM.

To determine whether the anti-tumor effects of NaB and RGFP966 depended on KCs, we administered these compounds to DT pre-treated R26^LSL-DTR mice and KC-DTR after surgery. R26^LSL-DTR mice treated with PBS served as the negative control (Figures S9A and S9E). NaB treatment significantly suppressed tumor growth and reduced metastatic burden in DT pre-treated R26^LSL-DTR mice; However, NaB-induced protective effect was abolished in DT pre-treated KC-DTR mice (Figures S9B–S9D). Similarly, administration of the RGFP966 also attenuated tumor progression in DT pre-treated R26^LSL-DTR mice but not DT pre-treated KC-DTR mice (Figures S9F–S9H), confirming that NaB and RGFP966 rely on the presence of KCs to exert their protective role against surgical stress-promoted CRLM.

Discussion

Surgery-induced immunologic disturbances can alter the host tumor environment, promoting disease recurrence.⁶³^,⁶⁴^,⁶⁵ Indeed, both systemic and local inflammation, along with their sustained responses after surgery (e.g., complications, infections), directly correlate with cancer outcomes for CRC ⁶⁶^,⁶⁷^,⁶⁸^,⁶⁹ and other malignancies.⁷⁰^,⁷¹ However, the mechanistic understanding of inflammation triggered by surgery in enhancing the risk of tumor recurrence is still in its infancy. This current study demonstrates that short-term aerobic exercise can durably reprogram the liver immune microenvironment. Specifically, we show that 4 weeks of PEx enhances the anti-tumor transcriptional profile of hepatic immune cells, preventing surgical stress-promoted CRLM. Notably, PEx did not reduce CRLM in the absence of surgical stress, emphasizing the context-dependent nature of this protective effect.

Exercise is a well-established, safe therapeutic intervention across the perioperative continuum for CRC patients, with proven benefits in reducing recurrence and mortality.⁷² In our study, in vivo imaging showed no difference in liver metastasis one week after tumor injection; however, metastasis progression was significantly reduced in the PEx group compared to sedentary controls after two weeks, which provides evidence that PEx is more likely to affect metastatic outgrowth than initial seeding. The intensity and duration of exercise are critical determinates of its efficacy.²⁰ For instance, moderate to vigorous physical activity has been shown to improve survival rates in metastatic CRC.⁷³ Additionally, maintaining physical activity equivalent to brisk walking for 150 min per week during the first postoperative year is associated with significantly improved disease-free survival in CRLM patients.⁷⁴ Based on these observations, our study adopted a moderate-intensity exercise regimen. We selected a 4-week PEx protocol for two primary reasons. First, our previous study demonstrated that PEx-induced reductions in surgical stress-promoted hepatic injury and inflammation were most pronounced after 4 weeks, with no further benefit observed at longer durations up to 16 weeks.²⁷ Second, most CRLM patients underwent neoadjuvant chemotherapy (NAC) prior to surgery, and previous study suggested an optimal interval of 2–4 weeks between NAC completion and surgery to maximize liver recovery and reduce postoperative complications.⁷⁵

In our study, the only experimental difference between the Sed and PEx groups was the implementation of the structured involuntary treadmill-based exercise regimen. We did not use home cage activity monitoring to quantify spontaneous movement; however, both groups were housed under identical conditions without running wheels or enrichment that could promote voluntary activity. Prior work indicates that structured exercise interventions, but not baseline physical activity, produce the systemic immuno-modulatory effects associated with decreased tumor growth in mice.⁷⁶ Moreover, voluntary exercise such as wheel running significantly increases total activity levels, whereas standard cage spontaneous activity remains low and stable across multiple mouse lines.⁷⁷

Exercise exerts its anti-cancer effects, in part, by modulating the number and function of immune cells.⁷³ Hepatic KCs are critical regulators of liver tumor development, restricting tumor growth during early metastatic seeding.⁷⁸ KCs are anatomically confined to liver sinusoids and generally remain at the periphery of metastatic lesions, making direct KC-tumor contact infrequent during metastasis.²⁹ This spatial restriction can create a KC-depleted “dark zone” around metastases that segregates tumor cells from both KCs and T cells.⁷⁸ Our data support a model in which PEx does not require KC infiltration into the tumor; instead, PEx reprograms sinusoidal KCs to serve as chemokine hubs that recruit and retain CXCR3⁺ CD8⁺ T cells at the sinusoid-tumor interface, where T cells can access and control metastatic outgrowth. Prior work shows that therapeutic disruption of MafB/c-Maf can reprogram KCs to detach from sinusoids and infiltrate tumors, reversing this immune-excluded niche.⁷⁸ While our findings do not require KC infiltration, they align with the broader concept that KC state and positioning are tractable levers for overcoming spatial immune barriers. Future spatial transcriptomics and imaging will map KC and T cell distances and test whether PEx remodels these spatial relationships in vivo. Our scRNA-seq analysis also revealed an increase in NK cell clusters in the liver of PEx mice compared with Sed mice, consistent with reports that exercise-induced cytokines and catecholamines enhance NK cell mobilization and infiltration.⁷⁹ While NK cells are well-recognized mediators of anti-tumor immunity in CRLM,⁸⁰ our findings emphasized a more prominent role for CD8⁺ T cells, which exhibited enhanced crosstalk with KCs, as indicated by increased CXCR3 expression following PEx.

CXCL9 is a chemokine closely associated with favorable prognoses in CRC patients, with elevated CXCL9 expression correlating with improved overall survival.⁸¹ Moreover, macrophage-derived CXCL9 is essential for anti-tumor immune responses following immunotherapy.³⁸ A key finding in our study is the identification of hepatic KCs as the predominant source of CXCL9 following PEx. CXCL9 not only recruits CXCR3⁺ CD8⁺ T cells but also directly enhances their cytotoxic activity by promoting the production of anti-tumor protease and cytokines in CD8⁺ T cells. We further demonstrated that serum from PEx mice and CRLM patients, as well as recombinant CXCL9, significantly enhances the CD8⁺ T cell anti-tumor activity, supporting the essential role of KC-CD8⁺ T cell crosstalk in mediating the protective effects of PEx. In addition, genetic or pharmacological disruption of the CXCL9-CXCR3 axis significantly exacerbated liver metastasis burden in both Sed and PEx mice. This aligns with prior evidence that CXCL9-CXCR3 signaling is indispensable for immune-mediated control of MC38 tumors, primarily through recruitment of anti-tumor CD8⁺ T cells and NK cells to the liver microenvironment.³⁸ Consistent with this, PEx failed to mitigate metastasis progression in CXCL9/CXCR3-deficient models, indicating that PEx does not compensate for loss of this fundamental chemotactic pathway. Together, these support that intact CXCL9-CXCR3 signaling is a non-redundant requirement for restraining liver metastases, independent of exercise-induced adaptations.

Exercise-induced protective effects are mediated by a complex network of factors that extend beyond metabolites. Although our metabolic analysis identified changes in butyrate levels following PEx, other exercise-induced factors, such as cytokines and secreted proteins, may contribute to anti-tumor functions. For instance, exercise induced acute IL-6 signaling may promote anti-tumor adaptive immunity by regulating T cell priming in lymphoid organs and inducting the migration of cytotoxic T cells to tumor-draining lymph nodes and tumor vessels.⁸²Exercise is also associated with enhanced secreted protein acidic and rich in cysteine secretion in humans and animal models; this matricellular protein regulates cell function and tissue remodeling, while inhibiting proliferation and promoting apoptosis of a mouse colon cancer cell line.⁸³

Beyond modulating immune cell interactions, exercise mitigates tumor growth by regulating inter-organ communication, with the gut-liver axis playing a pivotal role in this process.⁸⁴ We discovered that PEx induces alterations in the gut microbiome, specifically increasing the abundance of butyrate-producing Clostridia species. Butyrate, transported to the liver via the portal vein,⁵⁸ acts as an HDAC3 inhibitor, driving epigenetic reprogramming in hepatic KCs to promote an anti-tumor phenotype. While prior study has primarily focused on butyrate’s anti-proliferative effects in CRC cells,⁸⁵ our study demonstrates that butyrate can induce epigenetic modifications in KCs, thereby enhancing their ability to counteract surgical stress-promoted CRLM.

In conclusion, our study provides the intellectual framework for devising mechanism-based exercise therapies to improve surgical outcomes of cancer patients, including those who are exercise intolerant. We demonstrate that PEx elevates butyrate levels in KCs, inducing its reprogramming and subsequent CXCL9-CXCR3-mediated recruitment and activation of CD8⁺ T cells. Our findings open therapeutic avenues for integrating exercise into perioperative cancer care and demonstrate the potential for leveraging exercise-induced systemic reprogramming to enhance treatment efficacy.

Limitations of the study

One limitation of our study is that our animal model does not fully replicate the clinical setting. In our animal model, exercise was administered prior to surgical stress-promoted tumorigenesis, whereas in clinical practice, CRLM patients typically initiate PEx interventions after diagnosis. Despite this discrepancy, our current model was intentionally designed to investigate how PEx reprograms the hepatic immune microenvironment, particularly KCs, to enhance anti-tumor immunity during the perioperative window, a period recognized as highly susceptible to immune suppression and metastatic dissemination. A second limitation is the need to clarify whether exercise exerts a direct effect on CD8⁺ T cell anti-tumor activity. While our data suggest that PEx enhances CD8⁺ T cell function, further research is needed to determine if this is a primary effect or a consequence of upstream immune modulation. Additionally, PEx improved antitumor immunity in an immunogenic CRLM model but not in poorly immunogenic models, limiting immediate generalizability to the predominantly pMMR/MSS CRC population. Future work will evaluate PEx together with immunogenic-priming strategies to determine whether efficacy can be broadened in pMMR/MSS CRC. Finally, although CXCL9 is recognized as a favorable prognostic marker in CRC, our trial was not powered, given the modest sample size and limited follow-up, to determine whether perioperative CXCL9 levels predict postoperative CRLM recurrence. Pandemic-related disruptions (2020–2022) further affected accrual and adherence. Larger, multicenter cohorts with longer follow-up are warranted to establish whether PEx-induced, KC-linked CXCL9 and CXCR3⁺ CD8⁺ frequencies changes are associated with reduced recurrence and to inform development of pharmacologic strategies that mimic or potentiate the immunomodulatory effects of exercise.

Resource availability

Lead contact

For additional information or requests for resources and reagents, please contact the lead researcher, Hongji Zhang (jhn5wx@virginia.edu).

Materials availability

No new unique reagents were generated in this study.

Data and code availability

The single-cell RNA-seq raw data and files reported in this study have been deposited in the GEO database with accession number GEO: GSE290055. The metabolomics data have been deposited to MetaboLights repository with the study identifier MTBLS13419. The 16S rRNA sequencing data have been deposited at the NCBI SRA as PRJNA1372431.

This paper does not report original code.

Any additional information required to reanalyze the data reported in this work is available from the lead contact upon request.

Acknowledgments

This research was funded by the National Institutes of Health R01-CA214865 and R01-GM95566 to A.T., National Institutes of Health R01-GM137203 to H.H., and State funding within the University of Virginia Comprehensive Cancer Center “IDEA-Cancer pilot award” and “Cancer Therapeutics (CRX) pilot award” to H.Z. We thank Xinghua Liao and Pengyan Fa for their technical assistance. Graphics of experimental setups were created with BioRender.com.

Author contributions

All authors read and approved the final version of the manuscript. Conceptualization: H.Z. and A.T.; data curation: Yunwei Zhang, Yiyu Zhang, C.S., J.L., and H.Z.; validation: H.W., Y.W., Y.X., A.H., and M.E.M.; patient recruitment: A.E.; investigation: K.C.C.; formal analysis: Yunwei Zhang, C.S., and J.B.; funding acquisition: H.Z., A.T., and H.H.; project administration: H.Z. and A.T.; resources: H.Z. and A.T.; validation: C.S.; supervision: H.Z. and A.T.; writing—original draft: Yunwei Zhang, C.C., and H.Z.; and writing—review & editing: H.Z., H.H., A.T., and Yunwei Zhang.

Declaration of interests

The authors declare no competing interests.

STAR★Methods

Key resources table

REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies

CD45 FITC (Clone: 30-F11)	BioLegend	Cat# 103108; RRID: AB_312973
CD11b Brilliant Violet 421™ (Clone:M1/70)	BioLegend	Cat# 101251; RRID: AB_2562904
F4/80 Spark YG™ 593 (Clone:QA17A29)	BioLegend	Cat# 101251; RRID: AB_3105858
Ly-6G Alexa Fluor® 700 (Clone:1A8)	BioLegend	Cat# 127622; RRID: AB_10643269
Ly-6C Brilliant Violet 510™ (Clone:HK1.4)	BioLegend	Cat# 128033; RRID: AB_2562351
CD11c Brilliant Violet 650™ (Clone: N418)	BioLegend	Cat# 117339; RRID: AB_2562414
CLEC4F Alexa Fluor® 647 (Clone: 3E3F9)	BioLegend	Cat# 156804; RRID: AB_2814082
NK-1.1 Brilliant Violet 570™ (Clone: PK136)	BioLegend	Cat# 108733; RRID: AB_10896952
I-A/I-E Brilliant Violet 711™ (Clone: M5/114.15.2)	BioLegend	Cat# 107643; RRID: AB_2565976
CD183 (CXCR3) PerCP/Cyanine5.5 (Clone: CXCR3-173)	BioLegend	Cat# 126514; RRID: AB_1186015
CXCL9 (MIG) PE (Clone: MIG-2F5.5)	BioLegend	Cat# 515604; RRID: AB_2245489
Granzyme B PE (Clone:QA18A28)	BioLegend	Cat# 396406; RRID: AB_2801075
IFN-γ PE/Cyanine7 (Clone: XMG1.2)	BioLegend	Cat# 505826; RRID: AB_2295770
TNF-α Brilliant Violet 605™ (Clone: MP6-XT22)	BioLegend	Cat# 506329; RRID: AB_11123912
CD19 BUV563 (Clone: 1D3)	BD Biosciences	Cat# 749028; RRID: AB_2873425
CD3 BUV661 (Clone: 17A2)	BD Biosciences	Cat# 741562; RRID: AB_2870988
CD4 BUV615 (Clone: GK1.5)	BD Biosciences	Cat# 613006; RRID: AB_2870274
CD8a BUV496 (Clone: 53–6.7)	BD Biosciences	Cat# 750024; RRID: AB_2874242
CD84 BUV737 (Clone: 1D3/CD84)	BD Biosciences	Cat# 749569; RRID: AB_2873894
CD3 Monoclonal Antibody (Clone: 17A2)	Thermo Fisher Scientific	Cat# 16-0032-82, RRID: AB_468851
CD28 Monoclonal Antibody (Clone: 37.51)	Thermo Fisher Scientific	Cat# 16-0281-82, RRID: AB_468921
anti-mouse HDAC3 Antibody (Clone: Y415)	Abcam	Cat# ab32369, RRID: AB_732780
Anti-CXCL9 antibody (Clone: EPR26512-118)	Abcam	Cat# ab290643, RRID: AB_3665738
Anti-CXCR3 antibody (Clone: EPR25373-32)	Abcam	Cat# ab288437, RRID: AB_2943206
polyclonal Armenian hamster IgG (Clone: Polyclonal)	BioXCell	Cat# BE0091; RRID: AB_1107773
anti-mouse CXCL9 Ab (Clone: MIG-2F5.5)	BioXCell	Cat# BE0309; RRID: AB_2736989
anti-mouse CD8α Ab (Clone: 2.43)	BioXCell	Cat# BE0061; RRID: AB_1125541

Chemicals, peptides, and recombinant proteins

Cell Activation Cocktail	BioLegend	Cat# 423304
Brefeldin A Solution (1,000X)	BioLegend	Cat# 420601
Fixable Viability Dye eFluor™ 780	eBioScience	Cat# 65-0865-18
Diphtheria Toxin	Calbiochem	Cat# 322326
CXCR3 antagonist (AMG487)	MedChemExpress	Cat# HY-15319
Sodium Butyrate	Sigma-Aldrich	Cat# 567430
HDAC3 inhibitor (RGFP966)	MedChemExpress	Cat# HY-13909
Firefly Luciferase (FLuc)-IRES-Puro Lentivirus	Biosettia	Cat# GlowCell-14p-10
D-Luciferin	Gold Biotechnology	Cat# eLUCK-10G
10% paraformaldehyde	Stat Lab	Cat# 28600-5
Tris-HCl (pH 7.4)	Sigma-Aldrich	Cat# T2194
Sodium chloride solution	Sigma-Aldrich	Cat# 59222C
Magnesium chloride solution	Sigma-Aldrich	Cat# M1028
MACS® BSA Stock Solution	Miltenyi Biotec	Cat# 130-091-376
Tween 20	Bio-Rad Laboratories	Cat# 1662404
DL-Dithiothreitol solution	Sigma-Aldrich	Cat# 646563
Protector RNase Inhibitor	Sigma-Aldrich	Cat# 3335399001
Nuclease-free Water	Sigma-Aldrich	Cat# 3098
IGEPAL® CA-630	Sigma-Aldrich	Cat# I8896
Digitonin (5%)	Thermo Fisher Scientific	Cat# BN2006
Recombinant Mouse CXCL9/MIG Protein	Bio-Techne	Cat# 492-MM-010/CF
Recombinant Human CXCL9/MIG Protein	Bio-Techne	Cat# 392-MG-010/CF

Critical commercial assays

Mouse Granzyme B ELISA Kit	Abcam	Cat# ab238265
Mouse IFN-gamma ELISA Kit	Abcam	Cat# ab282874
Mouse TNF alpha ELISA Kit	Abcam	Cat# ab208348
Mouse IL-12 p70 ELISA Kit	Abcam	Cat# ab119531
Mouse IL-10 ELISA Kit	Abcam	Cat# ab255729
Mouse TGF beta 1 ELISA Kit	Abcam	Cat# ab119557
Mouse CXCL9 ELISA Kit	Abcam	Cat# ab203364
Butyric Acid ELISA Kit	LSBio	Cat# LS-F55824-1
Human Granzyme B ELISA Kit	Abcam	Cat# ab235635
Human IFN gamma ELISA Kit	Abcam	Cat# ab174443
Human TNF alpha ELISA Kit	Abcam	Cat# ab181421
Human CXCL9 ELISA Kit	Abcam	Cat# NBP262861
BD Pharmingen™ Transcription Factor Buffer Set	BD Biosciences	Cat# 562725
Tumor Dissociation Kit, mouse	Miltenyi Biotec	Cat# 130-096-730
Liver Dissociation Kit, mouse	Miltenyi Biotec	Cat# 130-105-807
CD45 MicroBeads, mouse	Miltenyi Biotec	Cat# 130-052-301
Anti-F4/80 MicroBeads UltraPure, mouse	Miltenyi Biotec	Cat# 130-110-443
CD8a⁺ T cell Isolation Kit, mouse	Miltenyi Biotec	Cat# 130-104-075
CD8a⁺ T cell Isolation Kit, human	Miltenyi Biotec	Cat# 130-096-495
CD163 MicroBead Kit, human	Miltenyi Biotec	Cat# 130-124-420
Cell Counting Kit-8 (CCK-8)	Selleck Chemicals	Cat# B34302

Deposited data

Raw and processed scRNA-seq data from sedentary and pre-operative exercise mice liver CD45⁺ immune cells	This manuscript	GEO: GSE290055
Metabolomics data from sedentary and pre-operative exercise mice liver Kupffer cells	This manuscript	MTBLS13419
16S rRNA sequencing data	This manuscript	PRJNA1372431

Experimental models: Cell lines

MC38	Sigma-Aldrich	Cat#: SCC172
CT26	ATCC	Cat#: CRL-2638

Experimental models: Organisms/strains

C57BL/6J	The Jackson Laboratory	RRID: IMSR_JAX:000664
BALB/cJ	The Jackson Laboratory	RRID: IMSR_JAX:000651
C57BL/6J-Clec4^{fem1(cre)Glass}/J	The Jackson Laboratory	RRID: IMSR_JAX:033296
C57BL/6-Gt (ROSA)26Sor^{tm1(HBEGF)Awai}/J	The Jackson Laboratory	RRID: IMSR_JAX:007900
B6.129P2-Cxcr3^tm1Dgen/J	The Jackson Laboratory	RRID: IMSR_JAX:005796
B6.129S4-Cxcl9^tm1Jmf/J	The Jackson Laboratory	RRID: IMSR_JAX:030285

Software and algorithms

FlowJo v10	Treestar	RRID: SCR_008520
BioRender	BioRender	RRID: SCR_018361
GraphPad Prism V9	GraphPad Software	RRID: SCR_002798
MetaboAnalyst 6.0	N/A	https://www.metaboanalyst.ca/MetaboAnalyst/ModuleView.xhtml

Open in a new tab

Experimental model and study participant details

Patient recruitment & sample acquisition

This randomized controlled trial (RCT), titled “The RIOT Trial: Re-Defining Frailty and Improving Outcomes with Prehabilitation for Pancreatic, Liver, or Gastric Cancer” (ClinicalTrials.gov ID: NCT04602026), was conducted at The Ohio State University Wexner Medical Center, Columbus, Ohio, United States. The trial protocol was approved by the Institutional Review Board (IRB #2020C0128). A total of 17 patients with CRLM scheduled for liver resection were enrolled in the study, with 11 assigned to the sedentary group and 6 to the PEx group. The cohort consisted of 9 male and 8 female, with a mean age of 55.6 ± 9.8 years (range: 35–69). Patients underwent safety screening, which included a fall risk assessment and a 2-min walk test, during their appointment with their physician. Based on clinical evaluation, 6 participants were assigned to home-based exercise regimens. These exercises were performed at home at least 3 times per week during the PEx period. Compliance was monitored through daily self-reported diaries, which were submitted at the time of surgery, post-operative visits, or by mail using a pre-stamped envelope provided to the patients. Weekly phone check-ins were conducted by study personnel to ensure adherence and address patient concerns. Written informed consent was obtained from all participants prior to blood collection. Blood samples were drawn at the enrollment and the day of surgery. Tumor and peritumoral liver tissue were collected following the standard biospecimen collection protocol. Participation in the study was voluntary, and subjects were informed that they could withdraw from the study at any time without penalty.

Mice

All murine studies utilized male mice aged 8–10 weeks. The male WT mice (C57BL/6J, Stock no.000664), BALB/cJ mice (Stock no.000651), Clec4f-Cre mice (C57BL/6J-Clec4f^{em1(cre)Glass}/J, Stock no.033296), B6-DTR mice (C57BL/6-Gt (ROSA)26Sor^{tm1(HBEGF)Awai}/J Stock no.007900), Cxcr3 KO mice (B6.129P2-Cxcr3^tm1Dgen/J, Stock no.005796) and Cxcl9 KO mice (B6.129S4-Cxcl9^tm1Jmf/J, Stock no.030285) were purchased from The Jackson Laboratory (U.S.A). Mice expressing diphtheria toxin receptors on KCs (further designated as KC-DTR mice) were produced by crossing Clec4f-Cre mice with B6-DTR mice. 8–10 weeks male mice were randomly assigned to a Sed or PEx group. For PEx, the mice ran on a motorized treadmill (Columbus Instrument, Columbus, OH) with a moderate exercise regimen of 12.5 meter per min for 60 min per day, 5 days per week for 4 weeks. The Sed and PEx mice were either co-housed or housed separately for 4 weeks. All animals were housed in the Center for Comparative Medicine (CCM) at the University of Virginia (UVA) under specified pathogen-free conditions. All animal protocols were approved by the UVA Animal Care and Use Committee and performed in adherence with National Institutes of Health (NIH) guidelines for the care and use of animals.

Tumor cell lines

MC38 and CT26 colon carcinoma cells expressing firefly luciferase (FLuc) and GFP genes were generated using Firefly Luciferase (FLuc)-IRES-Puro Lentivirus (Biosettia). MC38 and CT26 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 100 IU/mL Penicillin/Streptomycin at 37°C and 5% CO₂.

Method details

Surgical stress-promoted CRLM model

To establish surgical stress-promoted CRLM model, 4 weeks of Sed or PEx mice were injected intravenously via the portal vein with 2 × 10⁵ MC38-Luc-GFP cells (C57BL/6J) or 1×10⁵ CT26-Luc-GFP (BALB/cJ) suspended in 50 μL PBS. After the tumor cells circulated for 15 min, mice underwent a non-lethal hepatic segmental (70%) warm ischemia and reperfusion procedure, involving 1 h of ischemia followed by reperfusion. Mice were euthanized 3 weeks after tumor cells injection.²⁷^,³⁷

Treatment of tumor-bearing mice

In studies involving CXCL9 neutralization, mice received an intraperitoneal injection of 200 μg per mouse anti-CXCL9 Ab 1 day before surgery, followed by injected every 3 days.⁸⁶ To administer the CXCR3 antagonist, mice were injected with 5 mg/kg of the CXCR3 antagonist twice daily after surgery.⁸⁷ For recombinant CXCL9 treatment, Sed mice received an intraperitoneal injection of recombinant CXCL9 (500 ng) following surgery. For NaB treatment, Sed mice received daily intragastric administration of NaB (350 mg/kg) following surgery.⁸⁸ For HDAC3 inhibitor treatment, Sed mice received daily intraperitoneal injections of 50 mg/kg RGFP966 following surgery.⁸⁹

Antibiotics administration

The ABx cocktail was prepared as previously described.⁹⁰ Mice in the ABx pre-treated group were administered an antibiotic cocktail consisting of Ampicillin (0.1 g/kg), Neomycin (0.1 g/kg), Vancomycin (0.05 g/kg), and Metronidazole (0.1 g/kg) via oral gavage twice daily for 7 days before tumor cell injection then the surgical stress-promoted CRLM model was established.

KCs isolation and treatment

KCs were isolated from non-parenchymal cells (NPCs) and were further purified via subsequent flow cytometry sorting with anti-mouse CD45 FITC and CLEC4F AF647 staining for KCs using Sony MA900 Cell Sorter. Human liver immune cells were isolated by gradient centrifugation and KCs were further purified using CD163⁺ magnetic beads as previously described.⁹¹ For the anti-CXCL9 Ab pre-treatment, KCs (2×10⁶ cells per well) were seeded in 24-well plates, and 1 μg/mL of anti-CXCL9 neutralizing Ab⁹² was added to the medium for 24 h. Subsequently, the KCs were co-cultured with CD8⁺ T cells using transwell.

Depletion and adoptive transfer of KCs

KCs-depleted Sed or PEx KC-DTR mice served as recipients. KCs from KC- DTR mice were depleted by intraperitoneal injection of 200 ng DT before surgery.²⁹ Donor KCs were isolated from WT Sed or PEx mice livers as previously described.²⁷ A total of 2×10⁶ KCs from Sed or PEx donors were transferred intravenously into each Sed or PEx mice recipient mouse 24 h after KCs depletion. Recipient mice then received a tumor injection and underwent hepatic I/R model 12 h after the adoptive transfer of donor KCs.

Isolation of KCs’ nuclei

KCs’ nuclei were isolated using the Nuclei Isolation protocol (10X Genomics cat# CG000169)⁹³ under defined conditions. Briefly, after isolating the KCs, the supernatant was removed without disrupting the cell pellet, and the cells were suspended in 1 mL PBS +0.04% BSA. The mixture was gently pipetted 5 times and transferred to a 2-mL microcentrifuge tube. After centrifuged at 300 g for 5 min and removing the supernatant, the samples were resuspended in cold lysis buffer (10 mM Tris-HCl, 10 mM NaCl, 0.1% Tween 20, 3 mM MgCl₂, 0.1% NP-40, 0.01% Digitonin, 1% BSA, 1 mM DTT, and 1 U/μL RNase inhibitor), pipetting up and down 10 times. After 3 min of lysis on ice, 1 mL chilled Wash Buffer was added, the mixture was pipetted 5 times, and centrifuged at 500 g for 5 min at 4°C. Following 3 washes, nuclei were resuspended in diluted nuclei buffer, and nuclei concentration was measured by taking the mean of two separate counts using Trypan blue (Thermo Fisher) and the countess II instrument (Thermo Fisher). Less than 5% of live cells were observed when visualized with the countess.

CD8⁺ T cells isolation and treatment

Mouse liver and tumor CD8⁺ T cells and human CD8⁺ T cells from peripheral blood mononuclear cells were isolated using CD8⁺ T cell isolation kit⁹⁴^,⁹⁵ and cultured in RPMI-1640 medium with 10% FBS and 1% penicillin/streptomycin. Anti-CD3 (5 μg/mL) and anti-CD28 (5 μg/mL) antibodies were added for 24 h. For serum or recombinant CXCL9 stimulation, CD8⁺ T cells (2×10⁶ cells per well) were co-cultured with 100 μL of mouse or human serum, or 50 ng/mL of recombinant CXCL9 for 24 h.⁹⁶ For CXCR3 antagonist pre-treatment, CD8⁺ T cells (2×10⁶ cells per well) were co-cultured with 100 nM AMG 487 for 24 h.⁹⁷ Subsequently, the CD8⁺ T cells were co-cultured with KCs using transwell.

Adoptive transfer of CD8⁺ T cells

For CD8⁺ T cells adoptive transfer, WT Sed and PEx mice were given an intraperitoneal injection of anti-CD8 antibody (200 μg/mouse, BioXcell) on day −1. 2×10⁶ WT or Cxcr3 KO CD8⁺ T cells were then intravenously transferred into Sed or PEx mice. The depletion and adoptive transfer were repeated on day 5 and continued twice a week until 3 weeks after tumor injection.⁹⁸

IVIS

To monitor tumor growth, mice were anesthetized with inhaled isoflurane followed by i.p. of potassium luciferin (150 mg/kg in PBS). After 10 min, to allow for luciferin distribution, the mice were imaged using the Spectral Instruments Lago X bioluminescence according to the manufacturer’s instructions. The data were analyzed using Aura imaging software. Abdominal regions were manually selected and quantified for bioluminescence signal analysis (photons per second; p/s).⁹⁹

Flow cytometry

Mouse liver and tumor NPCs were isolated as previously described.²⁷ Single-cell suspensions were obtained using liver or tumor dissociation kits and then passed through a 70 μm cell strainer. Differential centrifugation separated NPCs from hepatocytes or tumor cells (50 g for 5 min, repeated twice). The supernatant was centrifuged again (300 g for 10 min) to obtain NPCs. Before intracellular staining, 2 × 10⁶ cells/mL NPCs were co-cultured with 4 μL of the Cell Activation Cocktail for 4 h. Mouse Fc receptors were blocked by incubating cells with Anti-Mouse CD16/CD32 in PBS for 30 min at 4°C. NPCs were incubated with fixable viability dye in PBS for 30 min at 4°C. Next, cells were surface stained, followed by intracellular staining using the BD Pharmingen Transcription Factor Buffer Set according to the manufacturer’s instructions. Following intracellular staining, cells were suspended in FACS buffer and analyzed on the Aurora 5L spectral flow cytometer (Cytek). Data were analyzed using FlowJo to obtain the percentage of liver and tumor immune cells.

ELISA

To determine CXCL9, Granzyme B, IFN-γ, TNF-α, IL-12, IL-10, and TGF-β, 50–100 μL of serum and 100 μL of the Antibody Cocktail were added to the corresponding ELISA kit wells and incubated for 1 h at room temperature. After washing the wells 3 times, 100 μL of TMB substrate was added and left to incubate in the dark for 10 min. Subsequently, 100 μL of Stop Solution was added, followed by shaking the plate for 1 min. Finally, the optical density (OD) at 450 nm was recorded.

Single-cell RNA-sequencing

Sample and library preparation

Sedentary and PEx mice bearing MC38 tumors were sacrificed 3 weeks after tumor injection. The liver was harvested and processed into single-cell suspensions as previously described. CD45⁺ immune cells were sorted using CD45⁺ beads from these single-cell suspensions, achieving a purity greater than 90%. Sample quality control and scRNA-seq were both performed by the University of Virginia. Following quality control checks for cell viability and concentration, aiming for a target capture of 6000 cells, samples were transferred to a pre-chilled 96-well plate, heat-sealed, and reverse transcription was carried out using a Veriti 96-well thermal cycler. Following reverse transcription, cDNA was extracted using Recovery Agent (10x) and purified with a Silane DynaBead clean-up as described in the user guide. The purified cDNA was then amplified for 12 cycles, diluted at a 4:1 ratio (elution buffer, Qiagen, to cDNA), and analyzed on a Bioanalyzer (Agilent Technologies) to measure cDNA concentration. cDNA libraries were prepared using the 10X Chromium Single Cell 3′ Library and Gel Bead Kit v3.1 (10X Genomics, Cat# PN-1000075).

Sequencing

The molar concentration of each library was determined based on the library size determined by a Bioanalyzer (Agilent Technologies) and qPCR amplification data (Kappa/Roche). Samples were then pooled and normalized to 10 nM and diluted to 2 nM with elution buffer (Qiagen) containing 0.1% Tween 20. Each 2 nM pool was denatured by mixing with equal volume of 0.1 N NaOH and incubating for 5 min at room temperature. The library pools were diluted to 20 pM using HT-1 (Illumina) and then further adjusted to a final loading concentration of 14 pM. A volume of 150 μL from the 14 pM pool was dispensed into each well of an 8-well strip tube and then loaded onto a cBot (Illumina) for cluster generation. The completed libraries were sequenced on a HiSeq 2500 at a depth of 300 million paired reads when targeting 10,000 cells, with the following run parameters: Read 1-26 cycles, Read 2-98 cycles, Index 1–8 cycles.

Gene expression analysis

Fastq files were generated using Cell Ranger mkfastq (10x Genomics) and were analyzed with the 10x Genomics Cell Ranger software v7.0.0. Mapping of the FASTQ files was performed using the mm10 reference genome (GRCm38.91). We performed preliminary data analysis using CellRanger count/aggr and generated a file containing barcode tables, gene tables, and gene expression matrices. Gene expression analysis was conducted using R (version 4.0.0) and the Seurat R package (version 3.1). Cells with 2,500 genes and a mitochondrial gene percentage of >10% were excluded. After data normalization, PCA with variable genes was used as the input, and significant principal components (PCs) were identified based on the jackStraw function.

Cell type annotation

Two-step clustering was used to determine cell types in this study. Broad cell identities were assigned through low-resolution clustering by the ‘FindClusters’ function at a resolution of 0.1 after removing doublets and following differential expression analysis by the ‘FindAllMarkers’ function with default parameters. A total of 9 clusters were generated. DEGs between the two clusters were identified using the Wilcoxon rank-sum test with an adjusted p-value threshold of <0.05. Heat maps were generated using the ComplexHeatmap function. KEGG and GO enrichment analyses were performed using the enrichGO and enrichKEGG functions implemented in the R package ClusterProfiler (version 3.16.1). Enriched pathways with an adjusted p-value of <0.05 were visualized using the dot plot function.

Ligand-receptor interaction analysis

To determine cell-cell interactions in the liver, we utilized CellPhoneDB (v2.1.7) to investigate ligand-receptor interactions between cells. We applied NicheNet (v2.0.4) to infer ligand-receptor interactions that potentially regulate gene expression changes in receiver cell populations. This approach identified vital interactions between different cell types in the liver of Sed and PEx mice.

KCs untargeted metabolomics analysis

KCs (2×10⁶ cells) from both Sed and PEx mice were harvested for Metabolomics. 3 weeks after tumor injection, both mice bearing MC38 tumors were sacrificed. The livers were harvested and processed into single-cell suspensions, and KCs were isolated as previously described. The cells were vortexed for 30 s, then sonicated for 30 min at 4°C. Subsequently, each sample was kept at −20°C for 1 h, and then the sample was centrifuged at 12,000 rpm and 4°C for 15 min. Finally, 200 μL of the supernatant and 5 μL of DL-o-Chlorophenylalanine (0.2 mg/mL) were transferred to vials for LC-MS analysis. Data analysis, including principal component analysis, pathway impact analysis, and heatmap generation, was performed using MetaboAnalyst 6.0 software.²⁷

16s RNA-sequencing

Fecal pellets (50 mg) from Sed and PEx mice before and after establishing the surgical stress-promoted CRLM model were collected and conducted 16s rRNA sequencing. DNA was extracted using either the ZymoBIOMICS-96 MagBead DNA Kit. Bacterial 16S rRNA gene sequencing focused on the V3-V4 region for targeted library preparation.¹⁰⁰ The library was prepared using a real-time PCR process to limit PCR chimera formation, followed by quantification, pooling, and cleaning. Control samples included positive controls for each DNA extraction and library preparation and negative controls to monitor bioburden. Sequencing was conducted on an Illumina Nextseq with a P1 reagent kit (600 cycles) and a 30% PhiX spike-in. Bioinformatics analysis was conducted using the DADA2 pipeline for error correction and taxonomy assignment with Uclust from Qiime v.1.9.1. Additional analyses were performed with internal scripts. For absolute abundance quantification, quantitative real-time PCR was used with a standard curve to calculate gene copies per microliter of DNA sample. The number of genome copies was determined by dividing the gene copy number by the assumed number of gene copies per genome.

Western blotting

Whole-cell protein lysates from KCs were used for western blotting. Membranes were incubated HDAC3 (1:500, Abcam), CXCL9 (1:500, Abcam), CXCR3 (1:500, Abcam), and β-actin (1:1000, Cell Signaling Technology) as an internal control.

Cell growth assays

The Cell-counting Kit-8 (CCK-8, Selleck Chemicals) assays were used to determine the effects of NaB or RGFP966 treatment on cell growth. Briefly, the cells were seeded in a 96-well plate at densities of 5,000 cells/well and incubated with 5mM NaB¹⁰¹ or 10uM RGFP966¹⁰² for the indicated time. At the end of treatment, CCK-8 reagent was added to each well at a ratio of 1:10. After incubation for 2 h at 37°C, the absorbance of each well was measured at a wavelength of 450 nm. The growth rates of different time point were normalized to the baseline of the 0-h time point of treatment.

Quantification and statistical analysis

Assessment of statistical significance between two groups was performed using an unpaired two-tailed Student’s t test and more than two groups using a one-way ANOVA with Tukey’s correction for multiple comparisons. All statistical tests were analyzed using the GraphPad Prism software (V.9). Data represents the mean ± SEM unless stated otherwise. The statistical significance symbols were used in the figures: ns, not significant (p > 0.05), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Additional resources

This clinical trial has been registered on https://clinicaltrials.gov(NCT04602026).

Published: February 9, 2026

Footnotes

Supplemental information can be found online at https://doi.org/10.1016/j.xcrm.2026.102589.

Contributor Information

Allan Tsung, Email: crf9aa@uvahealth.org.

Hongji Zhang, Email: jhn5wx@virginia.edu.

Supplemental information

Document S1. Figures S1–S9 and Table S1

mmc1.pdf^{(1.7MB, pdf)}

Document S2. Article plus supplemental information

mmc2.pdf^{(32.6MB, pdf)}

References

1.Siegel R.L., Wagle N.S., Cercek A., Smith R.A., Jemal A. Colorectal cancer statistics, 2023. CA Cancer J. Clin. 2023;73:233–254. doi: 10.3322/caac.21772. [DOI] [PubMed] [Google Scholar]
2.Zhou H., Liu Z., Wang Y., Wen X., Amador E.H., Yuan L., Ran X., Xiong L., Ran Y., Chen W., Wen Y. Colorectal liver metastasis: molecular mechanism and interventional therapy. Signal Transduct. Targeted Ther. 2022;7:70. doi: 10.1038/s41392-022-00922-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
3.Nordlinger B., Sorbye H., Glimelius B., Poston G.J., Schlag P.M., Rougier P., Bechstein W.O., Primrose J.N., Walpole E.T., Finch-Jones M., et al. Perioperative FOLFOX4 chemotherapy and surgery versus surgery alone for resectable liver metastases from colorectal cancer (EORTC 40983): long-term results of a randomised, controlled, phase 3 trial. Lancet Oncol. 2013;14:1208–1215. doi: 10.1016/s1470-2045(13)70447-9. [DOI] [PubMed] [Google Scholar]
4.Zhang Y., Liu G., Zeng Q., Wu W., Lei K., Zhang C., Tang M., Zhang Y., Xiang X., Tan L., et al. CCL19-producing fibroblasts promote tertiary lymphoid structure formation enhancing anti-tumor IgG response in colorectal cancer liver metastasis. Cancer Cell. 2024;42:1370–1385.e9. doi: 10.1016/j.ccell.2024.07.006. [DOI] [PubMed] [Google Scholar]
5.de Jong M.C., Pulitano C., Ribero D., Strub J., Mentha G., Schulick R.D., Choti M.A., Aldrighetti L., Capussotti L., Pawlik T.M. Rates and patterns of recurrence following curative intent surgery for colorectal liver metastasis: an international multi-institutional analysis of 1669 patients. Ann. Surg. 2009;250:440–448. doi: 10.1097/SLA.0b013e3181b4539b. [DOI] [PubMed] [Google Scholar]
6.Heinz M.C., Peters N.A., Oost K.C., Lindeboom R.G.H., van Voorthuijsen L., Fumagalli A., van der Net M.C., de Medeiros G., Hageman J.H., Verlaan-Klink I., et al. Liver Colonization by Colorectal Cancer Metastases Requires YAP-Controlled Plasticity at the Micrometastatic Stage. Cancer Res. 2022;82:1953–1968. doi: 10.1158/0008-5472.Can-21-0933. [DOI] [PMC free article] [PubMed] [Google Scholar]
7.Weitz J., Kienle P., Lacroix J., Willeke F., Benner A., Lehnert T., Herfarth C., von Knebel Doeberitz M. Dissemination of tumor cells in patients undergoing surgery for colorectal cancer. Clin. Cancer Res. 1998;4:343–348. [PubMed] [Google Scholar]
8.Rahbari N.N., Birgin E., Bork U., Mehrabi A., Reißfelder C., Weitz J. Anterior Approach vs Conventional Hepatectomy for Resection of Colorectal Liver Metastasis: A Randomized Clinical Trial. JAMA Surg. 2021;156:31–40. doi: 10.1001/jamasurg.2020.5050. [DOI] [PMC free article] [PubMed] [Google Scholar]
9.Hiller J.G., Perry N.J., Poulogiannis G., Riedel B., Sloan E.K. Perioperative events influence cancer recurrence risk after surgery. Nat. Rev. Clin. Oncol. 2018;15:205–218. doi: 10.1038/nrclinonc.2017.194. [DOI] [PubMed] [Google Scholar]
10.Quail D.F., Joyce J.A. Microenvironmental regulation of tumor progression and metastasis. Nat. Med. 2013;19:1423–1437. doi: 10.1038/nm.3394. [DOI] [PMC free article] [PubMed] [Google Scholar]
11.Cao Y., Zheng X., Hu Y., Li J., Huang B., Zhao N., Liu T., Cai K., Tian S. Levels of systemic inflammation response index are correlated with tumor-associated bacteria in colorectal cancer. Cell Death Dis. 2023;14:69. doi: 10.1038/s41419-023-05602-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
12.Maspero M., Yilmaz S., Cazzaniga B., Raj R., Ali K., Mazzaferro V., Schlegel A. The role of ischaemia-reperfusion injury and liver regeneration in hepatic tumour recurrence. JHEP Rep. 2023;5 doi: 10.1016/j.jhepr.2023.100846. [DOI] [PMC free article] [PubMed] [Google Scholar]
13.Chen Z., Zhang P., Xu Y., Yan J., Liu Z., Lau W.B., Lau B., Li Y., Zhao X., Wei Y., Zhou S. Surgical stress and cancer progression: the twisted tango. Mol. Cancer. 2019;18:132. doi: 10.1186/s12943-019-1058-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
14.Cho J.Y., Han H.S., Choi Y., Yoon Y.S., Kim S., Choi J.K., Jang J.S., Kwon S.U., Kim H. Association of Remnant Liver Ischemia With Early Recurrence and Poor Survival After Liver Resection in Patients With Hepatocellular Carcinoma. JAMA Surg. 2017;152:386–392. doi: 10.1001/jamasurg.2016.5040. [DOI] [PMC free article] [PubMed] [Google Scholar]
15.Nicoud I.B., Jones C.M., Pierce J.M., Earl T.M., Matrisian L.M., Chari R.S., Gorden D.L. Warm hepatic ischemia-reperfusion promotes growth of colorectal carcinoma micrometastases in mouse liver via matrix metalloproteinase-9 induction. Cancer Res. 2007;67:2720–2728. doi: 10.1158/0008-5472.Can-06-3923. [DOI] [PubMed] [Google Scholar]
16.van der Bilt J.D.W., Kranenburg O., Nijkamp M.W., Smakman N., Veenendaal L.M., Te Velde E.A., Voest E.E., van Diest P.J., Borel Rinkes I.H.M. Ischemia/reperfusion accelerates the outgrowth of hepatic micrometastases in a highly standardized murine model. Hepatology. 2005;42:165–175. doi: 10.1002/hep.20739. [DOI] [PubMed] [Google Scholar]
17.Greten F.R., Grivennikov S.I. Inflammation and Cancer: Triggers, Mechanisms, and Consequences. Immunity. 2019;51:27–41. doi: 10.1016/j.immuni.2019.06.025. [DOI] [PMC free article] [PubMed] [Google Scholar]
18.Tohme S., Simmons R.L., Tsung A. Surgery for Cancer: A Trigger for Metastases. Cancer Res. 2017;77:1548–1552. doi: 10.1158/0008-5472.Can-16-1536. [DOI] [PMC free article] [PubMed] [Google Scholar]
19.Hojman P., Gehl J., Christensen J.F., Pedersen B.K. Molecular Mechanisms Linking Exercise to Cancer Prevention and Treatment. Cell Metab. 2018;27:10–21. doi: 10.1016/j.cmet.2017.09.015. [DOI] [PubMed] [Google Scholar]
20.Zhang Y., Cao C., Li C., Witt R.G., Huang H., Tsung A., Zhang H. Physical exercise in liver diseases. Hepatology. 2024 doi: 10.1097/hep.0000000000000941. [DOI] [PMC free article] [PubMed] [Google Scholar]
21.Idorn M., Hojman P. Exercise-Dependent Regulation of NK Cells in Cancer Protection. Trends Mol. Med. 2016;22:565–577. doi: 10.1016/j.molmed.2016.05.007. [DOI] [PubMed] [Google Scholar]
22.Schmid D., Leitzmann M.F. Association between physical activity and mortality among breast cancer and colorectal cancer survivors: a systematic review and meta-analysis. Ann. Oncol. 2014;25:1293–1311. doi: 10.1093/annonc/mdu012. [DOI] [PubMed] [Google Scholar]
23.Clifford K., Woodfield J.C., Tait W., Campbell H.A., Baldi J.C. Association of Preoperative High-Intensity Interval Training With Cardiorespiratory Fitness and Postoperative Outcomes Among Adults Undergoing Major Surgery: A Systematic Review and Meta-Analysis. JAMA Netw. Open. 2023;6 doi: 10.1001/jamanetworkopen.2023.20527. [DOI] [PMC free article] [PubMed] [Google Scholar]
24.Heinz A., Hiller K. Run to survive-how preoperative exercise could prevent major surgical complications. Nat. Metab. 2021;3:732–734. doi: 10.1038/s42255-021-00403-w. [DOI] [PubMed] [Google Scholar]
25.Kurz E., Hirsch C.A., Dalton T., Shadaloey S.A., Khodadadi-Jamayran A., Miller G., Pareek S., Rajaei H., Mohindroo C., Baydogan S., et al. Exercise-induced engagement of the IL-15/IL-15Rα axis promotes anti-tumor immunity in pancreatic cancer. Cancer Cell. 2022;40:720–737.e5. doi: 10.1016/j.ccell.2022.05.006. [DOI] [PMC free article] [PubMed] [Google Scholar]
26.Koelwyn G.J., Zhuang X., Tammela T., Schietinger A., Jones L.W. Exercise and immunometabolic regulation in cancer. Nat. Metab. 2020;2:849–857. doi: 10.1038/s42255-020-00277-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
27.Zhang H., Chen T., Ren J., Xia Y., Onuma A., Wang Y., He J., Wu J., Wang H., Hamad A., et al. Pre-operative exercise therapy triggers anti-inflammatory trained immunity of Kupffer cells through metabolic reprogramming. Nat. Metab. 2021;3:843–858. doi: 10.1038/s42255-021-00402-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
28.Wen Y., Lambrecht J., Ju C., Tacke F. Hepatic macrophages in liver homeostasis and diseases-diversity, plasticity and therapeutic opportunities. Cell. Mol. Immunol. 2021;18:45–56. doi: 10.1038/s41423-020-00558-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
29.Thomas S.K., Wattenberg M.M., Choi-Bose S., Uhlik M., Harrison B., Coho H., Cassella C.R., Stone M.L., Patel D., Markowitz K., et al. Kupffer cells prevent pancreatic ductal adenocarcinoma metastasis to the liver in mice. Nat. Commun. 2023;14:6330. doi: 10.1038/s41467-023-41771-z. [DOI] [PMC free article] [PubMed] [Google Scholar]
30.Tsilimigras D.I., Brodt P., Clavien P.A., Muschel R.J., D'Angelica M.I., Endo I., Parks R.W., Doyle M., de Santibañes E., Pawlik T.M. Liver metastases. Nat. Rev. Dis. Primers. 2021;7:27. doi: 10.1038/s41572-021-00261-6. [DOI] [PubMed] [Google Scholar]
31.Ozga A.J., Chow M.T., Luster A.D. Chemokines and the immune response to cancer. Immunity. 2021;54:859–874. doi: 10.1016/j.immuni.2021.01.012. [DOI] [PMC free article] [PubMed] [Google Scholar]
32.Kimura Y., Inoue A., Hangai S., Saijo S., Negishi H., Nishio J., Yamasaki S., Iwakura Y., Yanai H., Taniguchi T. The innate immune receptor Dectin-2 mediates the phagocytosis of cancer cells by Kupffer cells for the suppression of liver metastasis. Proc. Natl. Acad. Sci. USA. 2016;113:14097–14102. doi: 10.1073/pnas.1617903113. [DOI] [PMC free article] [PubMed] [Google Scholar]
33.Li J., Liu X.G., Ge R.L., Yin Y.P., Liu Y.D., Lu W.P., Huang M., He X.Y., Wang J., Cai G., et al. The ligation between ERMAP, galectin-9 and dectin-2 promotes Kupffer cell phagocytosis and antitumor immunity. Nat. Immunol. 2023;24:1813–1824. doi: 10.1038/s41590-023-01634-7. [DOI] [PubMed] [Google Scholar]
34.Deng Z., Loyher P.L., Lazarov T., Li L., Shen Z., Bhinder B., Yang H., Zhong Y., Alberdi A., Massague J., et al. The nuclear factor ID3 endows macrophages with a potent anti-tumour activity. Nature. 2024;626:864–873. doi: 10.1038/s41586-023-06950-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
35.Yuan D., Huang S., Berger E., Liu L., Gross N., Heinzmann F., Ringelhan M., Connor T.O., Stadler M., Meister M., et al. Kupffer Cell-Derived Tnf Triggers Cholangiocellular Tumorigenesis through JNK due to Chronic Mitochondrial Dysfunction and ROS. Cancer Cell. 2017;31:771–789.e6. doi: 10.1016/j.ccell.2017.05.006. [DOI] [PMC free article] [PubMed] [Google Scholar]
36.Huang H.Y., Chen Y.Z., Zhao C., Zheng X.N., Yu K., Yue J.X., Ju H.Q., Shi Y.X., Tian L. Alternations in inflammatory macrophage niche drive phenotypic and functional plasticity of Kupffer cells. Nat. Commun. 2024;15:9337. doi: 10.1038/s41467-024-53659-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
37.Xia Y., He J., Zhang H., Wang H., Tetz G., Maguire C.A., Wang Y., Onuma A., Genkin D., Tetz V., et al. AAV-mediated gene transfer of DNase I in the liver of mice with colorectal cancer reduces liver metastasis and restores local innate and adaptive immune response. Mol. Oncol. 2020;14:2920–2935. doi: 10.1002/1878-0261.12787. [DOI] [PMC free article] [PubMed] [Google Scholar]
38.House I.G., Savas P., Lai J., Chen A.X.Y., Oliver A.J., Teo Z.L., Todd K.L., Henderson M.A., Giuffrida L., Petley E.V., et al. Macrophage-Derived CXCL9 and CXCL10 Are Required for Antitumor Immune Responses Following Immune Checkpoint Blockade. Clin. Cancer Res. 2020;26:487–504. doi: 10.1158/1078-0432.Ccr-19-1868. [DOI] [PubMed] [Google Scholar]
39.Kuhn N.F., Purdon T.J., van Leeuwen D.G., Lopez A.V., Curran K.J., Daniyan A.F., Brentjens R.J. CD40 Ligand-Modified Chimeric Antigen Receptor T Cells Enhance Antitumor Function by Eliciting an Endogenous Antitumor Response. Cancer Cell. 2019;35:473–488.e6. doi: 10.1016/j.ccell.2019.02.006. [DOI] [PMC free article] [PubMed] [Google Scholar]
40.Ghodke I., Remisova M., Furst A., Kilic S., Reina-San-Martin B., Poetsch A.R., Altmeyer M., Soutoglou E. AHNAK controls 53BP1-mediated p53 response by restraining 53BP1 oligomerization and phase separation. Mol. Cell. 2021;81:2596–2610.e7. doi: 10.1016/j.molcel.2021.04.010. [DOI] [PMC free article] [PubMed] [Google Scholar]
41.Lim R.J., Salehi-Rad R., Tran L.M., Oh M.S., Dumitras C., Crosson W.P., Li R., Patel T.S., Man S., Yean C.E., et al. CXCL9/10-engineered dendritic cells promote T cell activation and enhance immune checkpoint blockade for lung cancer. Cell Rep. Med. 2024;5 doi: 10.1016/j.xcrm.2024.101479. [DOI] [PMC free article] [PubMed] [Google Scholar]
42.Gao F., He S., Jin A. MiRNAs and lncRNAs in NK cell biology and NK/T-cell lymphoma. Genes Dis. 2021;8:590–602. doi: 10.1016/j.gendis.2020.08.006. [DOI] [PMC free article] [PubMed] [Google Scholar]
43.Chen D., Varanasi S.K., Hara T., Traina K., Sun M., McDonald B., Farsakoglu Y., Clanton J., Xu S., Garcia-Rivera L., et al. CTLA-4 blockade induces a microglia-Th1 cell partnership that stimulates microglia phagocytosis and anti-tumor function in glioblastoma. Immunity. 2023;56:2086–2104.e8. doi: 10.1016/j.immuni.2023.07.015. [DOI] [PMC free article] [PubMed] [Google Scholar]
44.Ma R.Y., Black A., Qian B.Z. Macrophage diversity in cancer revisited in the era of single-cell omics. Trends Immunol. 2022;43:546–563. doi: 10.1016/j.it.2022.04.008. [DOI] [PubMed] [Google Scholar]
45.Hinshaw D.C., Shevde L.A. The Tumor Microenvironment Innately Modulates Cancer Progression. Cancer Res. 2019;79:4557–4566. doi: 10.1158/0008-5472.Can-18-3962. [DOI] [PMC free article] [PubMed] [Google Scholar]
46.Liu R.Z., Choi W.S., Jain S., Xu X., Elsherbiny M.E., Glubrecht D.D., Tessier A.G., Easaw J.C., Fallone B.G., Godbout R. Stationary-to-migratory transition in glioblastoma stem-like cells driven by a fatty acid-binding protein 7-RXRα neurogenic pathway. Neuro Oncol. 2023;25:2177–2190. doi: 10.1093/neuonc/noad134. [DOI] [PMC free article] [PubMed] [Google Scholar]
47.Li Y., Qu P., Wu L., Li B., Du H., Yan C. Api6/AIM/Spα/CD5L overexpression in alveolar type II epithelial cells induces spontaneous lung adenocarcinoma. Cancer Res. 2011;71:5488–5499. doi: 10.1158/0008-5472.Can-10-4225. [DOI] [PMC free article] [PubMed] [Google Scholar]
48.Shen J., Zhou L., Ye K., Gong J., Wu F., Mo K., Zhu Y., Chen C., Zhan R. The role of SPI1/VSIG4/THBS1 on glioblastoma progression through modulation of the PI3K/AKT pathway. J. Adv. Res. 2025;71:487–500. doi: 10.1016/j.jare.2024.06.023. [DOI] [PMC free article] [PubMed] [Google Scholar]
49.Czech L., Mais C.-N., Kratzat H., Sarmah P., Giammarinaro P., Freibert S.-A., Esser H.F., Musial J., Berninghausen O., Steinchen W., et al. Inhibition of SRP-dependent protein secretion by the bacterial alarmone (p)ppGpp. Nat. Commun. 2022;13:1069. doi: 10.1038/s41467-022-28675-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
50.Giannou A.D., Kempski J., Shiri A.M., Lücke J., Zhang T., Zhao L., Zazara D.E., Cortesi F., Riecken K., Amezcua Vesely M.C., et al. Tissue resident iNKT17 cells facilitate cancer cell extravasation in liver metastasis via interleukin-22. Immunity. 2023;56:125–142.e12. doi: 10.1016/j.immuni.2022.12.014. [DOI] [PMC free article] [PubMed] [Google Scholar]
51.Zhao H., Wu L., Yan G., Chen Y., Zhou M., Wu Y., Li Y. Inflammation and tumor progression: signaling pathways and targeted intervention. Signal Transduct. Targeted Ther. 2021;6:263. doi: 10.1038/s41392-021-00658-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
52.Han J., Wu M., Liu Z. Dysregulation in IFN-γ signaling and response: the barricade to tumor immunotherapy. Front. Immunol. 2023;14 doi: 10.3389/fimmu.2023.1190333. [DOI] [PMC free article] [PubMed] [Google Scholar]
53.Tokunaga R., Zhang W., Naseem M., Puccini A., Berger M.D., Soni S., McSkane M., Baba H., Lenz H.J. CXCL9, CXCL10, CXCL11/CXCR3 axis for immune activation - A target for novel cancer therapy. Cancer Treat Rev. 2018;63:40–47. doi: 10.1016/j.ctrv.2017.11.007. [DOI] [PMC free article] [PubMed] [Google Scholar]
54.de Mingo Pulido Á., Gardner A., Hiebler S., Soliman H., Rugo H.S., Krummel M.F., Coussens L.M., Ruffell B. TIM-3 Regulates CD103(+) Dendritic Cell Function and Response to Chemotherapy in Breast Cancer. Cancer Cell. 2018;33:60–74.e6. doi: 10.1016/j.ccell.2017.11.019. [DOI] [PMC free article] [PubMed] [Google Scholar]
55.Li X., Egervari G., Wang Y., Berger S.L., Lu Z. Regulation of chromatin and gene expression by metabolic enzymes and metabolites. Nat. Rev. Mol. Cell Biol. 2018;19:563–578. doi: 10.1038/s41580-018-0029-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
56.Imhann F., Vich Vila A., Bonder M.J., Fu J., Gevers D., Visschedijk M.C., Spekhorst L.M., Alberts R., Franke L., van Dullemen H.M., et al. Interplay of host genetics and gut microbiota underlying the onset and clinical presentation of inflammatory bowel disease. Gut. 2018;67:108–119. doi: 10.1136/gutjnl-2016-312135. [DOI] [PMC free article] [PubMed] [Google Scholar]
57.Flemming A. Butyrate boosts microbicidal macrophages. Nat. Rev. Immunol. 2019;19:135. doi: 10.1038/s41577-019-0132-9. [DOI] [PubMed] [Google Scholar]
58.Mann E.R., Lam Y.K., Uhlig H.H. Short-chain fatty acids: linking diet, the microbiome and immunity. Nat. Rev. Immunol. 2024;24:577–595. doi: 10.1038/s41577-024-01014-8. [DOI] [PubMed] [Google Scholar]
59.Rivera-Chávez F., Zhang L.F., Faber F., Lopez C.A., Byndloss M.X., Olsan E.E., Xu G., Velazquez E.M., Lebrilla C.B., Winter S.E., Bäumler A.J. Depletion of Butyrate-Producing Clostridia from the Gut Microbiota Drives an Aerobic Luminal Expansion of Salmonella. Cell Host Microbe. 2016;19:443–454. doi: 10.1016/j.chom.2016.03.004. [DOI] [PMC free article] [PubMed] [Google Scholar]
60.Modoux M., Rolhion N., Lefevre J.H., Oeuvray C., Nádvorník P., Illes P., Emond P., Parc Y., Mani S., Dvorak Z., Sokol H. Butyrate acts through HDAC inhibition to enhance aryl hydrocarbon receptor activation by gut microbiota-derived ligands. Gut Microbes. 2022;14 doi: 10.1080/19490976.2022.2105637. [DOI] [PMC free article] [PubMed] [Google Scholar]
61.Angiolilli C., Kabala P.A., Grabiec A.M., Van Baarsen I.M., Ferguson B.S., García S., Malvar Fernandez B., McKinsey T.A., Tak P.P., Fossati G., et al. Histone deacetylase 3 regulates the inflammatory gene expression programme of rheumatoid arthritis fibroblast-like synoviocytes. Ann. Rheum. Dis. 2017;76:277–285. doi: 10.1136/annrheumdis-2015-209064. [DOI] [PMC free article] [PubMed] [Google Scholar]
62.Zheng H., Zhao W., Yan C., Watson C.C., Massengill M., Xie M., Massengill C., Noyes D.R., Martinez G.V., Afzal R., et al. HDAC Inhibitors Enhance T-Cell Chemokine Expression and Augment Response to PD-1 Immunotherapy in Lung Adenocarcinoma. Clin. Cancer Res. 2016;22:4119–4132. doi: 10.1158/1078-0432.Ccr-15-2584. [DOI] [PMC free article] [PubMed] [Google Scholar]
63.van der Bij G.J., Oosterling S.J., Beelen R.H.J., Meijer S., Coffey J.C., van Egmond M. The perioperative period is an underutilized window of therapeutic opportunity in patients with colorectal cancer. Ann. Surg. 2009;249:727–734. doi: 10.1097/SLA.0b013e3181a3ddbd. [DOI] [PubMed] [Google Scholar]
64.Pang L., Yeung O.W.H., Ng K.T.P., Liu H., Zhu J., Liu J., Yang X., Ding T., Qiu W., Wang Y., et al. Postoperative Plasmacytoid Dendritic Cells Secrete IFNα to Promote Recruitment of Myeloid-Derived Suppressor Cells and Drive Hepatocellular Carcinoma Recurrence. Cancer Res. 2022;82:4206–4218. doi: 10.1158/0008-5472.Can-22-1199. [DOI] [PubMed] [Google Scholar]
65.Ren J., He J., Zhang H., Xia Y., Hu Z., Loughran P., Billiar T., Huang H., Tsung A. Platelet TLR4-ERK5 Axis Facilitates NET-Mediated Capturing of Circulating Tumor Cells and Distant Metastasis after Surgical Stress. Cancer Res. 2021;81:2373–2385. doi: 10.1158/0008-5472.Can-20-3222. [DOI] [PMC free article] [PubMed] [Google Scholar]
66.Artinyan A., Orcutt S.T., Anaya D.A., Richardson P., Chen G.J., Berger D.H. Infectious postoperative complications decrease long-term survival in patients undergoing curative surgery for colorectal cancer: a study of 12,075 patients. Ann. Surg. 2015;261:497–505. doi: 10.1097/SLA.0000000000000854. [DOI] [PubMed] [Google Scholar]
67.Farid S.G., Aldouri A., Morris-Stiff G., Khan A.Z., Toogood G.J., Lodge J.P.A., Prasad K.R. Correlation between postoperative infective complications and long-term outcomes after hepatic resection for colorectal liver metastasis. Ann. Surg. 2010;251:91–100. doi: 10.1097/SLA.0b013e3181bfda3c. [DOI] [PubMed] [Google Scholar]
68.Gomez D., Morris-Stiff G., Wyatt J., Toogood G.J., Lodge J.P.A., Prasad K.R. Surgical technique and systemic inflammation influences long-term disease-free survival following hepatic resection for colorectal metastasis. J. Surg. Oncol. 2008;98:371–376. doi: 10.1002/jso.21103. [DOI] [PubMed] [Google Scholar]
69.Miki C., Tanaka K., Inoue Y., Araki T., Ohi M., Mohri Y., Uchida K., Kusunoki M. Perioperative host-tumor inflammatory interactions: a potential trigger for disease recurrence following a curative resection for colorectal cancer. Surg. Today. 2008;38:579–584. doi: 10.1007/s00595-007-3674-6. [DOI] [PubMed] [Google Scholar]
70.Goldfarb Y., Ben-Eliyahu S. Surgery as a risk factor for breast cancer recurrence and metastasis: mediating mechanisms and clinical prophylactic approaches. Breast Dis. 2006;26:99–114. doi: 10.3233/bd-2007-26109. [DOI] [PubMed] [Google Scholar]
71.Jamieson N.B., Glen P., McMillan D.C., McKay C.J., Foulis A.K., Carter R., Imrie C.W. Systemic inflammatory response predicts outcome in patients undergoing resection for ductal adenocarcinoma head of pancreas. Br. J. Cancer. 2005;92:21–23. doi: 10.1038/sj.bjc.6602305. [DOI] [PMC free article] [PubMed] [Google Scholar]
72.Rock C.L., Thomson C.A., Sullivan K.R., Howe C.L., Kushi L.H., Caan B.J., Neuhouser M.L., Bandera E.V., Wang Y., Robien K., et al. American Cancer Society nutrition and physical activity guideline for cancer survivors. CA Cancer J. Clin. 2022;72:230–262. doi: 10.3322/caac.21719. [DOI] [PubMed] [Google Scholar]
73.Wang Q., Zhou W. Roles and molecular mechanisms of physical exercise in cancer prevention and treatment. J. Sport Health Sci. 2021;10:201–210. doi: 10.1016/j.jshs.2020.07.008. [DOI] [PMC free article] [PubMed] [Google Scholar]
74.Brown J.C., Ma C., Shi Q., Niedzwiecki D., Zemla T., Couture F., Kuebler P., Kumar P., Hopkins J.O., Tan B., et al. Association between physical activity and the time course of cancer recurrence in stage III colon cancer. Br. J. Sports Med. 2023;57:965–971. doi: 10.1136/bjsports-2022-106445. [DOI] [PMC free article] [PubMed] [Google Scholar]
75.Huang Y., Jiang H., Xu L., Wu X., Wu J., Zhang Y. 2-4 weeks is the optimal time to operate on colorectal liver metastasis after neoadjuvant chemotherapy. Biosci. Trends. 2023;17:160–167. doi: 10.5582/bst.2022.01432. [DOI] [PubMed] [Google Scholar]
76.Emery A., Moore S., Turner J.E., Campbell J.P. Reframing How Physical Activity Reduces The Incidence of Clinically-Diagnosed Cancers: Appraising Exercise-Induced Immuno-Modulation As An Integral Mechanism. Front. Oncol. 2022;12 doi: 10.3389/fonc.2022.788113. [DOI] [PMC free article] [PubMed] [Google Scholar]
77.Copes L.E., Schutz H., Dlugosz E.M., Acosta W., Chappell M.A., Garland T., Jr. Effects of voluntary exercise on spontaneous physical activity and food consumption in mice: Results from an artificial selection experiment. Physiol. Behav. 2015;149:86–94. doi: 10.1016/j.physbeh.2015.05.025. [DOI] [PubMed] [Google Scholar]
78.Liu W., Zhou X., Yao Q., Chen C., Zhang Q., Ding K., Li L., Zeng Z. In situ expansion and reprogramming of Kupffer cells elicit potent tumoricidal immunity against liver metastasis. J. Clin. Investig. 2023;133 doi: 10.1172/jci157937. [DOI] [PMC free article] [PubMed] [Google Scholar]
79.Pedersen L., Idorn M., Olofsson G.H., Lauenborg B., Nookaew I., Hansen R.H., Johannesen H.H., Becker J.C., Pedersen K.S., Dethlefsen C., et al. Voluntary Running Suppresses Tumor Growth through Epinephrine- and IL-6-Dependent NK Cell Mobilization and Redistribution. Cell Metab. 2016;23:554–562. doi: 10.1016/j.cmet.2016.01.011. [DOI] [PubMed] [Google Scholar]
80.Wang Y., Zhong X., He X., Hu Z., Huang H., Chen J., Chen K., Zhao S., Wei P., Li D. Liver metastasis from colorectal cancer: pathogenetic development, immune landscape of the tumour microenvironment and therapeutic approaches. J. Exp. Clin. Cancer Res. 2023;42:177. doi: 10.1186/s13046-023-02729-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
81.Nagarsheth N., Peng D., Kryczek I., Wu K., Li W., Zhao E., Zhao L., Wei S., Frankel T., Vatan L., et al. PRC2 Epigenetically Silences Th1-Type Chemokines to Suppress Effector T-Cell Trafficking in Colon Cancer. Cancer Res. 2016;76:275–282. doi: 10.1158/0008-5472.Can-15-1938. [DOI] [PMC free article] [PubMed] [Google Scholar]
82.Orange S.T., Leslie J., Ross M., Mann D.A., Wackerhage H. The exercise IL-6 enigma in cancer. Trends Endocrinol. Metab. 2023;34:749–763. doi: 10.1016/j.tem.2023.08.001. [DOI] [PubMed] [Google Scholar]
83.Chow L.S., Gerszten R.E., Taylor J.M., Pedersen B.K., van Praag H., Trappe S., Febbraio M.A., Galis Z.S., Gao Y., Haus J.M., et al. Exerkines in health, resilience and disease. Nat. Rev. Endocrinol. 2022;18:273–289. doi: 10.1038/s41574-022-00641-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
84.Yan M., Man S., Sun B., Ma L., Guo L., Huang L., Gao W. Gut liver brain axis in diseases: the implications for therapeutic interventions. Signal Transduct. Targeted Ther. 2023;8:443. doi: 10.1038/s41392-023-01673-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
85.Song M., Chan A.T., Sun J. Influence of the Gut Microbiome, Diet, and Environment on Risk of Colorectal Cancer. Gastroenterology. 2020;158:322–340. doi: 10.1053/j.gastro.2019.06.048. [DOI] [PMC free article] [PubMed] [Google Scholar]
86.Klement J.D., Redd P.S., Lu C., Merting A.D., Poschel D.B., Yang D., Savage N.M., Zhou G., Munn D.H., Fallon P.G., Liu K. Tumor PD-L1 engages myeloid PD-1 to suppress type I interferon to impair cytotoxic T lymphocyte recruitment. Cancer Cell. 2023;41:620–636.e9. doi: 10.1016/j.ccell.2023.02.005. [DOI] [PMC free article] [PubMed] [Google Scholar]
87.Zhu G., Yan H.H., Pang Y., Jian J., Achyut B.R., Liang X., Weiss J.M., Wiltrout R.H., Hollander M.C., Yang L. CXCR3 as a molecular target in breast cancer metastasis: inhibition of tumor cell migration and promotion of host anti-tumor immunity. Oncotarget. 2015;6:43408–43419. doi: 10.18632/oncotarget.6125. [DOI] [PMC free article] [PubMed] [Google Scholar]
88.Ma X., Zhou Z., Zhang X., Fan M., Hong Y., Feng Y., Dong Q., Diao H., Wang G. Sodium butyrate modulates gut microbiota and immune response in colorectal cancer liver metastatic mice. Cell Biol. Toxicol. 2020;36:509–515. doi: 10.1007/s10565-020-09518-4. [DOI] [PubMed] [Google Scholar]
89.Deng S., Hu Q., Zhang H., Yang F., Peng C., Huang C. HDAC3 Inhibition Upregulates PD-L1 Expression in B-Cell Lymphomas and Augments the Efficacy of Anti-PD-L1 Therapy. Mol. Cancer Ther. 2019;18:900–908. doi: 10.1158/1535-7163.Mct-18-1068. [DOI] [PubMed] [Google Scholar]
90.Chambers L.M., Esakov Rhoades E.L., Bharti R., Braley C., Tewari S., Trestan L., Alali Z., Bayik D., Lathia J.D., Sangwan N., et al. Disruption of the Gut Microbiota Confers Cisplatin Resistance in Epithelial Ovarian Cancer. Cancer Res. 2022;82:4654–4669. doi: 10.1158/0008-5472.Can-22-0455. [DOI] [PMC free article] [PubMed] [Google Scholar]
91.Roca Suarez A.A., Plissonnier M.L., Michelet M., Dubois A., Heintz S., Saez-Palma M., Delphin M., Bordes I., Molle J., Diederichs A., et al. Protocol for isolating CD163(+) Kupffer cells from human liver resections. STAR Protoc. 2024;5 doi: 10.1016/j.xpro.2024.103359. [DOI] [PMC free article] [PubMed] [Google Scholar]
92.Wang Z., Zhang H., Liu R., Qian T., Liu J., Huang E., Lu Z., Zhao C., Wang L., Chu Y. Peyer's patches-derived CD11b(+) B cells recruit regulatory T cells through CXCL9 in dextran sulphate sodium-induced colitis. Immunology. 2018;155:356–366. doi: 10.1111/imm.12977. [DOI] [PMC free article] [PubMed] [Google Scholar]
93.Lareau C.A., Liu V., Muus C., Praktiknjo S.D., Nitsch L., Kautz P., Sandor K., Yin Y., Gutierrez J.C., Pelka K., et al. Mitochondrial single-cell ATAC-seq for high-throughput multi-omic detection of mitochondrial genotypes and chromatin accessibility. Nat. Protoc. 2023;18:1416–1440. doi: 10.1038/s41596-022-00795-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
94.Grebe K.M., Hickman H.D., Irvine K.R., Takeda K., Bennink J.R., Yewdell J.W. Sympathetic nervous system control of anti-influenza CD8+ T cell responses. Proc. Natl. Acad. Sci. USA. 2009;106:5300–5305. doi: 10.1073/pnas.0808851106. [DOI] [PMC free article] [PubMed] [Google Scholar]
95.Finney H.M., Akbar A.N., Lawson A.D.G. Activation of resting human primary T cells with chimeric receptors: costimulation from CD28, inducible costimulator, CD134, and CD137 in series with signals from the TCR zeta chain. J. Immunol. 2004;172:104–113. doi: 10.4049/jimmunol.172.1.104. [DOI] [PubMed] [Google Scholar]
96.Tian Y., Wen C., Zhang Z., Liu Y., Li F., Zhao Q., Yao C., Ni K., Yang S., Zhang Y. CXCL9-modified CAR T cells improve immune cell infiltration and antitumor efficacy. Cancer Immunol. Immunother. 2022;71:2663–2675. doi: 10.1007/s00262-022-03193-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
97.de Mingo Pulido Á., Gardner A., Hiebler S., Soliman H., Rugo H.S., Krummel M.F., Coussens L.M., Ruffell B. TIM-3 regulates CD103+ dendritic cell function and response to chemotherapy in breast cancer. Cancer Cell. 2018;33:60–74.e6. doi: 10.1016/j.ccell.2017.11.019. [DOI] [PMC free article] [PubMed] [Google Scholar]
98.Pham T.N., Spaulding C., Shields M.A., Metropulos A.E., Shah D.N., Khalafalla M.G., Principe D.R., Bentrem D.J., Munshi H.G. Inhibition of MNKs promotes macrophage immunosuppressive phenotype to limit CD8+ T cell antitumor immunity. JCI Insight. 2022;7 doi: 10.1172/jci.insight.152731. [DOI] [PMC free article] [PubMed] [Google Scholar]
99.Zhang H., Wang Y., Onuma A., He J., Wang H., Xia Y., Lal R., Cheng X., Kasumova G., Hu Z., et al. Neutrophils Extracellular Traps Inhibition Improves PD-1 Blockade Immunotherapy in Colorectal Cancer. Cancers (Basel) 2021;13 doi: 10.3390/cancers13215333. [DOI] [PMC free article] [PubMed] [Google Scholar]
100.Rintala A., Pietilä S., Munukka E., Eerola E., Pursiheimo J.-P., Laiho A., Pekkala S., Huovinen P. Gut microbiota analysis results are highly dependent on the 16S rRNA gene target region, whereas the impact of DNA extraction is minor. J. Biomol. Tech. 2017;28:19–30. doi: 10.7171/jbt.17-2801-003. [DOI] [PMC free article] [PubMed] [Google Scholar]
101.Wang W., Fang D., Zhang H., Xue J., Wangchuk D., Du J., Jiang L. Sodium Butyrate Selectively Kills Cancer Cells and Inhibits Migration in Colorectal Cancer by Targeting Thioredoxin-1. OncoTargets Ther. 2020;13:4691–4704. doi: 10.2147/ott.s235575. [DOI] [PMC free article] [PubMed] [Google Scholar]
102.Yu X., Yang F., Jiang H., Fan L. RGFP966 Suppresses Tumor Growth and Migration Through Inhibition of EGFR Expression in Hepatocellular Carcinoma Cells in vitro. Drug Des. Devel. Ther. 2020;14:121–128. doi: 10.2147/dddt.S234871. [DOI] [PMC free article] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Document S1. Figures S1–S9 and Table S1

mmc1.pdf^{(1.7MB, pdf)}

Document S2. Article plus supplemental information

mmc2.pdf^{(32.6MB, pdf)}

Data Availability Statement

This paper does not report original code.

Any additional information required to reanalyze the data reported in this work is available from the lead contact upon request.

[bib1] 1.Siegel R.L., Wagle N.S., Cercek A., Smith R.A., Jemal A. Colorectal cancer statistics, 2023. CA Cancer J. Clin. 2023;73:233–254. doi: 10.3322/caac.21772. [DOI] [PubMed] [Google Scholar]

[bib2] 2.Zhou H., Liu Z., Wang Y., Wen X., Amador E.H., Yuan L., Ran X., Xiong L., Ran Y., Chen W., Wen Y. Colorectal liver metastasis: molecular mechanism and interventional therapy. Signal Transduct. Targeted Ther. 2022;7:70. doi: 10.1038/s41392-022-00922-2. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib3] 3.Nordlinger B., Sorbye H., Glimelius B., Poston G.J., Schlag P.M., Rougier P., Bechstein W.O., Primrose J.N., Walpole E.T., Finch-Jones M., et al. Perioperative FOLFOX4 chemotherapy and surgery versus surgery alone for resectable liver metastases from colorectal cancer (EORTC 40983): long-term results of a randomised, controlled, phase 3 trial. Lancet Oncol. 2013;14:1208–1215. doi: 10.1016/s1470-2045(13)70447-9. [DOI] [PubMed] [Google Scholar]

[bib4] 4.Zhang Y., Liu G., Zeng Q., Wu W., Lei K., Zhang C., Tang M., Zhang Y., Xiang X., Tan L., et al. CCL19-producing fibroblasts promote tertiary lymphoid structure formation enhancing anti-tumor IgG response in colorectal cancer liver metastasis. Cancer Cell. 2024;42:1370–1385.e9. doi: 10.1016/j.ccell.2024.07.006. [DOI] [PubMed] [Google Scholar]

[bib5] 5.de Jong M.C., Pulitano C., Ribero D., Strub J., Mentha G., Schulick R.D., Choti M.A., Aldrighetti L., Capussotti L., Pawlik T.M. Rates and patterns of recurrence following curative intent surgery for colorectal liver metastasis: an international multi-institutional analysis of 1669 patients. Ann. Surg. 2009;250:440–448. doi: 10.1097/SLA.0b013e3181b4539b. [DOI] [PubMed] [Google Scholar]

[bib6] 6.Heinz M.C., Peters N.A., Oost K.C., Lindeboom R.G.H., van Voorthuijsen L., Fumagalli A., van der Net M.C., de Medeiros G., Hageman J.H., Verlaan-Klink I., et al. Liver Colonization by Colorectal Cancer Metastases Requires YAP-Controlled Plasticity at the Micrometastatic Stage. Cancer Res. 2022;82:1953–1968. doi: 10.1158/0008-5472.Can-21-0933. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib7] 7.Weitz J., Kienle P., Lacroix J., Willeke F., Benner A., Lehnert T., Herfarth C., von Knebel Doeberitz M. Dissemination of tumor cells in patients undergoing surgery for colorectal cancer. Clin. Cancer Res. 1998;4:343–348. [PubMed] [Google Scholar]

[bib8] 8.Rahbari N.N., Birgin E., Bork U., Mehrabi A., Reißfelder C., Weitz J. Anterior Approach vs Conventional Hepatectomy for Resection of Colorectal Liver Metastasis: A Randomized Clinical Trial. JAMA Surg. 2021;156:31–40. doi: 10.1001/jamasurg.2020.5050. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib9] 9.Hiller J.G., Perry N.J., Poulogiannis G., Riedel B., Sloan E.K. Perioperative events influence cancer recurrence risk after surgery. Nat. Rev. Clin. Oncol. 2018;15:205–218. doi: 10.1038/nrclinonc.2017.194. [DOI] [PubMed] [Google Scholar]

[bib10] 10.Quail D.F., Joyce J.A. Microenvironmental regulation of tumor progression and metastasis. Nat. Med. 2013;19:1423–1437. doi: 10.1038/nm.3394. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib11] 11.Cao Y., Zheng X., Hu Y., Li J., Huang B., Zhao N., Liu T., Cai K., Tian S. Levels of systemic inflammation response index are correlated with tumor-associated bacteria in colorectal cancer. Cell Death Dis. 2023;14:69. doi: 10.1038/s41419-023-05602-9. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib12] 12.Maspero M., Yilmaz S., Cazzaniga B., Raj R., Ali K., Mazzaferro V., Schlegel A. The role of ischaemia-reperfusion injury and liver regeneration in hepatic tumour recurrence. JHEP Rep. 2023;5 doi: 10.1016/j.jhepr.2023.100846. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib13] 13.Chen Z., Zhang P., Xu Y., Yan J., Liu Z., Lau W.B., Lau B., Li Y., Zhao X., Wei Y., Zhou S. Surgical stress and cancer progression: the twisted tango. Mol. Cancer. 2019;18:132. doi: 10.1186/s12943-019-1058-3. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib14] 14.Cho J.Y., Han H.S., Choi Y., Yoon Y.S., Kim S., Choi J.K., Jang J.S., Kwon S.U., Kim H. Association of Remnant Liver Ischemia With Early Recurrence and Poor Survival After Liver Resection in Patients With Hepatocellular Carcinoma. JAMA Surg. 2017;152:386–392. doi: 10.1001/jamasurg.2016.5040. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib15] 15.Nicoud I.B., Jones C.M., Pierce J.M., Earl T.M., Matrisian L.M., Chari R.S., Gorden D.L. Warm hepatic ischemia-reperfusion promotes growth of colorectal carcinoma micrometastases in mouse liver via matrix metalloproteinase-9 induction. Cancer Res. 2007;67:2720–2728. doi: 10.1158/0008-5472.Can-06-3923. [DOI] [PubMed] [Google Scholar]

[bib16] 16.van der Bilt J.D.W., Kranenburg O., Nijkamp M.W., Smakman N., Veenendaal L.M., Te Velde E.A., Voest E.E., van Diest P.J., Borel Rinkes I.H.M. Ischemia/reperfusion accelerates the outgrowth of hepatic micrometastases in a highly standardized murine model. Hepatology. 2005;42:165–175. doi: 10.1002/hep.20739. [DOI] [PubMed] [Google Scholar]

[bib17] 17.Greten F.R., Grivennikov S.I. Inflammation and Cancer: Triggers, Mechanisms, and Consequences. Immunity. 2019;51:27–41. doi: 10.1016/j.immuni.2019.06.025. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib18] 18.Tohme S., Simmons R.L., Tsung A. Surgery for Cancer: A Trigger for Metastases. Cancer Res. 2017;77:1548–1552. doi: 10.1158/0008-5472.Can-16-1536. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib19] 19.Hojman P., Gehl J., Christensen J.F., Pedersen B.K. Molecular Mechanisms Linking Exercise to Cancer Prevention and Treatment. Cell Metab. 2018;27:10–21. doi: 10.1016/j.cmet.2017.09.015. [DOI] [PubMed] [Google Scholar]

[bib20] 20.Zhang Y., Cao C., Li C., Witt R.G., Huang H., Tsung A., Zhang H. Physical exercise in liver diseases. Hepatology. 2024 doi: 10.1097/hep.0000000000000941. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib21] 21.Idorn M., Hojman P. Exercise-Dependent Regulation of NK Cells in Cancer Protection. Trends Mol. Med. 2016;22:565–577. doi: 10.1016/j.molmed.2016.05.007. [DOI] [PubMed] [Google Scholar]

[bib22] 22.Schmid D., Leitzmann M.F. Association between physical activity and mortality among breast cancer and colorectal cancer survivors: a systematic review and meta-analysis. Ann. Oncol. 2014;25:1293–1311. doi: 10.1093/annonc/mdu012. [DOI] [PubMed] [Google Scholar]

[bib23] 23.Clifford K., Woodfield J.C., Tait W., Campbell H.A., Baldi J.C. Association of Preoperative High-Intensity Interval Training With Cardiorespiratory Fitness and Postoperative Outcomes Among Adults Undergoing Major Surgery: A Systematic Review and Meta-Analysis. JAMA Netw. Open. 2023;6 doi: 10.1001/jamanetworkopen.2023.20527. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib24] 24.Heinz A., Hiller K. Run to survive-how preoperative exercise could prevent major surgical complications. Nat. Metab. 2021;3:732–734. doi: 10.1038/s42255-021-00403-w. [DOI] [PubMed] [Google Scholar]

[bib25] 25.Kurz E., Hirsch C.A., Dalton T., Shadaloey S.A., Khodadadi-Jamayran A., Miller G., Pareek S., Rajaei H., Mohindroo C., Baydogan S., et al. Exercise-induced engagement of the IL-15/IL-15Rα axis promotes anti-tumor immunity in pancreatic cancer. Cancer Cell. 2022;40:720–737.e5. doi: 10.1016/j.ccell.2022.05.006. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib26] 26.Koelwyn G.J., Zhuang X., Tammela T., Schietinger A., Jones L.W. Exercise and immunometabolic regulation in cancer. Nat. Metab. 2020;2:849–857. doi: 10.1038/s42255-020-00277-4. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib27] 27.Zhang H., Chen T., Ren J., Xia Y., Onuma A., Wang Y., He J., Wu J., Wang H., Hamad A., et al. Pre-operative exercise therapy triggers anti-inflammatory trained immunity of Kupffer cells through metabolic reprogramming. Nat. Metab. 2021;3:843–858. doi: 10.1038/s42255-021-00402-x. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib28] 28.Wen Y., Lambrecht J., Ju C., Tacke F. Hepatic macrophages in liver homeostasis and diseases-diversity, plasticity and therapeutic opportunities. Cell. Mol. Immunol. 2021;18:45–56. doi: 10.1038/s41423-020-00558-8. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib29] 29.Thomas S.K., Wattenberg M.M., Choi-Bose S., Uhlik M., Harrison B., Coho H., Cassella C.R., Stone M.L., Patel D., Markowitz K., et al. Kupffer cells prevent pancreatic ductal adenocarcinoma metastasis to the liver in mice. Nat. Commun. 2023;14:6330. doi: 10.1038/s41467-023-41771-z. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib30] 30.Tsilimigras D.I., Brodt P., Clavien P.A., Muschel R.J., D'Angelica M.I., Endo I., Parks R.W., Doyle M., de Santibañes E., Pawlik T.M. Liver metastases. Nat. Rev. Dis. Primers. 2021;7:27. doi: 10.1038/s41572-021-00261-6. [DOI] [PubMed] [Google Scholar]

[bib31] 31.Ozga A.J., Chow M.T., Luster A.D. Chemokines and the immune response to cancer. Immunity. 2021;54:859–874. doi: 10.1016/j.immuni.2021.01.012. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib32] 32.Kimura Y., Inoue A., Hangai S., Saijo S., Negishi H., Nishio J., Yamasaki S., Iwakura Y., Yanai H., Taniguchi T. The innate immune receptor Dectin-2 mediates the phagocytosis of cancer cells by Kupffer cells for the suppression of liver metastasis. Proc. Natl. Acad. Sci. USA. 2016;113:14097–14102. doi: 10.1073/pnas.1617903113. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib33] 33.Li J., Liu X.G., Ge R.L., Yin Y.P., Liu Y.D., Lu W.P., Huang M., He X.Y., Wang J., Cai G., et al. The ligation between ERMAP, galectin-9 and dectin-2 promotes Kupffer cell phagocytosis and antitumor immunity. Nat. Immunol. 2023;24:1813–1824. doi: 10.1038/s41590-023-01634-7. [DOI] [PubMed] [Google Scholar]

[bib34] 34.Deng Z., Loyher P.L., Lazarov T., Li L., Shen Z., Bhinder B., Yang H., Zhong Y., Alberdi A., Massague J., et al. The nuclear factor ID3 endows macrophages with a potent anti-tumour activity. Nature. 2024;626:864–873. doi: 10.1038/s41586-023-06950-4. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib35] 35.Yuan D., Huang S., Berger E., Liu L., Gross N., Heinzmann F., Ringelhan M., Connor T.O., Stadler M., Meister M., et al. Kupffer Cell-Derived Tnf Triggers Cholangiocellular Tumorigenesis through JNK due to Chronic Mitochondrial Dysfunction and ROS. Cancer Cell. 2017;31:771–789.e6. doi: 10.1016/j.ccell.2017.05.006. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib36] 36.Huang H.Y., Chen Y.Z., Zhao C., Zheng X.N., Yu K., Yue J.X., Ju H.Q., Shi Y.X., Tian L. Alternations in inflammatory macrophage niche drive phenotypic and functional plasticity of Kupffer cells. Nat. Commun. 2024;15:9337. doi: 10.1038/s41467-024-53659-7. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib37] 37.Xia Y., He J., Zhang H., Wang H., Tetz G., Maguire C.A., Wang Y., Onuma A., Genkin D., Tetz V., et al. AAV-mediated gene transfer of DNase I in the liver of mice with colorectal cancer reduces liver metastasis and restores local innate and adaptive immune response. Mol. Oncol. 2020;14:2920–2935. doi: 10.1002/1878-0261.12787. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib38] 38.House I.G., Savas P., Lai J., Chen A.X.Y., Oliver A.J., Teo Z.L., Todd K.L., Henderson M.A., Giuffrida L., Petley E.V., et al. Macrophage-Derived CXCL9 and CXCL10 Are Required for Antitumor Immune Responses Following Immune Checkpoint Blockade. Clin. Cancer Res. 2020;26:487–504. doi: 10.1158/1078-0432.Ccr-19-1868. [DOI] [PubMed] [Google Scholar]

[bib39] 39.Kuhn N.F., Purdon T.J., van Leeuwen D.G., Lopez A.V., Curran K.J., Daniyan A.F., Brentjens R.J. CD40 Ligand-Modified Chimeric Antigen Receptor T Cells Enhance Antitumor Function by Eliciting an Endogenous Antitumor Response. Cancer Cell. 2019;35:473–488.e6. doi: 10.1016/j.ccell.2019.02.006. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib40] 40.Ghodke I., Remisova M., Furst A., Kilic S., Reina-San-Martin B., Poetsch A.R., Altmeyer M., Soutoglou E. AHNAK controls 53BP1-mediated p53 response by restraining 53BP1 oligomerization and phase separation. Mol. Cell. 2021;81:2596–2610.e7. doi: 10.1016/j.molcel.2021.04.010. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib41] 41.Lim R.J., Salehi-Rad R., Tran L.M., Oh M.S., Dumitras C., Crosson W.P., Li R., Patel T.S., Man S., Yean C.E., et al. CXCL9/10-engineered dendritic cells promote T cell activation and enhance immune checkpoint blockade for lung cancer. Cell Rep. Med. 2024;5 doi: 10.1016/j.xcrm.2024.101479. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib42] 42.Gao F., He S., Jin A. MiRNAs and lncRNAs in NK cell biology and NK/T-cell lymphoma. Genes Dis. 2021;8:590–602. doi: 10.1016/j.gendis.2020.08.006. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib43] 43.Chen D., Varanasi S.K., Hara T., Traina K., Sun M., McDonald B., Farsakoglu Y., Clanton J., Xu S., Garcia-Rivera L., et al. CTLA-4 blockade induces a microglia-Th1 cell partnership that stimulates microglia phagocytosis and anti-tumor function in glioblastoma. Immunity. 2023;56:2086–2104.e8. doi: 10.1016/j.immuni.2023.07.015. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib44] 44.Ma R.Y., Black A., Qian B.Z. Macrophage diversity in cancer revisited in the era of single-cell omics. Trends Immunol. 2022;43:546–563. doi: 10.1016/j.it.2022.04.008. [DOI] [PubMed] [Google Scholar]

[bib45] 45.Hinshaw D.C., Shevde L.A. The Tumor Microenvironment Innately Modulates Cancer Progression. Cancer Res. 2019;79:4557–4566. doi: 10.1158/0008-5472.Can-18-3962. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib46] 46.Liu R.Z., Choi W.S., Jain S., Xu X., Elsherbiny M.E., Glubrecht D.D., Tessier A.G., Easaw J.C., Fallone B.G., Godbout R. Stationary-to-migratory transition in glioblastoma stem-like cells driven by a fatty acid-binding protein 7-RXRα neurogenic pathway. Neuro Oncol. 2023;25:2177–2190. doi: 10.1093/neuonc/noad134. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib47] 47.Li Y., Qu P., Wu L., Li B., Du H., Yan C. Api6/AIM/Spα/CD5L overexpression in alveolar type II epithelial cells induces spontaneous lung adenocarcinoma. Cancer Res. 2011;71:5488–5499. doi: 10.1158/0008-5472.Can-10-4225. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib48] 48.Shen J., Zhou L., Ye K., Gong J., Wu F., Mo K., Zhu Y., Chen C., Zhan R. The role of SPI1/VSIG4/THBS1 on glioblastoma progression through modulation of the PI3K/AKT pathway. J. Adv. Res. 2025;71:487–500. doi: 10.1016/j.jare.2024.06.023. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib49] 49.Czech L., Mais C.-N., Kratzat H., Sarmah P., Giammarinaro P., Freibert S.-A., Esser H.F., Musial J., Berninghausen O., Steinchen W., et al. Inhibition of SRP-dependent protein secretion by the bacterial alarmone (p)ppGpp. Nat. Commun. 2022;13:1069. doi: 10.1038/s41467-022-28675-0. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib50] 50.Giannou A.D., Kempski J., Shiri A.M., Lücke J., Zhang T., Zhao L., Zazara D.E., Cortesi F., Riecken K., Amezcua Vesely M.C., et al. Tissue resident iNKT17 cells facilitate cancer cell extravasation in liver metastasis via interleukin-22. Immunity. 2023;56:125–142.e12. doi: 10.1016/j.immuni.2022.12.014. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib51] 51.Zhao H., Wu L., Yan G., Chen Y., Zhou M., Wu Y., Li Y. Inflammation and tumor progression: signaling pathways and targeted intervention. Signal Transduct. Targeted Ther. 2021;6:263. doi: 10.1038/s41392-021-00658-5. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib52] 52.Han J., Wu M., Liu Z. Dysregulation in IFN-γ signaling and response: the barricade to tumor immunotherapy. Front. Immunol. 2023;14 doi: 10.3389/fimmu.2023.1190333. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib53] 53.Tokunaga R., Zhang W., Naseem M., Puccini A., Berger M.D., Soni S., McSkane M., Baba H., Lenz H.J. CXCL9, CXCL10, CXCL11/CXCR3 axis for immune activation - A target for novel cancer therapy. Cancer Treat Rev. 2018;63:40–47. doi: 10.1016/j.ctrv.2017.11.007. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib54] 54.de Mingo Pulido Á., Gardner A., Hiebler S., Soliman H., Rugo H.S., Krummel M.F., Coussens L.M., Ruffell B. TIM-3 Regulates CD103(+) Dendritic Cell Function and Response to Chemotherapy in Breast Cancer. Cancer Cell. 2018;33:60–74.e6. doi: 10.1016/j.ccell.2017.11.019. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib55] 55.Li X., Egervari G., Wang Y., Berger S.L., Lu Z. Regulation of chromatin and gene expression by metabolic enzymes and metabolites. Nat. Rev. Mol. Cell Biol. 2018;19:563–578. doi: 10.1038/s41580-018-0029-7. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib56] 56.Imhann F., Vich Vila A., Bonder M.J., Fu J., Gevers D., Visschedijk M.C., Spekhorst L.M., Alberts R., Franke L., van Dullemen H.M., et al. Interplay of host genetics and gut microbiota underlying the onset and clinical presentation of inflammatory bowel disease. Gut. 2018;67:108–119. doi: 10.1136/gutjnl-2016-312135. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib57] 57.Flemming A. Butyrate boosts microbicidal macrophages. Nat. Rev. Immunol. 2019;19:135. doi: 10.1038/s41577-019-0132-9. [DOI] [PubMed] [Google Scholar]

[bib58] 58.Mann E.R., Lam Y.K., Uhlig H.H. Short-chain fatty acids: linking diet, the microbiome and immunity. Nat. Rev. Immunol. 2024;24:577–595. doi: 10.1038/s41577-024-01014-8. [DOI] [PubMed] [Google Scholar]

[bib59] 59.Rivera-Chávez F., Zhang L.F., Faber F., Lopez C.A., Byndloss M.X., Olsan E.E., Xu G., Velazquez E.M., Lebrilla C.B., Winter S.E., Bäumler A.J. Depletion of Butyrate-Producing Clostridia from the Gut Microbiota Drives an Aerobic Luminal Expansion of Salmonella. Cell Host Microbe. 2016;19:443–454. doi: 10.1016/j.chom.2016.03.004. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib60] 60.Modoux M., Rolhion N., Lefevre J.H., Oeuvray C., Nádvorník P., Illes P., Emond P., Parc Y., Mani S., Dvorak Z., Sokol H. Butyrate acts through HDAC inhibition to enhance aryl hydrocarbon receptor activation by gut microbiota-derived ligands. Gut Microbes. 2022;14 doi: 10.1080/19490976.2022.2105637. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib61] 61.Angiolilli C., Kabala P.A., Grabiec A.M., Van Baarsen I.M., Ferguson B.S., García S., Malvar Fernandez B., McKinsey T.A., Tak P.P., Fossati G., et al. Histone deacetylase 3 regulates the inflammatory gene expression programme of rheumatoid arthritis fibroblast-like synoviocytes. Ann. Rheum. Dis. 2017;76:277–285. doi: 10.1136/annrheumdis-2015-209064. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib62] 62.Zheng H., Zhao W., Yan C., Watson C.C., Massengill M., Xie M., Massengill C., Noyes D.R., Martinez G.V., Afzal R., et al. HDAC Inhibitors Enhance T-Cell Chemokine Expression and Augment Response to PD-1 Immunotherapy in Lung Adenocarcinoma. Clin. Cancer Res. 2016;22:4119–4132. doi: 10.1158/1078-0432.Ccr-15-2584. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib63] 63.van der Bij G.J., Oosterling S.J., Beelen R.H.J., Meijer S., Coffey J.C., van Egmond M. The perioperative period is an underutilized window of therapeutic opportunity in patients with colorectal cancer. Ann. Surg. 2009;249:727–734. doi: 10.1097/SLA.0b013e3181a3ddbd. [DOI] [PubMed] [Google Scholar]

[bib64] 64.Pang L., Yeung O.W.H., Ng K.T.P., Liu H., Zhu J., Liu J., Yang X., Ding T., Qiu W., Wang Y., et al. Postoperative Plasmacytoid Dendritic Cells Secrete IFNα to Promote Recruitment of Myeloid-Derived Suppressor Cells and Drive Hepatocellular Carcinoma Recurrence. Cancer Res. 2022;82:4206–4218. doi: 10.1158/0008-5472.Can-22-1199. [DOI] [PubMed] [Google Scholar]

[bib65] 65.Ren J., He J., Zhang H., Xia Y., Hu Z., Loughran P., Billiar T., Huang H., Tsung A. Platelet TLR4-ERK5 Axis Facilitates NET-Mediated Capturing of Circulating Tumor Cells and Distant Metastasis after Surgical Stress. Cancer Res. 2021;81:2373–2385. doi: 10.1158/0008-5472.Can-20-3222. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib66] 66.Artinyan A., Orcutt S.T., Anaya D.A., Richardson P., Chen G.J., Berger D.H. Infectious postoperative complications decrease long-term survival in patients undergoing curative surgery for colorectal cancer: a study of 12,075 patients. Ann. Surg. 2015;261:497–505. doi: 10.1097/SLA.0000000000000854. [DOI] [PubMed] [Google Scholar]

[bib67] 67.Farid S.G., Aldouri A., Morris-Stiff G., Khan A.Z., Toogood G.J., Lodge J.P.A., Prasad K.R. Correlation between postoperative infective complications and long-term outcomes after hepatic resection for colorectal liver metastasis. Ann. Surg. 2010;251:91–100. doi: 10.1097/SLA.0b013e3181bfda3c. [DOI] [PubMed] [Google Scholar]

[bib68] 68.Gomez D., Morris-Stiff G., Wyatt J., Toogood G.J., Lodge J.P.A., Prasad K.R. Surgical technique and systemic inflammation influences long-term disease-free survival following hepatic resection for colorectal metastasis. J. Surg. Oncol. 2008;98:371–376. doi: 10.1002/jso.21103. [DOI] [PubMed] [Google Scholar]

[bib69] 69.Miki C., Tanaka K., Inoue Y., Araki T., Ohi M., Mohri Y., Uchida K., Kusunoki M. Perioperative host-tumor inflammatory interactions: a potential trigger for disease recurrence following a curative resection for colorectal cancer. Surg. Today. 2008;38:579–584. doi: 10.1007/s00595-007-3674-6. [DOI] [PubMed] [Google Scholar]

[bib70] 70.Goldfarb Y., Ben-Eliyahu S. Surgery as a risk factor for breast cancer recurrence and metastasis: mediating mechanisms and clinical prophylactic approaches. Breast Dis. 2006;26:99–114. doi: 10.3233/bd-2007-26109. [DOI] [PubMed] [Google Scholar]

[bib71] 71.Jamieson N.B., Glen P., McMillan D.C., McKay C.J., Foulis A.K., Carter R., Imrie C.W. Systemic inflammatory response predicts outcome in patients undergoing resection for ductal adenocarcinoma head of pancreas. Br. J. Cancer. 2005;92:21–23. doi: 10.1038/sj.bjc.6602305. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib72] 72.Rock C.L., Thomson C.A., Sullivan K.R., Howe C.L., Kushi L.H., Caan B.J., Neuhouser M.L., Bandera E.V., Wang Y., Robien K., et al. American Cancer Society nutrition and physical activity guideline for cancer survivors. CA Cancer J. Clin. 2022;72:230–262. doi: 10.3322/caac.21719. [DOI] [PubMed] [Google Scholar]

[bib73] 73.Wang Q., Zhou W. Roles and molecular mechanisms of physical exercise in cancer prevention and treatment. J. Sport Health Sci. 2021;10:201–210. doi: 10.1016/j.jshs.2020.07.008. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib74] 74.Brown J.C., Ma C., Shi Q., Niedzwiecki D., Zemla T., Couture F., Kuebler P., Kumar P., Hopkins J.O., Tan B., et al. Association between physical activity and the time course of cancer recurrence in stage III colon cancer. Br. J. Sports Med. 2023;57:965–971. doi: 10.1136/bjsports-2022-106445. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib75] 75.Huang Y., Jiang H., Xu L., Wu X., Wu J., Zhang Y. 2-4 weeks is the optimal time to operate on colorectal liver metastasis after neoadjuvant chemotherapy. Biosci. Trends. 2023;17:160–167. doi: 10.5582/bst.2022.01432. [DOI] [PubMed] [Google Scholar]

[bib76] 76.Emery A., Moore S., Turner J.E., Campbell J.P. Reframing How Physical Activity Reduces The Incidence of Clinically-Diagnosed Cancers: Appraising Exercise-Induced Immuno-Modulation As An Integral Mechanism. Front. Oncol. 2022;12 doi: 10.3389/fonc.2022.788113. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib77] 77.Copes L.E., Schutz H., Dlugosz E.M., Acosta W., Chappell M.A., Garland T., Jr. Effects of voluntary exercise on spontaneous physical activity and food consumption in mice: Results from an artificial selection experiment. Physiol. Behav. 2015;149:86–94. doi: 10.1016/j.physbeh.2015.05.025. [DOI] [PubMed] [Google Scholar]

[bib78] 78.Liu W., Zhou X., Yao Q., Chen C., Zhang Q., Ding K., Li L., Zeng Z. In situ expansion and reprogramming of Kupffer cells elicit potent tumoricidal immunity against liver metastasis. J. Clin. Investig. 2023;133 doi: 10.1172/jci157937. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib79] 79.Pedersen L., Idorn M., Olofsson G.H., Lauenborg B., Nookaew I., Hansen R.H., Johannesen H.H., Becker J.C., Pedersen K.S., Dethlefsen C., et al. Voluntary Running Suppresses Tumor Growth through Epinephrine- and IL-6-Dependent NK Cell Mobilization and Redistribution. Cell Metab. 2016;23:554–562. doi: 10.1016/j.cmet.2016.01.011. [DOI] [PubMed] [Google Scholar]

[bib80] 80.Wang Y., Zhong X., He X., Hu Z., Huang H., Chen J., Chen K., Zhao S., Wei P., Li D. Liver metastasis from colorectal cancer: pathogenetic development, immune landscape of the tumour microenvironment and therapeutic approaches. J. Exp. Clin. Cancer Res. 2023;42:177. doi: 10.1186/s13046-023-02729-7. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib81] 81.Nagarsheth N., Peng D., Kryczek I., Wu K., Li W., Zhao E., Zhao L., Wei S., Frankel T., Vatan L., et al. PRC2 Epigenetically Silences Th1-Type Chemokines to Suppress Effector T-Cell Trafficking in Colon Cancer. Cancer Res. 2016;76:275–282. doi: 10.1158/0008-5472.Can-15-1938. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib82] 82.Orange S.T., Leslie J., Ross M., Mann D.A., Wackerhage H. The exercise IL-6 enigma in cancer. Trends Endocrinol. Metab. 2023;34:749–763. doi: 10.1016/j.tem.2023.08.001. [DOI] [PubMed] [Google Scholar]

[bib83] 83.Chow L.S., Gerszten R.E., Taylor J.M., Pedersen B.K., van Praag H., Trappe S., Febbraio M.A., Galis Z.S., Gao Y., Haus J.M., et al. Exerkines in health, resilience and disease. Nat. Rev. Endocrinol. 2022;18:273–289. doi: 10.1038/s41574-022-00641-2. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib84] 84.Yan M., Man S., Sun B., Ma L., Guo L., Huang L., Gao W. Gut liver brain axis in diseases: the implications for therapeutic interventions. Signal Transduct. Targeted Ther. 2023;8:443. doi: 10.1038/s41392-023-01673-4. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib85] 85.Song M., Chan A.T., Sun J. Influence of the Gut Microbiome, Diet, and Environment on Risk of Colorectal Cancer. Gastroenterology. 2020;158:322–340. doi: 10.1053/j.gastro.2019.06.048. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib86] 86.Klement J.D., Redd P.S., Lu C., Merting A.D., Poschel D.B., Yang D., Savage N.M., Zhou G., Munn D.H., Fallon P.G., Liu K. Tumor PD-L1 engages myeloid PD-1 to suppress type I interferon to impair cytotoxic T lymphocyte recruitment. Cancer Cell. 2023;41:620–636.e9. doi: 10.1016/j.ccell.2023.02.005. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib87] 87.Zhu G., Yan H.H., Pang Y., Jian J., Achyut B.R., Liang X., Weiss J.M., Wiltrout R.H., Hollander M.C., Yang L. CXCR3 as a molecular target in breast cancer metastasis: inhibition of tumor cell migration and promotion of host anti-tumor immunity. Oncotarget. 2015;6:43408–43419. doi: 10.18632/oncotarget.6125. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib88] 88.Ma X., Zhou Z., Zhang X., Fan M., Hong Y., Feng Y., Dong Q., Diao H., Wang G. Sodium butyrate modulates gut microbiota and immune response in colorectal cancer liver metastatic mice. Cell Biol. Toxicol. 2020;36:509–515. doi: 10.1007/s10565-020-09518-4. [DOI] [PubMed] [Google Scholar]

[bib89] 89.Deng S., Hu Q., Zhang H., Yang F., Peng C., Huang C. HDAC3 Inhibition Upregulates PD-L1 Expression in B-Cell Lymphomas and Augments the Efficacy of Anti-PD-L1 Therapy. Mol. Cancer Ther. 2019;18:900–908. doi: 10.1158/1535-7163.Mct-18-1068. [DOI] [PubMed] [Google Scholar]

[bib90] 90.Chambers L.M., Esakov Rhoades E.L., Bharti R., Braley C., Tewari S., Trestan L., Alali Z., Bayik D., Lathia J.D., Sangwan N., et al. Disruption of the Gut Microbiota Confers Cisplatin Resistance in Epithelial Ovarian Cancer. Cancer Res. 2022;82:4654–4669. doi: 10.1158/0008-5472.Can-22-0455. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib91] 91.Roca Suarez A.A., Plissonnier M.L., Michelet M., Dubois A., Heintz S., Saez-Palma M., Delphin M., Bordes I., Molle J., Diederichs A., et al. Protocol for isolating CD163(+) Kupffer cells from human liver resections. STAR Protoc. 2024;5 doi: 10.1016/j.xpro.2024.103359. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib92] 92.Wang Z., Zhang H., Liu R., Qian T., Liu J., Huang E., Lu Z., Zhao C., Wang L., Chu Y. Peyer's patches-derived CD11b(+) B cells recruit regulatory T cells through CXCL9 in dextran sulphate sodium-induced colitis. Immunology. 2018;155:356–366. doi: 10.1111/imm.12977. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib93] 93.Lareau C.A., Liu V., Muus C., Praktiknjo S.D., Nitsch L., Kautz P., Sandor K., Yin Y., Gutierrez J.C., Pelka K., et al. Mitochondrial single-cell ATAC-seq for high-throughput multi-omic detection of mitochondrial genotypes and chromatin accessibility. Nat. Protoc. 2023;18:1416–1440. doi: 10.1038/s41596-022-00795-3. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib94] 94.Grebe K.M., Hickman H.D., Irvine K.R., Takeda K., Bennink J.R., Yewdell J.W. Sympathetic nervous system control of anti-influenza CD8+ T cell responses. Proc. Natl. Acad. Sci. USA. 2009;106:5300–5305. doi: 10.1073/pnas.0808851106. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib95] 95.Finney H.M., Akbar A.N., Lawson A.D.G. Activation of resting human primary T cells with chimeric receptors: costimulation from CD28, inducible costimulator, CD134, and CD137 in series with signals from the TCR zeta chain. J. Immunol. 2004;172:104–113. doi: 10.4049/jimmunol.172.1.104. [DOI] [PubMed] [Google Scholar]

[bib96] 96.Tian Y., Wen C., Zhang Z., Liu Y., Li F., Zhao Q., Yao C., Ni K., Yang S., Zhang Y. CXCL9-modified CAR T cells improve immune cell infiltration and antitumor efficacy. Cancer Immunol. Immunother. 2022;71:2663–2675. doi: 10.1007/s00262-022-03193-6. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib97] 97.de Mingo Pulido Á., Gardner A., Hiebler S., Soliman H., Rugo H.S., Krummel M.F., Coussens L.M., Ruffell B. TIM-3 regulates CD103+ dendritic cell function and response to chemotherapy in breast cancer. Cancer Cell. 2018;33:60–74.e6. doi: 10.1016/j.ccell.2017.11.019. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib98] 98.Pham T.N., Spaulding C., Shields M.A., Metropulos A.E., Shah D.N., Khalafalla M.G., Principe D.R., Bentrem D.J., Munshi H.G. Inhibition of MNKs promotes macrophage immunosuppressive phenotype to limit CD8+ T cell antitumor immunity. JCI Insight. 2022;7 doi: 10.1172/jci.insight.152731. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib99] 99.Zhang H., Wang Y., Onuma A., He J., Wang H., Xia Y., Lal R., Cheng X., Kasumova G., Hu Z., et al. Neutrophils Extracellular Traps Inhibition Improves PD-1 Blockade Immunotherapy in Colorectal Cancer. Cancers (Basel) 2021;13 doi: 10.3390/cancers13215333. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib100] 100.Rintala A., Pietilä S., Munukka E., Eerola E., Pursiheimo J.-P., Laiho A., Pekkala S., Huovinen P. Gut microbiota analysis results are highly dependent on the 16S rRNA gene target region, whereas the impact of DNA extraction is minor. J. Biomol. Tech. 2017;28:19–30. doi: 10.7171/jbt.17-2801-003. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib101] 101.Wang W., Fang D., Zhang H., Xue J., Wangchuk D., Du J., Jiang L. Sodium Butyrate Selectively Kills Cancer Cells and Inhibits Migration in Colorectal Cancer by Targeting Thioredoxin-1. OncoTargets Ther. 2020;13:4691–4704. doi: 10.2147/ott.s235575. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib102] 102.Yu X., Yang F., Jiang H., Fan L. RGFP966 Suppresses Tumor Growth and Migration Through Inhibition of EGFR Expression in Hepatocellular Carcinoma Cells in vitro. Drug Des. Devel. Ther. 2020;14:121–128. doi: 10.2147/dddt.S234871. [DOI] [PMC free article] [PubMed] [Google Scholar]

PERMALINK

Preoperative exercise induces anti-tumor Kupffer cells to prevent surgical stress-promoted colorectal cancer liver metastasis

Yunwei Zhang

Yiyu Zhang

Chengli Shen

Jiye Li

Han Wang

Ahmad Hamad

Chunyan Cao

Mohamad El Moheb

Yu Wang

Yujia Xia

Kaelyn C Cummins

Joal Beane

Aslam Ejaz

Hai Huang

Allan Tsung

Hongji Zhang

Summary

Graphical abstract

Highlights

Introduction

Results

PEx suppresses surgical stress-promoted highly immunogenic CRLM and remodels the liver immune microenvironment

Figure 1.

KCs are essential for alleviating surgical stress-promoted CRLM following PEx

Figure 2.

PEx enhances KC’s anti-tumor immunity by modulating cytokine secretion

PEx increases CXCL9 release from KCs and promotes CXCR3+ CD8+ T cells recruitment

Figure 3.

The CXCL9-CXCR3 axis is crucial for PEx-mediated protection against surgical stress-promoted CRLM

Figure 4.

CXCL9 enhances anti-tumor activity of CXCR3+ CD8+ T cells

Figure 5.

PEx-induced remodeling of the gut microbiota contributes to the alleviation of surgical stress-promoted CRLM

Figure 6.

PEx promotes CXCL9 expression in KCs via butyrate-mediated HDAC3 inhibition

Figure 7.

Discussion

Limitations of the study

Resource availability

Lead contact

Materials availability

Data and code availability

Acknowledgments

Author contributions

Declaration of interests

STAR★Methods

Key resources table

Experimental model and study participant details

Patient recruitment & sample acquisition

Mice

Tumor cell lines

Method details

Surgical stress-promoted CRLM model

Treatment of tumor-bearing mice

Antibiotics administration

KCs isolation and treatment

Depletion and adoptive transfer of KCs

Isolation of KCs’ nuclei

CD8+ T cells isolation and treatment

Adoptive transfer of CD8+ T cells

IVIS

Flow cytometry

ELISA

Single-cell RNA-sequencing

Sample and library preparation

Sequencing

Gene expression analysis

Cell type annotation

Ligand-receptor interaction analysis

KCs untargeted metabolomics analysis

16s RNA-sequencing

Western blotting

Cell growth assays

Quantification and statistical analysis

Additional resources

Footnotes

Contributor Information

Supplemental information

PEx increases CXCL9 release from KCs and promotes CXCR3⁺ CD8⁺ T cells recruitment

CXCL9 enhances anti-tumor activity of CXCR3⁺ CD8⁺ T cells

CD8⁺ T cells isolation and treatment

Adoptive transfer of CD8⁺ T cells