Colorectal Carcinoma Liver Metastases Research Articles

Are survival outcomes dependent on the tumor dose threshold of 139 Gy in patients with chemorefractory metastatic colorectal cancer treated with yttrium-90 radioembolization using glass particles? A real-world single-Centre study.

To compare the survival and objective response rate (ORR) of the patients receiving estimated tumor absorbed dose (ETAD) <140 Gy versus ETAD ≥140 Gy in patients with advanced chemorefractory colorectal carcinoma liver metastases (CRCLM) treated with yttrium-90 transarterial radioembolization (90Y TARE). Between August 2016 and August 2023 adult patients with unresectable, chemorefractory CRCLM treated with 90Y TARE using glass particles, were retrospectively enrolled. Primary outcomes were overall survival (OS) and hepatic progression free survival (hPFS). Secondary outcome was ORR. A total of 40 patients with a mean age of 66.2 ± 7.8 years met the inclusion criteria. Mean ETAD for the group 1 (ETAD <140 Gy) and group 2 (ETAD ≥140) were 131.2 ± 17.4 Gy versus 195 ± 45.6 Gy, respectively. The mean OS and hPFS for group 1 versus group 2 were 12 ± 10.3 months and 8.1 ± 9.3 months versus 9.3 ± 3 months and 7.1 ± 8.4 months, respectively and there were no significant differences (p = 0.181 and p = 0.366, respectively). ORR did not show significant difference between groups (p = 0.432). In real-world practice, no significant difference was found in OS, hPFS and ORR between patients who received ETAD <140 Gy versus ETAD ≥140 Gy in patients with CRCLM, in this series. This study demonstrated that increased tumour absorbed doses in radioembolization may not provide additional significant advantage for OS and hPFS for patients with CRCLM.

Abstract 3786: Genomic and transcriptomic characterization of peritoneal, lung and liver metastases of colorectal carcinoma reveals site-specific differences

Abstract Background: Metastasis in colorectal cancer (CRC) is a critical factor in patient outcomes, with 20% of patients affected at the time of diagnosis and up to 50% developing metachronous metastatic disease later on. Despite the growing importance of metastatic sites as independent predictors of overall survival (OS) and the ongoing development of new treatments, there has been limited effort to personalize systemic treatment based on specific metastatic locations. Aim: This study aims to compare the genomic landscape of different metastatic sites in CRC to detect novel diagnostic and therapeutic targets. Methods: DNA whole-genome-sequencing and RNA gene expression data from metastatic CRC samples were retrieved from the Hartwig Medical Foundation (HMF). Single nucleotide variants (SNVs), somatic copy number aberrations (SCNAs) and structural variants (SVs) were reported by HMF with their pipelines. In addition, focal amplifications and deletions were identified using GISTIC2 (v6.15.30). We included 464 microsatellite stable samples derived from liver metastases (n=369), peritoneal metastases (n=58) and lung metastases (n=37). We compared DNA mutation frequencies between tumors from different metastatic sites (two-sided Mann-Whitney tests) and performed differential gene expression and pathway enrichment analysis. Results: Liver metastases had the highest number of focal SCNAs per sample (median 33 vs 17 in peritoneum (p &amp;lt; 0.001); vs 18 in lung (p &amp;lt; 0.001)). Furthermore, we observed 39 site-specific SCNAs in liver, 9 in peritoneal and 15 in lung metastases. Of these, the most significant focal SCNAs were an amplification within 20q13.12 (79%) and deletion within 1p35.3 (64%) in peritoneal metastases and a deletion within 18q21.1 (84%) in lung metastases. Furthermore, peritoneal metastases displayed less genomic instability, featuring lower ploidy (median 2.65 vs 2.94 in liver (p=0.025); vs 3.15 in lung (p=0.025)) and lower aneuploidy scores (median 19 vs 26 in liver (p=0.016); vs 30 in lung (p=0.016)) while having no significant differences in tumor mutational burden or individual SNVs. Analysis of SVs revealed that peritoneal metastases exhibited fewer deletions (≥10kb; 1.3x fold change increase in liver and lung) and fewer duplications (≥1kb; 1.4x fold change increase in liver). Differential gene expression analysis uncovered higher expression of gene sets involved in the apical junction and the TGF-beta-signaling pathway in peritoneal metastases. Conclusion: Specific focal SCNA and differences in genomic instability exhibit an association with preferential organotropism in CRC metastasis and could offer potential targets for the development of new diagnostic and therapeutic approaches. Citation Format: Nerma Crnovrsanin, Soufyan Lakbir, Renske d. Wit, Steven L. Ketelaars, Niels F. Kok, Petur Snaebjornsson, Gerrit A. Meijer, Sanne Abeln, Remond J. Fijneman. Genomic and transcriptomic characterization of peritoneal, lung and liver metastases of colorectal carcinoma reveals site-specific differences [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3786.

Regional delivery of mesothelin-targeted chimeric antigen receptor T-cell effectively and safely targets colorectal cancer liver metastases in mice.

Liver metastasis is the major cause of colorectal cancer related death. Mesothelin (MSLN)-targeted chimeric antigen receptor (CAR) T-cell therapy has been illustrated effective and safe through regional delivery of breast cancer, ovarian cancer and malignant mesothelioma tumors. Herein, we investigated the safety, efficacy, and immune microenvironment of regional delivery of MSLN (CAR) T-cell in the treatment of colorectal carcinoma liver metastases (CRLM). Second-generation MSLN CAR T-cells were administered by portal vein (PV) or caudal vein (CV, systemic administration) delivery in an orthotopic MSLN+ CRLM nonobese diabetic (NOD)/severe combined immunodeficient (SCID)/γc-/- (NSG) mouse model. A total of 20 mice were randomly divided into control group, non-transduced T cell (NT)-CV group, NT-PV group, MSLN CAR T-cell CV (MSLN-CV) group, and MSLN CAR T-cell PV (MSLN-PV) group, with each group containing four mice to examine the safety and efficacy. The bioluminescence intensity (BLI) of tumor burden, tumor tissue macroscopic and microscopic observation were used to evaluate treatment efficacy. The safety was examined by body weight, survival time, and vital organ damage of mice. CAR T-cell infiltration and cytokine concentration were analyzed by flow cytometry, and immunostaining. The change of immune microenvironment between regional delivery and systemic delivery was investigated on an immune reconstructed CRLM patient-derived xenograft (PDX) model. Additionally, T cell subsets and immunosuppressive markers were examined. PV administration of 1×107/100 μL MSLN CAR T-cells in 20 NSG mice was well tolerated, and no overt toxicity was observed. The tumor burden in the PV group was obviously alleviated. The BLI was (0.73±0.52)×109 in PV group and (1.97±0.11)×109 in CV group (P<0.05), CD8+ granzyme B (GB)+ T cell percentage (MSLN-CV 4.42%±0.47% vs. MSLN-PV 13.5%±4.67%, P<0.01) and cytokine concentration were obviously increased in the MSLN-PV group. In the immune reconstituted CRLM PDX model, intratumor (IT) delivery of MSLN CAR T-cells exhibited much more infiltration of CD4+ and CD8+ T cells accompanied with elevated expression levels of PD-1, LAG-3, and TIM-3. Regional delivery of MSLN-targeted CAR T-cell therapy has encouraging results in the orthotopic CRLM NSG mouse model and PDX model, and converts the tumor microenvironment from cold to hot. This study may provide a new therapeutic approach for CRLM. Further clinical study is needed.

Bcl-xL mediates interferon-beta secretion by protease-activated receptor 2 deficiency through the mitochondrial permeability transition pore in colorectal cancer metastasis

Cellular plasticity and immune escape are synergistic drivers of tumor colonization in metastatic organs. Activation of protease-activated receptor 2 (PAR2) signaling promotes metastasis of colorectal carcinoma (CRC). The role of PAR2 in regulating the immune microenvironment and cancer progression remains unclear. We demonstrated that the regulation of liver metastasis by PAR2 requires a competent immune system. PAR2 knockdown enhanced liver infiltration of activated CD8+ T cells prior to metastatic foci formation in an interferon receptor-dependent manner. PAR2 depletion increased interferon (IFN)-β production via the cGAS-STING and RIG-1 pathways. PAR2 inhibition increased mitochondrial permeability and cytosolic accumulation of mitochondrial DNA, which was reversed by Bcl-xL expression. Strikingly, shRNA against PAR2 with an immune checkpoint blocker (ICB) acted synergistically to suppress liver metastasis. Analysis of single-cell sequence data and 24 paired samples confirmed the regulatory effect of PAR2 on the metastatic immune environment in human CRC. Therefore, PAR2 signaling is involved in stabilizing the mitochondrial membrane and regulating the immune microenvironment through IFN-β during liver metastasis in CRC. The synergistic effect of the PAR2 inhibitor and ICB provides a potential therapeutic strategy for metastatic CRC treatment.

Prediction of Local Tumor Progression After Microwave Ablation in Colorectal Carcinoma Liver Metastases Patients by MRI Radiomics and Clinical Characteristics-Based Combined Model: Preliminary Results

To investigate the predictability of local tumor progression (LTP) after microwave ablation (MWA) in colorectal carcinoma liver metastases (CRLM) patients by magnetic resonance imaging (MRI) radiomics and clinical characteristics-based combined model. Forty-two consecutive CRLM patients (67 tumors) with post-MWA complete response at 1st month MRI were included in this retrospective study. One hundred and eleven radiomics features were extracted for each tumor and for each phase by manual segmentation from pre-treatment MRI T2 fat-suppressed (Phase 2) and early arterial phase T1 fat-suppressed sequences (Phase 1). A clinical model was constructed using clinical data, two combined models were created with feature reduction and machine learning by combining clinical data and Phase 2 and Phase 1 radiomics features. The predicting performance for LTP development was investigated. LTP developed in 7 patients (16.6%) and 11 tumors (16.4%). In the clinical model, the presence of extrahepatic metastases before MWA was associated with a high probability of LTP (p < 0.001). The pre-treatment levels of carbohydrate antigen 19-9 and carcinoembryonic antigen were higher in the LTP group (p = 0.010, p = 0.020, respectively). Patients with LTP had statistically significantly higher radiomics scores in both phases (p < 0.001 for Phase 2 and p = 0.001 for Phase 1). The classification performance of the combined model 2, created by using clinical data and Phase 2-based radiomics features, achieved the highest discriminative performance in predicting LTP (p = 0,014; the area under curve (AUC) value 0.981 (95% CI 0.948-0.990). The combined model 1, created using clinical data and Phase 1-based radiomics features (AUC value 0,927 (95% CI 0.860-0.993, p < 0.001)) and the clinical model alone [AUC value of 0.887 (95% CI 0.807-0.967, p < 0.001)] had similar performance. Combined models based on clinical data and radiomics features obtained from T2 fat-suppressed and early arterial-phase T1 fat-suppressed MRI are valuable markers in predicting LTP after MWA in CRLM patients. Large-scale studies with internal and external validations are needed to come to a firm conclusion on the predictability of radiomics models in CRLM patients.

Exosomal circPABPC1 promotes colorectal cancer liver metastases by regulating HMGA2 in the nucleus and BMP4/ADAM19 in the cytoplasm

Liver metastasis is the leading cause of death in colorectal carcinoma (CRC). However, little is known about the mechanisms of transferring effector messages between the primary tumor and the site of metastasis. Exosomes provide a novel transfer message method, and exosomal circular RNAs (circRNAs) play critical regulatory roles in cancer biology. In this study, the results showed that the expression of circPABPC1 was aberrantly upregulated in CRC tissues and exosomes. Exosomal circPABPC1 was considered an oncogene by functional experimental analysis in vitro and in vivo. Mechanistically, circPABPC1 recruited KDM4C to the HMGA2 promoter, reduced its H3K9me3 modification and initiated the transcription process in the nucleus. Moreover, cytoplasmic circPABPC1 promoted CRC progression by protecting ADAM19 and BMP4 from miR-874-/miR-1292-mediated degradation. Our findings indicated that exosomal circPABPC1 is an essential regulator in CRC liver metastasis progression by promoting HMGA2 and BMP4/ADAM19 expression. CircPABPC1 is expected to be a novel biomarker and antimetastatic therapeutic target in CRC.

Tumoral and Parenchymal Morphological Assessment in Liver Metastases of Colorectal Carcinoma: Micrometastasis, Peritumoral Lymphocytes, Tumor Budding and Differentiation are Potential Prognostic Factors.

Introduction: Various potential prognostic histopathologic factors for colorectal carcinoma liver metastasis have been proposed. However, there is still no consensus on pathological reporting of colorectal carcinoma liver metastasis resection materials. The aim of this study was to investigate the relation between selected tumoral and parenchymal histopathologic features and prognostic factors for better characterization and prognostic prediction of the patients with colorectal carcinoma liver metastasis. Methods: Hematoxylin-eosin stained slides from 100 patients who underwent hepatic resection were evaluated. Pathologic characteristics; including number of tumor nodules, largest tumor size, status of surgical margin, tumor distance to closest margin, tumor necrosis, the presence of tumor capsule, tumor differentiation, perineural and lymphovascular invasion, micrometastasis, tumor budding, peritumoral lymphocytic infiltrate and parenchymal features including steatosis, steatohepatitis, lobular inflammation, confluent necrosis, hepatocyte ballooning, portal inflammation were assessed. For 49 patients who were treated with preoperative chemotherapy, tumor regression grade and chemotherapy-related parenchymal changes such as sinusoidal damage, venous obstruction, nodular regenerative hyperplasia, steatosis and steatohepatitis were also evaluated. Results: The presence of lymphovascular invasion (p < 0.001), micrometastasis (p=0.004), absent or mild peritumoral lymphocytic infiltration (p =0.013), high tumor budding score (p=0.033) and moderate/poor differentiation (p=0.022) were significantly associated with shorter overall survival. Lymphovascular invasion (p < 0.001) was an independent predictor of mortality in multivariate analysis. Conclusions: We conclude that tumor differentiation, lymphovascular invasion, micrometastasis, peritumoral lymphocytic reaction and tumor budding score are potential prognostic histopathological features and candidates for inclusion in pathology reports of colorectal carcinoma liver metastasis resections.

KIAA1522 is a new biomarker of promoting the tumorigenesis and distant metastasis of colorectal carcinoma

PurposeOur research was absorbed into exploring the expression, clinicopathological value, biological significance and signaling pathway of KIAA1522 in colorectal carcinoma and its distant metastasis. Materials and methodsThe expression of KIAA1522 and survival analysis in colorectal carcinoma (CRC) were assessed using GEPIA databases. Then we evaluated the expression of KIAA1522 immunohistochemically in tissue samples of 57 patients with colorectal carcinoma liver metastasis (CRLM). The correlations between the expression of KIAA1522, clinical significance and prognosis of these 57 patients with CRLM were analyzed. The migration and invasion of KIAA1522 were explored by western blotting, CCK-8, colony formation, flow cytometry, wound healing assays and transwell invasion in vitro and tail vein injection models in vivo. Then, transcriptome sequencing and gene set enrichment analysis was performed to identify the signaling pathways involved, while western blotting analysis and immunohistochemistry (IHC) were used to identify the expression of key genes in Notch signaling. ResultsKIAA1522 was overexpressed in CRLM tissues and colon cancer cell lines, and the expression of KIAA1522 in metastatic sites was positively correlated with that in primary sites. In addition, the overexpression of KIAA1522 is associated with poor clinicopathological features. Survival analysis showed that the overexpression of KIAA1522 predicted a low overall survival rate in patients with CRLM. Functional studies suggested that KIAA1522 promotes the proliferation, invasion and migration of colon carcinoma in vitro. KIAA1522 could promote distant metastasis of CRC in vivo. Moreover, KIAA1522 upregulated the Notch signaling pathway in colorectal cancer cell lines in vitro and lung metastatic nodes in vivo. ConclusionIn conclusion, it is suggested that the upregulation of KIAA1522 might promote the tumorigenicity and metastasis of colorectal carcinoma through Notch signaling pathway. KIAA1522 plays a carcinogenic role in the metastasis of colorectal carcinoma and might serve as a new molecular target for the treatment.

Impact of multidisciplinary tumor board (MTB) discussion on the outcome of patients with liver-limited metastasis from colorectal cancer (LMCRC).

50 Background: MTB is a valuable tool in complex cases, since they involve different medical fields aiming for a more comprehensive decision-making process. Our aims are to describe MTB decisions in complex LMCRC, to analyze decisions compliance and evaluate outcomes. Methods: This is a retrospective analysis of medical records of patients diagnosed with liver metastasis of colorectal carcinoma (LMCRC) discussed at a single center, weekly-basis MTB from January 2019 to December 2020. Descriptive statistics was used for demographic and treatment characteristics and adhesion to MTB decisions. Survival was estimated by Kaplan-Meier method and log-rank tests. Results: Out of 1033 cases discussed at MTB, 63 met the inclusion criteria. Median age was 56.5 years (range 32 – 81), 61.9% were male, ECOG was 0-1 in 96.8%. Regarding primary tumor sidedness: 81% and 19% were left and right, respectively. Synchronous metastasis was present in 69.8% and wild-type RAS was found in 52.4% and MSI-H in 4.8%. At time of MTB, 68.3% and 4.8% had realized a first or second-line chemotherapy (CT) respectively. No previous therapy was described in 19% and 7.9% had received adjuvant CT. Previous hepatic resections occurred in 20.6%. After MTB, liver resection was recommended for 44.5%. Preoperative CT followed by one or two-stage surgery for 23.8%. Non-surgical treatments were offered for 17.5% and palliative CT for 14.3%. Decisions from the MTB were implemented in 93.6%. Reason for not performing (N = 4) were: one for disease progression, one for bureaucratic restriction and two for other reasons. Median follow-up time was 17.2m. At date of analysis, 31 patients (49.2%) had relapsed or progressed and 8 had died. Restricting for patients who underwent MTB decision (N = 59), mPFS was 14.71m. In resected cases (N = 40), who were discussed at the onset of liver metastasis diagnosis (without CT in the metastatic setting) had mPFS not reached, while who were discussed after first or second-line had mPFS of 9.39m (p 0.002). R0 resections were reached in 92.5%. Evaluating the 30-day postoperative mortality, there was one death. mPFS for palliative CT was 6.83m. Conclusions: In our analysis the adherence to MTB recommendation was high (above 90%) and patients selected for surgery presented a prolonged PFS. Our data suggest that selection of patients for liver metastasis resection in a MTB discussion should occur early in the course of treatment.

HIST2H2BF Potentiates the Propagation of Cancer Stem Cells via Notch Signaling to Promote Malignancy and Liver Metastasis in Colorectal Carcinoma.

BackgroundGrowing evidence demonstrates that the initiation and progression of colorectal carcinoma (CRC) is related to the presence of cancer stem cells (CSCs). However, the mechanism through which the stem cell features of CRC cells are maintained is poorly understood. In this study, we identified the oncogenic histone cluster 2 H2B family member F (HIST2H2BF) and aimed to investigate the function of upregulated HIST2H2BF expression in maintaining the stem cell features of CRC cells, which accelerate the progression of CRC.MethodsHIST2H2BF expression was quantified using real-time polymerase chain reaction, immunohistochemistry, and western blotting. The correlation between CpG island methylation status and HIST2H2BF re-expression was assessed through bisulfite sequencing polymerase chain reaction, methylation-specific polymerase chain reaction, and 5-Aza-dC treatment. Functional assays were performed on CRC cells and mice to investigate the HIST2H2BF-induced stem cell-like and cancer properties of CRC. Using the Notch pathway inhibitor FLI-06, the regulatory effect of HIST2H2BF on downstream Notch signaling was confirmed.ResultsHIST2H2BF was highly expressed in CRC tissues and cell lines. The reactivation of HIST2H2BF in CRC stems at least in part from the hypomethylated CpG islands. CRC patients with high HIST2H2BF expression have poor survival outcomes. Functional studies have shown that HIST2H2BF promotes CSC phenotype, malignancy, and liver metastasis through the activation of Notch signaling in CRC. Blockage of the Notch pathway reduced the stem cell-like and cancer properties.ConclusionOur study suggests that HIST2H2BF upregulation enhances the CSC phenotype, malignancy, and liver metastasis through the activation of Notch signaling in CRC. These results identified a new perspective on the mechanism by which the stem cell features of CRC cells are maintained and highlighted the potential novel therapeutic targets for CRC.

Thermal Ablation of Colorectal Cancer Metastases Leads to Activation of Individual Cancer-Specific T Cell Immunity

Introduction: The underlying cellular mechanisms of successful immunotherapy are still insufficiently characterized. Especially, priming of tumor-specific T cells against relevant antigens and in situ tumor-recognition is crucial for therapy success in the development of active immunotherapeutic approaches, but remain a serious challenge. Thermal ablation of cancer, e.g. percutaneous radiofrequency ablation (RFA) is a well-established treatment option, particularly for patients with contraindications for surgical resection or in non-resectable primary or secondary liver cancers. RFA induces local tumor destruction by coagulation necrosis, leading to the release of danger signals (e.g. heat shock proteins) and cellular content that can induce and stimulate adaptive and innate immune responses, including tumor-antigen specific T cells. By breaking local immune tolerance, RFA may support the development of antigen-specific anti-tumor responses. Here, we analyzed patient-individual immune responses against autologous HLA-presented tumor antigens after RFA treatment.Materials and Methods: Patients (n=6) with liver metastases of colorectal carcinoma (CRC) in different liver segments were included. First, non-resectable manifestations were ablated using RFA. Subsequently, patients underwent metachronous liver surgery for resectable residual tumors. Using surgical specimens, immunopeptidome analyses of tumor and corresponding non-malignant liver tissues were performed by uHPLC-coupled tandem mass spectrometry (MS) after immunoprecipitation of HLA-ligands. Ligandome data were combined with results from whole transcriptome sequencing (WTS) in order to select for patient-specific HLA-ligands potentially relevant for the patient's individual tumor. Furthermore, identified peptides were compared with an extensive in house database of HLA ligands, derived from different non-transformed tissues including non-malignant colon (NMC) specimens. Peripheral blood mononuclear cells (PBMC) obtained at pre-defined time points before and after RFA and surgery were stimulated in vitro with these T cell epitope candidates. Functional T cell responses were characterized by intracellular cytokine staining (ICS).Results: Combining WTS data and analyses of autologous tumor-exclusive HLA-ligandomes, we selected 32 individual T cell epitope candidates for the respective patients. Antigen-specific T cells could be detected against approx. one third of the candidate epitopes. Both boosting of pre-existing as well as newly induced T cell responses against these epitopes could be detected after RFA. With regard to pre-existing immune responses, CD8+ and CD4+ T cell reactivities could be demonstrated against both HLA class I and class II epitopes. In 4/6 patients (66%) analyzed, de novo induced antigen-specific T cells could be detected after RFA. These reactivities comprised CD8+ as well as CD4+ T cells. Of particular interest, in one patient (diagnosed with mCRC) antigen-specific CD4+ T cells against a mutated HLA-class II peptide were detectable, which were induced by RFA. Of note, this epitope could only be detected on the transcriptome level but not directly as an HLA-ligand by MS.Conclusions: Employing this individualized approach, tumor-antigen specific T cells were more frequently detected as compared to our previous observations employing broadly expressed tumor-associated antigens. These findings suggest that RFA leads to induction of antigen-specific T cells more frequently than previously appreciated, and interestingly also encompasses responses against neo-antigens. The detected immune responses induced by RFA themselves may not be solely sufficient for effectively rejecting established tumors. However, these findings justify the clinical investigation of combinatorial therapies, using immune stimulatory agents or immune checkpoint inhibitors together with thermal ablation. DisclosuresNo relevant conflicts of interest to declare.

Organ-specific cholesterol metabolic aberration fuels liver metastasis of colorectal cancer.

Rationale: Metastasis, the development of secondary malignant growth at a distance from a primary tumor, is the main cause of cancer-associated death. However, little is known about how metastatic cancer cells adapt to and colonize in the new organ environment. Here we sought to investigate the functional mechanism of cholesterol metabolic aberration in colorectal carcinoma (CRC) liver metastasis.Methods: The expression of cholesterol metabolism-related genes in primary colorectal tumors (PT) and paired liver metastases (LM) were examined by RT-PCR. The role of SREBP2-dependent cholesterol biosynthesis pathway in cell growth and CRC liver metastasis were determined by SREBP2 silencing in CRC cell lines and experimental metastasis models including, intra-splenic injection models and liver orthotropic injection model. Growth factors treatment and co-culture experiment were performed to reveal the mechanism underlying the up-regulation of SREBP2 in CRC liver metastases. The in vivo efficacy of inhibition of cholesterol biosynthesis pathway by betulin or simvastatin were evaluated in experimental metastasis models.Results: In the present study, we identify a colorectal cancer (CRC) liver metastasis-specific cholesterol metabolic pathway involving the activation of SREBP2-dependent cholesterol biosynthesis, which is required for the colonization and growth of metastatic CRC cells in the liver. Inhibiting this cholesterol biosynthesis pathway suppresses CRC liver metastasis. Mechanically, hepatocyte growth factor (HGF) from liver environment activates SREBP2-dependent cholesterol biosynthesis pathway by activating c-Met/PI3K/AKT/mTOR axis in CRC cells.Conclusion: Our findings support the notion that CRC liver metastases show a specific cholesterol metabolic aberration. Targeting this cholesterol biosynthesis pathway could be a promising treatment for CRC liver metastasis.

Hallmark microRNA signature in liquid biopsy identifies hepatocellular carcinoma and differentiates it from liver metastasis.

Purpose: This study aims to develop a liquid biopsy assay to identify HCC and differentially diagnose hepatocellular carcinoma (HCC) from colorectal carcinoma (CRC) liver metastasis. Methods: Thirty-two microRNAs (“HallMark-32” panel) were designed to target the ten cancer hallmarks in HCC. Quantitative PCR and supervised machine learning models were applied to develop an HCC-specific diagnostic model. One hundred thirty-three plasma samples from intermediate-stage HCC patients, colorectal cancer (CRC) patients with liver metastasis, and healthy individuals were examined. Results: Six differentially expressed microRNAs (“Signature-Six” panel) were identified after comparing HCC and healthy individuals. The microRNA miR-221-3p, miR-223-3p, miR-26a-5p, and miR-30c-5p were significantly down-regulated in the plasma of HCC samples, while miR-365a-3p and miR-423-3p were significantly up-regulated. Machine learning models combined with HallMark-32 and Signature-Six panels demonstrated promising performance with an AUC of 0.85-0.96 (p ≤ 0.018) and 0.84-0.93 (p ≤ 0.021), respectively. Further modeling improvement by adjusting sample quality variation in the HallMark-32 panel boosted the accuracy to 95% ± 0.01 and AUC to 0.991 (95% CI 0.96-1, p = 0.001), respectively. Even in alpha fetoprotein (AFP)-negative (< 20ng/mL) HCC samples, HallMark-32 still achieved 100% sensitivity in identifying HCC. The Cancer Genome Atlas (TCGA, n=372) analysis demonstrated a significant association between HallMark-32 and HCC patient survival. Conclusion: To the best of our knowledge, this is the first report to utilize circulating miRNAs and machine learning to differentiate HCC from CRC liver metastasis. In this setting, HallMark-32 and Signature-Six are promising non-invasive tests for HCC differential diagnosis and distinguishing HCC from healthy individuals.

Follow-Up 18F-FDG PET/CT versus Contrast-Enhanced CT after Ablation of Liver Metastases of Colorectal Carcinoma-A Cost-Effectiveness Analysis.

Purpose: After a percutaneous ablation of colorectal liver metastases (CRLM), follow-up investigations to evaluate potential tumor recurrence are necessary. The aim of this study was to analyze whether a combined 18F-Fluordesoxyglucose positron emission tomography-computed tomography (18F-FDG PET/CT) scan is cost-effective compared to a contrast-enhanced computed tomography (CE-CT) scan for detecting local tumor progression. Materials and Methods: A decision model based on Markov simulations that estimated lifetime costs and quality-adjusted life years (QALYs) was developed. Model input parameters were obtained from the recent literature. Deterministic sensitivity analysis of diagnostic parameters based on a Monte-Carlo simulation with 30,000 iterations was performed. The willingness-to-pay (WTP) was set to $100,000/QALY. Results: In the base-case scenario, CE-CT resulted in total costs of $28,625.08 and an efficacy of 0.755 QALYs, whereas 18F-FDG PET/CT resulted in total costs of $29,239.97 with an efficacy of 0.767. Therefore, the corresponding incremental cost-effectiveness ratio (ICER) of 18F-FDG PET/CT was $50,338.96 per QALY indicating cost-effectiveness based on the WTP threshold set above. The results were stable in deterministic and probabilistic sensitivity analyses. Conclusion: Based on our model, 18F-FDG PET/CT can be considered as a cost-effective imaging alternative for follow-up investigations after percutaneous ablation of colorectal liver metastases.

CircCSPP1 Functions as a ceRNA to Promote Colorectal Carcinoma Cell EMT and Liver Metastasis by Upregulating COL1A1.

The aberrant regulation of circular RNAs (circRNAs), ring structures formed by exon or intron backsplicing, has been identified as a novel characteristic of multiple cancers. However, the role of circRNAs in colorectal carcinoma remains to be elucidated. In the present study, we investigated the mRNA level and the promoting effect of circRNA CSPP1 (circCSPP1) in colorectal carcinoma liver metastasis. By bioinformatic analysis of 10 paired samples of colorectal carcinoma and adjacent mucosal tissues, we identified circCSPP1 as a significantly upregulated circRNA in colorectal carcinoma tissues, and its upregulation was correlated with a higher M stage. The gain- and loss-of-function assays revealed that circCSPP1 promotes the migration and invasion of colorectal carcinoma cells in vitro and in vivo. Mechanistically, similar miRNA response elements are shared between circCSPP1 and COL1A1. We demonstrated that circCSPP1 upregulates the mRNA levels of COL1A1, which plays a pivotal role in the process of epithelial–mesenchymal transition (EMT), by competitively binding to miR-193a-5p and preventing miR-193a-5p from decreasing the expression of COL1A1. In conclusion, this finding indicates that circCSPP1 may act as a promising therapeutic target by regulating the EMT process in colorectal carcinoma via activation of the circCSPP1/miR-193a-5p/COL1A1 axis.

The value of late phase imaging with FDG-PET/CT in liver metastases of colorectal carcinoma

Aim: Our aim was investigated the role of late phase imaging with F18-FDG-PET/CT in colorectal carcinoma patients with liver metastases. Method: Dual phase FDG-PET/CT scan was retrospectively evaluated in colorectal carcinoma patients with liver metastases. Late phase imaging was acquired 92-253 minutes (mean 158.53±35.7 minutes) after the FDG injection. Sixty-eight metastatic lesions were determined in 37 patients. Mean lesion SUVmax and lesion-to non-tumorous liver tissue ratio were calculated and results of routine FDG-PET imaging were compared with late phase imaging. Results: Metastatic lesion sizes were 9 to230 mm (mean 3.71±3.7 cm). SUVmax values of the metastasis and non-tumorous liver SUVmax for routine and late-phase imaging were as follows; 7.19±3.8, 10.3±5.4; 2.98±0.7, 2.41±0.6 respectively. Metastatic liver lesions, SUVmax values were increased (p 0.01) and non-tumorous liver SUV max values were decreased (p 0.01) in late phase imaging. Compared to routine imaging, in late phase lesion to non-tumorous liver tissue was increased (p 0.001). Lesion retention index was 45.74± 31.8% and the non-tumorous liver index was decreased 18.63 ± 10.4%. Conclusion: The results of this study indicates that normal liver FDG uptake decreases in time and late phase imaging improves the tumor to normal tissue ratio and enable differentiation of metastatic liver lesions from normal liver.

Intravoxel Incoherent Motion of Colon Cancer Liver Metastases for the Assessment of Response to Antiangiogenic Treatment: Results from a Pilot Study

Objective: This study was aimed at evaluating the intravoxel incoherent motion (IVIM) parameter alterations of liver metastases of colorectal carcinoma (CRC) during antiangiogenic bevacizumab combination therapy. Methods: Twenty-five patients with CRC liver metastases treated with bevacizumab in combination with FOLFOX-or-FOLFIRI protocols were enrolled in the study. MRI was performed using a 1.5-tesla scanner pre-treatment (PT) and at 3, 6, and 9 months of therapy. Routine abdominal MRI sequences and an IVIM-DWI (diffusion-weighted imaging) sequence were obtained. The IVIM-DWI sequence was executed with 16 b-values varying from 0 to 1,400 s/mm². The mean values of apparent diffusion coefficient (ADC), true diffusion (D), pseudodiffusion (D*), and perfusion fraction (f) of each metastasis were obtained for all b-values, and the time-related changes were recorded to analyze the chronologic responses to antiangiogenic therapy. The RECIST 1.1 criteria were used for the evaluation of treatment response. Results: The diameters of the metastases diminished significantly at 9 months when compared with PT (p = 0.03). The D (p = 0.10) and ADC (p = 0.21) values of the metastases increased at 9 months of therapy. D* was the highest at 3 months (p =0.24); it decreased at 6 (p =0.97) and 9 months (p =0.87) of therapy. The f value had peaked at 3 months (p =0.51) and started to decrease thereafter. At 6 months, f decreased to the lowest values (p =0.12). Conclusion: IVIM parameters, particularly the perfusion fraction, may quantitatively reflect the response to antiangiogenic treatment. The antiangiogenic response manifests after 3 months of therapy before the RECIST-related response.

N6-methyladenosine modification of circNSUN2 facilitates cytoplasmic export and stabilizes HMGA2 to promote colorectal liver metastasis

Circular RNAs (circRNAs) have been implicated in cancer progression through largely unknown mechanisms. Herein, we identify an N6-methyladenosine (m6A) modified circRNA, circNSUN2, frequently upregulated in tumor tissues and serum samples from colorectal carcinoma (CRC) patients with liver metastasis (LM) and predicts poorer patient survival. The upregulated expression of circNSUN2 promotes LM in PDX metastasis models in vivo and accelerates cancer cells invasion in vitro. Importantly, N6-methyladenosine modification of circNSUN2 increases export to the cytoplasm. By forming a circNSUN2/IGF2BP2/HMGA2 RNA-protein ternary complex in the cytoplasm, circNSUN2 enhances the stability of HMGA2 mRNA to promote CRC metastasis progression. Clinically, the upregulated expressions of circNSUN2 and HMGA2 are more prevalent in LM tissues than in primary CRC tissues. These findings elucidate that N6-methyladenosine modification of circNSUN2 modulates cytoplasmic export and stabilizes HMGA2 to promote CRC LM, and suggest that circNSUN2 could represent a critical prognostic marker and/or therapeutic target for the disease.

CD133 + cell infusion in patients with colorectal liver metastases going to be submitted to a major liver resection (CELLCOL): A randomized open label clinical trial

BackgroundTreatment of liver metastases of colorectal carcinoma is surgical resection. However, only 10–15% of the patients in this context will be candidate for curative resection arising other 10–13% after response to neoadyuvant chemotherapy. In order to perform the liver metastases surgery, it is necessary to have a sufficient remnant liver volume (RLV) which allows maintaining an optimal liver function after resection.Studies on liver regeneration have determined that CD133 + stem cells are involved in liver hypertrophy developed after an hepatectomy with encouraging results. As presented in previous studies, CD133 + stem cells can be selected from peripheral blood after stimulation with G-CSF, being able to obtain a large number of them. We propose to treat patients who do not meet criteria for liver metastases surgery because of insufficient RLV (<40%) with CD133 + cells together with portal embolization, in order to achieve enough liver volume which avoids liver failure. Methods/Design: The aim of this study is to evaluate the effectiveness of preoperative PVE plus the administration of CD133 + mobilized from peripheral blood with G-CSF compared to PVE only. Secondary aims areto compare the grade of hypertrophy, speed and changes in liver function, anatomopathological study of hypertrophied liver, to determine the safety of the treatment and analysis of postoperative morbidity and surveillance. Study designProspective randomized longitudinal phase IIb clinical trial, open, to evaluate the efficacy of portal embolization (PVE) together with the administration of CD133 + cells obtained from peripheral blood versus PVE alone, in patients with hepatic metastasis of colorectal carcinoma (CCRHM). DiscussionThe number of CD133 + obtained from peripheral blood after G –CSF stimulation will be far greater than the number obtained with direct puncture of bone marrow. This will allow a greater intrahepatic infusion, which could have a direct impact on achieving a larger and quicker hypertrophy. Consequently, it will permit the treatment of a larger number of patients with an increase on their survival. Trial registrationClinicalTrials.gov, ID NCT03803241.

Histologic features and genomic alterations of primary colorectal adenocarcinoma predict growth patterns of liver metastasis.

BACKGROUNDDifferent histological growth patterns (HGPs) of colorectal carcinoma (CRC) liver metastasis are associated with patients’ prognosis and response to antiangiogenic therapy. However, the relationship between HGPs of liver metastasis and clinicopathological and genomic characteristics of primary cancer has not been well established.AIMTo assess whether certain clinicopathological and genomic features of primary CRC could predict the HGPs of liver metastasis.METHODSA total of 29 patients with paired resections of both primary CRC and liver metastasis were divided into two groups: A (15 cases with desmoplastic liver metastasis) and B (14 cases with replacement liver metastasis). Clinical information was obtained from patients’ charts. Mismatch repair proteins, BRAFV600E, and PD-L1 were evaluated by immunohistochemistry. Five cases were selected randomly from each group for whole exome sequencing (WES) analysis.RESULTSIn the primary tumor, expanding growth pattern, low tumor budding score (TBS), and Crohn’s disease-like response (CDR) were associated with desmoplastic liver metastasis and better overall survival, whereas infiltrating growth pattern alone of primary carcinoma could predict the replacement liver metastasis and worse overall survival (P < 0.05). On WES analysis, primary carcinoma with desmoplastic liver metastasis showed mutations in APC (4/5); TP53 (3/5); KRAS, PIK3CA, and FAT4 (2/5); BRCA-1, BRCA2, BRAF, and DNAH5 (1/5), whereas primary carcinoma with replacement liver metastasis showed mutations in APC and TP53 (3/5); KRAS, FAT4, DNH5, SMAD, ERBB2, ERBB3, LRP1, and SDK1 (1/5).CONCLUSIONThe HGPs, TBS, and CDR of primary CRC as well as the presence of specific genetic mutations such as those in PIK3CA could be used to predict the HGPs of liver metastasis, response to therapy, and patients’ prognosis.