Enhancing the understanding and management of metabolic dysfunction-associated steatotic liver disease in colorectal cancer patients: insights and future directions.

PurposeThe objective of this study was to compare the diagnostic performance of positron emission tomography (PET), PET/CT, CT and MRI as whole-body imaging modalities for the detection of local and/or distant recurrent disease in colorectal cancer (CRC) patients who have a (high) suspicion of recurrent disease, based on clinical findings or rise in carcinoembryonic antigen (CEA).MethodsA meta-analysis was undertaken. PubMed and Embase were searched for studies on the accuracy of whole-body imaging for patients with suspected local and/or distant recurrence of their CRC. Additionally, studies had to have included at least 20 patients with CRC and 2 × 2 contingency tables had to be provided or derivable. Articles evaluating only local recurrence or liver metastasis were excluded. Summary receiver-operating characteristic (ROC) curves were constructed from the data on sensitivity and specificity of individual studies and pooled estimates of diagnostic odds ratios (DORs) and areas under the ROC curve (AUCs) were calculated. To test for heterogeneity the Cochran Q test was used.ResultsFourteen observational studies were included which evaluated PET, PET/CT, CT and/or MRI. Study results were available in 12 studies for PET, in 5 studies for CT, in 5 studies for PET/CT and in 1 study for MRI. AUCs for PET, PET/CT and CT were 0.94 (0.90–0.97), 0.94 (0.87–0.98) and 0.83 (0.72–0.90), respectively. In patient based analyses PET/CT had a higher diagnostic performance than PET with an AUC of 0.95 (0.89–0.97) for PET/CT vs 0.92 (0.86–0.96) for PET.ConclusionBoth whole-body PET and PET/CT are very accurate for the detection of local and/or distant recurrent disease in CRC patients with a (high) suspicion of recurrent disease. CT has the lowest diagnostic performance. This difference is probably mainly due to the lower accuracy of CT for detection of extrahepatic metastases (including local recurrence). For clinical practice PET/CT might be the modality of choice when evaluating patients with a (high) suspicion of recurrent disease, because of its best performance in patient based analyses and confident prediction of disease status.

Introduction: Alcohol use is an independent risk factor for liver metastasis, a major cause of morbidity and mortality in colorectal cancer (CRC) patients. Serum CEA level is an established prognostic indicator in CRC, yet the correlation with behavioral factors such as alcohol use remains to be defined. In a single-center review, we evaluated alcohol use, gender, and CEA levels in predicting advanced disease in CRC patients. Methods: Retrospective analysis of UNMC patients diagnosed with CRC as the primary cancer between 2012-2019, stages I-IV, and age >19 with documentation of alcohol use. Univariable statistics were performed using Chi-Square and non-parametric tests. Associations between stage, gender, and alcohol use (some vs. none) and the log-transformed CEA outcome (either initial or rate of change) were assessed using linear regressions. Results: Alcohol use was found to be reported in 333 of 1243 CRC patients. The cohort was comprised of 192 male and 141 female subjects. Elevated CEA levels at CRC diagnosis were associated with increased all-cause mortality (33.0% for CEA > 3.4ng/ml vs 10.4% for CEA < 3.4ng/ml). Model analysis found that stage IV male alcohol users showed an increase in serial CEA levels compared to males who did not use alcohol, but this pattern was not observed among stage IV females. Conclusions: Males with a history of alcohol use may be at risk for advanced CRC disease suggesting the utility of serial serum CEA monitoring in these patients. A detailed alcohol use history should be obtained in all patients with CRC as it has prognostic value and may allow for early intervention. This analysis was limited by missing alcohol use data for the majority (73.2%) of CRC patients evaluated. A prospective study is warranted to define the implications of alcohol use and risk of CRC liver metastasis.

Objective: Existing methods for detecting circulating-tumor DNA (ctDNA) through fixed panels exhibit constrained sensitivity, while customized panels are time-consuming and rely on prerequisite tumor mutation profiles. Here, we aimed to introduce an adaptive noise cancellation approach for efficient Minimal Residuals Disease (MRD) detection in colorectal cancer patients employing a fixed panel under both tumor-informed and tumor-naïve situations. Methods: 108 plasma samples were collected from 52 colorectal cancer patients (11 relapse, 41 non-relapse) at various post-surgical timepoints. Comprehensive mutational profiling of plasma samples was conducted using a fixed MRD panel (SHIELDINGTM ULTRA) that encompasses hotspots in 2365 cancer-related genes. Background errors were removed by an adaptive noise cancellation algorithm relying on DNA fragment profiling, normal pool, and white blood cell background and intra-run plasma sampling. Variants were called both with (tumor-informed) and without (tumor-naïve) the present of tumor tissue. Results: Our longitudinal analysis of 108 plasma samples yielded 90.9% sensitivity, 97.6% specificity and 97.6% negative predictive value in the tumor-informed scenario. The ctDNA positive patients exhibited significantly higher risk than those with negative ctDNA status (Hazard Ratio = 87.2, p = 3.5 × 10^(-5)). For landmark analysis, 33 plasma samples (≤7 days post-operation) were assessed, yielding a remarkable performance of 66.7% sensitivity and 100% specificity. Our approach showed robustness under tumor-naïve scenario, illustrating 90.9% sensitivity, 92.7% specificity and 97.4% NPV for the longitudinal analysis. This corresponded to an increased risk of 49 times in the ctDNA positive patients (HR = 49.0, p = 2.2 × 10^(-4)). Notably, the results of tumor-naïve samples closely aligned with those of tumor-informed samples, showing a concordance of 92.6% and substantial agreement (Cohen’s Kappa = 0.691). Conclusion: Our method stands as an ultra-sensitive tool for identifying colorectal patients at high risk of recurrence. By employing an adaptive noise cancellation algorithm, our fixed panel approach demonstrates superiority over existing fixed panels in accuracy and proves to be a cost-effective alternative to personalized panels. Significantly, it extends potential clinical utilization towards tumor-naïve patients, which is not feasible for personalized panels. Citation Format: Hua Bao, Wanxiangfu Tang, Ningyou Li, Min Wu, Jinfeng Zhang, Baihan Zhu, Xue Wu, Yang Shao. Improved detection of minimal residual disease in colorectal cancer patients using adaptive noise cancellation algorithm [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 3685.

3581 Background: TS, the target enzym of 5-FU has a 28 bp polymorphism in the promoter region containing commonly double of triple tandem repeats of a 28 bp sequence (2R or 3R), and a 6bp ins/del polymorphism on the 3'UTR. Our aim was to study the influence of these TS polymorphisms on tumor response to adjuvant FUFA therapy in CRC patients. Methods: : 89 CRC patients receiving adjuvant FUFA were involved in the study. FUFA was given as bolus (Mayo regimen) or continuous infusion (de Gramont protocoll). TS polymorphisms were determined by PCR PAGE and RFLP method. Results: DFS was significantly longer (p=0,036 logrank test) in patients with +6/+6,+6/-6 genotypes than in –6/-6 genotype.OS was significantly longer (p=0,025) in 3/3 genotype than in 2/3and 2/2 genotype. In rectal cancer +/+ and +/- 3'UTR patients had longer OS than -/- patients (p=0,007).No significant correlation between gene polymorphisms, age, gender, and localization were found. Conclusions: Both TS polymorphisms studied do affect the clinical outcome of the disease in CRC patients treated with adjuvant FUFA chemotherapy. Supported by the NKFP1/48/2001 grant. No significant financial relationships to disclose.

Metachronous colorectal liver metastases (CRLM) had been the main cause of death in most colorectal cancer (CRC) patients after curative surgery. However, the clinical impact of the pre-existing metabolic dysfunction-associated steatotic liver disease (MASLD) on metachronous CRLM remains unclear, the present study aims to clarify this issue. This study retrospectively enrolled in 2155 CRC patients who underwent curative surgery at the Sixth Affiliated Hospital of Sun Yat-sen University from 2017 to 2018, including 300 cases with MASLD and 1855 cases of Control group. Cox regression and Kaplan-Meier curves were constructed to evaluate the influencing factors and cumulative incidence of recurrence (CIR) of metachronous CRLM. The pre-existing MASLD was an independent risk factor for metachronous CRLM in TNM stage 0-II CRC patients (HR, 2.696; 95% CI 1.553-4.681; p < 0.001), and this association persisted for both early (HR, 2.309; 95% CI 1.008-5.291; p = 0.048) and late (HR, 2.791; 95% CI 1.321-5.894; p = 0.007) metachronous CRLM. However, pre-existing MASLD was not independently associated with metachronous CRLM in TNM stage III CRC patients. In TNM stage 0-II CRC patients, the 5-year hepatic CIR rate in the MASLD group was significantly higher than that in the Control group (9.3% vs. 4.0%, p < 0.001). MASLD had no significant impact on metachronous extrahepatic metastases for TNM stage 0-II and stage III CRC patients. The pre-existing MASLD is an independent and important risk factor predicting hepatic recurrence for CRC patients of TNM stage 0-II after curative surgery.

Changing Nomenclature from Nonalcoholic Fatty Liver Disease to Metabolic Dysfunction-Associated Fatty Liver Disease – Not Only Premature But Also Confusing

Serum vascular endothelial growth factor has been associated with stage of disease in colorectal cancer patients. We investigated whether preoperative serum vascular endothelial growth factor can provide any relevant clinical prognostic information during long-term follow-up of colorectal cancer patients. Preoperative serum samples of 79 colorectal cancer patients and serum of 28 healthy controls were stored at -80 degrees C until later vascular endothelial growth factor analysis by enzyme-linked immunosorbent assay technique and carcinoembryogenic antigen concentration measurement were performed. There were three patient groups for comparison: 21 patients with overt liver metastases, 18 patients who developed recurrent disease after initial curative surgery, and 40 patients who remained disease-free for at least five years. We could not demonstrate any significant difference in serum vascular endothelial growth factor values between the patient groups and controls, nor between the three patient groups (Mann-Whitney U test). There was no relevant correlation between serum vascular endothelial growth factor and carcinoembryogenic antigen concentrations (Pearson r = 0.2; P = 0.07). Although vascular endothelial growth factor has been shown in previous studies to be a potent inducer of angiogenesis and metastases formation, the present data demonstrate that preoperative serum vascular endothelial growth factor concentration does not provide any relevant individual prognostic information in patients with colorectal cancer.

Background:Upregulation of the matrix metalloproteinases MMP-2 and MMP-9 in various cancers has been associated with worse survival of the patients.Methods:We assessed MMP-2 and MMP-9 levels in normal colorectal mucosa from colorectal cancer patients in relation to the course of the disease.Results:A high protein expression of MMP-2 as well as MMP-9 in normal mucosa was found to be correlated with worse 5-year survival. The combination of both parameters was an even stronger prognostic factor. These protein levels were found not to be related to the corresponding single nucleotide polymorphisms of MMP-2 (−1306C>T) and MMP-9 (−1562C>T). Multivariate analyses indicated that the MMP-2 and MMP-9 levels in normal mucosa are prognostic for survival, independent of TNM classification.Conclusion:MMP-2 and MMP-9 levels in normal mucosa are indicative of the course of disease in colorectal cancer patients.

Perioperative beta-adrenergic blockade and COX2 inhibition in colorectal cancer patients improves pro-metastatic indices in the excised tumor: EMT, tumor infiltrating lymphocytes (TILs), and gene regulatory pathways

Altered fatty acid metabolism, a hallmark of cancer, is an attractive target for therapeutic intervention. Fatty acid translocase (CD36), a multifunctional glycoprotein, has an important role in fatty acid metabolism: it serves as a fatty acid transporter. We have previously shown the importance of CD36 in promotion of colorectal cancer (CRC) cell proliferation and tumorigenesis and its role in compensation of de novo fatty acid synthesis inhibition. CD36 also plays important roles in migration, invasion and metastasis in gastric, ovarian and oral carcinomas. Cell lines with higher metastatic potential express higher levels of CD36; however, the role of CD36 in CRC metastasis has not been previously studied. The purpose of our study is (i) to determine the functional effect of CD36 on CRC metastasis, and (ii) to delineate the mechanistic pathways in which CD36 may regulate CRC metastasis. Methods: In this study we utilized HCT116 and HT29 CRC cell lines. The HT29 LuM3-GFP-Luc cell line was established from HT29 cells and has an increased propensity to metastasize to mouse lungs. The increase in metastatic capacity of HT29 LuM3 is associated with an increase in the expression of CD36. We utilized parental HT29-GFP-Luc and HT29 LuM3-GFP-Luc to overexpress and knockdown CD36, respectively, and we used a CD36 neutralizing antibody to block the activity of CD36 in HCT116 and HT29 LuM3 cells. To measure colonization, migration and invasion in vitro, we used soft agar colony formation assays, a wound healing assay, and spheroid and trans-well invasion assays. In vivo, we utilized the tail-vein injection model to study tumor cell colonization in mouse lungs, and the cecum injection model to study liver metastasis from the primary tumor. We analyzed samples with qRT-PCR, western blot and immunohistochemistry. Results: Knockdown of CD36 significantly reduced CRC cell invasion and colony formation in HCT116 and HT29 LuM3 in vitro. CRC cell invasion and colony formation increased with overexpression of CD36 in HCT116 cells. Overexpression of CD36 in established CRC cell lines increased markers of survival and invasion, such as phospho-Akt, P-Met, and MMP28. Most importantly, knockdown of CD36 in HT29 LuM3-GFP-Luc cells significantly reduced lung metastasis in vivo. Conclusions: CRC cells that have a higher propensity to initiate metastasis in vivo express higher levels of CD36. We showed that this upregulation of CD36 represents a unique advantage for CRC cells to initiate tumor colonies, migrate and invade both in vitro and in vivo. Thus, our study highlights the potential of CD36 as a viable target for metastatic disease in CRC patients. Citation Format: James Michael Drury, Piotr Rychahou, Heidi L. Weiss, Yekaterina Y. Zaytseva. CD36, a fatty acid translocase, promotes metastasis in CRC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2880.

In colorectal cancer (CRC), tumor buds (TB) are observed histologically as single tumor cell or small tumor cell clusters located mainly at the advancing tumor edge. TB are a marker of poor prognosis and correlate with metastatic disease in CRC patients. They often lack expression of CDX2 and overexpress markers involved in epithelial-mesenchymal transition (EMT). We evaluated the function of CDX2 in CRC proliferation and migration using CRISPR/Cas9 technology and demonstrated a possible link to tumor dissociation and tumor budding. Knocking out CDX2 in CRC cell lines significantly increased migration. Importantly, the observed phenotypes could be rescued by re-expressing CDX2 and by specific CRISPR synergistic activation mediator (SAM) of endogenous CDX2 in CDX2 low expressing CRC cell lines. Multiplex immunofluorescence (mIF) analysis of primary tumor regions compared to TB in a CDX2-positive CRC patient sample as well as patient derived xenografts (PDX) revealed significantly lower CDX2 expression and correlating E-cadherin levels in TB compared to primary tumor regions, in both models. Accordingly, increased invasiveness of CRC CDX2 knockout cells was seen in ex ovo xenografts. Taken together, our results provide further insight into the function of CDX2 in preventing CRC cell migration, tumor budding and tumor aggressiveness.

Colorectal cancer is one of the most common gastrointestinal malignancies and is also a disease of genetic heterogeneity. Our previous studies have shown that SPERT (sprermatid-associated protein) gene may be an underlying oncogene that is associated with the progression of the disease in colorectal cancer patients, and SPERT gene silencing can inhibit the proliferation of colorectal tumor cells and promote cell apoptosis. Here, we use the stably transfected human colorectal cancer cell line RKO to construct an animal xenograft model and study the effect of SPERT gene silencing on animal xenografts. The results showed that SPERT gene silencing can inhibit tumor growth in animals. In addition, through signaling pathway analysis, we found that the p38MAPK/HSP27 signaling pathway may be the molecular mechanism by which SPERT gene silencing inhibits the growth of xenograft tumors in nude mice. Combined with previous data, SPERT gene silencing has the same inhibitory effect on tumor growth in vitro and in vivo. These data suggest that SPERT gene may be a potential target for the treatment of colorectal cancer in clinic.

Is Occult Lymph Node Disease in Colorectal Cancer Patients Clinically Significant?: A Review of the Relevant Literature

This study was designed to investigate the efficiency of preoperative serum carcinoembryonic antigen (CEA) and carbohydrate cancer antigen (CA19-9) levels for diagnosing synchronous liver metastases and lymph node in colorectal carcinoma (CRC) patients. A total of 300 patients with histologically diagnosed CRC were included in this study between May 2014 and March 2015. The data were obtained prospectively from patient's medical records: medical history, demographics, tumor location, differentiation (grade), depth of the tumor (T), lymph node metastases (N), distant metastases (M), lymphatics, venous and perineural invasion, and disease stage. Tumor markers were measured with an electrochemiluminescent assay and the reference value was 5ng/ml for CEA and for Ca19-9, 37u/ml. There was A high statistically significant difference in the levels of serum CEA and CA19-9 between different disease stages of CRC (P<0.001). Regarding different T stages of CRC, We noticed a significant statistical difference in CEA (stage I 3.76±8.73; II 5.68±17.27, III 7.56±14.81, and IV 70.90±253.23) and CA 19-9 levels (stage I 9.65±11.03, II 9.83±11.09; III 19.58±36.91, and IV 228.9±985.38, respectively). The mean CEA and CA19-9 serum levels were significantly higher in patients with regional lymph nodes involvement (CEA 37.21±177.85 vs 4.79±9.90, CA19-9 119.51±687.71 VS 12.24±17.69, respectively, P<0.05) and in liver metastases (CEA 86.56±277.65 vs. 5.98±12.98, and CA19-9 273.27±1073.46 vs. 4.98±3142, respectively, with P<0.001) in comparison to patients without lymph node involvement and liver metastases. We noticed a cut-off value for lymph nodes involvement, for CEA and CA 19-9, 3.5 ng/mL and 7.5 U/mL, respectively. While, a cut-off value for the presence of synchronous liver metastases of these two markers was 3.5ng/mL AND 5.5 U/mL. Our study showed that tumor makers, CEA and CA19-9, can be used as diagnostic factors regarding the severity of CRC specifically to suggest metastatic disease in CRC.

4078 Background: HuA33 is a humanized antibody that targets the A33 antigen, which is highly expressed in intestinal epithelium and &gt;95% of human colon cancers, but not other normal tissues. Previous studies have shown huA33 can target and be retained in a metastatic tumor for 6 weeks, but eliminated from normal colonocytes within days. This phase 1 study used radio-labeled huA33 in combination with capecitabine chemotherapy to target chemoradiation to metastatic colorectal cancer. Methods: The primary objective was safety and tolerability of the combination of capecitabine and 131I-huA33. Pharmacokinetics, biodistribution, immunogenicity, and tumor response were also assessed. Eligibility included measurable metastatic colorectal cancer, adequate hematological and biochemical function, and informed consent. An outpatient scout 131I-huA33 dose was followed by a single therapy infusion one week later, when capecitabine was commenced. Dose escalation occurred over 5 dose levels. Patients were evaluated weekly, with tumor response assessment at the end of the12 week trial. Tumor targeting was assessed using gamma camera and single photon emission computerized tomography (SPECT) imaging. Results: 16 patients have enrolled with 2 currently on treatment, including one in the final dose cohort. Accrual will be completed by March 2007. The dose escalation protocol was amended following 2 dose limiting toxicities in the second cohort, but subsequent cohorts demonstrated good tolerability. Biodistribution analysis demonstrated excellent tumor targeting of the known tumor sites, expected transient bowel uptake, but no other normal tissue uptake. Maximal duration of stable disease is currently 3 years. Conclusions: 131I-huA33 achieves specific targeting of radiotherapy to sites of metastasis and can be safely combined with chemotherapy. This provides an opportunity to deliver chemoradiation specifically to metastatic disease in colorectal cancer patients. [Table: see text] No significant financial relationships to disclose.