Therapy In Colorectal Cancer Patients Research Articles

Abstract 1566: Cancer metastasis and stemness: The metastasis-inducer MACC1 regulates LGR5 to promote cancer stem cell-like properties in colorectal cancer

Abstract Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths worldwide. The high mortality is directly associated with metastatic disease which is thought to be initiated by colon cancer stem cells according to cancer stem cell (CSC) model. Consequently, early identification of those patients who are at high risk for metastasis is crucial for improved treatment and patient outcome. Metastasis-associated in colon cancer 1 (MACC1) is a novel prognostic biomarker for tumor progression and metastasis formation independent of tumor stage. We previously showed an involvement of MACC1 in cancer stemness in the mouse intestine of our transgenic mouse models. However, the expression of MACC1 in human CSCs and possible implications remain elusive. Here, we explored the molecular mechanisms by which MACC1 regulates stemness and CSC-associated invasive phenotype based on patient-derived 3D cell culture models (PD3D), patient-derived xenografts (PDX) and human CRC cell lines. We showed that CD44-enriched CSCs from PD3D models express significantly higher levels of MACC1 and display higher tumorigenicity in immunocompromised mice. Similarly, RNA sequencing performed on PD3D and PDX models demonstrated significantly increased MACC1 expression in ALDH1(+) CSCs, highlighting its involvement in cancer stemness. We further showed the correlation of MACC1 with CSC markers CD44, NANOG and LGR5 in PD3D models as well as established cell lines. Additionally, MACC1 increased stem cell gene expression, clonogenicity and sphere formation. Strikingly, we showed that MACC1 binds as a transcription factor to the LGR5 gene promoter uncovering the long-known CSC marker LGR5 as novel essential signaling mediator used by MACC1 to induce CSC-like properties in human CRC patients. Our in vitro findings were further substantiated by significant positive correlation of MACC1 and LGR5 in CRC cell lines as well as CRC patient tumors. Taken together, this study indicates that the metastasis-inducer MACC1 act as a cancer stem cell-associated marker. Interventional approaches targeting MACC1 would potentially improve further targeted therapies for colorectal cancer patients to eradicate CSCs, prevent cancer recurrence and distant metastasis formation. Citation Format: Müge Erdem, Kyung Hwan Lee, Markus Hardt, Joseph Regan, Dennis Kobelt, Wolfgang Walther, Christian Regenbrecht, Ulrike S. Stein. Cancer metastasis and stemness: The metastasis-inducer MACC1 regulates LGR5 to promote cancer stem cell-like properties in colorectal cancer [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 1566.

HUBUNGAN MUTASI GEN K-RAS TERHADAP RESPON TERAPI PADA PASIEN KANKER KOLOREKTAL DI RSUP PROF. DR. IGNG. NGOERAH TAHUN 2017-2019

K-RAS is one of the most frequently mutated genes found in colorectal cancer cases. The K-RAS gene has about 30%-40% contribution to the formation of colorectal cancer. This research is an analytic observational study with a cross-sectional design to explain the characteristics of patients with colorectal cancer at Prof. dr. IGNG. Ngoerah 2017-2019 and assessed the relationship between K-RAS gene mutations and response to therapy. The results of the chi-square test showing a significant relationship between the type of KRAS gene mutation and the response to therapy where p=0.035 (p<0.05). This shows that there is a significant relationship between mutations in the K-RAS gene in colorectal cancer and the response to therapy in colorectal cancer patients at Prof. Dr. IGNG. Ngoerah General Hospital in 2017-2019.

Management pattern and medication-related harms and its predictors in colorectal cancer patients: an institutional-based retrospective study.

Data on colorectal cancer (CRC) patients' thorough management practices and medication-related harms (MRH) are scarce. This study's aim was to investigate the MRHs in patients receiving CRC chemotherapy at the comprehensive specialized hospital of the University of Gondar (UoGCSH). A registry-based retrospective cohort study was conducted on CRC patients at the UoGCSH during 2017-2021. From February to May 2022, medical records were reviewed using a pretested data collection tool to collect socio-demographic and disease-related characteristics, MRHs, and medication regimens. MRHs occurrence and adverse drug reactions (ADRs) severity were assessed using standard guidelines and protocols. Version 16/MP of STATA for Windows was used for the analysis. Independent predictors of MRHs were investigated using logistic regression analysis. A p-value ≤0.05 was used to determine an independent variable's statistical significance. One hundred forty three CRC patients were included, with a mean age of 49.9 ± 14.5 years. About 32.9% and 33.6% had stage II and III cancer, respectively. Significant patients had co-morbidities (15.4%) and complications (13.3%). Fluorouracil (5-FU)-based regimens were given to more than half (56%) of the patients. MRHs were found in 53.1% of the patients, with a mean of 2.45 ± 1.37 MRHs. The most common MRHs were the need for additional drug therapy, sub-therapeutic dose, DDIs, and ADRs. Being on stage IV (AOR = 27.7, 95% CI = 3.85-199.38, p = 0.001), having co-morbidity (AOR = 7.42, 95% CI = 1.80-30.59, p = 0.018) and having complication (AOR = 11.04, 95% CI = 1.72-70.95, p = 0.011) and treated with five or more drugs (AOR = 2.54, 95% CI = 1.07-6.07, p = 0.035) were independent predictors of MRHs. A fluorouracil-based treatment regimen was most frequently used. MRHs were found in nearly half of CRC patients. Furthermore, MRHs were significantly associated with cancer stage, comorbidity and complication status, and the number of medications used. Because MRHs are common, improving clinical pharmacy services is critical for optimizing drug therapy in CRC patients.

METTL3/IGF2BP2 axis affects the progression of colorectal cancer by regulating m6A modification of STAG3

Colorectal cancer (CRC) is among the commonest malignant tumors of humans. Existing evidence has linked the poor prognosis of CRC with high expression of stromal antigen 3 (STAG3), but, the exact biological effect of STAG3 in CRC is still unclear. The aim of this research is to reveal the biological function and molecular mechanism of STAG3 in CRC. To investigate the differential expression of STAG3 in CRC tissues and cell lines compared to normal colon tissues and cell lines, Western blot (WB) and quantitative real-time PCR (qRT-PCR) techniques were utilized. STAG3 N6-methyladenosine (m6A) modification level were identified using m6A RNA immunoprecipitation (MeRIP). Additionally, the functional roles of methyltransferase-like protein 3 (METTL3) and insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) in CRC were explored by manipulating their levels via knockdown or overexpression. Cell proliferation was evaluated through Cell Counting Kit 8 (CCK-8) and clone formation experiments, while cell migration was assessed through wound healing experiments. Furthermore, cell apoptosis was detected using flow cytometry, and the protein expressions associated with proliferation and apoptosis were detected using WB. To identify the specific binding of target genes, RIP and pull-down assays were employed. Finally, the biological function of STAG3 in vivo was investigated through a xenotransplantation mouse tumor model. In CRC tissues and cell lines, STAG3 was up-regulated and accompanied by m6A methylation. Additionally, the expression of METTL3 was found to be upregulated in CRC tissues. Knocking down METTL3 resulted in a decrease in both the m6A level and protein expression of STAG3, inhibited cell proliferation and migration while promoting apoptosis, which were restored through STAG3 overexpression. Furthermore, online prediction indicated the interaction between STAG3 mRNA and IGF2BP2 protein, which was further verified by RIP experiments. IGF2BP2 downregulation led to decreased STAG3 protein expression, cell proliferation, and migration, but increased apoptosis. However, these impacts were reversed by STAG3 overexpression. Finally, subcutaneous tumor experiments conducted in nude mice also confirmed that METTL3 regulated CRC progression through STAG3 in vivo. The METTL3/IGF2BP2/STAG3 axis affects CRC progression in an m6A modification-dependent manner. This may guide targeted therapy in CRC patients.

Hsa_circRNA_102051 regulates colorectal cancer proliferation and metastasis by mediating Notch pathway

BackgroundThe purpose of this study was to investigate the role of hsa_circRNA_102051 in colorectal cancer (CRC) and its effect on the stemness of tumor cells.MethodsCircRNA microarray was under analysis to screen differentially expressed novel circRNAs in the pathology of CRC. Quantitative real-time PCR was used to detect the relative RNA expression in CRC cells and samples. The effects of hsa_circRNA_102051 on biological functions in CRC cells were accessed both in vitro and in vivo. FISH, RIP and luciferase reporter assay were conducted to confirm the regulatory correlations between hsa_circRNA_102051 and miR-203a, as well as miR-203a and BPTF. Xenograft models were applied to further verify the impacts and fluctuations of hsa_circRNA_102051/miR-203a/BPTF. Moreover, the mechanism how hsa_circRNA_102051 affected the Notch signals was also elucidated.ResultsHsa_circRNA_102051 was up-regulated in CRC tissues and cell lines, capable to promote the growth and invasion of CRC. In addition, hsa_circRNA_102051 could enhance stemness of CRC cells. BPTF was identified as downstream factors of hsa_circRNA_102051, and miR-203a was determined directly targeting both hsa_circRNA_102051 and BPTF as an intermediate regulator. Hsa_circRNA_102051 in CRC could block miR-203a expression, and subsequently activated BPTF. Hsa_circRNA_102051/miR-203a/BPTF axis modulated stemness of CRC cells by affecting Notch pathway.ConclusionsOur findings provided new clues that hsa_circRNA_102051 might be a potential predictive or prognostic factor in CRC, which induced the fluctuation of downstream miR-203a/BPTF, and subsequently influenced tumor growth, activities and stemness. Thereinto, the Notch signals were also involved. Hence, the hsa_circRNA_102051/miR-203a/BPTF axis could be further explored as a therapeutic target for anti-metastatic therapy in CRC patients.

Clinicopathological correlations between colorectal cancer and genetic mutations

Objective. Colorectal cancer is an oncological pathology that, unfortunately, has increased in terms of incidence in recent years. The presence of KRAS and BRAF mutations in colorectal cancer has significant clinical implications. As a result we want to conduct research that analyzes the impact of these mutations on patients diagnosed with colorectal cancer and also to observe the clinicopathological differences between mutant and wild-type tumors. Material and methods. We conducted a retrospective study in the period 2018-2022, including 118 patients diagnosed with colorectal cancer. The patients were subsequently divided into two groups equal in number of patients, depending on the presence or absence of mutations. Outcomes. After analyzing the data we were able to identify several differences between the two groups, regarding the histopathological type - mucinous correlated with the mutant tumors, the degree of infiltration of the locoregional lymph nodes (more N+ cases in the mutant group), the location of the primary tumor (right colon within the mutant tumors, the rectosigmoid region in the wild-type group), the location of secondary tumors (pulmonary ones with a triple incidence in the mutant group). Conclusions. The study of genetic mutations and their role in colorectal cancer has provided valuable insights into the underlying mechanisms of this complex disease. It is an ever-evolving field that promises to have a profound impact on patient care, ultimately leading us toward more effective prevention, early detection, and personalized therapies for colorectal cancer patients. By leveraging genetic information, clinicians can optimize treatment plans, minimize side effects, and increase the chances of successful outcomes for individual patients.

Rebound increase in microRNA levels at the end of 5-FU-based therapy in colorectal cancer patients

Treatment with 5-fluorouracil (5-FU) based therapy is still used for colorectal cancer (CRC). Epigenetics has become a focus of study in cancer because of its reversibility besides its known regulatory functions. In this study, we will monitor the change in microRNAs (miRNAs) levels with 5-FU-based therapy at baseline and after 3 and 6 months of treatment to be correlated with their prognostic potential. The expression levels of 5 miRNAs, namely miRNA223-3p, miRNA20a-5p, miRNA17-5p, miRNA19a-3p, and miRNA7-5p, were measured in the peripheral blood of 77 CRC patients, along with the expression of 3 proteins PTEN, ERK, and EGFR. At baseline, CRC patients had significantly higher levels of circulating miRNAs than healthy controls. This level was reduced after 3 months of 5-FU-based therapy, then increased after 6 months significantly in responder patients compared to non-responders. MiRNA19a-3p showed that significant pattern of change in the subgroups of patients with high ERK, EGFR, and PTEN protein levels, and its 6 months level after 5-FU-based therapy showed significance for the hazard of increased risk of disease recurrence and progression.

Bacillus subtilis supernatant improves the efficacy of radiation therapy in rat intestinal epithelial cells by upregulation of bax and caspase-3 genes.

Colorectal Cancer (CC) is among the most prevalent cancers in elderly persons. Radiotherapy is usually prescribed as CC develops, however, radiation beams indiscriminately affect normal cells. Previous studies nominated that probiotics and their metabolites can be used to minimize the side effects of radiotherapy. Hereby, the aim of this study was to investigate the probable correlation between cell-free supernatant of Bacillus subtilis and radiation response in normal and cancerous cell lines. IEC-18 and SW-48 cells were treated with different concentrations of B. subtilis supernatant. To evaluate the effect of probiotic treatments under radiation and the normal situation, the cytotoxicity of the treatments was measured using the MTT method. The cell cycle status was analyzed by flow cytometry. The expression levels of Bax, Bcl-2, and Caspase 3 genes were also determined by real-time (RT) PCR. B. subtilis supernatant increased the viability of normal cells under radiation treatment, although this effect was not significant. 40% v/v of this mixture could amplify the lethal effect of radiation and decreased the viability of cancer cells. SW-48 cells that received 40% v/v of the supernatant had a significantly higher rate of apoptosis. Probiotic supernatant effectively induced the expression of proapoptotic Bax and Caspase 3 genes. Presented results confirmed that the supernatant of B. subtilis can be supposed as a clue to improve the efficacy of radiation therapy in CC patients as it increased the sensitivity of cancerous cells and protected normal epithelial cells from detrimental effects of radiation.

A Systematic Review of Complementary Therapies in Colorectal cancer patients: Summarizing the Current Global Options

Colorectal cancer patients have been experiencing various side effects related to the therapies, both physically and physiologically. Complementary medicine was used to help patients alleviate complaints and increase their quality of life. This systematic review aimed to sort out complementary medicines that could be advised to be used for colorectal cancer patients. Search limitations include the population of colorectal cancer patients, published between 2015 and 2021 and written in English. We found that traditional Chinese medicine, aromatherapy massage, acupuncture, and mindfulness-based stress reduction tests were beneficial in improving patients’ physical and physiological complaints. However, dosing and herb-herb interactions in Chinese traditional medicine and the beneficial persistency effect of other modalities needed further investigation, especially in colorectal cancer patient therapy. Complementary modalities are generally safe; however, some therapies still warrant further investigation to confirm safety and beneficial consistency.

Association between adjuvant therapy and survival in colorectal cancer patients according to metabolic Warburg-subtypes

PurposeTumor location and tumor node metastasis (TNM) stage guide treatment decisions in colorectal cancer (CRC) patients. However, patients with the same disease stage do not benefit equally from adjuvant therapy. Hence, there remains an urgent clinical need to identify prognostic and/or predictive biomarker(s) to personalize treatment decisions. In this exploratory study, we investigated whether our previously defined metabolic Warburg-subtypes can predict which CRC patients might derive survival benefit from adjuvant therapy.MethodsInformation regarding treatment (surgery only: n = 1451; adjuvant radiotherapy: n = 82; or adjuvant chemotherapy: n = 260) and Warburg-subtype (Warburg-low: n = 485, -moderate: n = 641, or –high: n = 667) was available for 1793 CRC patients from the Netherlands Cohort Study (NLCS). Kaplan–Meier curves and Cox regression models were used to investigate survival benefit from adjuvant therapy compared to surgery-only for the different Warburg-subtypes.ResultsPatients with Warburg-moderate CRC (HRCRC-specific 0.64; 95% CI 0.47–0.86, HRoverall 0.61; 95% CI 0.47–0.80), and possibly Warburg-high CRC (HRCRC-specific 0.86; 95% CI 0.65–1.14, HRoverall 0.82; 95% CI 0.64–1.05), had survival benefit from adjuvant therapy. No survival benefit was observed for patients with Warburg-low CRC (HRCRC-specific 1.07; 95% CI 0.76–1.52, HRoverall 0.95; 95% CI 0.70–1.30). There was a significant interaction between Warburg-subtype and adjuvant therapy for CRC-specific survival (p = 0.049) and overall survival (p = 0.035).ConclusionOur results suggest that Warburg-subtypes may predict survival benefit from adjuvant therapy in CRC patients. A survival benefit from adjuvant therapy was observed for patients with Warburg-moderate and possibly Warburg-high CRC, but not for patients with Warburg-low CRC. Future prospective studies are necessary to validate our findings.

Resistin-like beta reduction is associated to low survival rate and is downregulated by adjuvant therapy in colorectal cancer patients

Colorectal Cancer (CRC) is one of the most common cancers accounting for 1.8 million new cases worldwide every year. Therefore, the identification of new potential therapeutic targets represents a continuous challenge to improve survival and quality of CRC patient’s life. We performed a microarray analysis dataset consisting of colon biopsies of healthy subjects (HS) and CRC patients. These results were further confirmed in a clinical setting evaluating a series of CRC patients to assess the expression of Resistin-Like Beta (RETNLB) and to correlate it with their clinical data. Our results showed a significant reduction of RETNLB expression in CRC biopsies compared to the HS mucosa. Furthermore, such reduction was significantly associated with the TNM grade and patients’ age. Furthermore, a significantly positive correlation was found within mutated subjects for KRAS, TP53, and BRAF. In particular, patients with poor prognosis at 5 years exhibited RETNLB lower levels. In-silico analysis data were confirmed by histochemical analysis in a series of CRC patients recruited by our group. The results obtained provided that RETNLB low levels are associated with an unfavorable prognosis in CRC patients and its expression is also dependent on adjuvant therapy. Further studies are warranted in order to evaluate the molecular mechanisms underlying the role of RETNLB in CRC progression.

Tumor-Associated Neutrophils in Colorectal Cancer Development, Progression and Immunotherapy.

Simple SummaryTumor-associated neutrophils (TANs) may differentiate into different patterns under the stimulation of different factors, and they play a dual role in the occurrence and progression of tumors in direct or indirect ways. The existing immune checkpoint inhibitors (ICIs) are effective in a small number of microsatellite-instability-high (MSI-H) or mismatch-repair-deficient (dMMR) colorectal cancer (CRC) patients, but they are still not suitable for microsatellite-stability (MSS) CRC patients. As an important component of the tumor immune microenvironment, TANs may overturn the current situation of immunotherapy for CRC. This review systematically summarizes the key regulatory role of TANs in the carcinogenesis, proliferation and metastasis of CRC, the prognostic value of TANs for CRC patients and the new immunotherapy strategies based on TANs as a target, providing an important reference for TANs as new target for CRC immunotherapy.The colorectal-cancer (CRC) incidence rate and mortality have remained high for several years. In recent years, immune-checkpoint-inhibitor (ICI) therapy has rapidly developed. However, it is only effective in a few CRC patients with microsatellite-instability-high (MSI-H) or mismatch-repair-deficient (dMMR) CRC. How to improve the efficiency of ICI therapy in CRC patients with microsatellite stability (MSS) remains a huge obstacle. Tumor-associated neutrophils (TANs), which are similar to macrophages, also have N1 and N2 phenotypes. They can be recruited and polarized through different cytokines or chemokines, and then play an antitumor or tumor-promoting role. In CRC, we find that the prognostic significance of TANs is still controversial. In this review, we describe the antitumor regulation of TANs, and their mechanism of promoting tumor progression by boosting the transformation of inflammation into tumors, facilitating tumor-cell proliferation, metastasis and angiogenesis. The targeting of TANs combined with ICIs may be a new treatment model for CRC. Relevant animal experiments have shown good responses, and clinical trials have also been carried out in succession. TANs, as “assistants” of ICI treatment, may become the key to the success of CRC immunotherapy, although no significant results have been obtained.

The Prediction of Necroptosis-Related lncRNAs in Prognosis and Anticancer Therapy of Colorectal Cancer.

Background Colorectal cancer is one of the most common gastrointestinal malignancies globally. Necroptosis has been proved to play a role in the occurrence and development of the tumor, which makes it a new target for molecular therapy. However, the role of necroptosis in colorectal cancer remains unknown yet. Our study aims to build a prognostic signature of necroptosis-related lncRNAs (nrlncRNAs) to predict the outcomes of patients with colorectal cancer and facilitate in anticancer therapy. Method We obtained RNA-seq and clinical data of colorectal adenocarcinoma from the TCGA database and got prognosis-related nrlncRNAs by univariate regression analysis. Then, we carried out the LASSO regression and multivariate regression analysis to build the prognostic signature, whose predictive ability was tested by the Kaplan-Meier as well as ROC curves and verified by the internal cohort. Moreover, we divided the cohort into 2 groups based on median of risk scores: high- and low-risk groups. By analyzing the difference in the tumor microenvironment, microsatellite instability, and tumor mutation burden between the two groups, we explored the potential chemotherapy and immunotherapy drugs. Results We screened out 9 nrlncRNAs and built a prognostic signature based on them. With its good prognostic ability, the risk scores can act as an independent prognostic factor for patients with colorectal cancer. The overall survival rate of patients in high-risk group was significantly higher than the low-risk one. Furthermore, risk scores can also give us hints about the tumor microenvironment and facilitate in predicting the response to the CTLA-4 blocker treatment and other chemotherapeutic agents with potential efficacy such as cisplatin and staurosporine. Conclusions In conclusion, our prognostic signature of necroptosis-related lncRNAs can facilitate in predicting the prognosis and response to the anticancer therapy of colorectal cancer patients.

Nutritional Support in Patients with Colorectal Cancer: State of Knowledge and Perspectives

Many cancer patients have a nutritional risk. Nutritional support therapy plays an important role in cancer treatment, especially for the patients with digestive system cancer. Nutritional support therapy for tumor patients should be based on the specific saturations. For example, types of tumors, degree of differentiation, and nutritional status of the patients need to be evaluated in detail, and then propose the specific implementation approach. For the colorectal cancer patients with promising treatment prospects, nutritional support should be standardized and make every effort to improve the safety and effectiveness of treatment. This article reviews the research progress on nutritional support therapy for colorectal cancer patients thereby promoting the standardized development of nutritional support therapy and better guiding the clinical nutritional support practice. In addition, this paper discussed the feasibility of a combination of nutritional and exercise interventions to efficiently improve muscle and strength, physical function, nutritional status, fatigue, and quality of life in colorectal cancer patients. Establishing a standard process for nutritional support should become an interdisciplinary task. Standard procedures and quality control should also be specified to ensure the effectiveness of nutritional therapy. The current knowledge, and perspectives of application of nutritional support in patients with colorectal cancer in this paper could provide a good reference for the future research of cancer diagnosis and treatment.

A Tumor‐Organoid‐Based Precision Medicine Platform for the Prediction of Drug Sensitivity of Colorectal Cancer

AbstractInvestigation of clinical actionable targets is crucial for the individual treatment of patients with advanced colorectal cancer (CRC). A complete platform integrating the molecular profiling and prediction of drug reactivity is currently scarce. An integrated precision medicine approach which combines high‐throughput sequencing platform with microfluidic organoid drug testing system is established. Capture sequencing identifies 6 drug targetable genes and 13 candidate drugs for a stage III colorectal cancer patient. The drug sensitivity is confirmed by using patient‐derived organoids (PDOs) model and microfluidic drug testing system. With this integrated precision medicine approach, the optimal drugs including Afatinib, Dacomitinib, and Osimertinib are successfully identified. The platform provides a practical method to guide the clinical therapies for colorectal cancer patients.

Crosstalk between PI3K/Akt and Wnt/β-catenin pathways promote colorectal cancer progression regardless of mutational status

ABSTRACT The PI3K/Akt and Wnt/β-catenin pathways play an important role in the acquisition of the malignant phenotype in cancer. However, there are few data regarding the role of the interplay between both pathways in colorectal cancer (CRC) progression. The mutational status and the clinicopathological characteristics of PI3K/Akt and Wnt/β-catenin pathways were accessed by bioinformatic analysis whereas that the impact of the interplay between the activity of both pathways to explain tumorigenic potential was performed in vitro using IGF-1 and Wnt3a treatments in CRC cell models. The mutational status of these pathways did not influence the survival of CRC patients, but an association between clinicopathological characteristics in patients with mutations in one, but not in both pathways was observed. A potentiating effect on the activation of both pathways and enhanced cellular migration and proliferation was observed when both pathways were activated simultaneously with IGF-1 and Wnt3a. In addition, these effects were hindered after pretreatment with LY294002, a specific PI3K inhibitor, suggesting some dependence between these two signaling cascades. Our findings show that, regardless of mutational status, there is an interplay between the activity of PI3K/Akt and Wnt/β-catenin pathways that contributes to events related to CRC progression and that the reversal of such events using a PI3K inhibitor highlights the value of targeting these pathways for potential directed therapies in CRC patients.

Acneiform eruptions with combination targeted cancer therapy in colorectal cancer patients.

Epidermal growth factor receptor inhibitors (EGFRI) can be used with pathway inhibitors, including mitogen-activated protein kinase kinase inhibitors (MEKIs), BRAF inhibitors (BRAFIs), and checkpoint inhibitors such as programmed death-ligand 1 (PD-L1) and programmed cell death protein 1 (PD-1) to treat colorectal cancer. These can precipitate treatment-resistant acneiform eruptions, prompting dose modification or discontinuation. Predicting the likelihood of severe rash development and crafting effective treatments may promote adherence to life-saving chemotherapy. An Institutional Review Board-approved retrospective chart review of patients with colorectal cancer treated with EGFRI or MEKI in combination with HER2, BRAF, PI3K, or checkpoint inhibitors between January 1, 2016, and January 1, 2020, was performed. Surrogates for rash severity were investigated, including lower extremity involvement, utilization of oral steroids or retinoids, dose modification, and incidence of superinfection. Of 122 patients treated with combination therapy, 105 developed a rash, and 87 developed an acneiform eruption. Common combinations included MEKI/PD-LI, EGFRI/MEKI, and MEKI/PD-1I. Patients treated with EGFRI/MEKI developed the most severe rashes (p = 0.02). Lower extremity involvement was more frequent with EGFRI/MEKI compared to alternative combinations (p = 0.05). Drug holiday correlated with all rash severity surrogates, including rash grade, lower extremity involvement, oral steroid or retinoid use, and incidence of superinfection. Use of oral steroids or retinoids was associated with development of superinfection (p = 0.002). Prophylactic tetracycline use did not impact rash severity or rash incidence. This is the first descriptive analysis to characterize acneiform eruptions for patients with colorectal cancer on combination cancer therapy. Approximately 85% of patients developed a cutaneous toxicity with what appears to be synergistic effects of EGFRI and MEKI combination therapy causing the most severe eruptions. Superinfection rate correlated to systemic therapy use beyond oral tetracyclines. Further investigation into the utility of prophylactic oral tetracyclines in this population is needed.

Abstract 1324: IL-36 signalling enhances a pro-tumorigenic phenotype in colon cancer cells with cancer cell growth restricted by administration of the IL-36R antagonist

Abstract Background & Aims: The IL-1 subfamily, IL-36, is increasingly being implicated as a potent cytokine family involved in chronic inflammatory conditions such as psoriasis and IBD. Recent work has also suggested IL-36 cytokines may act as a potential adjuvant in immunotherapy regimens to augment the anti-cancer immune response. Given the pluripotent nature of IL-1 cytokines and the relationship between inflammation and tumorigenesis, we investigated the effects of IL-36 signalling in colorectal cancer (CRC). Methods: IL-36 family expression in CRC progression was investigated by the NCBI GEO transcriptomic database, and then by immunohistochemistry (IHC) and qRT-PCR. Pro-tumorigenic properties such as cancer cell migration, invasion and proliferation were investigated in 2D and 3D models. In vivo models were performed using Balb-c mice which were subcutaneously inoculated with CT26 cells and treated with IP injections of recombinant IL-36R agonists (IL-36α, β, γ) or antagonist (IL-36RN). Tumour tissues were excised and studied by flow cytometry and IHC analysis. Additionally, CRISPR-Cas9 mediated IL-36R KO CT26 cells were generated and subcutaneously injected into Balb-c mice, with changes in tumour growth and immune cell infiltrate investigated. Results: IL-36R expression was shown to significantly increase with stage of disease in the adenoma-carcinoma sequence, including distant metastases. Additionally, expression of all family members was increased in tumour tissue relative to adjacent normal tissue in a separate colon cancer cohort. In vitro IL-36R signalling on cancer cells augmented a pro-tumorigenic phenotype by induction of pro-tumorigenic cytokines/chemokine production, increased cellular migration, invasion and proliferation of tumour cells. In vivo inhibition of IL-36R signalling by both administration of recombinant IL-36RN and through deletion of the IL-36R gene in CT26 cells, resulted in a significant reduction in tumour growth. This reduction was associated with a significant decrease in tumour cell proliferation, as well as an increase in CD8+ T cell infiltrate in IL-36R KO groups. Of note, administration of IL-36R agonists also resulted in reduced tumour growth, albeit to a lesser extent, associated with an increase in CD4+ and CD8+ T cell infiltration. Conclusions: This data indicates that IL-36 family members, similar to other IL-1 family members, have dual functions in colon cancer. In addition, this data suggests that targeting IL-36R signalling may be a useful targeted therapy for CRC patients with IL-36R+ tumour cells. Citation Format: Kevin J. Baker, Charlotte O'Donnell, Micheal O'Riordain, Elizabeth Brint, Maura Bendix, Aileen Houston. IL-36 signalling enhances a pro-tumorigenic phenotype in colon cancer cells with cancer cell growth restricted by administration of the IL-36R antagonist [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1324.

IL-36 signalling enhances a pro-tumorigenic phenotype in colon cancer cells with cancer cell growth restricted by administration of the IL-36R antagonist.

The IL-36 cytokines are a recently described subset of the IL-1 family of cytokines, shown to play a role in the pathogenesis of intestinal diseases such as Inflammatory Bowel Disease (IBD). Given the link between IBD and colitis –associated cancer, as well as the involvement of other IL-1 family members in intestinal tumorigenesis, the aim of this work was to investigate whether IL-36 cytokines play a role in the pathogenesis of colon cancer. Whilst research to date has focused on the role of IL-36 family members in augmenting the immune response to induce tumour rejection, very little remains known about IL-36R signalling in tumour cells in this context. In this study we demonstrate that expression of IL-36 family member mRNA and protein are significantly increased in colorectal cancer tissue compared to adjacent non-tumour. In vitro assays showed stimulation of colon cancer cell lines with IL-36R agonists resulted in the activation of the pro-tumorigenic phenotypes of increased cellular migration, invasion and proliferation in both 2D and 3D models. In addition, the IL-36 cytokines induced strong expression of pro-inflammatory chemokines in both human and murine cell lines. Intraperitoneal injection of IL-36Ra significantly reduced tumour burden using the subcutaneous CT26 tumour model in syngeneic Balb/mice, and this was associated with a decrease in Ki-67 expression by tumour cells in the IL-36Ra- treated group relative to untreated, suggesting the inhibition of the pro-proliferative signalling of IL-36 agonists resulted in the decreased tumour size. Moreover, colon cancer cells lacking the IL-36R also showed reduced tumour growth and reduced Ki-67 expression in vivo. Taken together, this data suggests that targeting IL-36R signalling may be a useful targeted therapy for colorectal cancer patients with IL-36R+ tumour cells.

Pre-Diagnosis Aspirin Use Has No Effect on Overall Survival in Patients With Colorectal Cancer: A Study of a Multi-Racial Population.

IntroductionAspirin has been associated with a reduction in mortality in patients diagnosed with colorectal cancer (CRC). A possible mechanism for this is related to the programmed cell death 1 (PD-1) immune checkpoint pathway. Aspirin may have a synergistic effect with PD-1 inhibitors via inhibition of prostaglandin E2 (PGE2) production, which can reverse the ability of tumor cells to evade the immune system. This appears to be strongest in cancers that express PI3 kinase (PI3K) signaling activity, which aspirin downregulates. However, the benefit of pre-diagnosis aspirin use on CRC overall survival (OS) and cancer-specific survival is still controversial, and most studies have been performed in racially homogenous populations. Our study examines the effect of pre-diagnosis aspirin therapy on OS in a racially diverse group of patients with CRC.MethodsThis is a retrospective chart review of 782 patients diagnosed with CRC from January 2007 to December 2020. Kaplan-Meier curve was created to study the association of aspirin exposure compared to no exposure on OS. In addition, univariate and multivariate binary logistic regression analyses were done to investigate potential predictors of survival.ResultsOf the 782 patients with CRC, 55.1% were males, 22.2% whites, 58.5% Asians, and 17.7% Pacific-Islanders. Moreover, 38.4% of the patients had a history of aspirin use, 79% of them used it for more than one year. There were more patients with hypertension (HTN), hyperlipidemia (HLD), diabetes mellitus (DM), and chronic kidney disease (CKD) among those with a history of aspirin use. There was no difference in one, three, and five-year OS among aspirin users compared to non-users, p-value = 0.63. Age, grade, and stage were potential predictors of worsened OS. However, treatment with chemotherapy and CKD were potential predictors of worsened OS on univariate analysis only. No significant association was noticed with gender, tumor location, or other associated comorbidities.ConclusionThe effect of pre-diagnosis aspirin use on CRC survival is not clear. In this retrospective analysis of a racially diverse population of CRC patients, we found that aspirin use was not associated with improved OS. Therefore, physicians should be careful about using aspirin as adjuvant therapy in CRC patients until high-quality prospective data are available, given the potential associated complications.