{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T11:30:31Z","timestamp":1786447831047,"version":"build-2736575974"},"reference-count":44,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T00:00:00Z","timestamp":1786406400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"VCOM FY25 Delta Collaborative Research Program","award":["VCOM1032612ULM"],"award-info":[{"award-number":["VCOM1032612ULM"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Background\/Objectives: Colorectal cancer (CRC) is the second most common cancer-causing death in the United States. The Mediterranean diet, rich in extra-virgin olive oil (EVOO), is associated with a lower risk of colorectal cancer (CRC). Earlier studies reported S-(\u2212)-oleocanthal (OC), the major EVOO phenolic, to be effective in CRC progression and recurrence suppression in a subcutaneous xenograft model by targeting the SMYD2\u2013EZH2\/c-MET signaling axis and favorably modulating gut microbiota (GM). However, lead validation in an orthotopically xenografted model that mimics the tumor microenvironment is yet to be achieved. The GM contribution to OC anti-CRC activity remains unknown. Methods: We used an orthotopic intra-cecally xenografted nude mouse model to validate OC anti-KRAS-mutant CRC activity and assessed the contribution of OC GM modulation to this activity using an oral antibiotic depletion strategy. Results: Daily oral 10 mg\/kg OC impressively suppressed KRASG13D-mutant CRC HCT-116-Luc progression and metastasis more effectively than intraperitoneal administration 3\u00d7\/week. By contrast, GM depletion with a broad-spectrum antibiotics cocktail (ABC) partially attenuated this activity, suggesting GM contribution to OC anti-CRC activity. Thus, fecal microbiota transplantation (FMT) was conducted using fresh daily oral fecal GM treatments collected from 20 mg\/kg OC-dosed nude mouse donors. A week before FMT dosing, orthotopic HCT-116-Luc tumor-bearing recipient mice were subjected to GM depletion using daily oral ABC dosing. Recipient mice treated with FMT from OC-treated donors exhibited dramatic &gt;99% reductions in primary and near-complete suppression of multi-organ metastatic tumor burden versus FMT controls. Conclusions: Collectively, oral OC is validated as an effective anti-KRASG13D-mutant CRC lead. Future clinical trials can validate its anti-CRC potential in a human model.<\/jats:p>","DOI":"10.3390\/nu18162623","type":"journal-article","created":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T10:32:16Z","timestamp":1786444336000},"page":"2623","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Lead Validation of the Olive Phenolic S-(\u2212)-Oleocanthal for Effective Control of KRASG13D-Mutant Colorectal Cancer Progression and Metastasis"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-4223-3432","authenticated-orcid":false,"given":"Md Towhidul Islam","family":"Tarun","sequence":"first","affiliation":[{"name":"School of Basic Pharmaceutical and Toxicological Sciences, College of Pharmacy, University of Louisiana at Monroe, 1800 Bienville Drive, Monroe, LA 71201, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hassan Y.","family":"Ebrahim","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, Discipline of Pharmacology, Edward Via College of Osteopathic Medicine, Monroe, LA 71203, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1456-4064","authenticated-orcid":false,"given":"Khalid A.","family":"El Sayed","sequence":"additional","affiliation":[{"name":"School of Basic Pharmaceutical and Toxicological Sciences, College of Pharmacy, University of Louisiana at Monroe, 1800 Bienville Drive, Monroe, LA 71201, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Siegel, R.L., Kratzer, T.B., Wagle, N.S., Sung, H., and Jemal, A. (2026). Cancer statistics, 2026. CA Cancer J. Clin., 76.","DOI":"10.3322\/caac.70043"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Siegel, R.L., Wagle, N.S., Star, J., Kratzer, T.B., Smith, R.A., and Jemal, A. (2026). Colorectal cancer statistics, 2026. CA Cancer J. Clin., 76.","DOI":"10.3322\/caac.70067"},{"key":"ref_3","first-page":"372","article-title":"Clinical management of metastatic colorectal cancer in the era of precision medicine","volume":"72","author":"Ciardiello","year":"2022","journal-title":"CA Cancer J. Clin."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"7087","DOI":"10.3748\/wjg.v23.i39.7087","article-title":"Detection of KRASG12D in colorectal cancer stool by droplet digital PCR","volume":"23","author":"Alexandre","year":"2017","journal-title":"World J. Gastroenterol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1038\/s41698-025-01166-3","article-title":"KRAS-targeted therapies in colorectal cancer: A systematic analysis of mutations, inhibitors, and clinical trials","volume":"9","author":"Mulet","year":"2025","journal-title":"npj Precis. Oncol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1620","DOI":"10.1038\/s41388-023-02676-9","article-title":"Feedback activation of EGFR\/wild-type RAS signaling axis limits KRASG12D inhibitor efficacy in KRASG12D-mutated colorectal cancer","volume":"42","author":"Feng","year":"2023","journal-title":"Oncogene"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Takeda, M., Yoshida, S., Inoue, T., Sekido, Y., Hata, T., Hamabe, A., Ogino, T., Miyoshi, N., Uemura, M., and Yamamoto, H. (2025). The role of KRAS mutations in colorectal cancer: Biological insights, clinical implications, and future therapeutic perspectives. Cancers, 17.","DOI":"10.3390\/cancers17030428"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Tarun, M.T.I., Elsayed, H.E., Ebrahim, H.Y., and El Sayed, K.A. (2025). The olive oil phenolic S-(-)-oleocanthal suppresses colorectal cancer progression and recurrence by modulating SMYD2-EZH2 and c-MET activation. Nutrients, 17.","DOI":"10.3390\/nu17030397"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"282","DOI":"10.3892\/ol.2025.15028","article-title":"Role of SMYD2 in gastrointestinal cancer progression (Review)","volume":"29","author":"Xia","year":"2025","journal-title":"Oncol. Lett."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2325","DOI":"10.1038\/s12276-024-01326-8","article-title":"SMYD family in cancer: Epigenetic regulation and molecular mechanisms of cancer proliferation, metastasis, and drug resistance","volume":"56","author":"Han","year":"2024","journal-title":"Exp. Mol. Med."},{"key":"ref_11","first-page":"997","article-title":"SMYD2 suppresses APC2 expression to activate the Wnt\/\u03b2-catenin pathway and promotes epithelial-mesenchymal transition in colorectal cancer","volume":"10","author":"Meng","year":"2020","journal-title":"Am. J. Cancer Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1482","DOI":"10.1016\/j.celrep.2019.10.004","article-title":"Regulation of EZH2 by SMYD2-mediated lysine methylation is implicated in tumorigenesis","volume":"29","author":"Zeng","year":"2019","journal-title":"Cell Rep."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1038\/s41419-018-0347-x","article-title":"Lysine methyltransferase SMYD2 promotes triple negative breast cancer progression","volume":"9","author":"Li","year":"2018","journal-title":"Cell Death Dis."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"744","DOI":"10.1038\/s41388-024-02945-1","article-title":"Lysine methyltransferase SMYD2 enhances androgen receptor signaling to modulate CRPC cell resistance to enzalutamide","volume":"43","author":"Li","year":"2024","journal-title":"Oncogene"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1038\/s41419-021-04483-0","article-title":"SMYD2 targets RIPK1 and restricts TNF-induced apoptosis and necroptosis to support colon tumor growth","volume":"13","author":"Yu","year":"2022","journal-title":"Cell Death Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"12704","DOI":"10.18632\/oncotarget.7260","article-title":"The relationship between EZH2 expression and microRNA-31 in colorectal cancer and the role in evolution of the serrated pathway","volume":"7","author":"Kurihara","year":"2016","journal-title":"Oncotarget"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Li, C., Song, J., Guo, Z., Gong, Y., Zhang, T., Huang, J., Cheng, R., Yu, X., Li, Y., and Chen, L. (2022). EZH2 Inhibitors suppress colorectal cancer by regulating macrophage polarization in the tumor microenvironment. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.857808"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Yu, H., Li, X.-X., Han, X., Chen, B.-X., Zhang, X.-H., Gao, S., Xu, D.-Q., Wang, Y., Gao, Z.-K., and Yu, L. (2023). Fecal microbiota transplantation inhibits colorectal cancer progression: Reversing intestinal microbial dysbiosis to enhance anti-cancer immune responses. Front. Microbiol., 14.","DOI":"10.3389\/fmicb.2023.1126808"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1186\/s40168-022-01289-w","article-title":"Gut dysbiosis promotes prostate cancer progression and docetaxel resistance via activating NF-\u03baB-IL6-STAT3 axis","volume":"10","author":"Zhong","year":"2022","journal-title":"Microbiome"},{"key":"ref_20","first-page":"43","article-title":"Commensal microbiota in the digestive tract: A review of its roles in carcinogenesis and radiotherapy","volume":"19","author":"Dong","year":"2021","journal-title":"Cancer Biol. Med."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Chen, J., Zhang, P., Zhao, Y., Zhao, J., Wu, X., Zhang, R., Cha, R., Yao, Q., and Gao, Y. (2022). Nitroreductase-instructed supramolecular assemblies for microbiome regulation to enhance colorectal cancer treatments. Sci. Adv., 8.","DOI":"10.1126\/sciadv.add2789"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"113846","DOI":"10.1016\/j.celrep.2024.113846","article-title":"Roseburia intestinalis sensitizes colorectal cancer to radiotherapy through the butyrate\/OR51E1\/RALB axis","volume":"43","author":"Dong","year":"2024","journal-title":"Cell Rep."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Kasagga, A., Hawrami, C., Ricci, E., Hailu, K.T., Salib, K., Nandeesha, S.S., and Hamid, P. (2024). Gut microbiota and colorectal cancer: An umbrella review of methodological trends and clinical correlations. Cureus, 16.","DOI":"10.7759\/cureus.54210"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Farr\u00e0s, M., Martinez-Gili, L., Portune, K., Arranz, S., Frost, G., Tondo, M., and Blanco-Vaca, F. (2020). Modulation of the gut microbiota by olive oil phenolic compounds: Implications for lipid metabolism, immune system, and obesity. Nutrients, 12.","DOI":"10.3390\/nu12082200"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Borz\u00ec, A.M., Biondi, A., Basile, F., Luca, S., Vicari, E.S.D., and Vacante, M. (2018). Olive oil effects on colorectal cancer. Nutrients, 11.","DOI":"10.3390\/nu11010032"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"10324","DOI":"10.1039\/D1FO02033A","article-title":"Main drivers of (poly)phenol effects on human health: Metabolite production and\/or gut microbiota-associated metabotypes?","volume":"12","author":"Selma","year":"2021","journal-title":"Food Funct."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1016\/j.semcancer.2023.08.001","article-title":"Mediterranean diet and olive oil, microbiota, and obesity-related cancers. From mechanisms to prevention","volume":"Volume 95","author":"Cano","year":"2023","journal-title":"Seminars in Cancer Biology"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Barber, T.M., Kabisch, S., Pfeiffer, A.F., and Weickert, M.O. (2023). The effects of the Mediterranean diet on health and gut microbiota. Nutrients, 15.","DOI":"10.3390\/nu15092150"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Jannati, S., Patel, A., Patnaik, R., and Banerjee, Y. (2025). Oleocanthal as a multifunctional anti-cancer agent: Mechanistic insights, advanced delivery strategies, and synergies for precision oncology. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26125521"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1013","DOI":"10.1055\/s-0030-1270724","article-title":"(\u2013)-Oleocanthal as a c-Met inhibitor for the control of metastatic breast and prostate cancers","volume":"77","author":"Elnagar","year":"2011","journal-title":"Planta Med."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Qusa, M.H., Abdelwahed, K.S., Siddique, A.B., and El Sayed, K.A. (2021). Comparative gene signature of (\u2013)-oleocanthal formulation treatments in heterogeneous triple negative breast tumor models: Oncological therapeutic target insights. Nutrients, 13.","DOI":"10.3390\/nu13051706"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Siddique, A.B., Ayoub, N.M., Tajmim, A., Meyer, S.A., Hill, R.A., and El Sayed, K.A. (2019). (\u2013)-Oleocanthal prevents breast cancer locoregional recurrence after primary tumor surgical excision and neoadjuvant targeted therapy in orthotopic nude mouse models. Cancers, 11.","DOI":"10.3390\/cancers11050637"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Siddique, A.B., Ebrahim, H.Y., Tajmim, A., King, J.A., Abdelwahed, K.S., Abd Elmageed, Z.Y., and El Sayed, K.A. (2022). Oleocanthal attenuates metastatic castration-resistant prostate cancer progression and recurrence by targeting SMYD2. Cancers, 14.","DOI":"10.3390\/cancers14143542"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Tarun, M.T.I., Ebrahim, H.Y., Dawud, D., Elmageed, Z.Y.A., Corey, E., and El Sayed, K.A. (2025). The olive phenolic S-(-)-oleocanthal as a novel intervention for neuroendocrine prostate cancers: Therapeutic and molecular insights. Nutrients, 17.","DOI":"10.3390\/nu17243947"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Tarun, M.T.I., Ebrahim, H.Y., and El Sayed, K.A. (2025). The olive phenolic S-(-)-hydroxyoleocanthal attenuates neuroendocrine prostate cancer via modulation of EPHA3-centered oncogenic network. Cancers, 18.","DOI":"10.3390\/cancers18010026"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Qusa, M.H., Abdelwahed, K.S., Hill, R.A., and El Sayed, K.A. (2023). S-(\u2212)-oleocanthal ex vivo modulatory effects on gut microbiota. Nutrients, 15.","DOI":"10.3390\/nu15030618"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Siddique, A.B., Ebrahim, H., Mohyeldin, M., Qusa, M., Batarseh, Y., Fayyad, A., Tajmim, A., Nazzal, S., Kaddoumi, A., and El Sayed, K.A. (2019). Novel liquid-liquid extraction and self-emulsion methods for simplified isolation of extra-virgin olive oil phenolics with emphasis on (\u2212)-oleocanthal and its oral anti-breast cancer activity. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0214798"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1077","DOI":"10.2353\/ajpath.2007.060773","article-title":"Orthotopic microinjection of human colon cancer cells in nude mice induces tumor foci in all clinically relevant metastatic sites","volume":"170","author":"Espina","year":"2007","journal-title":"Am. J. Pathol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Lee, S.B., Jeon, H.J., Hyun, H., and Jeon, Y.H. (2025). Establishment of an Orthotopic and Metastatic Colorectal Cancer Mouse Model Using a Tissue Adhesive-Based Implantation Method. Cancers, 17.","DOI":"10.3390\/cancers17132266"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"943","DOI":"10.1080\/1061186X.2018.1509983","article-title":"Development of patient-derived orthotopic xenografts from metastatic colorectal cancer in nude mice","volume":"27","author":"Roque","year":"2019","journal-title":"J. Drug Target."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"136","DOI":"10.4103\/JMAU.JMAU_51_18","article-title":"In vivo bioluminescence-based monitoring of liver metastases from colorectal cancer: An experimental model","volume":"7","author":"Magistri","year":"2019","journal-title":"J. Microsc. Ultrastruct."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"27","DOI":"10.4103\/0976-0105.177703","article-title":"A simple practice guide for dose conversion between animals and human","volume":"7","author":"Nair","year":"2016","journal-title":"J. Basic Clin. Pharm."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Siddique, A.B., King, J.A., Meyer, S.A., Abdelwahed, K., Busnena, B., and El Sayed, K. (2020). Safety evaluations of single dose of the olive secoiridoid S-(\u2013)-oleocanthal in Swiss albino mice. Nutrients, 12.","DOI":"10.3390\/nu12020314"},{"key":"ref_44","first-page":"100357","article-title":"Dose-dependent evaluation of chronic oleocanthal on metabolic phenotypes and organ toxicity in 5xFAD mice","volume":"8","author":"Yang","year":"2025","journal-title":"Pharmacol. Res. Nat. Prod."}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/18\/16\/2623\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T10:42:09Z","timestamp":1786444929000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/18\/16\/2623"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,11]]},"references-count":44,"journal-issue":{"issue":"16","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["nu18162623"],"URL":"https:\/\/doi.org\/10.3390\/nu18162623","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,11]]}}}