{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,10]],"date-time":"2026-03-10T04:40:21Z","timestamp":1773117621053,"version":"3.50.1"},"reference-count":36,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,3,9]],"date-time":"2026-03-09T00:00:00Z","timestamp":1773014400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000090","name":"Department of Defense Peer Reviewed Cancer Research Program CDMRP PRCRP","doi-asserted-by":"publisher","award":["W81XWH-21-1-0575"],"award-info":[{"award-number":["W81XWH-21-1-0575"]}],"id":[{"id":"10.13039\/100000090","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Cancer Institute of the National Institutes of Health Cancer Center Support","award":["P30CA012197"],"award-info":[{"award-number":["P30CA012197"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Background\/Objectives:\u00a0TP53 mutation or deletion status is important for determining cellular responses to DNA-damaging drugs. Oxaliplatin (OXA) is combined with the fluoropyrimidine (FP) drug 5-fluorouracil (5-FU) in the FOLFOX regimen used to treat advanced colorectal cancer (CRC). However, the effects of TP53 deletion on 5-FU + OXA synergy are not well known. We investigated potential synergy between OXA and 5-FU and compared it with OXA synergy with a novel polymeric FP, CF10, in four cell lines harboring either wild-type (WT) or TP53-null status. Methods: Using CompuSyn and the highest single agent (HSA) models, we compared synergy between CF10 and OXA (COXA) and between 5-FU and OXA (FOXA). Cell cycle analysis was performed, as was Western blot quantification of canonical DNA damage pathway proteins. Likewise, immunofluorescent and confocal analysis allowed us to compare topoisomerase 1 cleavage complex and double-strand DNA break formation. Results: COXA synergy displayed minimal TP53 dependence with greatly improved potency compared to FOXA. COXA synergy resulted from OXA increasing: (i) Topoisomerase 1 (Top1) cleavage complex formation; (ii) DNA double-strand breaks (DSBs), and (iii) Checkpoint Kinase 1 and 2 (p-Chk1\/2) phosphorylation, consistent with increased replication stress. Additionally, increased S-phase entry in TP53-null cells enhanced synergy between CF10, 5-FU, and OXA as S-phase drugs. Conclusions: Our results demonstrate that OXA synergizes with CF10 more effectively than with 5-FU through enhanced replication stress in both WT and TP53-null cells by causing greater Top1-mediated DNA double-strand breaks. Our studies provide a foundation for further testing of this combination in an orthotopic liver metastatic setting and eventual clinical development.<\/jats:p>","DOI":"10.3390\/cancers18050882","type":"journal-article","created":{"date-parts":[[2026,3,9]],"date-time":"2026-03-09T15:49:42Z","timestamp":1773071382000},"page":"882","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["CF10 Displays Improved Synergy with Oxaliplatin in TP53-Null and Wild-Type CRC Cells from Increased Top1cc and Replication Stress"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-7048-2614","authenticated-orcid":false,"given":"Taylor M.","family":"Young","sequence":"first","affiliation":[{"name":"Department of Cancer Biology, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rida","family":"Moumouni","sequence":"additional","affiliation":[{"name":"Department of Cancer Biology, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Akanksha","family":"Behl","sequence":"additional","affiliation":[{"name":"Department of Cancer Biology, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Upasana","family":"Das","sequence":"additional","affiliation":[{"name":"Department of Cancer Biology, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1883-3791","authenticated-orcid":false,"given":"William H.","family":"Gmeiner","sequence":"additional","affiliation":[{"name":"Department of Cancer Biology, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/S1470-2045(24)00600-4","article-title":"Colorectal cancer incidence trends in younger versus older adults: An analysis of population-based cancer registry data","volume":"26","author":"Sung","year":"2025","journal-title":"Lancet Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1177\/10732748231202470","article-title":"Comparative Analysis of First-Line FOLFOX Treatment with and Without Anti-VEGF Therapy in Metastatic Colorectal Carcinoma: A Real-World Data Study","volume":"30","author":"Brenner","year":"2023","journal-title":"Cancer Control"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1318","DOI":"10.1186\/s40064-016-2976-9","article-title":"Adjuvant FOLFOX treatment for stage III colon cancer: How many cycles are enough?","volume":"5","author":"Tsai","year":"2016","journal-title":"Springerplus"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1111\/jns.12432","article-title":"Platinum accumulation in oxaliplatin-induced peripheral neuropathy","volume":"26","author":"Wei","year":"2021","journal-title":"J. Peripher. Nerv. Syst."},{"key":"ref_5","first-page":"133","article-title":"FOLFOX and FOLFIRI Use in Stage IV Colon Cancer: Analysis of SEER-Medicare Data","volume":"18","author":"Neugut","year":"2019","journal-title":"Clin. Color. Cancer"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"5080","DOI":"10.1200\/JCO.22.02773","article-title":"Leucovorin and Fluorouracil with or Without Oxaliplatin as First-Line Treatment in Advanced Colorectal Cancer","volume":"41","author":"Figer","year":"2023","journal-title":"J. Clin. Oncol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2006","DOI":"10.18632\/oncotarget.28075","article-title":"Pan-drug and drug-specific mechanisms of 5-FU, irinotecan (CPT-11), oxaliplatin, and cisplatin identified by comparison of transcriptomic and cytokine responses of colorectal cancer cells","volume":"12","author":"Carlsen","year":"2021","journal-title":"Oncotarget"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"101778","DOI":"10.1016\/j.xcrm.2024.101778","article-title":"An RNA damage response network mediates the lethality of 5-FU in colorectal cancer","volume":"5","author":"Chen","year":"2024","journal-title":"Cell Rep. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1172\/JCI6863","article-title":"Disruption of p53 in human cancer cells alters the responses to therapeutic agents","volume":"104","author":"Bunz","year":"1999","journal-title":"J. Clin. Investig."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1016\/j.bcp.2007.05.001","article-title":"p53 dependent and independent sensitivity to oxaliplatin of colon cancer cells","volume":"74","author":"Toscano","year":"2007","journal-title":"Biochem. Pharmacol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"772","DOI":"10.1093\/annonc\/mdp383","article-title":"Impact of p53 expression and microsatellite instability on stage III colon cancer disease-free survival in patients treated by 5-fluorouracil and leucovorin with or without oxaliplatin","volume":"21","author":"Zaanan","year":"2010","journal-title":"Ann. Oncol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2332","DOI":"10.1158\/1535-7163.MCT-14-0748","article-title":"The 1,2-Diaminocyclohexane Carrier Ligand in Oxaliplatin Induces p53-Dependent Transcriptional Repression of Factors Involved in Thymidylate Biosynthesis","volume":"14","author":"Kiyonari","year":"2015","journal-title":"Mol. Cancer Ther."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1317","DOI":"10.1038\/s41598-022-05243-6","article-title":"Predicting clinical outcomes of cancer patients with a p53 deficiency gene signature","volume":"12","author":"Schaafsma","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3561","DOI":"10.1182\/blood-2011-06-362442","article-title":"Unique dual targeting of thymidylate synthase and topoisomerase1 by FdUMP[10] results in high efficacy against AML and low toxicity","volume":"119","author":"Pardee","year":"2012","journal-title":"Blood"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1200\/PO.19.00237","article-title":"Clinical Outcomes in Patients with Colon Cancer with Microsatellite Instability of Sporadic or Familial Origin Treated with Adjuvant FOLFOX with or Without Cetuximab: A Pooled Analysis of the PETACC8 and N0147 Trials","volume":"4","author":"Zaanan","year":"2020","journal-title":"JCO Precis. Oncol."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Gmeiner, W.H. (2020). Fluoropyrimidine Modulation of the Anti-Tumor Immune Response-Prospects for Improved Colorectal Cancer Treatment. Cancers, 12.","DOI":"10.3390\/cancers12061641"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"257","DOI":"10.20517\/cdr.2022.136","article-title":"Review of 5-FU resistance mechanisms in colorectal cancer: Clinical significance of attenuated on-target effects","volume":"6","author":"Gmeiner","year":"2023","journal-title":"Cancer Drug Resist."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Okechukwu, C.C., Ma, X., Sah, N., Mani, C., Palle, K., and Gmeiner, W.H. (2024). Enhanced Therapeutic Efficacy of the Nanoscale Fluoropyrimidine Polymer CF10 in a Rat Colorectal Cancer Liver Metastasis Model. Cancers, 16.","DOI":"10.3390\/cancers16071360"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Okechukwu, C.C., Ma, X., Li, W., D\u2019Agostino, R., Rees, M.G., Ronan, M.M., Roth, J.A., and Gmeiner, W.H. (2025). CF10 Displayed Improved Activity Relative to 5-FU in a Mouse CRLM Model Under Conditions of Physiological Folate. Cancers, 17.","DOI":"10.3390\/cancers17172739"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"771","DOI":"10.1002\/1097-0215(20000915)87:6<771::AID-IJC2>3.0.CO;2-V","article-title":"Folate depletion increases sensitivity of solid tumor cell lines to 5-fluorouracil and antifolates","volume":"87","author":"Backus","year":"2000","journal-title":"Int. J. Cancer"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Peters, G.J., Kathmann, I., Giovannetti, E., Smid, K., Assaraf, Y.G., and Jansen, G. (2024). The role of l-leucovorin uptake and metabolism in the modulation of 5-fluorouracil efficacy and antifolate toxicity. Front. Pharmacol., 15.","DOI":"10.3389\/fphar.2024.1450418"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Fardous, A.M., and Heydari, A.R. (2023). Uncovering the Hidden Dangers and Molecular Mechanisms of Excess Folate: A Narrative Review. Nutrients, 15.","DOI":"10.3390\/nu15214699"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2315","DOI":"10.1038\/nprot.2006.339","article-title":"Clonogenic assay of cells in vitro","volume":"1","author":"Franken","year":"2006","journal-title":"Nat. Protoc."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1158\/0008-5472.CAN-09-1947","article-title":"Drug combination studies and their synergy quantification using the Chou-Talalay method","volume":"70","author":"Chou","year":"2010","journal-title":"Cancer Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"a026070","DOI":"10.1101\/cshperspect.a026070","article-title":"p53 in the DNA-Damage-Repair Process","volume":"6","author":"Williams","year":"2016","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"4804","DOI":"10.18632\/oncotarget.3103","article-title":"p53 activates G1 checkpoint following DNA damage by doxorubicin during transient mitotic arrest","volume":"6","author":"Hyun","year":"2014","journal-title":"Oncotarget"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1038\/cdd.2017.169","article-title":"How does p53 induce apoptosis and how does this relate to p53-mediated tumour suppression?","volume":"25","author":"Aubrey","year":"2018","journal-title":"Cell Death Differ."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1007\/978-1-62703-236-0_4","article-title":"p53 and cell cycle effects after DNA damage","volume":"962","author":"Senturk","year":"2013","journal-title":"Methods Mol. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1016\/S0006-2952(97)00541-8","article-title":"Cellular interactions of 5-fluorouracil and the camptothecin analogue CPT-11 (irinotecan) in a human colorectal carcinoma cell line","volume":"55","author":"Guichard","year":"1998","journal-title":"Biochem. Pharmacol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1158\/1535-7163.MCT-20-0516","article-title":"Improved Antitumor Activity of the Fluoropyrimidine Polymer CF10 in Preclinical Colorectal Cancer Models through Distinct Mechanistic and Pharmacologic Properties","volume":"20","author":"Gmeiner","year":"2021","journal-title":"Mol. Cancer Ther."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1652","DOI":"10.3390\/biom5031652","article-title":"Inhibition of Topoisomerase (DNA) I (TOP1): DNA Damage Repair and Anticancer Therapy","volume":"5","author":"Xu","year":"2015","journal-title":"Biomolecules"},{"key":"ref_32","first-page":"255","article-title":"ATM Is Required for the Repair of Oxaliplatin-Induced DNA Damage in Colorectal Cancer","volume":"17","author":"Bakkenist","year":"2018","journal-title":"Clin. Color. Cancer"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1038\/s42003-022-04055-8","article-title":"Rewiring glucose metabolism improves 5-FU efficacy in p53-deficient\/KRAS(G12D) glycolytic colorectal tumors","volume":"5","author":"Ludikhuize","year":"2022","journal-title":"Commun. Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1093\/jmcb\/mjy075","article-title":"Mutant p53 in colon cancer","volume":"11","author":"Nakayama","year":"2019","journal-title":"J. Mol. Cell Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2421584","DOI":"10.1080\/15384047.2024.2421584","article-title":"The polymeric fluoropyrimidine CF10 overcomes limitations of 5-FU in pancreatic ductal adenocarcinoma cells through increased replication stress","volume":"25","author":"Finan","year":"2024","journal-title":"Cancer Biol. Ther."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2933","DOI":"10.1158\/0008-5472.CAN-18-2807","article-title":"Inhibition of Ataxia-Telangiectasia Mutated and RAD3-Related (ATR) Overcomes Oxaliplatin Resistance and Promotes Antitumor Immunity in Colorectal Cancer","volume":"79","author":"Andrade","year":"2019","journal-title":"Cancer Res."}],"container-title":["Cancers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6694\/18\/5\/882\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,9]],"date-time":"2026-03-09T16:05:03Z","timestamp":1773072303000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6694\/18\/5\/882"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,9]]},"references-count":36,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,3]]}},"alternative-id":["cancers18050882"],"URL":"https:\/\/doi.org\/10.3390\/cancers18050882","relation":{},"ISSN":["2072-6694"],"issn-type":[{"value":"2072-6694","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,9]]}}}