Cleavage of CAD by caspase-3 determines the cancer cell fate during chemotherapy

Cleavage of CAD by caspase-3 determines the cancer cell fate during chemotherapy | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Cleavage of CAD by caspase-3 determines the cancer cell fate during chemotherapy Xuehui Hong, Jingsong Ma, Jiabao Zhao, Chen-Song Zhang, Jinshui Tan, and 16 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4826976/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 30 May, 2025 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract Metabolic heterogeneity resulting from the intrinsic heterogeneity of human tumors has been shown to cause a myriad of adverse outcomes of tumor therapy, including resistance to chemotherapy, but the mechanisms inside remain largely unknown. Here, we found that the de novo pyrimidine synthesis pathway, which we previously identified as crucial for the proliferation of gastric and colon cancer cells, determines the chemosensitivity of these cancer types. In the chemosensitive cells, chemotherapeutic drugs such as 5-FU promoted the degradation of CAD, an enzyme that is rate-limiting for pyrimidine synthesis, leading to the apoptosis of these cells. We also found that CAD needed to be cleaved, by activated caspase-3 on its Asp1371 residue, before its degradation. Upregulation of CAD, either by overexpression or by a mutation on the Asp1371 to block the caspase-3 cleavage, conferred tumor chemoresistance in both mouse xenograft models and the Cldn18-ATK mouse gastric tumor models. Importantly, mutations related to Asp1371 of CAD were found in gastric tumor samples of patients who underwent unsuccessful neoadjuvant chemotherapy. By searching for a compound that capable for degradating the CAD-Asp1371 mutant, we discovered that a compound called RMY-186 can be used to treat chemoresistant tumors related to CAD mutations, resulting in a significant improvement in the effectiveness of chemotherapy. We have, therefore, revealed the vulnerability of de novo pyrimidine synthesis during chemotherapy, which can be targeted to overcome chemotherapeutic resistance and augment the antitumor efficacy of genotoxic chemotherapy agents. Biological sciences/Cancer/Gastrointestinal cancer/Gastric cancer Biological sciences/Cancer/Gastrointestinal cancer/Colorectal cancer/Colon cancer Biological sciences/Cancer/Tumour heterogeneity Biological sciences/Cancer/Cancer metabolism Biological sciences/Cancer/Tumour biomarkers Tumor heterogeneity Pyrimidine metabolism CAD Caspase-3 Chemosensitivity Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryFigures.pdf Cite Share Download PDF Status: Published Journal Publication published 30 May, 2025 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4826976","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":338246975,"identity":"63319bc3-cbf4-4237-810e-4f5d8429dcb1","order_by":0,"name":"Xuehui 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