Mechanistic Study and Target Identification of NNK-Induced Head and Neck Squamous Cell Carcinoma via Systems Bioinformatics, Network Toxicology, and Machine Learning

Mechanistic Study and Target Identification of NNK-Induced Head and Neck Squamous Cell Carcinoma via Systems Bioinformatics, Network Toxicology, and Machine Learning | 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 Research Article Mechanistic Study and Target Identification of NNK-Induced Head and Neck Squamous Cell Carcinoma via Systems Bioinformatics, Network Toxicology, and Machine Learning Ziwei Dai, Junqi Su, Xiaofeng Shan, Yifan Kang, Qiushi Feng, Zixuan You, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8260887/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: Head and neck squamous cell carcinoma (HNSCC) is an aggressive malignancy strongly associated with tobacco use. The tobacco-specific nitrosamine NNK is a potent carcinogen, but its molecular mechanisms, particularly its impact on the tumor immune microenvironment and drug resistance in HNSCC, are not fully understood. This study aimed to identify key molecular targets and pathways involved in NNK-induced HNSCC. Methods: We employed an in silico framework integrating systems bioinformatics, network toxicology, and machine learning. Potential targets of NNK and HNSCC-related genes were collected from multiple databases. Core genes were identified from transcriptomic data (GSE107591, GSE138206) through differential expression analysis and machine learning algorithms (XGBoost, Boruta). The stability of the NNK-target interaction was validated using molecular docking and 100 ns molecular dynamics simulations. Immune infiltration and drug sensitivity analyses were also performed. Results: We identified 71 core genes linked to both NNK and HNSCC, from which eight key genes, including monoamine oxidase B (MAOB), were pinpointed as crucial for HNSCC progression. Functional analysis revealed their involvement in the MAPK signaling pathway. MAOB was strongly correlated with an immunosuppressive microenvironment, characterized by increased pro-tumor M2 macrophages and Tregs. Molecular simulations confirmed a stable, high-affinity binding between NNK and MAOB (-7.5 kcal/mol). High MAOB expression was also associated with resistance to MEK inhibitors and Dasatinib. Conclusion: Our computational analysis identifies MAOB as a key mediator in NNK-induced HNSCC, playing a dual role in promoting immune evasion and drug resistance. These findings establish MAOB as a promising therapeutic target and offer new strategies for overcoming treatment resistance in smoking-related HNSCC. Head and Neck Squamous Cell Carcinoma (HNSCC) NNK Network Toxicology Machine Learning Molecular Dynamics Simulation Full Text Additional Declarations No competing interests reported. Supplementary Files GraphicalAbstract.tif Cite Share Download PDF Status: Posted 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-8260887","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":562173920,"identity":"bd2486ad-0b30-4f1c-973f-ca1197ec0cc7","order_by":0,"name":"Ziwei Dai","email":"","orcid":"","institution":"Peking University School and Hospital of Stomatology","correspondingAuthor":false,"prefix":"","firstName":"Ziwei","middleName":"","lastName":"Dai","suffix":""},{"id":562173921,"identity":"1527e85a-64ec-49f8-a16e-3a3e166c7314","order_by":1,"name":"Junqi Su","email":"","orcid":"","institution":"Peking University School and Hospital of 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Dynamics Simulation","lastPublishedDoi":"10.21203/rs.3.rs-8260887/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8260887/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eHead and neck squamous cell carcinoma (HNSCC) is an aggressive malignancy strongly associated with tobacco use. The tobacco-specific nitrosamine NNK is a potent carcinogen, but its molecular mechanisms, particularly its impact on the tumor immune microenvironment and drug resistance in HNSCC, are not fully understood. This study aimed to identify key molecular targets and pathways involved in NNK-induced HNSCC.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods: \u003c/strong\u003eWe employed an in silico framework integrating systems bioinformatics, network toxicology, and machine learning. Potential targets of NNK and HNSCC-related genes were collected from multiple databases. Core genes were identified from transcriptomic data (GSE107591, GSE138206) through differential expression analysis and machine learning algorithms (XGBoost, Boruta). The stability of the NNK-target interaction was validated using molecular docking and 100 ns molecular dynamics simulations. Immune infiltration and drug sensitivity analyses were also performed.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eWe identified 71 core genes linked to both NNK and HNSCC, from which eight key genes, including monoamine oxidase B (MAOB), were pinpointed as crucial for HNSCC progression. Functional analysis revealed their involvement in the MAPK signaling pathway. MAOB was strongly correlated with an immunosuppressive microenvironment, characterized by increased pro-tumor M2 macrophages and Tregs. Molecular simulations confirmed a stable, high-affinity binding between NNK and MAOB (-7.5 kcal/mol). High MAOB expression was also associated with resistance to MEK inhibitors and Dasatinib.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion: \u003c/strong\u003eOur computational analysis identifies MAOB as a key mediator in NNK-induced HNSCC, playing a dual role in promoting immune evasion and drug resistance. These findings establish MAOB as a promising therapeutic target and offer new strategies for overcoming treatment resistance in smoking-related HNSCC.\u003c/p\u003e","manuscriptTitle":"Mechanistic Study and Target Identification of NNK-Induced Head and Neck Squamous Cell Carcinoma via Systems Bioinformatics, Network Toxicology, and Machine Learning","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-19 19:30:23","doi":"10.21203/rs.3.rs-8260887/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"d048a6c3-6933-4fbf-a18b-ebae44f5d700","owner":[],"postedDate":"December 19th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-02-25T09:27:51+00:00","versionOfRecord":[],"versionCreatedAt":"2025-12-19 19:30:23","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8260887","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8260887","identity":"rs-8260887","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}