Drugging the Medullary Thyroid Cancer Surfaceome with T Cell Engagers Targeting CEA, GFRA4 and DLL3 as Monotherapy and In Combination with Tyrosine Kinase Inhibitors

Drugging the Medullary Thyroid Cancer Surfaceome with T Cell Engagers Targeting CEA, GFRA4 and DLL3 as Monotherapy and In Combination with Tyrosine Kinase Inhibitors | 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 Drugging the Medullary Thyroid Cancer Surfaceome with T Cell Engagers Targeting CEA, GFRA4 and DLL3 as Monotherapy and In Combination with Tyrosine Kinase Inhibitors Tim Andrew Erickson This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7942399/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 Medullary thyroid cancer (MTC) is a rare form of thyroid cancer, and the definitive treatment is surgical resection. For patients who are not cured by surgery or for patients who present with distant metastatic disease, no curative therapy exists. T cell engagers (TCEs) are an emerging class of biologics that simultaneously bind to tumor cell surface antigens and the CD3 domain of T cells, thereby redirecting endogenous T cells to lyse tumor cells. TCEs can mediate durable tumor regression, including complete responses, and are now approved to treat multiple cancer types outside of MTC. RNASeq analysis of 30 MTC tumors, cross-referencing with public databases and an extensive literature review were used to identify CEA, DLL3 and GFRA4 as promising tumor antigens to target with TCEs. Expression of these antigens was then validated on MTC cell lines. Herein, I describe the discovery, application and development of MTC-targeted TCEs (MTCEs) recognizing the MTC tumor antigens CEA, DLL3 and GFRA4. In vitro, MTCEs mediate potent target-dependent and T cell-dependent cytotoxicity against MTC cell lines at concentrations of just 10 ng/mL. MTCEs also induce target-dependent IFNg secretion from T cells, with no measurable IFNg secretion observed in the presence of antigen-negative cell lines. Further, MTCEs are functionally compatible with the FDA-approved tyrosine kinase inhibitors selpercatinib and cabozantinib, and combination therapy numerically enhances cytotoxicity. These preclinical data provide strong rationale for continued development of MTCEs, which may one day revolutionize the treatment of metastatic MTC. Biological sciences/Cancer Biological sciences/Drug discovery Health sciences/Oncology Full Text Additional Declarations Competing interest reported. By virtue of being the founder of ThyThera, there may be a competing financial interest. The author attests to the accuracy of the data, and in the future may submit patents based on presented and future experiments. While results have been uploaded to preprint servers, a peer-reviewed publication could potentially enhance future funding opportunities through grants or private capital markets. As tarlatamab is already FDA-approved, there is no declared financial interest in any tarlatamab findings. The anti-GFRA4 and anti-CEA TCEs described have novel amino acid sequences and are therefore novel molecular entities. 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. 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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-7942399","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":539146387,"identity":"84bda450-75ba-436f-a23c-3e23b019d914","order_by":0,"name":"Tim Andrew Erickson","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/UlEQVRIiWNgGAWjYDACZjApAYRA8IHhAAOEhQfwIGthnJFwgIGHoBYYA6SQmYcYLfbs3AkMH/dYyEv2Hz722PbHncT90s3HPvxgsJPTbcDlMN4NjDOeSRjOlkhLN85JeJbYI3MseWYPQ7Kx2QHcWph5DkgkyEnwmEnnJBxO7JHIMQa690DiNnxa/oC08J8xk7YAa8n/zPiHkBYGoBZphhwzaQaILczMeG05zLvhYM8BCcOZM9LSJHvSDhv33EgzZpYxwO0X9v6zGx/8OFAnL3H+8DGJHzaHZdtnJD9mfFNhJ4dLCwigSDk2gCkD3MoxgD0JakfBKBgFo2CEAAAiIlaeKrS1eQAAAABJRU5ErkJggg==","orcid":"","institution":"ThyThera, LLC","correspondingAuthor":true,"prefix":"","firstName":"Tim","middleName":"Andrew","lastName":"Erickson","suffix":""}],"badges":[],"createdAt":"2025-10-25 23:24:21","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7942399/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7942399/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":95110264,"identity":"3dc85607-6e08-4d81-8dab-46e5e32d9e44","added_by":"auto","created_at":"2025-11-04 11:45:36","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1150410,"visible":true,"origin":"","legend":"","description":"","filename":"DruggingMTCSurfaceomeScientificReports.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7942399/v1/8f02218ea1171c59f19a2300.pdf"},{"id":95110263,"identity":"3dcc6fcb-250d-42ab-94a8-3f8b349b58af","added_by":"auto","created_at":"2025-11-04 11:45:35","extension":"json","order_by":1,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":4991,"visible":true,"origin":"","legend":"","description":"","filename":"bfe42aad7ec54f5e890bb415366d18bd.json","url":"https://assets-eu.researchsquare.com/files/rs-7942399/v1/7d11d1bb9b6c076b53b20d02.json"},{"id":108093669,"identity":"5a8b00f7-4dae-405c-b617-c0bc5f9d5b46","added_by":"auto","created_at":"2026-04-29 09:41:50","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":932147,"visible":true,"origin":"","legend":"","description":"","filename":"DruggingMTCSurfaceomeScientificReports.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7942399/v1_covered_81988dbc-01ff-43a3-b79e-b08807192ce6.pdf"}],"financialInterests":"Competing interest reported. By virtue of being the founder of ThyThera, there may be a competing financial interest. The author attests to the accuracy of the data, and in the future may submit patents based on presented and future experiments. While results have been uploaded to preprint servers, a peer-reviewed publication could potentially enhance future funding opportunities through grants or private capital markets. As tarlatamab is already FDA-approved, there is no declared financial interest in any tarlatamab findings. The anti-GFRA4 and anti-CEA TCEs described have novel amino acid sequences and are therefore novel molecular entities.","formattedTitle":"Drugging the Medullary Thyroid Cancer Surfaceome with T Cell Engagers Targeting CEA, GFRA4 and DLL3 as Monotherapy and In Combination with Tyrosine Kinase Inhibitors","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"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},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-7942399/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7942399/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Medullary thyroid cancer (MTC) is a rare form of thyroid cancer, and the definitive treatment is surgical resection. For patients who are not cured by surgery or for patients who present with distant metastatic disease, no curative therapy exists. T cell engagers (TCEs) are an emerging class of biologics that simultaneously bind to tumor cell surface antigens and the CD3 domain of T cells, thereby redirecting endogenous T cells to lyse tumor cells. TCEs can mediate durable tumor regression, including complete responses, and are now approved to treat multiple cancer types outside of MTC. RNASeq analysis of 30 MTC tumors, cross-referencing with public databases and an extensive literature review were used to identify CEA, DLL3 and GFRA4 as promising tumor antigens to target with TCEs. Expression of these antigens was then validated on MTC cell lines. Herein, I describe the discovery, application and development of MTC-targeted TCEs (MTCEs) recognizing the MTC tumor antigens CEA, DLL3 and GFRA4. In vitro, MTCEs mediate potent target-dependent and T cell-dependent cytotoxicity against MTC cell lines at concentrations of just 10 ng/mL. MTCEs also induce target-dependent IFNg secretion from T cells, with no measurable IFNg secretion observed in the presence of antigen-negative cell lines. Further, MTCEs are functionally compatible with the FDA-approved tyrosine kinase inhibitors selpercatinib and cabozantinib, and combination therapy numerically enhances cytotoxicity. 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