Identifying TMEM127-deficient pheochromocytomas/paragangliomas via RET overexpression by immunohistochemistry

Identifying TMEM127-deficient pheochromocytomas/paragangliomas via RET overexpression by immunohistochemistry | 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 Identifying TMEM127-deficient pheochromocytomas/paragangliomas via RET overexpression by immunohistochemistry Cynthia Estrada-Zuniga, Rui Liang, Bethany Landry, Andrea Alvarez, and 15 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8553860/v2 This work is licensed under a CC BY 4.0 License Status: Posted Version 2 posted You are reading this latest preprint version Show more versions Abstract Context : Pheochromocytomas and paragangliomas (PPGLs) are rare, genetically diverse tumors originating from the adrenal medulla or extra-adrenal paraganglia, respectively. It is critically important to establish the pathogenic status of genetic variants, especially for the 35-40% patients carrying a germline change, as it impacts patient management and family surveillance. However, determining the clinical impact of variants of uncertain significance (VUS) remains challenging and requires additional testing. This is especially relevant for genes for which limited functional information is available, such as the TMEM127 gene. We recently reported that loss of TMEM127 promotes RET accumulation by reducing its degradation. Objective : Here, we evaluated RET expression by immunohistochemistry (IHC) as a potential aid to highlight TMEM127 dysfunction in PPGLs carrying TMEM127 germline variants. Methods : We performed RET IHC in 104 formalin-fixed and paraffin-embedded (FFPE) sections of clinically and genetically diverse PPGLs and applied histochemical scoring (HS) for membrane (MH-S) and cytoplasm (CH-S) staining. Results : Tumors driven by TMEM127 variants carried the highest RET expression scores (151.8±62), predominantly MH-S, when compared with all other PPGL genotypes, including those with RET pathogenic disruptions (69.9±96.8, adjusted p=0.03) or tumors of undefined genotype (40.8±69, adjusted p=0.0001). RET membrane immunoreactivity also distinguished PPGLs carrying TMEM127 germline variants with likely damaging effects from non-disrupting variants. Conclusions : These findings point to increased RET membrane expression as a promising biomarker for loss-of-function TMEM127 variants in PPGLs. If validated in independent cohorts, RET IHC could be an effective tool for assessing the functional implications of PPGLs from patients carrying TMEM127 VUS. TMEM127 RET pheochromocytomas paragangliomas immunohistochemistry germline variant biomarker VUS Full Text Additional Declarations The authors declare no competing interests. Supplementary Files EstradaZunigasupplfigures.pdf Cite Share Download PDF Status: Posted Version 2 posted You are reading this latest preprint version Show more versions 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-8553860","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":577446757,"identity":"e8ebf953-9d82-41b0-8ca3-b67c32af0a13","order_by":0,"name":"Cynthia Estrada-Zuniga","email":"","orcid":"","institution":"The University of Texas Health Science Center at San Antonio","correspondingAuthor":false,"prefix":"","firstName":"Cynthia","middleName":"","lastName":"Estrada-Zuniga","suffix":""},{"id":577446758,"identity":"9e749cc9-457c-4ba6-a3ad-aa0a6feaf677","order_by":1,"name":"Rui Liang","email":"","orcid":"","institution":"The University of 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immunohistochemistry","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"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":"TMEM127, RET, pheochromocytomas, paragangliomas, immunohistochemistry, germline variant, biomarker, VUS","lastPublishedDoi":"10.21203/rs.3.rs-8553860/v2","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8553860/v2","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eContext\u003c/strong\u003e: Pheochromocytomas and paragangliomas (PPGLs) are rare, genetically diverse tumors originating from the adrenal medulla or extra-adrenal paraganglia, respectively. It is critically important to establish the pathogenic status of genetic variants, especially for the 35-40% patients carrying a germline change, as it impacts patient management and family surveillance. However, determining the clinical impact of variants of uncertain significance (VUS) remains challenging and requires additional testing. This is especially relevant for genes for which limited functional information is available, such as the \u003cem\u003eTMEM127\u003c/em\u003e gene. We recently reported that loss of \u003cem\u003eTMEM127\u003c/em\u003e promotes RET accumulation by reducing its degradation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eObjective\u003c/strong\u003e: Here, we evaluated RET expression by immunohistochemistry (IHC) as a potential aid to highlight TMEM127 dysfunction in PPGLs carrying \u003cem\u003eTMEM127\u003c/em\u003e germline variants.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e: We performed RET IHC in 104 formalin-fixed and paraffin-embedded (FFPE) sections of clinically and genetically diverse PPGLs and applied histochemical scoring (HS) for membrane (MH-S) and cytoplasm (CH-S) staining.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e: Tumors driven by \u003cem\u003eTMEM127\u003c/em\u003evariants carried the highest RET expression scores (151.8±62), predominantly MH-S, when compared with all other PPGL genotypes, including those with \u003cem\u003eRET \u003c/em\u003epathogenic disruptions (69.9±96.8, adjusted p=0.03) or tumors of undefined genotype (40.8±69, adjusted p=0.0001). RET membrane immunoreactivity also distinguished PPGLs carrying \u003cem\u003eTMEM127 \u003c/em\u003egermline variants with likely damaging effects from non-disrupting variants.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions\u003c/strong\u003e: These findings point to increased RET membrane expression as a promising biomarker for loss-of-function \u003cem\u003eTMEM127\u003c/em\u003evariants in PPGLs. If validated in independent cohorts, RET IHC could be an effective tool for assessing the functional implications of PPGLs from patients carrying \u003cem\u003eTMEM127\u003c/em\u003e VUS.\u003c/p\u003e","manuscriptTitle":"Identifying TMEM127-deficient pheochromocytomas/paragangliomas via RET overexpression by immunohistochemistry","msid":"","msnumber":"","nonDraftVersions":[{"code":2,"date":"2026-02-19 21:45:10","doi":"10.21203/rs.3.rs-8553860/v2","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}},{"code":1,"date":"2026-01-20 12:22:54","doi":"10.21203/rs.3.rs-8553860/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":"5c4731a8-66a1-4f20-8798-c86b2697a13a","owner":[],"postedDate":"February 19th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-02-02T22:24:16+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-19 21:45:10","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v2","identity":"rs-8553860","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8553860","identity":"rs-8553860","version":["v2"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}