Assessment of Cell Surface Targets in Metastatic Prostate Cancer: Expression Landscape and Molecular Correlates

Assessment of Cell Surface Targets in Metastatic Prostate Cancer: Expression Landscape and Molecular Correlates | 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 Help Center Sign In Submit a Preprint Cite Share Download PDF Article Assessment of Cell Surface Targets in Metastatic Prostate Cancer: Expression Landscape and Molecular Correlates Michael Haffner, Azra Ajkunic, Erolcan Sayar, Martine Roudier, and 23 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3745991/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 17 May, 2024 Read the published version in npj Precision Oncology → Version 1 posted 11 You are reading this latest preprint version Abstract Therapeutic approaches targeting proteins on the surface of cancer cells have emerged as an important strategy for precision oncology. To fully capitalize on the potential impact of drugs targeting surface proteins, detailed knowledge about the expression patterns of the target proteins in tumor tissues is required. In castration-resistant prostate cancer (CRPC), agents targeting prostate-specific membrane antigen (PSMA) have demonstrated clinical activity. However, PSMA expression is lost in a significant number of CRPC tumors, and the identification of additional cell surface targets is necessary in order to develop new therapeutic approaches. Here, we performed a comprehensive analysis of the expression and co-expression patterns of trophoblast cell-surface antigen 2 (TROP2), delta-like ligand 3 (DLL3), and carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5) in CRPC samples from a rapid autopsy cohort. We show that DLL3 and CEACAM5 exhibit the highest expression in neuroendocrine prostate cancer (NEPC), while TROP2 is expressed across different CRPC molecular subtypes, except for NEPC. We observed variable intra-tumoral and inter-tumoral heterogeneity and no dominant metastatic site predilections for TROP2, DLL3, and CEACAM5. We further show that AR amplifications were associated with higher expression of PSMA and TROP2 but lower DLL3 and CEACAM5 levels. Conversely, PSMA and TROP2 expression was lower in RB1-altered tumors. In addition to genomic alterations, we demonstrate a tight correlation between epigenetic states, particularly histone H3 lysine 27 methylation (H3K27me3) at the transcriptional start site and gene body of TACSTD2 (encoding TROP2), DLL3, and CEACAM5, and their respective protein expression in CRPC patient-derived xenografts. Collectively, these findings provide novel insights into the patterns and determinants of expression of TROP2, DLL3, and CEACAM5 with important implications for the clinical development of cell surface targeting agents in CRPC. Biological sciences/Cancer/Urological cancer/Prostate cancer Biological sciences/Cancer/Cancer therapy/Targeted therapies Figures Figure 1 Figure 2 Figure 3 Full Text Additional Declarations (Not answered) Supplementary Files SupplementalMaterial.pdf Cite Share Download PDF Status: Published Journal Publication published 17 May, 2024 Read the published version in npj Precision Oncology → Version 1 posted Editorial decision: revise 19 Jan, 2024 Review # 3 received at journal 15 Jan, 2024 Review # 2 received at journal 11 Jan, 2024 Reviewer # 3 agreed at journal 09 Jan, 2024 Review # 1 received at journal 08 Jan, 2024 Reviewer # 2 agreed at journal 02 Jan, 2024 Reviewer # 1 agreed at journal 21 Dec, 2023 Reviewers invited by journal 16 Dec, 2023 Editor assigned by journal 14 Dec, 2023 Submission checks completed at journal 14 Dec, 2023 First submitted to journal 12 Dec, 2023 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. 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Also discoverable on Platform About Our Team In Review Editorial Policies 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-3745991","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":260322919,"identity":"659f91d8-d5ad-4abf-8b79-a568e342ea76","order_by":0,"name":"Michael 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1","display":"","copyAsset":false,"role":"figure","size":1828015,"visible":true,"origin":"","legend":"\u003cp\u003eDistribution and co-expression patterns of DLL3, CEACAM5, PSMA, and TROP2 expressions across different molecular subtypes of mCRPC. A. Representative images of cell \u0026nbsp;surface antigen expressions (determined by IHC) across different molecular subtypes (AR+/NE- \u0026nbsp;[green], AR-/NE+ [yellow], AR+/NE+ [red], and AR-/NE- [blue]). Molecular subtypes were defined by expression of AR signaling markers (AR, NKX3.1) and NE markers (SYP, INSM1) as described previously 20 . Box plots show the distribution of B. DLL3, C. CEACAM5, and D. TROP2 expressions based on H-score in the UW-TAN cohort (N=753). Box and dot colors indicate molecular phenotypes as above. E. Top, micrographs of PSMA and TROP2 in AR+/NE- tumors. Bottom, donut chart shows \u0026nbsp;the distribution of PSMA and TROP2 reactivity. F. Top, micrographs of DLL3 and CEACAM5 in AR- /NE+ tumors. Bottom, donut chart shows the distribution of DLL3 and CEACAM5 reactivity. (See Supplementary Table 2 for all co-expression profiles). Scale bars denote 50 μm.\u003c/p\u003e","description":"","filename":"Figures131.png","url":"https://assets-eu.researchsquare.com/files/rs-3745991/v1/7afb84395121897ba8b86d80.png"},{"id":48491480,"identity":"4e141fd5-8c2e-42fb-ac42-5ec3697aa2f4","added_by":"auto","created_at":"2023-12-19 20:49:10","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":130293,"visible":true,"origin":"","legend":"\u003cp\u003eAnatomic site distribution and inter- and intra-tumoral heterogeneity of TROP2, \u0026nbsp;DLL3 and CEACAM5 expression in mCRPC. A. Distribution of DLL3, TROP2, and CEACAM5 \u0026nbsp;protein expression across different organ sites based on IHC H-scores. Dot colors indicate \u0026nbsp;molecular phenotypes. Each dot represents a tumor sample; the color codes indicate the molecular \u0026nbsp;subtype (AR+/NE- [green], AR-/NE+ [yellow], AR+/NE+ [red], and AR-/NE- [blue]). B. Inter- and \u0026nbsp;intra-tumoral heterogeneity of TROP2, PSMA, CEACAM5 and DLL3 expression. Mean (95% \u0026nbsp;confidence interval) hypergeometric expression heterogeneity indices across different metastatic \u0026nbsp;sites in a given patient (inter-tumoral heterogeneity, red) and within a metastatic site (intratumoral heterogeneity, gray). Dot and box plots showing the distribution of C. DLL3, D. TROP2, \u0026nbsp;and E. CEACAM5 protein expression IHC H-scores in 52 cases from the UW-TAN cohort. Each \u0026nbsp;dot represents a tumor sample; the color codes indicate the molecular subtype (AR+/NE- [green], \u0026nbsp;AR-/NE+ [yellow], AR+/NE+ [red], and AR-/NE- [blue]). Gray shadings show interquartile ranges. \u0026nbsp;Percentages show the frequencies of cell surface antigens in cases with uniformly low/negative \u0026nbsp;expression (all sites H-score \u0026lt;20), heterogeneous expression (both H-scores \u0026lt;20 and H-score ≥20) \u0026nbsp;and uniformly high expression (all sites H-scores ≥20).\u003c/p\u003e","description":"","filename":"Figures132.png","url":"https://assets-eu.researchsquare.com/files/rs-3745991/v1/df845045deb80dbd06201560.png"},{"id":48491481,"identity":"cf837332-aa1d-4fc1-994a-cde25d6a2844","added_by":"auto","created_at":"2023-12-19 20:49:10","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":129627,"visible":true,"origin":"","legend":"\u003cp\u003eGenetic and epigenetic determinants of TROP2, PSMA, DLL3 and CEACAM5 expression in CRPC. A. Mosaic plots show the frequencies of TROP2, PSMA, DLL3, and \u0026nbsp;CEACAM5 protein expression determined by IHC (1, expressed; 0, not expressed) as a function of \u0026nbsp;13 the genomic status of AR, CHD1, PTEN, RB1, and TP53 (1, altered; 0, not altered) in 44 cases of \u0026nbsp;the UW-TAN cohort. B. Mosaic plots show the frequencies of TACSTD2, FOLH1, DLL3 and \u0026nbsp;CEACAM5 mRNA expression determined by RNA-seq (1, expressed; 0, not expressed) as a \u0026nbsp;function of the genomic status of AR, BRCA2, CHD1, PTEN, RB1, SPOP, and TP53 (1, altered; 0, \u0026nbsp;not altered) in 99 cases of the SU2C-WCDT. C. Representative H3K27ac (gray) and H3K27me3 \u0026nbsp;ChIP-seq tracks from AR+/NE- (LuCaP 77 and LuCaP 78) and AR-/NE+ (LuCaP 93 and LuCaP 145.1) PDX lines. Note the inverse differential enrichment pattern of H3K27ac and H3K27me3 \u0026nbsp;(yellow box) in the upstream regulatory regions of TACSTD2, DLL3, and CEACAM5.\u003c/p\u003e","description":"","filename":"Figures133.png","url":"https://assets-eu.researchsquare.com/files/rs-3745991/v1/b0e355d162858409f92987d6.png"},{"id":56697466,"identity":"9dd5ef71-0001-4a94-ad3a-10c75a6ad4db","added_by":"auto","created_at":"2024-05-18 07:09:46","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1186505,"visible":true,"origin":"","legend":"","description":"","filename":"Ajkunicetal.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3745991/v1_covered_61e1e237-3cd4-4431-a883-cc11830db763.pdf"},{"id":48491483,"identity":"2f0d17b3-0f67-48c7-99e7-6cc0e3acd093","added_by":"auto","created_at":"2023-12-19 20:49:10","extension":"pdf","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":1203881,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementalMaterial.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3745991/v1/fdba0a10517084db3f3ab8ae.pdf"}],"financialInterests":"(Not answered)","formattedTitle":"\u003cp\u003eAssessment of Cell Surface Targets in Metastatic Prostate Cancer: Expression Landscape and Molecular Correlates\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"npj-precision-oncology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"npjprecisiononcology","sideBox":"Learn more about [npj Precision Oncology](http://www.nature.com/npjprecisiononcology/)","snPcode":"41698","submissionUrl":"https://submission.springernature.com/new-submission/41698/3","title":"npj Precision Oncology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"NPJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-3745991/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3745991/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Therapeutic approaches targeting proteins on the surface of cancer cells have emerged as an important strategy for precision oncology. To fully capitalize on the potential impact of drugs targeting surface proteins, detailed knowledge about the expression patterns of the target proteins in tumor tissues is required. In castration-resistant prostate cancer (CRPC), agents targeting prostate-specific membrane antigen (PSMA) have demonstrated clinical activity. However, PSMA expression is lost in a significant number of CRPC tumors, and the identification of additional cell surface targets is necessary in order to develop new therapeutic approaches. Here, we performed a comprehensive analysis of the expression and co-expression patterns of trophoblast cell-surface antigen 2 (TROP2), delta-like ligand 3 (DLL3), and carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5) in CRPC samples from a rapid autopsy cohort. We show that DLL3 and CEACAM5 exhibit the highest expression in neuroendocrine prostate cancer (NEPC), while TROP2 is expressed across different CRPC molecular subtypes, except for NEPC. We observed variable intra-tumoral and inter-tumoral heterogeneity and no dominant metastatic site predilections for TROP2, DLL3, and CEACAM5. We further show that AR amplifications were associated with higher expression of PSMA and TROP2 but lower DLL3 and CEACAM5 levels. Conversely, PSMA and TROP2 expression was lower in RB1-altered tumors. In addition to genomic alterations, we demonstrate a tight correlation between epigenetic states, particularly histone H3 lysine 27 methylation (H3K27me3) at the transcriptional start site and gene body of TACSTD2 (encoding TROP2), DLL3, and CEACAM5, and their respective protein expression in CRPC patient-derived xenografts. Collectively, these findings provide novel insights into the patterns and determinants of expression of TROP2, DLL3, and CEACAM5 with important implications for the clinical development of cell surface targeting agents in CRPC.","manuscriptTitle":"Assessment of Cell Surface Targets in Metastatic Prostate Cancer: Expression Landscape and Molecular Correlates","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-12-19 20:49:05","doi":"10.21203/rs.3.rs-3745991/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"revise","date":"2024-01-19T10:41:38+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"This content is not available.","date":"2024-01-16T04:47:34+00:00","index":3,"fulltext":"This content is not available."},{"type":"editorInvitedReview","content":"This content is not available.","date":"2024-01-11T20:18:25+00:00","index":2,"fulltext":"This content is not available."},{"type":"reviewerAgreed","content":"This content is not available.","date":"2024-01-09T23:06:39+00:00","index":3,"fulltext":"This content is not available."},{"type":"editorInvitedReview","content":"This content is not available.","date":"2024-01-08T23:26:11+00:00","index":1,"fulltext":"This content is not available."},{"type":"reviewerAgreed","content":"This content is not available.","date":"2024-01-02T21:55:17+00:00","index":2,"fulltext":"This content is not available."},{"type":"reviewerAgreed","content":"This content is not available.","date":"2023-12-21T11:27:42+00:00","index":1,"fulltext":"This content is not available."},{"type":"reviewersInvited","content":"","date":"2023-12-16T15:47:00+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2023-12-14T14:00:54+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2023-12-14T14:00:40+00:00","index":"","fulltext":""},{"type":"submitted","content":"npj Precision Oncology","date":"2023-12-13T01:27:46+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"npj-precision-oncology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"npjprecisiononcology","sideBox":"Learn more about [npj Precision Oncology](http://www.nature.com/npjprecisiononcology/)","snPcode":"41698","submissionUrl":"https://submission.springernature.com/new-submission/41698/3","title":"npj Precision Oncology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"NPJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"80084194-2f4f-4d63-9d4f-bfd9e3b5d8b2","owner":[],"postedDate":"December 19th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":27557577,"name":"Biological sciences/Cancer/Urological cancer/Prostate cancer"},{"id":27557578,"name":"Biological sciences/Cancer/Cancer therapy/Targeted therapies"}],"tags":[],"updatedAt":"2024-05-18T07:09:37+00:00","versionOfRecord":{"articleIdentity":"rs-3745991","link":"https://doi.org/10.1038/s41698-024-00599-6","journal":{"identity":"npj-precision-oncology","isVorOnly":false,"title":"npj Precision Oncology"},"publishedOn":"2024-05-17 04:00:00","publishedOnDateReadable":"May 17th, 2024"},"versionCreatedAt":"2023-12-19 20:49:05","video":"","vorDoi":"10.1038/s41698-024-00599-6","vorDoiUrl":"https://doi.org/10.1038/s41698-024-00599-6","workflowStages":[]},"version":"v1","identity":"rs-3745991","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3745991","identity":"rs-3745991","version":["v1"]},"buildId":"wLkW0s4AflPzk-lpfg-fK","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}