HDAC inhibitors induce HLA class I molecules through the SOX10-IRF1 axis in clear cell sarcoma cells

HDAC inhibitors induce HLA class I molecules through the SOX10-IRF1 axis in clear cell sarcoma cells | 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 HDAC inhibitors induce HLA class I molecules through the SOX10-IRF1 axis in clear cell sarcoma cells Minh T. Nguyen, Ryota Kikuchi, Soshi Nishibu, Yue Zhou, Hiroshi Moritake, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4427899/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 Although immune checkpoint inhibitors (ICIs) are an effective treatment for clear cell sarcoma (CCS), a rare melanocytic sarcoma with a poor prognosis, their efficacies are still limited. Therefore, a novel therapeutic strategy is required to improve the efficacy of ICIs. We previously reported that HDAC inhibitors increased melanoma immunogenicity through the SOX10-IRF1 pathway and may improve the efficacy of ICIs for melanoma. We herein demonstrated that the inhibition of HDAC induced the expression of HLA class I molecules through IRF1 in CCS cells, similar to melanoma. The suppression of SOX10 by siRNA induced the expression of HLA class I molecules. In addition, the isoform-specific inhibition of HDAC1/3 induced the expression of another IRF1 downstream molecule, PD-L1 in CCS cells in concert with the suppression of SOX10. Furthermore, the knockdown of IRF1 impaired the induction of PD-L1 expression in CCS cells. Therefore, the inhibition of HDAC1/3 has potential as a novel strategy to increase immunogenicity and as combination therapy with ICIs for CCS and melanoma. Biological sciences/Cancer/Sarcoma Biological sciences/Cancer/Skin cancer Histone deacetylase inhibitors SOX10 IRF1 PD-L1 clear cell sarcoma Figures Figure 1 Figure 2 Figure 3 Figure 4 Full Text Additional Declarations No competing interests reported. Supplementary Files SupplementalFigure1.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-4427899","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":307918317,"identity":"cb4a00d1-7496-4f10-b66f-c322e389cc90","order_by":0,"name":"Minh T. Nguyen","email":"","orcid":"","institution":"University of Toyama","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Minh","middleName":"T.","lastName":"Nguyen","suffix":""},{"id":307918318,"identity":"fea48bf8-5471-4689-b5be-0f291ae413d6","order_by":1,"name":"Ryota Kikuchi","email":"","orcid":"","institution":"University of Toyama","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ryota","middleName":"","lastName":"Kikuchi","suffix":""},{"id":307918319,"identity":"62f675fe-bbe8-4b89-bab2-07b2f4821055","order_by":2,"name":"Soshi Nishibu","email":"","orcid":"","institution":"University of Toyama","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Soshi","middleName":"","lastName":"Nishibu","suffix":""},{"id":307918320,"identity":"76bf250c-39b3-4d3f-ad51-388c718917ef","order_by":3,"name":"Yue Zhou","email":"","orcid":"","institution":"University of Toyama","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yue","middleName":"","lastName":"Zhou","suffix":""},{"id":307918321,"identity":"834f2be6-8a03-4c96-b2a0-acf3b226fc67","order_by":4,"name":"Hiroshi Moritake","email":"","orcid":"","institution":"University of Miyazaki","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hiroshi","middleName":"","lastName":"Moritake","suffix":""},{"id":307918322,"identity":"7b263bde-ad45-4a0d-937c-7debfb74e8dc","order_by":5,"name":"Takuro Nakamura","email":"","orcid":"","institution":"Tokyo Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Takuro","middleName":"","lastName":"Nakamura","suffix":""},{"id":307918323,"identity":"3a090412-0e7c-4b17-80ac-61dffc9aafce","order_by":6,"name":"Hidetatsu Outani","email":"","orcid":"","institution":"Osaka University Graduate School of Medicine","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hidetatsu","middleName":"","lastName":"Outani","suffix":""},{"id":307918324,"identity":"d4759498-924d-4b69-b4e9-39f1214910c5","order_by":7,"name":"Ryuji Hayashi","email":"","orcid":"","institution":"Toyama University Hospital","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ryuji","middleName":"","lastName":"Hayashi","suffix":""},{"id":307918325,"identity":"229c6e0f-5eca-4e65-9e51-224679942303","order_by":8,"name":"Hiroaki Sakurai","email":"","orcid":"","institution":"University of Toyama","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hiroaki","middleName":"","lastName":"Sakurai","suffix":""},{"id":307918326,"identity":"1cd316d8-3d72-4cf7-9989-eb1711681ca5","order_by":9,"name":"Satoru Yokoyama","email":"data:image/png;base64,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","orcid":"","institution":"University of Toyama","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Satoru","middleName":"","lastName":"Yokoyama","suffix":""}],"badges":[],"createdAt":"2024-05-16 02:23:29","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4427899/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4427899/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":57327273,"identity":"727ecd35-b6f6-4f46-848e-97cb5331b053","added_by":"auto","created_at":"2024-05-29 07:34:39","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":911041,"visible":true,"origin":"","legend":"\u003cp\u003eSOX10 negatively regulates the expression of IRF1 and HLA class I\u003c/p\u003e\n\u003cp\u003emolecules in clear cell sarcoma.\u003c/p\u003e\n\u003cp\u003eA, The expression of SOX10 and MITF in clear cell sarcoma cells was assessed by\u003c/p\u003e\n\u003cp\u003eWestern blotting. B and C, Hewga-CCS and MP-CCS-SY cells transfected with siCNTL\u003c/p\u003e\n\u003cp\u003eor siSOX10 (#9 or #10) for 48 hours were subjected to Western blotting (C) or flow\u003c/p\u003e\n\u003cp\u003ecytometry (C). Representative flow cytometry graphs are shown.\u003c/p\u003e","description":"","filename":"Figure1031524.png","url":"https://assets-eu.researchsquare.com/files/rs-4427899/v1/18dc74a0685bce42456e61f6.png"},{"id":57327272,"identity":"7252da8f-45ee-4de7-8fee-d1aff952e9a6","added_by":"auto","created_at":"2024-05-29 07:34:39","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":121009,"visible":true,"origin":"","legend":"\u003cp\u003eHDAC1/3 inhibition induces the expression of HLA class I molecules in\u003c/p\u003e\n\u003cp\u003eHewga-CCS cells.\u003c/p\u003e\n\u003cp\u003eA, Hewga-CCS cells were treated with each HDAC inhibitor, vorinostat (2 µM),\u003c/p\u003e\n\u003cp\u003ericolinostat (10 µM), or RGFP109 (10 µM) for 24 hours and then subjected to flow\u003c/p\u003e\n\u003cp\u003ecytometry. Representative flow cytometry graphs are shown. B, Hewga-CCS cells were\u003c/p\u003e\n\u003cp\u003etreated with TMP269 (10 µM) and BRD73954 (10 µM). Other conditions were similar to\u003c/p\u003e\n\u003cp\u003ethose in figure 2A.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e","description":"","filename":"Figure2031524.png","url":"https://assets-eu.researchsquare.com/files/rs-4427899/v1/606042d69ac8c4f79527958f.png"},{"id":57327275,"identity":"444cda00-a10b-4f31-b92a-187206dbc5d4","added_by":"auto","created_at":"2024-05-29 07:34:39","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":198760,"visible":true,"origin":"","legend":"\u003cp\u003eHDAC1/3 inhibition induces the expression of PD-L1 and HLA class I\u003c/p\u003e\n\u003cp\u003emolecules in various clear cell sarcomas.\u003c/p\u003e\n\u003cp\u003eEach clear cell sarcoma cell line was treated with RGFP109 (10 µM) for 24 hours and\u003c/p\u003e\n\u003cp\u003ethen subjected to flow cytometry. Representative flow cytometry graphs are shown.\u003c/p\u003e","description":"","filename":"Figure3031524.png","url":"https://assets-eu.researchsquare.com/files/rs-4427899/v1/795aeede5b75e69d21d0a13b.png"},{"id":57327274,"identity":"0a61fda9-078e-4610-a8f6-44be56946e31","added_by":"auto","created_at":"2024-05-29 07:34:39","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":370400,"visible":true,"origin":"","legend":"\u003cp\u003eIRF1 is indispensable for the induction of PD-L1 expression by HDAC1/3\u003c/p\u003e\n\u003cp\u003einhibition.\u003c/p\u003e\n\u003cp\u003eA. Hewga-CCS cells were treated with RGFP109 at the indicated dose for 24 hours and\u003c/p\u003e\n\u003cp\u003ethen subjected to Western blotting. B, Hewga-CCS cells were transfected with siCNTL\u003c/p\u003e\n\u003cp\u003eor siIRF1 for 24 hours. Cells were then treated with RGFP109 (10 µM) for 24 hours and\u003c/p\u003e\n\u003cp\u003esubjected to flow cytometry. Representative flow cytometry graphs are shown. C.\u003c/p\u003e\n\u003cp\u003eHewga-CCS cells were pretreated with baricitinib (0.5 µM) for 30 min and then treated\u003c/p\u003e\n\u003cp\u003ewith RGFP109 (10 µM) for 24 hours. Cells were subjected to flow cytometry.\u003c/p\u003e\n\u003cp\u003eRepresentative flow cytometry graphs are shown.\u003c/p\u003e","description":"","filename":"Figure4031524.png","url":"https://assets-eu.researchsquare.com/files/rs-4427899/v1/ceb81ab9a67beccdb933a8af.png"},{"id":64180863,"identity":"1053f3da-7ede-46c3-b916-359e37c4298e","added_by":"auto","created_at":"2024-09-09 14:44:51","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2120371,"visible":true,"origin":"","legend":"","description":"","filename":"MinhetalSciRep051724.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4427899/v1_covered_12412e61-2c11-48de-b33f-91e2106d4100.pdf"},{"id":57327276,"identity":"aad99b4d-389e-4c6f-91ca-ff627f1d8876","added_by":"auto","created_at":"2024-05-29 07:34:39","extension":"tif","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":6211139,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementalFigure1.tif","url":"https://assets-eu.researchsquare.com/files/rs-4427899/v1/9dbe3d4e3774a7806e806d84.tif"}],"financialInterests":"No competing interests reported.","formattedTitle":"HDAC inhibitors induce HLA class I molecules through the SOX10-IRF1 axis in clear cell sarcoma cells","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":"Histone deacetylase inhibitors, SOX10, IRF1, PD-L1, clear cell sarcoma","lastPublishedDoi":"10.21203/rs.3.rs-4427899/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4427899/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Although immune checkpoint inhibitors (ICIs) are an effective treatment for clear cell sarcoma (CCS), a rare melanocytic sarcoma with a poor prognosis, their efficacies are still limited. Therefore, a novel therapeutic strategy is required to improve the efficacy of ICIs. We previously reported that HDAC inhibitors increased melanoma immunogenicity through the SOX10-IRF1 pathway and may improve the efficacy of ICIs for melanoma. We herein demonstrated that the inhibition of HDAC induced the expression of HLA class I molecules through IRF1 in CCS cells, similar to melanoma. The suppression of SOX10 by siRNA induced the expression of HLA class I molecules. In addition, the isoform-specific inhibition of HDAC1/3 induced the expression of another IRF1 downstream molecule, PD-L1 in CCS cells in concert with the suppression of SOX10. Furthermore, the knockdown of IRF1 impaired the induction of PD-L1 expression in CCS cells. 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