Dynamic Targetable Extracellular Vesicle Surface Proteins Monitor Depth of Response to CAR T-Cell Therapy

Dynamic Targetable Extracellular Vesicle Surface Proteins Monitor Depth of Response to CAR T-Cell Therapy | 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 Dynamic Targetable Extracellular Vesicle Surface Proteins Monitor Depth of Response to CAR T-Cell Therapy Yazhen Zhu, Chen Zhao, Lei Qiu, Ning An, Yong Ju, Jacqueline Yang, and 30 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8913641/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Extracellular vesicles (EVs) represent a promising liquid biopsy platform in multiple myeloma (MM). We developed an MM EV Surface Protein Assay to quantify and dynamically monitor four MM EV subpopulations defined by targetable MM surface proteins (BCMA, CD38, GPRC5D, and CD319) across 336 serial blood samples from 45 relapsed/refractory MM (RRMM) patients treated with anti-BCMA chimeric antigen receptor (CAR) T-cell therapy. All four MM EV subpopulations significantly decreased in 43 patients with initial response, while BCMA⁺, GPRC5D⁺, and CD319⁺ MM EVs increased in 19 patients with progression, and antigen escape was detected by BCMA⁺ MM EVs. MM EV subpopulations differentiated minimal residual disease (MRD) status and complemented MRD for detecting early relapse before clinical progression. Notably, CD319⁺ MM EVs were early predictors of progression-free and overall survival in MRD-negative patients. This assay enables noninvasive monitoring of deep response, progression, and antigen escape, and stratifies survival in MRD-negative patients with RRMM. Health sciences/Biomarkers/Predictive markers Health sciences/Oncology/Cancer/Tumour biomarkers Full Text Additional Declarations Yes there is potential Competing Interest. HRT is a co-founder and shareholder in Cytolumina Technologies Corp, Pulsar Therapeutics, and Eximius Diagnostics Corp. Y Zhu is a co-founder and shareholder in Eximius Diagnostics Corp. Written informed consent was obtained from participants when required. The study was approved by IRB. Supplementary Files SupplementaryMaterialsNC.pdf Supplementary Materials Cite Share Download PDF Status: Under Review 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-8913641","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":605019973,"identity":"a68211ea-6d09-4493-b423-e4a502f2601e","order_by":0,"name":"Yazhen Zhu","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABDUlEQVRIie3QMUvDQBTA8VeEy3Ih64XY73Cla+lnuSOQLsbFxUkyvUm7Z+h3iBRqxysBu6S7pUuyZNGCboramkTccqGjw/2Hu+NxPzgOwGT6h3EFpNoOAqwIgP4Oe6peom5CBFDVEFZPO4kb/REmTiSOtS7ztyW95Ntnme+Xoxtgfq7eYdRPVDshVEwGccau+C6cD2ZZwIAFfHULwVBLQDx6NnKZ7MJFdUgrcgGKQiq1xCnQ+0Yhk+3m4dPGY0NWX3DUE+YTr4dKJk/24sxG1ZCUguogJXHvKhJn4dydoe8iLXl6zv1hrH3YpGQfeJDT9eb+dY9jx7H8oni5HvenGtL2IXX81Osmk8lkausHz8JjrnPW27UAAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0002-2130-8085","institution":"University of California, Los Angeles","correspondingAuthor":true,"prefix":"","firstName":"Yazhen","middleName":"","lastName":"Zhu","suffix":""},{"id":605019974,"identity":"4812451b-1fe7-4e45-b3b9-ef561d25e613","order_by":1,"name":"Chen Zhao","email":"","orcid":"","institution":"University of California, Los Angeles","correspondingAuthor":false,"prefix":"","firstName":"Chen","middleName":"","lastName":"Zhao","suffix":""},{"id":605019975,"identity":"5e2033cc-22c7-4ed9-962b-6974bebcb8b4","order_by":2,"name":"Lei Qiu","email":"","orcid":"","institution":"Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences","correspondingAuthor":false,"prefix":"","firstName":"Lei","middleName":"","lastName":"Qiu","suffix":""},{"id":605019976,"identity":"febc2732-a32e-41f7-a0a6-fb3665532620","order_by":3,"name":"Ning An","email":"","orcid":"","institution":"Huazhong University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Ning","middleName":"","lastName":"An","suffix":""},{"id":605019977,"identity":"e8b8aeb4-ed67-4ca6-9ffb-085756e95fe2","order_by":4,"name":"Yong Ju","email":"","orcid":"","institution":"University of California, Los Angeles","correspondingAuthor":false,"prefix":"","firstName":"Yong","middleName":"","lastName":"Ju","suffix":""},{"id":605019978,"identity":"c8ee4c6f-5b0b-48c3-954c-207d1c72d292","order_by":5,"name":"Jacqueline Yang","email":"","orcid":"","institution":"UCLA","correspondingAuthor":false,"prefix":"","firstName":"Jacqueline","middleName":"","lastName":"Yang","suffix":""},{"id":605019979,"identity":"1c477e26-5d96-4e53-b9b1-74219d0238bd","order_by":6,"name":"Hyoyong Kim","email":"","orcid":"https://orcid.org/0000-0002-8931-8298","institution":"University of California, Los Angeles","correspondingAuthor":false,"prefix":"","firstName":"Hyoyong","middleName":"","lastName":"Kim","suffix":""},{"id":605019980,"identity":"e46390d4-f809-441f-9181-cd65f7596641","order_by":7,"name":"Ryan Zhang","email":"","orcid":"","institution":"University of California, Los Angeles","correspondingAuthor":false,"prefix":"","firstName":"Ryan","middleName":"","lastName":"Zhang","suffix":""},{"id":605019981,"identity":"ae2d1fbf-e497-4656-bee7-ccbc95f5340c","order_by":8,"name":"Junseok Lee","email":"","orcid":"https://orcid.org/0000-0002-3709-1386","institution":"University of California, Los Angeles","correspondingAuthor":false,"prefix":"","firstName":"Junseok","middleName":"","lastName":"Lee","suffix":""},{"id":605019982,"identity":"91cc956d-8f2d-412a-a02a-63c42487550c","order_by":9,"name":"Liang-Yan Chen","email":"","orcid":"","institution":"UCLA","correspondingAuthor":false,"prefix":"","firstName":"Liang-Yan","middleName":"","lastName":"Chen","suffix":""},{"id":605019983,"identity":"3be570d6-5c29-4aa4-8f25-fbca27de2af5","order_by":10,"name":"Yaya Xu","email":"","orcid":"","institution":"UCLA","correspondingAuthor":false,"prefix":"","firstName":"Yaya","middleName":"","lastName":"Xu","suffix":""},{"id":605019984,"identity":"9af2576f-3e3c-4ddd-bcf8-c09971bb4763","order_by":11,"name":"Yue Ma","email":"","orcid":"","institution":"UCLA","correspondingAuthor":false,"prefix":"","firstName":"Yue","middleName":"","lastName":"Ma","suffix":""},{"id":605019985,"identity":"99050353-8d94-4b5f-8fec-aa4e2c80b609","order_by":12,"name":"Sophie Pan","email":"","orcid":"","institution":"UCLA","correspondingAuthor":false,"prefix":"","firstName":"Sophie","middleName":"","lastName":"Pan","suffix":""},{"id":605019986,"identity":"d170efb5-8d76-41fd-9c58-8ea913e445fe","order_by":13,"name":"Pranay Sashikanth","email":"","orcid":"","institution":"UCLA","correspondingAuthor":false,"prefix":"","firstName":"Pranay","middleName":"","lastName":"Sashikanth","suffix":""},{"id":605019987,"identity":"d78af7b3-2eb8-4bea-b219-98a3f855e53f","order_by":14,"name":"Jina Kim","email":"","orcid":"","institution":"Cedars-Sinai Medical Center","correspondingAuthor":false,"prefix":"","firstName":"Jina","middleName":"","lastName":"Kim","suffix":""},{"id":605019988,"identity":"a91fad64-3e77-44fd-8c79-0ac1c5bd089d","order_by":15,"name":"Di Wang","email":"","orcid":"","institution":"Huazhong University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Di","middleName":"","lastName":"Wang","suffix":""},{"id":605019989,"identity":"ecd63bb2-a7a4-486f-971d-5423db9e27e7","order_by":16,"name":"Peiling Zhang","email":"","orcid":"https://orcid.org/0000-0002-1567-7458","institution":"Huazhong University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Peiling","middleName":"","lastName":"Zhang","suffix":""},{"id":605019990,"identity":"efe171d3-c749-4913-8832-9dbdade63b15","order_by":17,"name":"Jianlin Hu","email":"","orcid":"","institution":"Huazhong University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Jianlin","middleName":"","lastName":"Hu","suffix":""},{"id":605019991,"identity":"b6b1adb4-4fc1-468e-be9f-3bd9ca4dff4b","order_by":18,"name":"Yang Gao","email":"","orcid":"","institution":"Huazhong University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Yang","middleName":"","lastName":"Gao","suffix":""},{"id":605019992,"identity":"aeaedbc5-56b1-4a0a-9fb7-be8886329bc6","order_by":19,"name":"Yuhan Bao","email":"","orcid":"","institution":"Huazhong University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Yuhan","middleName":"","lastName":"Bao","suffix":""},{"id":605019993,"identity":"9db6b484-a74e-4d8b-9372-322a9ec0cf5c","order_by":20,"name":"Yanhua Tu","email":"","orcid":"","institution":"Nanjing IASO Medical Technology Co. Ltd.","correspondingAuthor":false,"prefix":"","firstName":"Yanhua","middleName":"","lastName":"Tu","suffix":""},{"id":605019994,"identity":"9380f9ac-ea74-471b-96a3-1a61e16db407","order_by":21,"name":"Li Wang","email":"","orcid":"","institution":"Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences","correspondingAuthor":false,"prefix":"","firstName":"Li","middleName":"","lastName":"Wang","suffix":""},{"id":605019995,"identity":"0dbb59a0-c3aa-48cb-b981-cd6936eaf92f","order_by":22,"name":"Zhili Wang","email":"","orcid":"","institution":"Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences","correspondingAuthor":false,"prefix":"","firstName":"Zhili","middleName":"","lastName":"Wang","suffix":""},{"id":605019996,"identity":"d2f647e1-afb8-4367-b932-317fc59b07c6","order_by":23,"name":"Zhicheng Zhang","email":"","orcid":"","institution":"Huazhong University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Zhicheng","middleName":"","lastName":"Zhang","suffix":""},{"id":605019997,"identity":"134565e0-042a-4488-b2fb-b9d124e73233","order_by":24,"name":"Edwin Posadas","email":"","orcid":"","institution":"Cedars-Sinai Medical Center","correspondingAuthor":false,"prefix":"","firstName":"Edwin","middleName":"","lastName":"Posadas","suffix":""},{"id":605019998,"identity":"758fd74c-8d76-4a9e-9260-c61b518605ca","order_by":25,"name":"Sungyong You","email":"","orcid":"https://orcid.org/0000-0003-3513-1783","institution":"Cedars-Sinai Medical Center","correspondingAuthor":false,"prefix":"","firstName":"Sungyong","middleName":"","lastName":"You","suffix":""},{"id":605019999,"identity":"3b5a29c0-025b-4283-b03b-d94623598078","order_by":26,"name":"Tasha Lin","email":"","orcid":"https://orcid.org/0000-0003-4281-4152","institution":"University of California, Los Angeles","correspondingAuthor":false,"prefix":"","firstName":"Tasha","middleName":"","lastName":"Lin","suffix":""},{"id":605020000,"identity":"c005c51a-776c-46b4-bcd8-dc003c748388","order_by":27,"name":"Dinesh Rao","email":"","orcid":"https://orcid.org/0000-0002-0794-9337","institution":"University of California, Los Angeles","correspondingAuthor":false,"prefix":"","firstName":"Dinesh","middleName":"","lastName":"Rao","suffix":""},{"id":605020001,"identity":"81ea7e15-f538-45b1-8696-326b6c46f540","order_by":28,"name":"Sarah Larson","email":"","orcid":"","institution":"UCLA","correspondingAuthor":false,"prefix":"","firstName":"Sarah","middleName":"","lastName":"Larson","suffix":""},{"id":605020002,"identity":"31ee3b5e-d44b-4bc4-9047-73930b8d45e1","order_by":29,"name":"Renjun Pei","email":"","orcid":"https://orcid.org/0000-0002-9353-3935","institution":"Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences","correspondingAuthor":false,"prefix":"","firstName":"Renjun","middleName":"","lastName":"Pei","suffix":""},{"id":605020003,"identity":"6720a567-c503-4c65-8e86-ad7d7f6eb2b0","order_by":30,"name":"Hsian-Rong Tseng","email":"","orcid":"https://orcid.org/0000-0003-0942-5905","institution":"University of California, Los Angeles","correspondingAuthor":false,"prefix":"","firstName":"Hsian-Rong","middleName":"","lastName":"Tseng","suffix":""},{"id":605020004,"identity":"c084e03e-0fae-45bf-8189-dadca2338f75","order_by":31,"name":"Na Sun","email":"","orcid":"","institution":"Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences","correspondingAuthor":false,"prefix":"","firstName":"Na","middleName":"","lastName":"Sun","suffix":""},{"id":605020005,"identity":"41abb7db-f004-4c70-b683-511d4dc128db","order_by":32,"name":"Chunrui Li","email":"","orcid":"https://orcid.org/0000-0001-5134-7133","institution":"Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China;","correspondingAuthor":false,"prefix":"","firstName":"Chunrui","middleName":"","lastName":"Li","suffix":""},{"id":605020006,"identity":"c8e4f4d3-df91-46fb-a191-3937c4c9d571","order_by":33,"name":"Jingjing He","email":"","orcid":"","institution":"UCLA","correspondingAuthor":false,"prefix":"","firstName":"Jingjing","middleName":"","lastName":"He","suffix":""},{"id":605020007,"identity":"82b9177c-9522-46fe-bca8-7f37446be6cf","order_by":34,"name":"Wanxing Chai-Ho","email":"","orcid":"","institution":"UCLA","correspondingAuthor":false,"prefix":"","firstName":"Wanxing","middleName":"","lastName":"Chai-Ho","suffix":""},{"id":605020008,"identity":"b9629a93-555b-4a75-bb7f-2f02d1ed662f","order_by":35,"name":"Emily Ren","email":"","orcid":"","institution":"UCLA","correspondingAuthor":false,"prefix":"","firstName":"Emily","middleName":"","lastName":"Ren","suffix":""}],"badges":[],"createdAt":"2026-02-19 04:22:29","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8913641/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8913641/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":105562635,"identity":"ccb8cab6-9739-49b6-bc31-da60e6a52961","added_by":"auto","created_at":"2026-03-27 12:43:53","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2557421,"visible":true,"origin":"","legend":"Article File","description":"","filename":"ManuscriptNC.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8913641/v1_covered_0035e9ed-984b-4b22-8437-caaf5044753e.pdf"},{"id":104871351,"identity":"9ecf54b1-cded-4ff2-8535-5f7df5564072","added_by":"auto","created_at":"2026-03-18 08:08:13","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1677217,"visible":true,"origin":"","legend":"Supplementary Materials","description":"","filename":"SupplementaryMaterialsNC.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8913641/v1/9a8f0126d1ec8ec4167685d8.pdf"}],"financialInterests":"\u003cp\u003e\u003cstrong\u003eYes\u003c/strong\u003e there is potential Competing Interest. HRT is a co-founder and shareholder in Cytolumina Technologies Corp, Pulsar Therapeutics, and Eximius Diagnostics Corp. Y Zhu is a co-founder and shareholder in Eximius Diagnostics Corp.\u003c/p\u003e\n\u003cp\u003eWritten informed consent was obtained from participants when required. The study was approved by IRB.\u003c/p\u003e","formattedTitle":"Dynamic Targetable Extracellular Vesicle Surface Proteins Monitor Depth of Response to CAR T-Cell Therapy","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"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":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-8913641/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8913641/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Extracellular vesicles (EVs) represent a promising liquid biopsy platform in multiple myeloma (MM). We developed an MM EV Surface Protein Assay to quantify and dynamically monitor four MM EV subpopulations defined by targetable MM surface proteins (BCMA, CD38, GPRC5D, and CD319) across 336 serial blood samples from 45 relapsed/refractory MM (RRMM) patients treated with anti-BCMA chimeric antigen receptor (CAR) T-cell therapy. All four MM EV subpopulations significantly decreased in 43 patients with initial response, while BCMA⁺, GPRC5D⁺, and CD319⁺ MM EVs increased in 19 patients with progression, and antigen escape was detected by BCMA⁺ MM EVs. MM EV subpopulations differentiated minimal residual disease (MRD) status and complemented MRD for detecting early relapse before clinical progression. Notably, CD319⁺ MM EVs were early predictors of progression-free and overall survival in MRD-negative patients. This assay enables noninvasive monitoring of deep response, progression, and antigen escape, and stratifies survival in MRD-negative patients with RRMM.","manuscriptTitle":"Dynamic Targetable Extracellular Vesicle Surface Proteins Monitor Depth of Response to CAR T-Cell Therapy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-03-18 08:08:05","doi":"10.21203/rs.3.rs-8913641/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"nature-communications","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"NCOMMS","sideBox":"Learn more about [Nature Communications](http://www.nature.com/ncomms/)","snPcode":"","submissionUrl":"https://mts-ncomms.nature.com/","title":"Nature Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature Communications","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"bbf76486-8bca-4c01-8730-6b3109d1ee42","owner":[],"postedDate":"March 18th, 2026","published":true,"recentEditorialEvents":[{"type":"editorInvitedReview","content":"This content is not available.","date":"2026-05-09T19:24:12+00:00","index":4,"fulltext":"This content is not available."},{"type":"reviewerAgreed","content":"This content is not available.","date":"2026-04-29T18:21:19+00:00","index":5,"fulltext":"This content is not available."}],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":64386049,"name":"Health sciences/Biomarkers/Predictive markers"},{"id":64386050,"name":"Health sciences/Oncology/Cancer/Tumour biomarkers"}],"tags":[],"updatedAt":"2026-03-18T08:08:05+00:00","versionOfRecord":[],"versionCreatedAt":"2026-03-18 08:08:05","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8913641","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8913641","identity":"rs-8913641","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}