The Generation of an ocular anti-inflammatory biotherapeutic to enhance wound healing.

The Generation of an ocular anti-inflammatory biotherapeutic to enhance wound healing. | 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 The Generation of an ocular anti-inflammatory biotherapeutic to enhance wound healing. Anthony St. Leger, Jackie Shane, Matthew Evans, Yannis Rigas, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4354377/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 Microbes exist at and colonize mucosal surfaces striking a balance with the host immune system, so that these microbes can thrive on host tissues without causing pathology. Because of this, mucosal barrier-colonizing bacteria can be leveraged to act as long-term delivery vehicles for naturally derived therapeutics. Here, we use a mouse model of corneal wound healing to show that the eye-colonizing bacterium, Corynebacterium mastitidis (C. mast) can be engineered to produce and secrete bioactive murine anti-inflammatory interleukin (mIL)-10. Specifically, we used transposon mutagenesis to identify a native C. mast-specific secretion signal that was used to direct C. mast to secrete mIL-10. Mini-transposons were generated to deliver secretion capable mIL-10 to the bacterial genome. After screening, two isolates were identified that can: 1) colonize the eye, 2) produce and secrete mIL-10, and 3) enhance wound healing in an IL-10-dependent manner. This proof of concept illustrates that eye-colonizing bacteria can be engineered to deliver therapeutics to the ocular surface for the alleviation of ocular surface disease(s). Biological sciences/Immunology/Immunotherapy/Immunosuppression Biological sciences/Microbiology/Bacteria/Bacterial techniques and applications Biological sciences/Genetics/Gene expression Health sciences/Medical research/Translational research Full Text Additional Declarations There is NO Competing Interest. Supplementary Files Sourcedatafile.xlsx Related Manuscript File 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-4354377","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":300717801,"identity":"d3fc7975-ad68-4546-86a4-0d40079d51a5","order_by":0,"name":"Anthony St. Leger","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABB0lEQVRIiWNgGAWjYHACNijN3AAi5RiYITw8OpihWtgYwVqMGZiZQSwStCQ2MBDQwj+7/9iDjzsOM8jPb2x8+KXiXvp2dv7jDxgqrBMbcGiRuHOY3XDmmcMMBscYm41lzhTn7mwG2XImHacWhhvJbNK8bUAtbIxt0pJtCbkbDgO1MLYdxqlFHqZFvo2x/bfkv4R0A7CWf7i1GMC0MBxjbGP82JCQANHSgFuL4Y1kM8mZbek8BscSm6UZjiUYAv1iOCPhWLoxLi1yNxKfSXxss5aTbz588OOPmgR5c/6DDz58qLGWxel9CGjmAZHMINIAxErArxwE6sAk4w+YllEwCkbBKBgFSAAAcANX/o8F7wQAAAAASUVORK5CYII=","orcid":"","institution":"University of Pittsburgh School of Medicine","correspondingAuthor":true,"prefix":"","firstName":"Anthony","middleName":"St.","lastName":"Leger","suffix":""},{"id":300717802,"identity":"bc220119-6894-4803-9729-17d39d7866f3","order_by":1,"name":"Jackie Shane","email":"","orcid":"","institution":"University of Pittsburgh","correspondingAuthor":false,"prefix":"","firstName":"Jackie","middleName":"","lastName":"Shane","suffix":""},{"id":300717803,"identity":"9e2ff02e-94f6-40e5-8fac-8e1eb44e7206","order_by":2,"name":"Matthew Evans","email":"","orcid":"","institution":"University of Pittsburgh","correspondingAuthor":false,"prefix":"","firstName":"Matthew","middleName":"","lastName":"Evans","suffix":""},{"id":300717804,"identity":"c13c61ff-fcf3-418e-a858-6cde0f2eb7e6","order_by":3,"name":"Yannis Rigas","email":"","orcid":"","institution":"University of Pittsburgh","correspondingAuthor":false,"prefix":"","firstName":"Yannis","middleName":"","lastName":"Rigas","suffix":""},{"id":300717805,"identity":"2ceec426-2fb4-4db2-b738-5df1a0b2c178","order_by":4,"name":"Robert Shanks","email":"","orcid":"","institution":"University of Pittsburgh","correspondingAuthor":false,"prefix":"","firstName":"Robert","middleName":"","lastName":"Shanks","suffix":""}],"badges":[],"createdAt":"2024-05-01 12:55:06","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4354377/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4354377/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":62568803,"identity":"cfd04ae4-2816-42ae-b104-efb40b9ce829","added_by":"auto","created_at":"2024-08-16 02:41:53","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":465821,"visible":true,"origin":"","legend":"","description":"","filename":"singlesubmissionfile.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4354377/v1_covered_9cb53c6c-ba6b-45f7-b246-fa484a62f093.pdf"},{"id":62568454,"identity":"90dd6112-72f0-43c7-acf8-ff6a3a00e648","added_by":"auto","created_at":"2024-08-16 02:33:51","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":39775,"visible":true,"origin":"","legend":"Related Manuscript File","description":"","filename":"Sourcedatafile.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4354377/v1/3e4ee707758e192184a063d9.xlsx"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"The Generation of an ocular anti-inflammatory biotherapeutic to enhance wound healing.","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"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":"npj-biofilms-and-microbiomes","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"npjbiofilms","sideBox":"Learn more about [npj Biofilms and Microbiomes](http://www.nature.com/npjbiofilms/)","snPcode":"41522","submissionUrl":"https://submission.springernature.com/new-submission/41522/3","title":"npj Biofilms and Microbiomes","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"NPJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-4354377/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4354377/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Microbes exist at and colonize mucosal surfaces striking a balance with the host immune system, so that these microbes can thrive on host tissues without causing pathology. Because of this, mucosal barrier-colonizing bacteria can be leveraged to act as long-term delivery vehicles for naturally derived therapeutics. Here, we use a mouse model of corneal wound healing to show that the eye-colonizing bacterium, Corynebacterium mastitidis (C. mast) can be engineered to produce and secrete bioactive murine anti-inflammatory interleukin (mIL)-10. Specifically, we used transposon mutagenesis to identify a native C. mast-specific secretion signal that was used to direct C. mast to secrete mIL-10. Mini-transposons were generated to deliver secretion capable mIL-10 to the bacterial genome. After screening, two isolates were identified that can: 1) colonize the eye, 2) produce and secrete mIL-10, and 3) enhance wound healing in an IL-10-dependent manner. This proof of concept illustrates that eye-colonizing bacteria can be engineered to deliver therapeutics to the ocular surface for the alleviation of ocular surface disease(s).","manuscriptTitle":"The Generation of an ocular anti-inflammatory biotherapeutic to enhance wound healing.","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-08-16 02:33:47","doi":"10.21203/rs.3.rs-4354377/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"npj-biofilms-and-microbiomes","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"npjbiofilms","sideBox":"Learn more about [npj Biofilms and Microbiomes](http://www.nature.com/npjbiofilms/)","snPcode":"41522","submissionUrl":"https://submission.springernature.com/new-submission/41522/3","title":"npj Biofilms and Microbiomes","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"NPJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"887a501d-5cd1-4dd9-b59b-742b7da48417","owner":[],"postedDate":"August 16th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":31730935,"name":"Biological sciences/Immunology/Immunotherapy/Immunosuppression"},{"id":31730936,"name":"Biological sciences/Microbiology/Bacteria/Bacterial techniques and applications"},{"id":31730937,"name":"Biological sciences/Genetics/Gene expression"},{"id":31730938,"name":"Health sciences/Medical research/Translational research"}],"tags":[],"updatedAt":"2024-08-16T02:33:47+00:00","versionOfRecord":[],"versionCreatedAt":"2024-08-16 02:33:47","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4354377","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4354377","identity":"rs-4354377","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}