Type I interferon-mediated autoinflammation in two unrelated patients due to a proximal intronic splice site variant in DNASE2

Type I interferon-mediated autoinflammation in two unrelated patients due to a proximal intronic splice site variant in DNASE2 | 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 Type I interferon-mediated autoinflammation in two unrelated patients due to a proximal intronic splice site variant in DNASE2 Oskar Schnappauf, Hongying Wang, Qing Zhou, Vasily Burlakov, Anna Kozlova, and 14 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7039031/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 Purpose Type I interferonopathies are Mendelian inborn errors of immunity caused by defective clearance or recognition of nucleic acids, resulting in chronic type I interferon (IFN) activation. DNase II is a lysosomal endonuclease critical for degrading DNA during erythropoiesis and apoptosis. Biallelic loss-of-function variants in DNASE2 cause a rare autoinflammatory disorder. We aimed to characterize the clinical, genetic, and functional impact of a novel DNASE2 splice site variant in two unrelated patients. Methods We performed exome and RNA sequencing to identify and validate variants. A minigene assay assessed splicing effects. DNase II activity was tested in cell lysates. Structural modeling, cytokine profiling, and immune stimulation assays were conducted to evaluate pathway activation and therapeutic response. Results Both patients carried a homozygous DNASE2 splice site variant (c.511 + 5G > A), resulting in exon 4 skipping and impaired DNase II activity. They presented at birth with rash, thrombocytopenia, and anemia, later developing neutropenia, arthritis, failure to thrive, and systemic inflammation. One patient also harbored a homozygous USP43 variant that reduced USP43 expression and enhanced IFN signaling. Both patients showed increased JAK-STAT activation and responded clinically to JAK1/2 inhibition with ruxolitinib. Conclusion We report a novel proximal intronic splice variant in DNASE2 associated with DNase II deficiency and type I interferon–mediated autoinflammation. Our findings expand the genotypic and phenotypic spectrum of this disorder, highlight the role of intronic variants in rare immune diseases, and support JAK inhibition as a targeted treatment approach. Type I interferonopathy DNASE2 Autoinflammation Splice site variant JAK-STAT signaling Full Text Additional Declarations No competing interests reported. Supplementary Files ManuscriptSchnappaufetalSupplememtaryinformation.docx 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-7039031","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":491892968,"identity":"6b01d5e4-b1e0-4e29-87ae-6ac0d8646968","order_by":0,"name":"Oskar 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DNase II is a lysosomal endonuclease critical for degrading DNA during erythropoiesis and apoptosis. Biallelic loss-of-function variants in \u003cem\u003eDNASE2\u003c/em\u003e cause a rare autoinflammatory disorder. We aimed to characterize the clinical, genetic, and functional impact of a novel \u003cem\u003eDNASE2\u003c/em\u003e splice site variant in two unrelated patients.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e\u003cp\u003eWe performed exome and RNA sequencing to identify and validate variants. A minigene assay assessed splicing effects. DNase II activity was tested in cell lysates. Structural modeling, cytokine profiling, and immune stimulation assays were conducted to evaluate pathway activation and therapeutic response.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003eBoth patients carried a homozygous \u003cem\u003eDNASE2\u003c/em\u003e splice site variant (c.511\u0026thinsp;+\u0026thinsp;5G\u0026thinsp;\u0026gt;\u0026thinsp;A), resulting in exon 4 skipping and impaired DNase II activity. They presented at birth with rash, thrombocytopenia, and anemia, later developing neutropenia, arthritis, failure to thrive, and systemic inflammation. One patient also harbored a homozygous \u003cem\u003eUSP43\u003c/em\u003e variant that reduced USP43 expression and enhanced IFN signaling. Both patients showed increased JAK-STAT activation and responded clinically to JAK1/2 inhibition with ruxolitinib.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e\u003cp\u003eWe report a novel proximal intronic splice variant in \u003cem\u003eDNASE2\u003c/em\u003e associated with DNase II deficiency and type I interferon\u0026ndash;mediated autoinflammation. Our findings expand the genotypic and phenotypic spectrum of this disorder, highlight the role of intronic variants in rare immune diseases, and support JAK inhibition as a targeted treatment approach.\u003c/p\u003e","manuscriptTitle":"Type I interferon-mediated autoinflammation in two unrelated patients due to a proximal intronic splice site variant in DNASE2","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-31 22:14:16","doi":"10.21203/rs.3.rs-7039031/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":"081ad131-f28c-4288-a214-484ae542a998","owner":[],"postedDate":"July 31st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-01-06T13:24:48+00:00","versionOfRecord":[],"versionCreatedAt":"2025-07-31 22:14:16","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7039031","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7039031","identity":"rs-7039031","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}