Functional and structural impact of novel SERPINA1 variants on Alpha-1 Antitrypsin Deficiency

Functional and structural impact of novel SERPINA1 variants on Alpha-1 Antitrypsin Deficiency | 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 Functional and structural impact of novel SERPINA1 variants on Alpha-1 Antitrypsin Deficiency Nerea Matamala, Lourdes Osaba, Estrella Drobnic, Sara Gil-Martín, and 19 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6654733/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 4 You are reading this latest preprint version Abstract Background. Mutations in the SERPINA1 gene can result in alpha-1 antitrypsin deficiency (AATD), which may be associated with lung or liver injury. Although the S and Z alleles account for over 95% of cases of AATD, a wide variety of rare variants have been linked to deficiency and dysfunction, while other variants are associated with normal alpha-1 antitrypsin (AAT) levels and activity. Here, we present the identification and characterization of thirteen rare SERPINA1 variants discovered during the genetic diagnosis of AATD by the Progenika diagnostic network. Methods. The new variants were identified by sequencing the exons of SERPINA1 gene in cases with discrepancies between AAT serum levels and initial genotyping. In order to determine their pathogenic impact, the variants were expressed in a cellular model and evaluated for AAT secretion, intracellular accumulation and elastase inhibitory activity. In addition, protein structural mapping of the variants and analysis of positioning and residue/atomic contacts were performed. Results. The in silico and functional in vitro analysis allowed us to classify these AAT variants as six deficient (p.Val234Glu, p.Val242_Pro243insLeu, p.Leu291Phe, p.Ala308Ser, p.Pro393Thr and p.Pro393Arg), one dysfunctional (p.Thr96Ile), three normal (p.Ser71Arg, p.Ala349Pro and p.Asp365Glu) and three null alleles (p.Gln33*, p.Gln285* and p.Leu310Phefs*14). Conclusions. Functional assays and protein structural information are useful tools in the characterization of novel variants of the SERPINA1 gene. The newly characterized mutations expand the number of SERPINA1 variants with proven pathogenic effects, facilitating the diagnoses of future cases of AATD. alpha1 antitrypsin deficiency mutations diagnosis SERPINA1 Full Text Cite Share Download PDF Status: Under Review Version 1 posted Reviewers agreed at journal 09 Jul, 2025 Reviewers invited by journal 24 Jun, 2025 Editor assigned by journal 14 May, 2025 First submitted to journal 13 May, 2025 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-6654733","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":475490397,"identity":"2f04c2ec-4673-4eff-8732-8f2278413bbf","order_by":0,"name":"Nerea Matamala","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAz0lEQVRIiWNgGAWjYBADGQb2xgYGBgMJHmJUMwKVMvAw8BwkWYtEApEOMhc7/PzBzxw7Hv6Zj5s//CiwALqw/QFeLZaz0wwbe7cl80jcTmyT7AE5jOeMAV4tBrcTDBt4tzHzMAC1MIP9IpGD32EGt9M/Nv7dVs8jf/Ng82ewFvnn+B1mcDvHsJl322EegxuMDdIQWxgIOSyncLbstuM8hmegfmHjySGkJX3Dx7fbquXkjh9//OHHnzp7fvbj+B2GCdhIVD8KRsEoGAWjAAsAAPRvQdx9x3nGAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0002-3082-2414","institution":"ISCIII: Instituto de Salud Carlos III","correspondingAuthor":true,"prefix":"","firstName":"Nerea","middleName":"","lastName":"Matamala","suffix":""},{"id":475490398,"identity":"2e232ce6-4dcd-4f0b-ada6-83ca80846ac1","order_by":1,"name":"Lourdes Osaba","email":"","orcid":"","institution":"Progenika Biopharma","correspondingAuthor":false,"prefix":"","firstName":"Lourdes","middleName":"","lastName":"Osaba","suffix":""},{"id":475490399,"identity":"11f18f30-fdc9-4d41-8432-6015a9218820","order_by":2,"name":"Estrella Drobnic","email":"","orcid":"","institution":"Grifols SA","correspondingAuthor":false,"prefix":"","firstName":"Estrella","middleName":"","lastName":"Drobnic","suffix":""},{"id":475490400,"identity":"aee14805-be01-4c36-b3b9-a9b5f1305963","order_by":3,"name":"Sara Gil-Martín","email":"","orcid":"","institution":"ISCIII: Instituto de Salud Carlos III","correspondingAuthor":false,"prefix":"","firstName":"Sara","middleName":"","lastName":"Gil-Martín","suffix":""},{"id":475490401,"identity":"abcc65fa-8a36-4779-8bb7-583f31f435be","order_by":4,"name":"Antonio J. Martín-Galiano","email":"","orcid":"","institution":"ISCIII: Instituto de Salud Carlos III","correspondingAuthor":false,"prefix":"","firstName":"Antonio","middleName":"J.","lastName":"Martín-Galiano","suffix":""},{"id":475490402,"identity":"c3f82121-55a4-4f29-ad69-9d5a9cfc818a","order_by":5,"name":"Virginia Almadana-Pacheco","email":"","orcid":"","institution":"Hospital Universitario Virgen Macarena","correspondingAuthor":false,"prefix":"","firstName":"Virginia","middleName":"","lastName":"Almadana-Pacheco","suffix":""},{"id":475490403,"identity":"71327348-de03-4c63-a0d0-bf51689fd5cc","order_by":6,"name":"Maria Jesús Cazorla","email":"","orcid":"","institution":"Hospital Universitario Virgen de las Nieves","correspondingAuthor":false,"prefix":"","firstName":"Maria","middleName":"Jesús","lastName":"Cazorla","suffix":""},{"id":475490404,"identity":"1fe3e63a-571d-4762-b145-160132f3ca2c","order_by":7,"name":"Fabio Costa","email":"","orcid":"","institution":"Hospital de Clinicas","correspondingAuthor":false,"prefix":"","firstName":"Fabio","middleName":"","lastName":"Costa","suffix":""},{"id":475490405,"identity":"1965a42c-2bcb-4a55-a283-f35fbf11ee0b","order_by":8,"name":"Caroline Souza Sokoloski","email":"","orcid":"","institution":"Yedikule Chest Diseases and Thoracic Surgery Training and Research Hospital","correspondingAuthor":false,"prefix":"","firstName":"Caroline","middleName":"Souza","lastName":"Sokoloski","suffix":""},{"id":475490406,"identity":"faeb16f8-5545-4b93-b712-85d51703e5a3","order_by":9,"name":"Mustafa Çörtük","email":"","orcid":"","institution":"Yedikule Chest Diseases and Thoracic Surgery Training and Research Hospital","correspondingAuthor":false,"prefix":"","firstName":"Mustafa","middleName":"","lastName":"Çörtük","suffix":""},{"id":475490407,"identity":"61809911-db0d-4106-ba24-4ec2219195bd","order_by":10,"name":"Mireia Serra-Fortuny","email":"","orcid":"","institution":"Hospital Verge de la Cinta","correspondingAuthor":false,"prefix":"","firstName":"Mireia","middleName":"","lastName":"Serra-Fortuny","suffix":""},{"id":475490408,"identity":"236a2745-2ba6-4e78-baf1-eaca49aec91d","order_by":11,"name":"Juan Ortiz-de Saracho","email":"","orcid":"","institution":"Hospital del Bierzo Ponferrada: Hospital del Bierzo","correspondingAuthor":false,"prefix":"","firstName":"Juan","middleName":"Ortiz-de","lastName":"Saracho","suffix":""},{"id":475490409,"identity":"9d945018-42b1-433c-a785-428fbacddc6b","order_by":12,"name":"Juan-Antonio Lloret-Queraltó","email":"","orcid":"","institution":"Hospital of Viladecans: Hospital de Viladecans","correspondingAuthor":false,"prefix":"","firstName":"Juan-Antonio","middleName":"","lastName":"Lloret-Queraltó","suffix":""},{"id":475490410,"identity":"328196be-d6a5-41bc-a35c-10c330a1400e","order_by":13,"name":"Nuria Rodriguez-Lázaro","email":"","orcid":"","institution":"Consorci Sanitari Alt Penedès-Garraf: Consorci Sanitari Alt Penedes-Garraf","correspondingAuthor":false,"prefix":"","firstName":"Nuria","middleName":"","lastName":"Rodriguez-Lázaro","suffix":""},{"id":475490411,"identity":"7f3e6815-943d-4ce6-a84c-88c1e59af3ac","order_by":14,"name":"Francisco Javier Michel-de la Rosa","email":"","orcid":"","institution":"Hospital de Donostia: Hospital Universitario de Donostia","correspondingAuthor":false,"prefix":"","firstName":"Francisco","middleName":"Javier Michel-de la","lastName":"Rosa","suffix":""},{"id":475490412,"identity":"4e5c70ed-6fc9-4280-a6a2-ea032b78afff","order_by":15,"name":"Juan Luis Rodríguez-Hermosa","email":"","orcid":"","institution":"Hospital Clínico San Carlos: Hospital Clinico San Carlos","correspondingAuthor":false,"prefix":"","firstName":"Juan","middleName":"Luis","lastName":"Rodríguez-Hermosa","suffix":""},{"id":475490413,"identity":"154b355e-5ab3-49ef-b91e-5be012100f2f","order_by":16,"name":"Leonardo Saldaña-Pérez","email":"","orcid":"","institution":"Hospital Universitario de Getafe","correspondingAuthor":false,"prefix":"","firstName":"Leonardo","middleName":"","lastName":"Saldaña-Pérez","suffix":""},{"id":475490414,"identity":"fba3e426-71fd-4248-97a6-7e38a2be5264","order_by":17,"name":"Elif Akıncı","email":"","orcid":"","institution":"Ankara University: Ankara Universitesi","correspondingAuthor":false,"prefix":"","firstName":"Elif","middleName":"","lastName":"Akıncı","suffix":""},{"id":475490415,"identity":"4ebdddf6-d9c4-4650-abec-05adf15c3588","order_by":18,"name":"Javier Alonso","email":"","orcid":"","institution":"ISCIII: Instituto de Salud Carlos III","correspondingAuthor":false,"prefix":"","firstName":"Javier","middleName":"","lastName":"Alonso","suffix":""},{"id":475490416,"identity":"177fde29-2e4f-42c3-803c-6884eaaa9814","order_by":19,"name":"Carlos Benítez-Buelga","email":"","orcid":"","institution":"ISCIII: Instituto de Salud Carlos III","correspondingAuthor":false,"prefix":"","firstName":"Carlos","middleName":"","lastName":"Benítez-Buelga","suffix":""},{"id":475490417,"identity":"bfd0a745-956a-4f94-9c46-f1507416c414","order_by":20,"name":"Gema Gómez-Mariano","email":"","orcid":"","institution":"ISCIII: Instituto de Salud Carlos III","correspondingAuthor":false,"prefix":"","firstName":"Gema","middleName":"","lastName":"Gómez-Mariano","suffix":""},{"id":475490418,"identity":"10b54ad8-3e1e-46db-a558-2a416620bfbb","order_by":21,"name":"José Luis López-Campos","email":"","orcid":"","institution":"Hospital Universitario Virgen del Rocío: Hospital Universitario Virgen del Rocio","correspondingAuthor":false,"prefix":"","firstName":"José","middleName":"Luis","lastName":"López-Campos","suffix":""},{"id":475490419,"identity":"bd330ef4-8961-4c8f-82da-e5dcfa423a72","order_by":22,"name":"Beatriz Martínez-Delgado","email":"","orcid":"","institution":"ISCIII: Instituto de Salud Carlos III","correspondingAuthor":false,"prefix":"","firstName":"Beatriz","middleName":"","lastName":"Martínez-Delgado","suffix":""}],"badges":[],"createdAt":"2025-05-13 11:04:47","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6654733/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6654733/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":85429873,"identity":"14d376bc-3819-4a4e-997f-039cfa44fa60","added_by":"auto","created_at":"2025-06-25 18:02:14","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1202064,"visible":true,"origin":"","legend":"","description":"","filename":"SERPINA1newvariantsJTranslMed20250512.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6654733/v1_covered_3bceabca-53f2-4dbc-9f0a-00a3425ff600.pdf"}],"financialInterests":"","formattedTitle":"Functional and structural impact of novel SERPINA1 variants on Alpha-1 Antitrypsin Deficiency","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":"journal-of-translational-medicine","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jtrm","sideBox":"Learn more about [Journal of Translational Medicine](http://translational-medicine.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/jtrm/default.aspx","title":"Journal of Translational Medicine","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"alpha1 antitrypsin deficiency, mutations, diagnosis, SERPINA1","lastPublishedDoi":"10.21203/rs.3.rs-6654733/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6654733/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBackground. Mutations in the SERPINA1 gene can result in alpha-1 antitrypsin deficiency (AATD), which may be associated with lung or liver injury. Although the S and Z alleles account for over 95% of cases of AATD, a wide variety of rare variants have been linked to deficiency and dysfunction, while other variants are associated with normal alpha-1 antitrypsin (AAT) levels and activity. Here, we present the identification and characterization of thirteen rare SERPINA1 variants discovered during the genetic diagnosis of AATD by the Progenika diagnostic network.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eMethods. The new variants were identified by sequencing the exons of SERPINA1 gene in cases with discrepancies between AAT serum levels and initial genotyping. In order to determine their pathogenic impact, the variants were expressed in a cellular model and evaluated for AAT secretion, intracellular accumulation and elastase inhibitory activity. In addition, protein structural mapping of the variants and analysis of positioning and residue/atomic contacts were performed.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eResults. The in silico and functional in vitro analysis allowed us to classify these AAT variants as six deficient (p.Val234Glu, p.Val242_Pro243insLeu, p.Leu291Phe, p.Ala308Ser, p.Pro393Thr and p.Pro393Arg), one dysfunctional (p.Thr96Ile), three normal (p.Ser71Arg, p.Ala349Pro and p.Asp365Glu) and three null alleles (p.Gln33*, p.Gln285* and p.Leu310Phefs*14).\u003c/p\u003e\n\u003cp\u003eConclusions. Functional assays and protein structural information are useful tools in the characterization of novel variants of the SERPINA1 gene. The newly characterized mutations expand the number of SERPINA1 variants with proven pathogenic effects, facilitating the diagnoses of future cases of AATD.\u003c/p\u003e","manuscriptTitle":"Functional and structural impact of novel SERPINA1 variants on Alpha-1 Antitrypsin Deficiency","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-25 17:54:08","doi":"10.21203/rs.3.rs-6654733/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2025-07-09T12:05:03+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-06-24T04:01:13+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-05-14T13:40:48+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Translational Medicine","date":"2025-05-13T07:04:22+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"journal-of-translational-medicine","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jtrm","sideBox":"Learn more about [Journal of Translational Medicine](http://translational-medicine.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/jtrm/default.aspx","title":"Journal of Translational Medicine","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"d98bf7ea-bd7c-4da1-b7db-90fbe81d4b0d","owner":[],"postedDate":"June 25th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2025-06-25T17:54:08+00:00","versionOfRecord":[],"versionCreatedAt":"2025-06-25 17:54:08","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6654733","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6654733","identity":"rs-6654733","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}