OTULIN-related conditions: Report of a new case and review of the literature using GenIA

OTULIN-related conditions: Report of a new case and review of the literature using GenIA | 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 OTULIN-related conditions: Report of a new case and review of the literature using GenIA Andrés Caballero-Oteyza, Laura Crisponi, Xiao P. Peng, Hongying Wang, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3950863/v2 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 01 Jun, 2024 Read the published version in Clinical Immunology → Version 2 posted You are reading this latest preprint version Show more versions Abstract OTULIN encodes an eponymous linear deubiquitinase (DUB), which through the regulation of M1-Ub dynamics, is essential for controlling inflammation as a negative regulator of the canonical NF-𝛋B signaling pathway. Biallelic loss-of-function (LOF) mutations in OTULIN cause an autosomal recessive condition named Otulin-Related Autoinflammatory Syndrome (ORAS), also known as Otulipenia or AutoInflammation, Panniculitis, and Dermatosis Syndrome (AIPDS). Monoallelic OTULIN LOF, also known as OTULIN Haploinsufficiency (OHI) or Immunodeficiency 107 (IMD107), has been linked to an incompletely penetrant, dominantly inherited susceptibility to invasive Staphylococcal infections. At the same time, a recent novel ORAS-like inflammatory syndrome was described in association with a heterozygous missense mutation that appears to exert dominant negative effects. In this manuscript, we report the identification of a novel homozygous missense mutation, c.595T>A; p.(Trp199Arg), in a Moroccan infant with an ORAS phenotype. We go on to systematically review the literature for OTULIN-related human disease phenotypes by using the GenIA database to collect, extract and harmonize all clinical, laboratory and functional data for published patients and variants. Our comprehensive synthesis of genotypic, phenotypic, and mechanistic data enables a more in-depth view of the diverse mechanisms and pathways by which the OTULIN pathogenic variants may lead to human immune disease. This review may help variant classification activities and the drafting of diagnostic and management guidelines; but it also identifies outstanding knowledge gaps and raises additional questions for future investigation. Systematic review OTULIN ORAS IMD107 OTULIN haploinsufficiency autoinflammation immunodeficiency ubiquitin NF-𝛋B GenIA human genetics Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Full Text Additional Declarations The authors declare no competing interests. Supplementary Files SuppFig1.png Figure S1. Data collection, curation and extraction using GenIA. Flowchart diagram showing the flow of information through the different phases of collection, transformation, storage, query, export and data analysis enabled by the GenIA data structure and used for our systematic review. SuppFig3.png Figure S3. Pedigrees of all 16 families with members affected by OTULIN-related diseases (ORAS, DN-ORAS, IMD107). Each family cohort is assigned a unique letter and each individual in the family is assigned a number associated with that letter along with the known disease-related variant(s) they carry. Symbols are filled according to each individual’s clinical status. SuppFig4.png Figure S4. Functional characterization of OTULIN dysfunction in patient cells. Matrix/grid of rows with specific assays on the y-axis and individuals (using the same codes found in their pedigrees and the supplementary tables) on the x-axis. This figure needs to be examined together with the details provided in Table S3 for accurate interpretation of the results. FigS23Dmodel.pdf Figure S2. In silico 3D modeling of OUTLIN’s wildtype and OTULIN’s W199R variant. (A) Adaptive Poisson-Boltzmann Solver (APSB) analysis performed on wild type (WT) and mutated (W199R) OTULIN. Results are shown using a color-coded surface, with red circles indicating the position of Trp 199 and Arg 199. (B) Root-mean-square fluctuation (RMSF) graph of WT (blue) vs W199R (orange). The red arrow indicates the amino acid (aa)281-286 loop. (C) Representation of OTULIN (purple cartoon), the aa281-286 loop (yellow sticks), the catalytic CORE (white sticks) and the interaction with polyUb (cyan cartoon). (D) Western blot of HEK293 protein lysates after 48h of transfection with LUBAC plasmids (equal amount mix of HOIP, HOIL-1, SHARPIN), OTULIN WT or mutant plasmids, empty vector, together with equal amounts of NEMO and HA-tag Ub plasmid. (E) Agarose gel showing PCR amplification products of total RNA from HEK 293T cells, which was retro-transcribed into cDNA. HEK cells were either untransfected or were transiently expressing the OTULIN wild-type or OTULIN-W199R plasmids. GAPDH amplification was used as a loading control. The “No RT” negative control PCR used the same cDNA samples but without RT enzyme. (F) WB quantification of mean fluorescence intensity of Ub-HA (see Figure 1F) shown as mean plus standard deviation of 4 independent experiments Otulinmssuppv7final.pdf Supp. material - case report Cite Share Download PDF Status: Published Journal Publication published 01 Jun, 2024 Read the published version in Clinical Immunology → Version 2 posted You are reading this latest preprint version Show more versions 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. 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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-3950863","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":273053524,"identity":"4b4a1c34-389f-436d-80e6-c58b98e05e82","order_by":0,"name":"Andrés Caballero-Oteyza","email":"data:image/png;base64,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","orcid":"","institution":"Cluster of Excellence \"RESIST - Resolving Infection Susceptibility\"","correspondingAuthor":true,"prefix":"","firstName":"Andrés","middleName":"","lastName":"Caballero-Oteyza","suffix":""},{"id":273053525,"identity":"c58f68ac-4182-4d51-b856-5720c157dac4","order_by":1,"name":"Laura Crisponi","email":"","orcid":"","institution":"National Research Council","correspondingAuthor":false,"prefix":"","firstName":"Laura","middleName":"","lastName":"Crisponi","suffix":""},{"id":273053526,"identity":"28b9f3e7-89ce-43b3-b105-a5cb163ba5de","order_by":2,"name":"Xiao P. Peng","email":"","orcid":"","institution":"Johns Hopkins University","correspondingAuthor":false,"prefix":"","firstName":"Xiao","middleName":"P.","lastName":"Peng","suffix":""},{"id":273053527,"identity":"af774df5-8dcf-469c-b7a7-6eea9d884101","order_by":3,"name":"Hongying Wang","email":"","orcid":"","institution":"National Human Genome Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Hongying","middleName":"","lastName":"Wang","suffix":""},{"id":273053528,"identity":"bff6d58f-2c7f-435e-84ce-efa74cbb6058","order_by":4,"name":"Pavla Mrovecova","email":"","orcid":"","institution":"University Medical Center Freiburg","correspondingAuthor":false,"prefix":"","firstName":"Pavla","middleName":"","lastName":"Mrovecova","suffix":""},{"id":273053529,"identity":"349f370f-c2fd-4d1d-a917-0253c588c676","order_by":5,"name":"Stefania Olla","email":"","orcid":"","institution":"National Research Council","correspondingAuthor":false,"prefix":"","firstName":"Stefania","middleName":"","lastName":"Olla","suffix":""},{"id":273053530,"identity":"eb02509a-8565-4845-99c2-8652ca42bacf","order_by":6,"name":"Chiara Siguri","email":"","orcid":"","institution":"National Research Council","correspondingAuthor":false,"prefix":"","firstName":"Chiara","middleName":"","lastName":"Siguri","suffix":""},{"id":273053531,"identity":"94b5ebdf-8386-4144-b2bb-1bb5fce54ffa","order_by":7,"name":"Farida Marnissi","email":"","orcid":"","institution":"University of Hassan II Casablanca","correspondingAuthor":false,"prefix":"","firstName":"Farida","middleName":"","lastName":"Marnissi","suffix":""},{"id":273053532,"identity":"213aed51-f9fc-42cf-a873-bd1e00c2c4f7","order_by":8,"name":"Zineb Jouhadi","email":"","orcid":"","institution":"University of Hassan II Casablanca","correspondingAuthor":false,"prefix":"","firstName":"Zineb","middleName":"","lastName":"Jouhadi","suffix":""},{"id":273053533,"identity":"8cd86e3f-0498-41b1-bc12-44ac3bb72d0b","order_by":9,"name":"Ivona Aksentijevich","email":"","orcid":"","institution":"National Human Genome Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Ivona","middleName":"","lastName":"Aksentijevich","suffix":""},{"id":273053534,"identity":"8dc9f4e4-53ff-41ac-9650-5604db167540","order_by":10,"name":"Bodo Grimbacher","email":"","orcid":"","institution":"University Medical Center Freiburg","correspondingAuthor":false,"prefix":"","firstName":"Bodo","middleName":"","lastName":"Grimbacher","suffix":""},{"id":273053535,"identity":"dd7e4347-419b-4ed5-b723-7247bea1cdbf","order_by":11,"name":"Michele Proietti","email":"","orcid":"","institution":"Hannover Medical School","correspondingAuthor":false,"prefix":"","firstName":"Michele","middleName":"","lastName":"Proietti","suffix":""}],"badges":[],"createdAt":"2024-02-12 11:35:03","currentVersionCode":2,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-3950863/v2","doiUrl":"https://doi.org/10.21203/rs.3.rs-3950863/v2","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1016/j.clim.2024.110292","type":"published","date":"2024-06-01T08:56:53+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":52306330,"identity":"ee0d74c6-edba-4bd3-a425-d04c96054ff6","added_by":"auto","created_at":"2024-03-08 19:44:23","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":9581482,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCase presentation\u003c/strong\u003e. (\u003cstrong\u003eA\u003c/strong\u003e) Pedigree for our patient case with parental segregation of the novel \u003cem\u003eOTULIN\u003c/em\u003e mutation shown. The black-filled symbol represents the affected patient, while the white symbol represents unaffected parents. (\u003cstrong\u003eB\u003c/strong\u003e) Images from the deceased patient harboring the novel homozygous OTULIN mutation. (\u003cstrong\u003eC\u003c/strong\u003e) Sanger sequencing electropherograms showing the nucleotide sequence change and below, the predicted codon and amino acid sequence change in the OTULIN protein. (\u003cstrong\u003eD\u003c/strong\u003e) \u003cem\u003eIn silico\u003c/em\u003e 3D modeling of the missense change: interactions established between Trp199 or Arg199 and the residues of OTULIN. Hydrophobic interactions are depicted in red, aromatic in blue, pi-pi in orange, carbon-pi in magenta, metsulfur-pi in yellow, amide-ring in hot pink, and hydrogen bonds in salmon. (\u003cstrong\u003eE\u003c/strong\u003e) Western blotting of protein extracts from HEK293T cells transfected with an empty plasmid, LUBAC plasmids (equal amounts of HOIP, HOIL-1, SHARPIN), OTULIN wild type (WT) or with a mutant plasmid (W199R or L272P) using antibodies against OTULIN, SHARPIN, HOIL-1, HOIP. (\u003cstrong\u003eF\u003c/strong\u003e) WB of co-immunoprecipitation assay using protein extracts from HEK293 cells shown in (E) that additionally express HA-tagged Ubiquitin and NEMO. NEMO was used as bait to pull down the complex. Immunoblot shows the presence and relative abundance of NEMO, OTULIN and Ub chains. (\u003cstrong\u003eG\u003c/strong\u003e) Quantification of protein expression relative to GAPDH and to OTULIN-WT levels from WB images in (E) and Figure S2Dusing ImageJ software. See Figure S2F for quantification of ubiquitination (Ub-HA) (\u003cstrong\u003eH\u003c/strong\u003e) Dual-luciferase assay on the HEK293T cells used in (E) additionally transfected with equal amounts of NF-kB driven luciferase reporter plasmid/renilla control plasmid, after 18 hr in culture. The fold change of Firefly luciferase versus Renilla luciferase was normalized to cells transfected with an empty vector. Results of three independent experiments are shown. Error bars depict standard deviations from triplicate samples\u003c/p\u003e","description":"","filename":"Fig1case.png","url":"https://assets-eu.researchsquare.com/files/rs-3950863/v2/77507cd2402aba3e9ea53ce3.png"},{"id":52307350,"identity":"c2b68ab1-bc38-4d91-a0f5-8812cd28da87","added_by":"auto","created_at":"2024-03-08 19:52:23","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1426372,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSystematic literature review of OTULIN disease-causing variants.\u003c/strong\u003e (\u003cstrong\u003eA\u003c/strong\u003e) Total number of individuals (patients and family members) according to the number of times they were reported or mentioned in an article. Below, total number of disease-associated OTULIN variants found in GenIA vs OMIM and ClinVar. (\u003cstrong\u003eB\u003c/strong\u003e) Upset plot shows how many patients had genetic, clinical, functional, and lab data available across all articles. It also shows how many patients had a combination of clinical and genetic data; clinical, genetic and lab data; or all 4 datasets available. (\u003cstrong\u003eC\u003c/strong\u003e) Schematic representation of all OTULIN disease-causing variants displayed along OTULIN’s gene/cDNA and protein sequences. Variants associated with ORAS/DN-ORAS are shown above the respective cDNA and protein sequences and below variants associated with OHI. Each dot represents a patient.\u003c/p\u003e","description":"","filename":"Fig2variants.png","url":"https://assets-eu.researchsquare.com/files/rs-3950863/v2/7b8e1561a3c0bea8db3ca75f.png"},{"id":52307348,"identity":"114aae84-67e1-41e6-a303-928681389bdb","added_by":"auto","created_at":"2024-03-08 19:52:23","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":3455499,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eClinical and management data\u003c/strong\u003e. (\u003cstrong\u003eA\u003c/strong\u003e) Schematic representation of the cardinal symptoms found to be present or absent in OTULIN-related diseases (ORAS, DN-ORAS and OHI). Human figure template \u003ca href=\"https://commons.wikimedia.org/wiki/Human_body_diagrams\"\u003eborrowed\u003c/a\u003e and modified for the purposes of this paper. (\u003cstrong\u003eB\u003c/strong\u003e) Graph showing the ages at which the different ORAS patients began to present clinically (AFM, age at first manifestation), the ages at which they were genetically studied or diagnosed (ADx), the ages at which they died (ADeath), and the age at which one patient received HSCT. Circles indicate females, and triangles indicate males. (\u003cstrong\u003eC\u003c/strong\u003e) Graph showing disease penetrance in OHI for confirmed heterozygotes or confirmed plus presumed/obligate heterozygotes for pathogenic variants in \u003cem\u003eOTULIN\u003c/em\u003e. (\u003cstrong\u003eD\u003c/strong\u003e) Matrix showing the different therapies ORAS patients received and their respective responses/outcomes. \u003cem\u003eUnspecified \u003c/em\u003emeans that outcome was not explicitly mentioned by authors.\u003c/p\u003e","description":"","filename":"Fig3clinical.png","url":"https://assets-eu.researchsquare.com/files/rs-3950863/v2/0d7f36f05faea2fba8046ae0.png"},{"id":52307351,"identity":"f3c38af2-8966-41b0-8564-9c11bd2d49c8","added_by":"auto","created_at":"2024-03-08 19:52:23","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":1341041,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eIn vitro\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e (or \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ein silico\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e) functional consequences of OTULIN mutants.\u003c/strong\u003e Matrix showing the assays used and respective outcomes for all reported \u003cem\u003eOTULIN\u003c/em\u003e variants, both naturally occurring and artificially generated. In bold are those variants with some causal association to human OTULIN-related diseases. A box containing multiple colors indicates that multiple experimental data points were generated for an assay, either in different studies or in the same study using different conditions. For the full details associated with each assay, please see Table S4.\u003c/p\u003e","description":"","filename":"Fig4functionalvars.png","url":"https://assets-eu.researchsquare.com/files/rs-3950863/v2/82878d0cc668cbe64d2e500a.png"},{"id":52306334,"identity":"945047fe-d995-48a1-ad95-e02ea8bcc904","added_by":"auto","created_at":"2024-03-08 19:44:23","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1123991,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eOTULIN function and dysfunction\u003c/strong\u003e. Simplified and schematic representation of OTULIN’s role as a DUB within the NF-𝛋B signaling pathway in WT and ORAS cells. Defective OTULIN cannot remove Met1-Ub chains from I𝛋Bs, and this leads to increased phosphorylation of I𝛋Kalpha/beta, I𝛋B-alpha, and p65-NF-𝛋B following TNF signaling, with consequent activation of NF-𝛋B signaling. Presumed pathway alterations in mutant cells are shown in bold.\u003c/p\u003e","description":"","filename":"Fig5pathway.png","url":"https://assets-eu.researchsquare.com/files/rs-3950863/v2/4d5c72eac629117db5336ce0.png"},{"id":58930132,"identity":"d723e5a0-e03f-46d3-9997-96deaa800976","added_by":"auto","created_at":"2024-06-24 08:58:08","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":11793352,"visible":true,"origin":"","legend":"","description":"","filename":"Otulinmsmainv7final.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3950863/v2_covered_8ed44b76-477f-46ac-b954-9f5d0ca48832.pdf"},{"id":52306326,"identity":"bc816c77-dbb4-412f-99e9-07e71d840417","added_by":"auto","created_at":"2024-03-08 19:44:23","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":182026,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure S1. Data collection, curation and extraction using GenIA. \u003c/strong\u003eFlowchart diagram showing the flow of information through the different phases of collection, transformation, storage, query, export and data analysis enabled by the GenIA data structure and used for our systematic review.\u003c/p\u003e","description":"","filename":"SuppFig1.png","url":"https://assets-eu.researchsquare.com/files/rs-3950863/v2/7dc13cb846b7c3e146b4dd93.png"},{"id":52306325,"identity":"4065944a-9363-41c2-b723-b55932a7e303","added_by":"auto","created_at":"2024-03-08 19:44:23","extension":"png","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":79209,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure S3. Pedigrees of all 16 families with members affected by OTULIN-related diseases (ORAS, DN-ORAS, IMD107). \u003c/strong\u003eEach family cohort is assigned a unique letter and each individual in the family is assigned a number associated with that letter along with the known disease-related variant(s) they carry. Symbols are filled according to each individual’s clinical status.\u003c/p\u003e","description":"","filename":"SuppFig3.png","url":"https://assets-eu.researchsquare.com/files/rs-3950863/v2/935148d8b0306ff85c9d4f2b.png"},{"id":52307495,"identity":"439acb95-cb59-4de8-b4bd-412b2b48e0aa","added_by":"auto","created_at":"2024-03-08 20:00:23","extension":"png","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":118514,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure S4. Functional characterization of OTULIN dysfunction in patient cells. \u003c/strong\u003eMatrix/grid of rows with specific assays on the y-axis and individuals (using the same codes found in their pedigrees and the supplementary tables) on the x-axis. This figure needs to be examined together with the details provided in Table S3 for accurate interpretation of the results.\u003c/p\u003e","description":"","filename":"SuppFig4.png","url":"https://assets-eu.researchsquare.com/files/rs-3950863/v2/20a8681fbd226264a5ced60e.png"},{"id":52307496,"identity":"b21f85dc-d527-414b-8bf9-c681132c3cf4","added_by":"auto","created_at":"2024-03-08 20:00:23","extension":"pdf","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":956281,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure S2. \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eIn silico\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e 3D modeling of OUTLIN’s wildtype and OTULIN’s W199R variant.\u003c/strong\u003e (\u003cstrong\u003eA\u003c/strong\u003e) Adaptive Poisson-Boltzmann Solver (APSB) analysis performed on wild type (WT) and mutated (W199R) OTULIN. Results are shown using a color-coded surface, with red circles indicating the position of Trp 199 and Arg 199. (\u003cstrong\u003eB\u003c/strong\u003e) Root-mean-square fluctuation (RMSF) graph of WT (blue) vs W199R (orange). The red arrow indicates the amino acid (aa)281-286 loop. (\u003cstrong\u003eC\u003c/strong\u003e) Representation of OTULIN (purple cartoon), the aa281-286 loop (yellow sticks), the catalytic CORE (white sticks) and the interaction with polyUb (cyan cartoon). (\u003cstrong\u003eD\u003c/strong\u003e) Western blot of HEK293 protein lysates after 48h of transfection with LUBAC plasmids (equal amount mix of HOIP, HOIL-1, SHARPIN), OTULIN WT or mutant plasmids, empty vector, together with equal amounts of NEMO and HA-tag Ub plasmid. (\u003cstrong\u003eE\u003c/strong\u003e) Agarose gel showing PCR amplification products of total RNA from HEK 293T cells, which was retro-transcribed into cDNA. HEK cells were either untransfected or were transiently expressing the OTULIN wild-type or OTULIN-W199R plasmids. GAPDH amplification was used as a loading control. The “No RT” negative control PCR used the same cDNA samples but without RT enzyme. (F) WB quantification of mean fluorescence intensity of Ub-HA (see Figure 1F) shown as mean plus standard deviation of 4 independent experiments\u003c/p\u003e","description":"","filename":"FigS23Dmodel.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3950863/v2/d222ad14ef2b45aec302f248.pdf"},{"id":52306331,"identity":"1a3266f9-417f-4286-af0a-4191dd78d400","added_by":"auto","created_at":"2024-03-08 19:44:23","extension":"pdf","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":447229,"visible":true,"origin":"","legend":"\u003cp\u003eSupp. material - case report\u003c/p\u003e","description":"","filename":"Otulinmssuppv7final.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3950863/v2/4e422a6d249ec907c1425563.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"OTULIN-related conditions: Report of a new case and review of the literature using GenIA","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"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":"Systematic review, OTULIN, ORAS, IMD107, OTULIN haploinsufficiency, autoinflammation, immunodeficiency, ubiquitin, NF-𝛋B, GenIA, human genetics","lastPublishedDoi":"10.21203/rs.3.rs-3950863/v2","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3950863/v2","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cem\u003eOTULIN\u003c/em\u003e encodes an eponymous linear deubiquitinase (DUB), which through the regulation of M1-Ub dynamics, is essential for controlling inflammation as a negative regulator of the canonical NF-𝛋B signaling pathway. Biallelic loss-of-function (LOF) mutations in \u003cem\u003eOTULIN\u003c/em\u003e cause an autosomal recessive condition named Otulin-Related Autoinflammatory Syndrome (ORAS), also known as Otulipenia or AutoInflammation, Panniculitis, and Dermatosis Syndrome (AIPDS). Monoallelic \u003cem\u003eOTULIN\u003c/em\u003e LOF, also known as OTULIN Haploinsufficiency (OHI) or Immunodeficiency 107 (IMD107), has been linked to an incompletely penetrant, dominantly inherited susceptibility to invasive Staphylococcal infections. At the same time, a recent novel ORAS-like inflammatory syndrome was described in association with a heterozygous missense mutation that appears to exert dominant negative effects. In this manuscript, we report the identification of a novel homozygous missense mutation, c.595T\u0026gt;A; p.(Trp199Arg), in a Moroccan infant with an ORAS phenotype. We go on to systematically review the literature for OTULIN-related human disease phenotypes by using the \u003ca href=\"http://www.geniadb.net/\"\u003eGenIA database\u003c/a\u003eto collect, extract and harmonize all clinical, laboratory and functional data for published patients and variants. Our comprehensive synthesis of genotypic, phenotypic, and mechanistic data enables a more in-depth view of the diverse mechanisms and pathways by which the \u003cem\u003eOTULIN\u003c/em\u003e pathogenic variants may lead to human immune disease. This review may help variant classification activities and the drafting of diagnostic and management guidelines; but it also identifies outstanding knowledge gaps and raises additional questions for future investigation.\u003c/p\u003e","manuscriptTitle":"OTULIN-related conditions: Report of a new case and review of the literature using GenIA","msid":"","msnumber":"","nonDraftVersions":[{"code":2,"date":"2024-03-08 19:44:18","doi":"10.21203/rs.3.rs-3950863/v2","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}},{"code":1,"date":"2024-02-15 15:52:12","doi":"10.21203/rs.3.rs-3950863/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":"f3ab4893-217e-48fc-a25e-5b73b14fbbf0","owner":[],"postedDate":"March 8th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-06-24T08:56:53+00:00","versionOfRecord":{"articleIdentity":"rs-3950863","link":"https://doi.org/10.1016/j.clim.2024.110292","journal":{"identity":"clinical-immunology","isVorOnly":true,"title":"Clinical Immunology"},"publishedOn":"2024-06-01 08:56:53","publishedOnDateReadable":"June 1st, 2024"},"versionCreatedAt":"2024-03-08 19:44:18","video":"","vorDoi":"10.1016/j.clim.2024.110292","vorDoiUrl":"https://doi.org/10.1016/j.clim.2024.110292","workflowStages":[]},"version":"v2","identity":"rs-3950863","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3950863","identity":"rs-3950863","version":["v2"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}