Genetic analysis of a pedigree with hereditary coagulation factor XII deficiency

Genetic analysis of a pedigree with hereditary coagulation factor XII 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 Case Report Genetic analysis of a pedigree with hereditary coagulation factor XII deficiency Weiwei Fang, Bile Chen, Anqin Zou, Fei Xu, Langyi Qin, Lihong Yang, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4978926/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 22 Jan, 2025 Read the published version in Annals of Hematology → Version 1 posted 9 You are reading this latest preprint version Abstract Objective: Analyze the clinical phenotype and gene mutations of a family with hereditary FXII deficiency, and preliminarily explore its molecular pathogenic mechanism. Methods: The routine coagulation indicators and related coagulation factors were measured.. Thromboelastography and thrombin generation tests simulated coagulation and anticoagulation states in vitro and in vivo. PCR direct sequencing was utilized to analyze all exons and flanking sequences of the F12 gene in the proband, confirming suspected mutations through reverse sequencing, and identifying corresponding mutation sites in family members. Using ClustalX-2.1-win to analyze the conservation of the variant, and employing online software to predict the pathogenicity of mutations. Results: The proband exhibited significantly prolonged APTT (169.1 seconds) and a pronounced decrease in FXII:C to 1.0%. Thromboelastography testing indicated a diminished function of the endogenous coagulation system, while thrombin generation testing revealed a normal ability for thrombin production in the proband. Gene sequencing revealed that the proband harbored a deletion mutation c.303_304delCA in exon 5 and a substitution mutation c.800+1G>A in intron 8. All three bioinformatics software indicated that the mutations were pathogenic and could lead to the production of a terminator, potentially altering the structure and function of the protein. Conclusion: The deletion mutation c.303_304delCA and substitution mutation c.800+1G>A are associated with a decreased in FXII levels in this family, with the c.800+1G>A mutation being the first reported mutation worldwide. Coagulation FXII deficiency Genetic analysis Bioinformatics Gene mutation Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction FXII is a serine protease synthesized in the liver, playing a pivotal role in regulating both the blood coagulation and fibrinolysis systems. FXII participates in fibrinolysis through activation of the kinin-forming and kinin-releasing enzyme systems, with a protein structure closely resembling that of fibrinolysis system proteins [ 1 ] . Therefore, the lack of FXII leads to increased thrombosis rather than increased bleeding. Hereditary FXII deficiency is an autosomal inherited disease caused by mutations in the gene encoding FXII protein. While most patients display autosomal recessive inheritance, some also show dominant inheritance [ 2 ] . Patients with hereditary FXII deficiency typically do not manifest clinical symptoms associated with other coagulation factor deficiencies, such as spontaneous bleeding or severe post-injury bleeding. [ 3 ] This condition is often identified during preoperative screening. This study aims to identify the pathogenic gene mutation site and preliminarily explore its molecular pathogenic mechanism by conducting coagulation index and gene mutation detection on a family with hereditary FXII deficiency. Material and methods Patients The proband, a 24-year-old female, is two months pregnant. During routine antenatal check-ups, routine coagulation tests showed significantly prolonged APTT. Further coagulation factor testing revealed a significant decrease in FXII:C, while other coagulation indicators were within the normal range. but there was no clinical evidence of bleeding. A pedigree investigation was conducted on six individuals spanning four generations (Fig. 1 ). All members had normal liver and kidney function, with no history of spontaneous bleeding or thrombosis. 100 healthy subjects were recruited for this study as healthy controls, comprising 58 males and 42 females, with an average age of 35 years (range: 19–63 years). None of them had a history of abnormal bleeding or thrombosis tendency, and they did not have liver or kidney diseases. Ethics Our study was approved by the Ethics Committee of the First Affiliated Hospital of Wenzhou Medical University (KY2022-R193). Written informed consent was obtained from all participants prior to their involvement in the study. Blood Samples Collect 2.7 mL of peripheral venous blood from the subject, anticoagulated with 0.109 mol/L sodium citrate (ratio 1:9), and centrifuge at 3000 r/min for 15 minutes. The upper layer of platelet depleted plasma is used for coagulation index detection, which is completed within 2 hours. The lower layer of blood cells is used for genomic DNA extraction, PCR amplification, and sequencing. Coagulation routine testing PT, APTT, FIB, coagulation factors, and other coagulation indicators were measured using the one-stage coagulation method, while D-D was measured using immunoturbidimetry. All coagulation tests were conducted on the French Stago STA-R fully automatic blood coagulation analyzer with the original manufacturer's reagents.All procedures are strictly conducted in accordance with the reagent instructions. F12 gene sequencing Extract genomic DNA from the proband and family members using the NP968-G fully automatic nucleic acid extractor and its accompanying reagents (Xi'an Tianlong Company). Amplify the 12 exons, flanking sequences, 5' and 3' non-coding regions of the F12 gene from the proband using specific primer sequences and amplification conditions. Sanger sequencing was performed on the amplified product using the 3730xl s/n 1524-034 sequencer (ABI, USA). The sequencing results were compared with the F12 gene sequence (NM:000505.3) using Chromas software to identify mutation sites. After confirming the mutation site, amplify the corresponding exons of the family members accordingly. Bioinformatics analysis We performed online prediction analysis on the mutations using MutationTaster ( https://www.mutationtaster.org ) and the Franklin database ( https://franklin.genoox.com ). Thromboela-stogram Extract 2 mL of peripheral venous blood, add kaolin, mix well, and let it stand; After activation,insert the sample cup into the TCA6000 thromboela-stography instrument to automatically record TEG images and parameters, mainly including coagulation reaction time (R), coagulation formation time (K), alpha angle (α angle, Angle), and maximum thrombus amplitude (MA). R refers to the latency period from the start of the coagulation system to the formation of fibrin clots, K refers to the time from the start of coagulation to the amplitude of 20 mm in the recording image, and Angle refers to the time from the start of coagulation to the amplitude of 20 mm in the recording image. The formation rate of fibrin clots, MA refers to the final strength of fibrin clots. [ 4 ] Thrombin generation test According to reference [ 5 ] , a calibrated automatic thrombin generation method (CAT) is used to assess thrombin generation. We conducted experiments using the reagent kit and analytical tools provided by Thrombinoscope BV (Rijswijk, Netherlands) with the Fluoroskan Ascent FL reader (Thermo Electron and Fisher Scientific, Waltham, Massachusetts, USA). The measured parameters included delay time (lag time), thrombin potential (ETP), peak (peak), and time to peak (time to peak) to assess the total amount of thrombin produced by the subjects. Results Coagulation routine test results The proband's APTT was extended to 169.1 seconds (reference range: 29.0–43.0 seconds), and FXII:C decreased to 1% (reference range: 72–113%). Meanwhile, the APTT of the proband's grandfather, father, mother, and daughter all showed slight prolongation, with FXII: C levels of 45%, 41%, 51%, and 24%, respectively. All other indicators of the family members were within normal ranges. (Table) Thromboela-stogram and Thrombin generation test The thromboelastogram results indicated a notable increase in the R value, with all other parameters within the normal reference range. The thrombin production test results showed that the proband's production curve was generally consistent with the reference, with all parameters closely aligned. (Fig. 2 ) Genetic analysis Gene sequence analysis revealed that the proband harbored a compound heterozygous mutation: a deletion mutation c.303_304delCA in exon 5 and a substitution mutation c.800 + 1G > A in intron 8. The proband's father carries a heterozygous deletion mutation c.303_304delCA, while the proband's grandfather, mother, and daughter carry a heterozygous substitution mutation c.800 + 1G > A. These test results indicate that the proband inherited the two mutations from the father and mother, respectively. (Fig. 3 ) Bioinformatics analysis MutationTaster and the Franklin database predict that both mutations are deleterious. Two RNA splicing prediction models for this mutation were established on the RDDC platform developed by the Guangzhou Rare Disease Gene Therapy Alliance. (Fig. 4 ) Discussion In early coagulation theory, FXII was considered one of the components in contact with the coagulation system, activating FXII, which in turn activates FXI, FX, FIX, and FXI through the endogenous coagulation pathway to enter the common coagulation pathway and exert coagulation effects. Therefore, defects in FXII can lead to prolonged APTT [ 6 – 7 ] . However, since coagulation reactions in the body primarily occur through the extrinsic pathway, FXII deficiency alone does not lead to bleeding phenomena. On the contrary, due to the significant role FXII plays in activating the fibrinolytic system, defects or activation disorders may decrease fibrinolytic activity in the body, potentially leading to thrombotic diseases. In recent years, studies have shown that FXII plays an important role in thrombosis and is not essential in endogenous coagulation processes [ 8 ] . In our study, the total R value of the thrombelastogram result of the proband significantly increased, indicating that the patient's comprehensive coagulation state is in a hypocoagulable state, with overall weak activity of the endogenous coagulation system’s coagulation factors, which is consistent with the prolonged APTT and the extremely low FXII activity. However, the above experiments were conducted in vitro to simulate the activation of the endogenous coagulation pathway and may not fully reflect the actual coagulation process in the body [ 9 ] . Therefore, we conducted thrombin generation test to simulate the dynamic changes in thrombin production under the combined influence of the body’s coagulation and anticoagulation systems. [ 10 ] The results showed that the proband's thrombin generation ability was essentially consistent with that of the normal control, which also explains why the proband did not experience any bleeding or coagulation abnormalities on a daily basis or during pregnancy clinical manifestations. Online analysis tools predict that both mutations are deleterious. The c.303_104delCA mutation causes a frameshift, substituting serine at position 101 with proline (His101Pro), and generating a premature termination codon in exon 6, leading to the production of a truncated protein. It is speculated that this site may have serious effects on coagulation and fibrinolysis functions, as has been reported [ 11 ] . The c.800 + 1G > A mutation occurs at the first position downstream of intron 8, precisely at the canonical splice site GT-AG domain, and is regarded as a pathogenic variant. We established an RNA splicing prediction model for this mutation using the RDDC platform developed by Guangzhou Rare Disease Gene Therapy Alliance. The results showed that the mutation disrupts the original donor splice site, affecting the normal mRNA splicing process. There may be two splicing models: one that deletes 166 bp causing exon skipping, and another that inserts 85 bp causing intron retention. Both ultimately result in frameshift mutations and premature termination. The protein translated after the mutation is rapidly degraded due to its high instability. It is also possible that abnormally cleaved mRNA cannot be transported into the cytoplasm, leading to a lack of protein translation [ 12 ] . The c.800 + 1G > A mutation has not been reported in Query human gene polymorphism database( https://www.ncbi.nlm.nih.gov/snp/ ) and the Human Gene Mutation Database( http://www.hgmd.cf.ac.uk/ac/index.php ). In summary, we identified a family with coagulation factor XII deficiency, where the proband carried compound heterozygous mutations: c.303_304 delCA inherited from her father and c.800 + 1G > A inherited from her mother. Both mutations can impact the gene structure by introducing different termination signals, thereby affecting the normal function of the protein. This results in varying degrees of decreased FXII activity among family members. The c.800 + 1G > A mutation is reported here for the first time. However, its precise molecular pathogenesis requires further research. The thrombelastogram and thrombin generation test explain why the proband had low FXII activity levels despite showing no clinical manifestations. Abbreviations FXII：Coagulation factor XII PCR: polymerase chain reaction FXII:C：FXII activity APTT：activated partial thromboplastin time PT：prothrombin time FIB：Fibrinogen D-D：D-Dimer TEG：thromboela-stogram Declarations Acknowledgements We appreciate the patient and her family members for their cooperation. All authors declared no competing interests that could be perceived as influencing the findings or conclusions.We are grateful to the patient and her family members for their cooperation. All authors stated that they had no interests which might be perceived as posing a conflict or bias. Disclaimer statements Disclosure of interest: The authors have no relevant financial or nonfinancial interests to disclose. Contributors : All authors have made significant contributions to the study's conception and design, data acquisition, analysis and interpretation, as well as to the drafting and critical revision of the manuscript for important intellectual content. Furthermore, all authors have provided their final approval for the submitted version of the article. Funding This work was supported by Key Laboratory of Clinical Laboratory Diagnosis and Translational Research of Zhejiang Province(2022E10022) and the Wenzhou Science and Technology Bureau Project (Y2023508). References BJORKQVIST J, NICKEL K F, STAVROU E, et al. In vivo activation and functions of the protease factor XII[J]. Thromb Haemost, 2014, 112(5): 868-875. Fernandes HD, Newton S, Rodrigues JM. Factor XII deficiency mimicking bleeding diathesis: a unique presentation and diagnostic pitfall. Cureus 2018;10(06):e2817. Yang L,Wang Y,Zhou J,et al.Identification of genetic defects un derlying FXII deficiency in four unrelated Chinese patients.Acta Haematol,2016,135(4):238-240. Trinh,TH,Pao,LP.Pivalizza,EG.Thrombelastograph platelet mapping during hyperfibrinolysis[J].J Cardiothorac Vasc Anesth,2020,34(6):1708-1710. Deng Y, Zhu J, Gong Y, Yi X, Zhou L, Xie Y, et al. Hereditary factor V deficiency from heterozygous mutations with a novel variant p.Pro798Leufs∗13 in the F5 gene. Blood Coagul Fibrinolysis. 2021 Oct 1;32(7):513-518. Matsukuma E, Gotoh Y, Kuroyanagi Y,et al .A case of atypical hemolytic uremic syndrome due to anti-factor H antibody in a patient presenting with a factor XII deficiency identified two novel mutations[J].Clinical & Experimental Nephrology, 2011, 15(2):269-274. Thomas Renné.The procoagulant and proinflammatory plasma contact system[J].Seminars in Immunopathology, 2012, 34(1):31-41. KWON M J, KIM H J, LEE K O, et al. Molecular genetic analysis of Korean patients with coagulation factor XII deficiency[J]. Blood Coagul Fibrinolysis, 2010, 21(4): 308-312. Depasse F,Binder NB,Mueller J,et al. Thrombin generation assays are versatile tools in blood coagulation analysis:A review of technical features,and applications from research to laboratory routine[J]. J Thromb Haemost,2021,19(12):2907-2917. Santagostino E,Mancuso ME,Tripodi A,et al.Severe hemophilia with mild bleeding phenotype:molecular haracterization and global coagulation profile[J].JThromb Haemost,2010,8(4):737-743. Xie H,Lu M,Yang X et al. Gene analysis in four inherited coagulation FⅫ deficiency pedigree [J]. Chinese Journal of Hematology, 2013, 3­­4(3): 5. Ding Q, Wu W, Fu Q, et al. Novel aberrant splicings caused by a splice site mutation (IVS1a+5g>a) in F7 gene[J]. Thromb Haemost, 2005,93(6): 1077-1081. Table Table. Phenotypes andgenotypes of the inheritedcoagulation factor XII deficiencyfamily Patient PT (s) APTT (s) FIB (g/L) D-D (g/L) LAC FⅧ:C (%) FⅨ:C (%) FⅪ:C (%) FⅫ:C (%) Genetic mutations Proband 14.8 169.1 2.43 0.48 1.05 93 76 87 1 c.303_304 del CA c.800+1G>A Grandfather 12.9 44.5 3.86 0.41 1.09 128 86 104 45 c.800+1G>A Father 13.4 48.2 2.67 0.26 1.01 106 95 89 41 c.303_304 del CA Mother 13.9 43.56 2.54 0.32 1.04 112 87 96 51 c.800+1G>A Husband 13.5 35.6 2.58 0.27 1.08 120 90 105 96 Wild-type Daughter 14.2 44.1 2.24 0.35 1.02 120 88 100 24 c.800+1G>A Normal range 12.6-14.4 29.0-43.0 2.00-4.00 <0.50 0.80-1.20 78~128 75~128 82~118 72~113 Wild-type Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 22 Jan, 2025 Read the published version in Annals of Hematology → Version 1 posted Editorial decision: Revision requested 23 Sep, 2024 Reviews received at journal 23 Sep, 2024 Reviews received at journal 23 Sep, 2024 Reviewers agreed at journal 13 Sep, 2024 Reviewers agreed at journal 13 Sep, 2024 Reviewers invited by journal 12 Sep, 2024 Editor assigned by journal 28 Aug, 2024 Submission checks completed at journal 28 Aug, 2024 First submitted to journal 26 Aug, 2024 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. 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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-4978926","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Case Report","associatedPublications":[],"authors":[{"id":358194839,"identity":"326e07aa-0c26-4cf4-b871-8c827910c070","order_by":0,"name":"Weiwei Fang","email":"","orcid":"","institution":"Wenzhou TCM Hospital of Zhejiang Chinese Medical University Wenzhou","correspondingAuthor":false,"prefix":"","firstName":"Weiwei","middleName":"","lastName":"Fang","suffix":""},{"id":358194840,"identity":"71a2545c-112d-47fa-bce0-e0e652012b02","order_by":1,"name":"Bile Chen","email":"","orcid":"","institution":"Department of Clinical Laboratory, 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Zhou","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAx0lEQVRIie3QsQrCMBCA4ZRApqtdIy3WR4gEMhV8EJdAoJuDm0OGgGBHV30TR0OgLgVfIdAX6Co42NVBGjeHfPP9cHcIRdEfIuXd9s+Xhgxj54OSGQXFE9MW84bULChZUCTyxOCKPWBJwxbLjeK7KwHuADGkq810UljbX7oChEtvHrX11kwmSCoGhIzJTLLEuKBEUCAY+AEYDUuoFHl6xMBwcAKdWp27FqgbnyxDbimbxvphr9fZyTk/6Go6+SR/G4+iKIq+eQNdXTnh4J+RlgAAAABJRU5ErkJggg==","orcid":"","institution":"Department of Clinical Laboratory, Key Laboratory of Clinical Laboratory Diagnosis and Translational Research of Zhejiang Province, the First Affiliated Hospital of Wenzhou Medical University","correspondingAuthor":true,"prefix":"","firstName":"Xingxing","middleName":"","lastName":"Zhou","suffix":""}],"badges":[],"createdAt":"2024-08-26 15:09:12","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4978926/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4978926/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s00277-025-06205-4","type":"published","date":"2025-01-22T15:57:17+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":66887173,"identity":"20ca1107-51a9-44f5-ae0b-1669df7862ef","added_by":"auto","created_at":"2024-10-17 13:54:04","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":180666,"visible":true,"origin":"","legend":"\u003cp\u003ePedigree investigation for the family\u003c/p\u003e","description":"","filename":"Figure1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4978926/v1/40728898ae23543bab2f12df.jpg"},{"id":66887170,"identity":"ec8f5c99-304e-46a5-8eba-ba869e41ceaf","added_by":"auto","created_at":"2024-10-17 13:54:04","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":410754,"visible":true,"origin":"","legend":"\u003cp\u003eThrombela-stogram results of the proband(A), thrombin generation test results of the proband (B)\u003c/p\u003e","description":"","filename":"Figure2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4978926/v1/114fd9e7b6f1d1f2160a8967.jpg"},{"id":66887769,"identity":"577905d8-2517-4d4a-88af-6ce4b50a54a0","added_by":"auto","created_at":"2024-10-17 14:02:04","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":649250,"visible":true,"origin":"","legend":"\u003cp\u003eChromatogram of DNA sequencing.\u003c/p\u003e\n\u003cp\u003e(A) is a heterozygous c.303_304 delCA sequencing map,\u003c/p\u003e\n\u003cp\u003e(B) is a wild-type forward sequencing of c.303_304 delCA ,\u003c/p\u003e\n\u003cp\u003e(C) is a heterozygous c.800+1G\u0026gt;A sequencing map,\u003c/p\u003e\n\u003cp\u003e(D) a wild-type forward sequencing of c.800+1G\u0026gt;A .\u003c/p\u003e\n\u003cp\u003eThe position of the mutational base is indicated with an arrow.\u003c/p\u003e","description":"","filename":"Figure3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4978926/v1/3c778dbc062a4b55fab4b4a2.jpg"},{"id":66887171,"identity":"faeb5f70-aa1f-4ebb-aa6a-4097cfc387ff","added_by":"auto","created_at":"2024-10-17 13:54:04","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":407418,"visible":true,"origin":"","legend":"\u003cp\u003eRNA splicing prediction model\u003c/p\u003e","description":"","filename":"Figure4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4978926/v1/78b46244c7b244205095fa9e.jpg"},{"id":74858558,"identity":"fcd2481e-7f31-423e-ab23-0d67be0c6fe8","added_by":"auto","created_at":"2025-01-27 16:11:32","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2227707,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4978926/v1/e45fb30c-0e18-404e-949d-0c00dda58c6a.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Genetic analysis of a pedigree with hereditary coagulation factor XII deficiency","fulltext":[{"header":"Introduction","content":"\u003cp\u003eFXII is a serine protease synthesized in the liver, playing a pivotal role in regulating both the blood coagulation and fibrinolysis systems. FXII participates in fibrinolysis through activation of the kinin-forming and kinin-releasing enzyme systems, with a protein structure closely resembling that of fibrinolysis system proteins\u003csup\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]\u003c/sup\u003e. Therefore, the lack of FXII leads to increased thrombosis rather than increased bleeding. Hereditary FXII deficiency is an autosomal inherited disease caused by mutations in the gene encoding FXII protein. While most patients display autosomal recessive inheritance, some also show dominant inheritance \u003csup\u003e[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]\u003c/sup\u003e. Patients with hereditary FXII deficiency typically do not manifest clinical symptoms associated with other coagulation factor deficiencies, such as spontaneous bleeding or severe post-injury bleeding.\u003csup\u003e[\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]\u003c/sup\u003e This condition is often identified during preoperative screening.\u003c/p\u003e \u003cp\u003eThis study aims to identify the pathogenic gene mutation site and preliminarily explore its molecular pathogenic mechanism by conducting coagulation index and gene mutation detection on a family with hereditary FXII deficiency.\u003c/p\u003e"},{"header":"Material and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePatients\u003c/h2\u003e \u003cp\u003eThe proband, a 24-year-old female, is two months pregnant. During routine antenatal check-ups, routine coagulation tests showed significantly prolonged APTT. Further coagulation factor testing revealed a significant decrease in FXII:C, while other coagulation indicators were within the normal range. but there was no clinical evidence of bleeding. A pedigree investigation was conducted on six individuals spanning four generations (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). All members had normal liver and kidney function, with no history of spontaneous bleeding or thrombosis.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e100 healthy subjects were recruited for this study as healthy controls, comprising 58 males and 42 females, with an average age of 35 years (range: 19\u0026ndash;63 years). None of them had a history of abnormal bleeding or thrombosis tendency, and they did not have liver or kidney diseases.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eEthics\u003c/h2\u003e \u003cp\u003e Our study was approved by the Ethics Committee of the First Affiliated Hospital of Wenzhou Medical University (KY2022-R193). Written informed consent was obtained from all participants prior to their involvement in the study.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eBlood Samples\u003c/h2\u003e \u003cp\u003eCollect 2.7 mL of peripheral venous blood from the subject, anticoagulated with 0.109 mol/L sodium citrate (ratio 1:9), and centrifuge at 3000 r/min for 15 minutes. The upper layer of platelet depleted plasma is used for coagulation index detection, which is completed within 2 hours. The lower layer of blood cells is used for genomic DNA extraction, PCR amplification, and sequencing.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eCoagulation routine testing\u003c/h2\u003e \u003cp\u003ePT, APTT, FIB, coagulation factors, and other coagulation indicators were measured using the one-stage coagulation method, while D-D was measured using immunoturbidimetry. All coagulation tests were conducted on the French Stago STA-R fully automatic blood coagulation analyzer with the original manufacturer's reagents.All procedures are strictly conducted in accordance with the reagent instructions.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eF12 gene sequencing\u003c/h2\u003e \u003cp\u003eExtract genomic DNA from the proband and family members using the NP968-G fully automatic nucleic acid extractor and its accompanying reagents (Xi'an Tianlong Company). Amplify the 12 exons, flanking sequences, 5' and 3' non-coding regions of the F12 gene from the proband using specific primer sequences and amplification conditions. Sanger sequencing was performed on the amplified product using the 3730xl s/n 1524-034 sequencer (ABI, USA). The sequencing results were compared with the F12 gene sequence (NM:000505.3) using Chromas software to identify mutation sites. After confirming the mutation site, amplify the corresponding exons of the family members accordingly.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eBioinformatics analysis\u003c/h2\u003e \u003cp\u003eWe performed online prediction analysis on the mutations using MutationTaster (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.mutationtaster.org\u003c/span\u003e\u003cspan address=\"https://www.mutationtaster.org\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) and the Franklin database (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://franklin.genoox.com\u003c/span\u003e\u003cspan address=\"https://franklin.genoox.com\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eThromboela-stogram\u003c/h2\u003e \u003cp\u003eExtract 2 mL of peripheral venous blood, add kaolin, mix well, and let it stand; After activation,insert the sample cup into the TCA6000 thromboela-stography instrument to automatically record TEG images and parameters, mainly including coagulation reaction time (R), coagulation formation time (K), alpha angle (α angle, Angle), and maximum thrombus amplitude (MA). R refers to the latency period from the start of the coagulation system to the formation of fibrin clots, K refers to the time from the start of coagulation to the amplitude of 20 mm in the recording image, and Angle refers to the time from the start of coagulation to the amplitude of 20 mm in the recording image. The formation rate of fibrin clots, MA refers to the final strength of fibrin clots. \u003csup\u003e[\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]\u003c/sup\u003e\u003c/p\u003e \u003cdiv id=\"Sec10\" class=\"Section3\"\u003e \u003ch2\u003eThrombin generation test\u003c/h2\u003e \u003cp\u003eAccording to reference \u003csup\u003e[\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]\u003c/sup\u003e, a calibrated automatic thrombin generation method (CAT) is used to assess thrombin generation. We conducted experiments using the reagent kit and analytical tools provided by Thrombinoscope BV (Rijswijk, Netherlands) with the Fluoroskan Ascent FL reader (Thermo Electron and Fisher Scientific, Waltham, Massachusetts, USA). The measured parameters included delay time (lag time), thrombin potential (ETP), peak (peak), and time to peak (time to peak) to assess the total amount of thrombin produced by the subjects.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eCoagulation routine test results\u003c/h2\u003e \u003cp\u003eThe proband's APTT was extended to 169.1 seconds (reference range: 29.0\u0026ndash;43.0 seconds), and FXII:C decreased to 1% (reference range: 72\u0026ndash;113%). Meanwhile, the APTT of the proband's grandfather, father, mother, and daughter all showed slight prolongation, with FXII: C levels of 45%, 41%, 51%, and 24%, respectively. All other indicators of the family members were within normal ranges. (Table)\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eThromboela-stogram and Thrombin generation test\u003c/h2\u003e \u003cp\u003eThe thromboelastogram results indicated a notable increase in the R value, with all other parameters within the normal reference range. The thrombin production test results showed that the proband's production curve was generally consistent with the reference, with all parameters closely aligned. (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e)\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eGenetic analysis\u003c/h2\u003e \u003cp\u003eGene sequence analysis revealed that the proband harbored a compound heterozygous mutation: a deletion mutation c.303_304delCA in exon 5 and a substitution mutation c.800\u0026thinsp;+\u0026thinsp;1G\u0026thinsp;\u0026gt;\u0026thinsp;A in intron 8. The proband's father carries a heterozygous deletion mutation c.303_304delCA, while the proband's grandfather, mother, and daughter carry a heterozygous substitution mutation c.800\u0026thinsp;+\u0026thinsp;1G\u0026thinsp;\u0026gt;\u0026thinsp;A. These test results indicate that the proband inherited the two mutations from the father and mother, respectively. (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e)\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eBioinformatics analysis\u003c/h2\u003e \u003cp\u003eMutationTaster and the Franklin database predict that both mutations are deleterious. Two RNA splicing prediction models for this mutation were established on the RDDC platform developed by the Guangzhou Rare Disease Gene Therapy Alliance. (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e)\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn early coagulation theory, FXII was considered one of the components in contact with the coagulation system, activating FXII, which in turn activates FXI, FX, FIX, and FXI through the endogenous coagulation pathway to enter the common coagulation pathway and exert coagulation effects. Therefore, defects in FXII can lead to prolonged APTT \u003csup\u003e[\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]\u003c/sup\u003e. However, since coagulation reactions in the body primarily occur through the extrinsic pathway, FXII deficiency alone does not lead to bleeding phenomena. On the contrary, due to the significant role FXII plays in activating the fibrinolytic system, defects or activation disorders may decrease fibrinolytic activity in the body, potentially leading to thrombotic diseases. In recent years, studies have shown that FXII plays an important role in thrombosis and is not essential in endogenous coagulation processes \u003csup\u003e[\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eIn our study, the total R value of the thrombelastogram result of the proband significantly increased, indicating that the patient's comprehensive coagulation state is in a hypocoagulable state, with overall weak activity of the endogenous coagulation system\u0026rsquo;s coagulation factors, which is consistent with the prolonged APTT and the extremely low FXII activity. However, the above experiments were conducted in vitro to simulate the activation of the endogenous coagulation pathway and may not fully reflect the actual coagulation process in the body \u003csup\u003e[\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]\u003c/sup\u003e. Therefore, we conducted thrombin generation test to simulate the dynamic changes in thrombin production under the combined influence of the body\u0026rsquo;s coagulation and anticoagulation systems. \u003csup\u003e[\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]\u003c/sup\u003eThe results showed that the proband's thrombin generation ability was essentially consistent with that of the normal control, which also explains why the proband did not experience any bleeding or coagulation abnormalities on a daily basis or during pregnancy clinical manifestations.\u003c/p\u003e \u003cp\u003eOnline analysis tools predict that both mutations are deleterious. The c.303_104delCA mutation causes a frameshift, substituting serine at position 101 with proline (His101Pro), and generating a premature termination codon in exon 6, leading to the production of a truncated protein. It is speculated that this site may have serious effects on coagulation and fibrinolysis functions, as has been reported \u003csup\u003e[\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]\u003c/sup\u003e. The c.800\u0026thinsp;+\u0026thinsp;1G\u0026thinsp;\u0026gt;\u0026thinsp;A mutation occurs at the first position downstream of intron 8, precisely at the canonical splice site GT-AG domain, and is regarded as a pathogenic variant. We established an RNA splicing prediction model for this mutation using the RDDC platform developed by Guangzhou Rare Disease Gene Therapy Alliance. The results showed that the mutation disrupts the original donor splice site, affecting the normal mRNA splicing process. There may be two splicing models: one that deletes 166 bp causing exon skipping, and another that inserts 85 bp causing intron retention. Both ultimately result in frameshift mutations and premature termination. The protein translated after the mutation is rapidly degraded due to its high instability. It is also possible that abnormally cleaved mRNA cannot be transported into the cytoplasm, leading to a lack of protein translation \u003csup\u003e[\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]\u003c/sup\u003e. The c.800\u0026thinsp;+\u0026thinsp;1G\u0026thinsp;\u0026gt;\u0026thinsp;A mutation has not been reported in Query human gene polymorphism database(\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ncbi.nlm.nih.gov/snp/\u003c/span\u003e\u003cspan address=\"https://www.ncbi.nlm.nih.gov/snp/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) and the Human Gene Mutation Database(\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.hgmd.cf.ac.uk/ac/index.php\u003c/span\u003e\u003cspan address=\"http://www.hgmd.cf.ac.uk/ac/index.php\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn summary, we identified a family with coagulation factor XII deficiency, where the proband carried compound heterozygous mutations: c.303_304 delCA inherited from her father and c.800\u0026thinsp;+\u0026thinsp;1G\u0026thinsp;\u0026gt;\u0026thinsp;A inherited from her mother. Both mutations can impact the gene structure by introducing different termination signals, thereby affecting the normal function of the protein. This results in varying degrees of decreased FXII activity among family members. The c.800\u0026thinsp;+\u0026thinsp;1G\u0026thinsp;\u0026gt;\u0026thinsp;A mutation is reported here for the first time. However, its precise molecular pathogenesis requires further research. The thrombelastogram and thrombin generation test explain why the proband had low FXII activity levels despite showing no clinical manifestations.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eFXII：Coagulation factor XII\u003c/p\u003e\n\u003cp\u003ePCR: polymerase chain reaction\u003c/p\u003e\n\u003cp\u003eFXII:C：FXII activity\u003c/p\u003e\n\u003cp\u003eAPTT：activated partial thromboplastin time\u003c/p\u003e\n\u003cp\u003ePT：prothrombin time\u003c/p\u003e\n\u003cp\u003eFIB：Fibrinogen\u003c/p\u003e\n\u003cp\u003eD-D：D-Dimer\u003c/p\u003e\n\u003cp\u003eTEG：thromboela-stogram\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe appreciate the patient and her family members for their cooperation. All authors declared no competing interests that could be perceived as influencing the findings or conclusions.We are grateful to the patient and her family members for their cooperation. All authors stated that they had no interests which might be perceived as posing a conflict or bias.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDisclaimer statements\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eDisclosure of interest: The authors have no relevant financial or nonfinancial interests to disclose.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eContributors\u003c/strong\u003e: All authors have made significant contributions to the study's conception and design, data acquisition, analysis and interpretation, as well as to the drafting and critical revision of the manuscript for important intellectual content. Furthermore, all authors have provided their final approval for the submitted version of the article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThis work was supported by Key Laboratory of Clinical Laboratory Diagnosis and Translational Research of Zhejiang Province(2022E10022) and the Wenzhou Science and Technology Bureau Project (Y2023508).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eBJORKQVIST J, NICKEL K F, STAVROU E, et al. In vivo activation and functions of the protease factor XII[J]. Thromb Haemost, 2014, 112(5): 868-875.\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eFernandes HD, Newton S, Rodrigues JM. Factor XII deficiency mimicking bleeding diathesis: a unique presentation and diagnostic pitfall. Cureus 2018;10(06):e2817.\u003c/li\u003e\n \u003cli\u003eYang L,Wang Y,Zhou J,et al.Identification of genetic defects un derlying FXII deficiency in four unrelated Chinese patients.Acta Haematol,2016,135(4):238-240.\u003c/li\u003e\n \u003cli\u003eTrinh,TH,Pao,LP.Pivalizza,EG.Thrombelastograph platelet mapping during hyperfibrinolysis[J].J Cardiothorac Vasc Anesth,2020,34(6):1708-1710.\u003c/li\u003e\n \u003cli\u003eDeng Y, Zhu J, Gong Y, Yi X, Zhou L, Xie Y,\u0026nbsp;et al. Hereditary factor V deficiency from heterozygous mutations with a novel variant p.Pro798Leufs\u0026lowast;13 in the F5 gene. Blood Coagul Fibrinolysis. 2021 Oct 1;32(7):513-518.\u003c/li\u003e\n \u003cli\u003eMatsukuma E, Gotoh Y, Kuroyanagi Y,et al .A case of atypical hemolytic uremic syndrome due to anti-factor H antibody in a patient presenting with a factor XII deficiency identified two novel mutations[J].Clinical \u0026amp; Experimental Nephrology, 2011, 15(2):269-274.\u003c/li\u003e\n \u003cli\u003eThomas Renn\u0026eacute;.The procoagulant and proinflammatory plasma contact system[J].Seminars in Immunopathology, 2012, 34(1):31-41.\u003c/li\u003e\n \u003cli\u003eKWON M J, KIM H J, LEE K O, et al. Molecular genetic analysis of Korean patients with coagulation factor XII deficiency[J]. Blood Coagul Fibrinolysis, 2010, 21(4): 308-312.\u003c/li\u003e\n \u003cli\u003eDepasse F,Binder NB,Mueller J,et al. Thrombin generation assays are versatile tools in blood coagulation analysis:A review of technical features,and applications from research to laboratory routine[J]. J Thromb Haemost,2021,19(12):2907-2917.\u003c/li\u003e\n \u003cli\u003eSantagostino E,Mancuso ME,Tripodi A,et al.Severe hemophilia with mild bleeding phenotype:molecular haracterization and global coagulation profile[J].JThromb Haemost,2010,8(4):737-743.\u003c/li\u003e\n \u003cli\u003eXie H,Lu M,Yang X et al. Gene analysis in four inherited coagulation FⅫ\u0026nbsp;deficiency pedigree [J]. Chinese Journal of Hematology, 2013, 3\u0026shy;\u0026shy;4(3): 5.\u003c/li\u003e\n \u003cli\u003eDing Q, Wu W, Fu Q, et al. Novel aberrant splicings caused by a splice site mutation (IVS1a+5g\u0026gt;a) in F7 gene[J]. Thromb Haemost, 2005,93(6): 1077-1081.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Table","content":"\u003cp\u003e\u003cstrong\u003eTable. Phenotypes andgenotypes of the inheritedcoagulation factor XII deficiencyfamily\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"Left\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"784\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"10.814249363867685%\"\u003e\n \u003cp\u003ePatient\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.669211195928753%\"\u003e\n \u003cp\u003ePT\u003c/p\u003e\n \u003cp\u003e(s)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.669211195928753%\"\u003e\n \u003cp\u003eAPTT\u003c/p\u003e\n \u003cp\u003e(s)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.669211195928753%\"\u003e\n \u003cp\u003eFIB\u003c/p\u003e\n \u003cp\u003e(g/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.979643765903308%\"\u003e\n \u003cp\u003eD-D\u003c/p\u003e\n \u003cp\u003e(g/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.669211195928753%\"\u003e\n \u003cp\u003eLAC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.396946564885496%\"\u003e\n \u003cp\u003eFⅧ:C\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.251908396946565%\"\u003e\n \u003cp\u003eFⅨ:C\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.251908396946565%\"\u003e\n \u003cp\u003eFⅪ:C\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.124681933842239%\"\u003e\n \u003cp\u003eFⅫ:C\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.50381679389313%\"\u003e\n \u003cp\u003eGenetic mutations\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"10.814249363867685%\"\u003e\n \u003cp\u003eProband\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.669211195928753%\"\u003e\n \u003cp\u003e14.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.669211195928753%\"\u003e\n \u003cp\u003e169.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.669211195928753%\"\u003e\n \u003cp\u003e2.43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.979643765903308%\"\u003e\n \u003cp\u003e0.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.669211195928753%\"\u003e\n \u003cp\u003e1.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.396946564885496%\"\u003e\n \u003cp\u003e93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.251908396946565%\"\u003e\n \u003cp\u003e76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.251908396946565%\"\u003e\n \u003cp\u003e87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.124681933842239%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.50381679389313%\"\u003e\n \u003cp\u003ec.303_304 del CA c.800+1G\u0026gt;A\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"10.814249363867685%\"\u003e\n \u003cp\u003eGrandfather\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.669211195928753%\"\u003e\n \u003cp\u003e12.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.669211195928753%\"\u003e\n \u003cp\u003e44.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.669211195928753%\"\u003e\n \u003cp\u003e3.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.979643765903308%\"\u003e\n \u003cp\u003e0.41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.669211195928753%\"\u003e\n \u003cp\u003e1.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.396946564885496%\"\u003e\n \u003cp\u003e128\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.251908396946565%\"\u003e\n \u003cp\u003e86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.251908396946565%\"\u003e\n \u003cp\u003e104\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.124681933842239%\"\u003e\n \u003cp\u003e45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.50381679389313%\"\u003e\n \u003cp\u003ec.800+1G\u0026gt;A\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd 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\u003c/table\u003e\n\u003c/div\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"annals-of-hematology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"aohe","sideBox":"Learn more about [Annals of Hematology](http://link.springer.com/journal/277)","snPcode":"277","submissionUrl":"https://submission.nature.com/new-submission/277/3","title":"Annals of Hematology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Coagulation FXII deficiency, Genetic analysis, Bioinformatics, Gene mutation","lastPublishedDoi":"10.21203/rs.3.rs-4978926/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4978926/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eObjective: \u003c/strong\u003eAnalyze the clinical phenotype and gene mutations of a family with hereditary FXII deficiency, and preliminarily explore its molecular pathogenic mechanism.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e The routine coagulation indicators and related coagulation factors were measured.. Thromboelastography and thrombin generation tests simulated coagulation and anticoagulation states in vitro and in vivo. PCR direct sequencing was utilized to analyze all exons and flanking sequences of the F12 gene in the proband, confirming suspected mutations through reverse sequencing, and identifying corresponding mutation sites in family members. Using ClustalX-2.1-win to analyze the conservation of the variant, and employing online software to predict the pathogenicity of mutations.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e The proband exhibited significantly prolonged APTT (169.1 seconds) and a pronounced decrease in FXII:C to 1.0%. Thromboelastography testing indicated a diminished function of the endogenous coagulation system, while thrombin generation testing revealed a normal ability for thrombin production in the proband. Gene sequencing revealed that the proband harbored a deletion mutation c.303_304delCA in exon 5 and a substitution mutation c.800+1G\u0026gt;A in intron 8. All three bioinformatics software indicated that the mutations were pathogenic and could lead to the production of a terminator, potentially altering the structure and function of the protein.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e The deletion mutation c.303_304delCA and substitution mutation c.800+1G\u0026gt;A are associated with a decreased in FXII levels in this family, with the c.800+1G\u0026gt;A mutation being the first reported mutation worldwide.\u003c/p\u003e","manuscriptTitle":"Genetic analysis of a pedigree with hereditary coagulation factor XII deficiency","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-10-17 13:53:58","doi":"10.21203/rs.3.rs-4978926/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-09-24T01:42:59+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-09-23T19:18:50+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-09-23T15:12:12+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"157654706383194061750260992094427643286","date":"2024-09-14T02:17:36+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"154605882764193399124625011210307826494","date":"2024-09-13T05:19:24+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-09-12T23:00:38+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-08-28T10:47:28+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-08-28T10:44:28+00:00","index":"","fulltext":""},{"type":"submitted","content":"Annals of Hematology","date":"2024-08-26T15:07:58+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"annals-of-hematology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"aohe","sideBox":"Learn more about [Annals of Hematology](http://link.springer.com/journal/277)","snPcode":"277","submissionUrl":"https://submission.nature.com/new-submission/277/3","title":"Annals of Hematology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"baf4591f-2ddf-4988-b534-3d7ce4e0da4e","owner":[],"postedDate":"October 17th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-01-27T16:05:35+00:00","versionOfRecord":{"articleIdentity":"rs-4978926","link":"https://doi.org/10.1007/s00277-025-06205-4","journal":{"identity":"annals-of-hematology","isVorOnly":false,"title":"Annals of Hematology"},"publishedOn":"2025-01-22 15:57:17","publishedOnDateReadable":"January 22nd, 2025"},"versionCreatedAt":"2024-10-17 13:53:58","video":"","vorDoi":"10.1007/s00277-025-06205-4","vorDoiUrl":"https://doi.org/10.1007/s00277-025-06205-4","workflowStages":[]},"version":"v1","identity":"rs-4978926","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4978926","identity":"rs-4978926","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}