Using Mendelian randomization and bioinformatics methods to explore the causal effect of hypothyroidism on urticaria

Using Mendelian randomization and bioinformatics methods to explore the causal effect of hypothyroidism on urticaria | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Using Mendelian randomization and bioinformatics methods to explore the causal effect of hypothyroidism on urticaria Qianqian Chen, Ting Yang, Shifeng Qiu, Min Xiao This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4252621/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Observational studies connect hypothyroidism with urticaria, but causality is unclear. This study explores genetic variations in hypothyroidism and their potential influence on urticaria risk.We collected statistical data from open genome-wide association studies in the Integrative Epidemiology Unit (IEU OpenGwas) project, analyzed genetic data from patients with hypothyroidism and urticaria to study their causal relationship using various methods such as the inverse variance weighting (IVW) method, MR-Egger method, weighted median estimator (WME) method, and single-nucleotide polymorphism (SNP) analysis through gene ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, and protein-protein interaction (PPI) network evaluation.Our research identified SNPs that significantly elevate urticaria risk in hypothyroidism. GO analysis highlighted crucial genes regulating immune cell function, adhesion, growth, and cytokine production. KEGG analysis demonstrated enrichment of these genes in immune response and autoimmune pathways. PPI network analysis pinpointed key genes involved in these processes. The study provides strong genetic evidence that there is a causal relationship between hypothyroidism and an increased incidence of urticaria. This finding is expected to help develop more effective treatment strategies for urticaria patients. Biological sciences/Computational biology and bioinformatics Health sciences/Medical research/Genetics research Health sciences/Medical research/Epidemiology Mendelian randomization study SNPs hypothyroidism urticaria Figures Figure 1 Figure 2 Figure 3 Figure 4 1. Introduction Urticaria is a common disease characterized by localized edema reactions caused by dilation and increased permeability of small blood vessels in the skin and mucous membranes. Typical symptoms include pale red patches and itching on the skin, which usually subside on their own in a short period of time [ 1 ] . Currently, more than 70% of urticaria patients cannot find the exact cause, but immune responses induced by genetic, physical, endocrine metabolism, and environmental factors are considered important steps in its pathogenesis [ 2 ] . Urticaria occurs in one out of every five people worldwide. The disease is easy to diagnose, but due to its persistence, recurrence, and often developing into chronic spontaneous skin allergic reactions, as well as various pathogenic factors, there are certain difficulties in treatment, which easily burden economic and social life. A common cause of urticaria is autoimmune diseases. Current research shows that the proportion of thyroid dysfunction in people with chronic urticaria is significantly increased [ 3 ] . Some studies have shown that patients with chronic urticaria have high affinity receptors for Immunoglobulin E (IgE) FcepsilonRI, anti-IgE, and anti-thyroid antibodies. Early treatment with thyroid drugs may help alleviate chronic spontaneous urticaria (CSU) in patients with both autoimmune thyroid disease and CSU [ 4 – 6 ] .The relationship between urticaria and hypothyroidism is worth studying. Mendelian Randomization (MR) is an effective causal inference analysis method that deduces the causal relationship between exposure and outcome based on genetic variation, and largely avoids confounding bias in traditional epidemiological studies [ 7 – 8 ] . Currently, the relationship between urticaria and hypothyroidism remains unclear. This study employs MR analysis to explore potential causal relationships between the two from a genetic perspective, aiming to discover new therapeutic strategies and preventive measures. 2. Methodology 2.1 Research Design Two-sample MR was used to study the causal relationship between hypothyroidism and urticaria. MR is an effective method to derive the causal relationship between exposure and outcome using genetic variation as an instrumental variable (SNPs). The instrumental variable must satisfy three core assumptions: ①there is a strong correlation between SNPs and exposure factors;②SNPs only affect outcomes through exposure factors (exclusivity hypothesis);③SNPs and confounding factors affecting the "exposure-outcome" relationship are independent of each other [ 9 ] .The analysis workflow in this bidirectional study is shown in Fig. 1 . 2.2 Data source The datasets used in this MR analysis are all from publicly available IEU OpenGwas data and are all from European populations, reducing potential biases caused by significant differences in genetic material among populations. Summary statistical data on GWAS exposures (hypothyroidism, ebi-a-GCST90018862) and outcomes (urticaria, fin-b-L12_URTICARIA) were obtained through https://gwas.mrcieu.ac.uk/access . These summary data have obtained informed consent and ethical approval, so no additional ethical approval or informed consent is required for this study. 2.3 Instrumental variable selection To satisfy the correlation hypothesis in MR, we set the genome-wide significance threshold to p < 5 × 10 − 8 to identify instrumental variables significantly correlated with exposure. To avoid linkage disequilibrium, two parameters kb = 10,000 and r2 = 0.01 were used for clustered SNPs. When checking the exposure and outcome Statistical Data, palindrome sequences and incompatible SNPs were removed; at the same time, SNPs related to exposure that could not be matched in the GWAS results statistical data were also excluded. Finally, F statistic was used for SNPs correlation assessment (F > 10) to avoid bias caused by weak instrumental variables [ 10 – 11 ] . 2.4 Statistical analysis MR analysis methods are performed using the "TwoSampleMR" package in the R version of the 4.3.1. (IVW) method is the primary causal effect estimation in MR research and has strong causal relationship detection ability [ 12 ] . MR-Egger method and WME method test the pleiotropy of the results. In order to evaluate the stability of the results and whether there is bias, we use MR-egger intercept to test whether multiple IVs have horizontal pleiotropy, whether the evaluation results are robust, and whether the conclusions are reliable [ 13 ] .By observing the values of Cochran's Q test and MR-PRESSO, evaluate whether there is potential bias in the evaluation results. When P > 0.05, there is no heterogeneity in the data, and when there is heterogeneity, random effects IVW is used. The "leave-one-out" method removes SNPs one by one and performs MR analysis to evaluate whether there is a certain instrumental variable that seriously affects the outcome variable. In addition, MR Steiger tests the causal direction of verification. 2.5. Enrichment analysis of SNP-associated genes and PPI network construction By integrating SNPs from exposure and result tables, ensure their directionality and effect size are consistent. Use the core solo polymorphism database ( https://www.ncbi.nlm.nih.gov/snp/ ) of the US National Human Genome Research Institute to label the corresponding gene name for each rsID. GO and KEGG pathway enrichment analysis was performed using the clusterProfiler package, and the R package ggplot2 [3.3.6] was used for visualization. The threshold screening satisfied FDR (qvalue) < 0.25, and the corrected p-value 0.4) to obtain the PPI network diagram. The PPI network was visualized using Cytoscape 3.10.1 software [ 14 – 15 ] . 3. Results 3.1 Hypothyroidism is significantly associated with an increased risk of urticaria When conducting a two-sample MR analysis of the association between hypothyroidism and urticaria, the selected genetic SNPs were selected based on their check point significance (p 10) (supplementary table 1). After screening, 85 SNPs met the criteria for inclusion in the analysis. Different methods were used for Mendelian randomization analysis, and the results showed that the SNPs of hypothyroidism significantly increased the risk of urticaria, as shown in Figure 2a: IVW method (OR = 1.11, 95% CI: 1.06-1.17, p = 2.78 x 10 ^ (-5)); MR Egger (OR = 1.13, 95% CI: 1.01-1.26, p = 3.38 x 10 ^ (-2)); WME median (OR = 1.09, 95% CI: 1.01-1.18, p = 3.16 x 10^(-2)). Sensitivity analysis results are listed in Table 1. IVW analysis (p = 0.35) and MR-Egger analysis (p = 0.32) were performed in Cochran's Q test, and no significant heterogeneity was found between IVs. MR-Egger intercept (p = 0.82) and MR-PRESSO (p = 0.35) analysis showed no potential confounding factors and horizontal pleiotropy. Leave-one-out analysis confirmed the robustness and reliability of MR analysis, excluding any significant impact of SNP on the overall results (Figures 2b and 2c). Table 1 Sensitivity analysis of the MR study on the associations between hypothyroidism and the risk of urticaria. MR:Mendelian randomization Exposures Outcome Heterogeneity test Horizontal pleiotropy test Hypothyroiddism Urticaria method Q Q_df Q_pval method se pval Inverse variance weighted 88.40 84 0.35 egger_intercept 6.01×10^(-3) 0.82 MR Egger 88.35 83 0.32 MR-PRESSO 0.35 3.2 GO and KEGG enrichment analysis By harmonizing exposure and outcome data, we obtained SNPs with consistent effect alleles and effect amounts, and searched for the genes corresponding to each rsID number (Supplementary Table 2). GO analysis showed that core genes play important roles in regulating immune cell activation, intercellular adhesion, cell proliferation, and cytokine production. These genes are mainly enriched in positive regulation of leukocyte activation, positive regulation of lymphocyte activation, positive regulation of cell activation, regulation of leukocyte cell-cell adhesion, regulation of T cell activation, leukocyte cel-cell adhesion, regulation of cell-cell adhesion, mononuclear cell proliferation, ymphocyte proliferation, leukocyte proliferation, positive regulation of cytokine production. (Figure 3a, Supplementary Data Table 3) KEGG analysis showed that core genes are mainly enriched in Th1 and Th2 cell differentiation, Th17 cell differentiation, Influenza A, Influtory bowel disease, T cell receptor signaling pathway, Hematopoietic cell lineage, Autoimmune thyroid disease. (Figure 3b, Supplementary Data Table 4) 3.3 Construction of PPI network Using the STRING database to construct a protein-protein interaction (PPI) network and exclude isolated node genes. In this network, the depth of node color reflects the strength of gene association(Figure4). The analysis results show that PTPN22 , CTLA4 , CD247 , TBX21 , SH2B3 , IL2RA , TYK2 , STAT4 , and TLR3 play important roles in the association mechanism between hypothyroidism and urticaria. 4. Discussion Urticaria is a common skin disease characterized by pruritic erythema and localized skin lesions, closely related to angioedema, and be divided into acute and chronic types. Among them, chronic urticaria be related to autoimmune diseases such as hypothyroidism [16–17] . Studies have shown that most CSU patients have elevated levels of thyroid peroxidase antibody(TPO-Ab) and thyroglobulin antibody (TG-Ab), and CSU patients with hypothyroidism are more likely to be older women with specific biochemical markers [ 18 – 19 ] . In addition, A systematic review study showed that Immunoglobulin G (IgG) levels of anti-thyroid autoantibodies (AAbs) and thyroid peroxidase (TPO) are often elevated in CSU patients, especially in adult patients, and thyroid dysfunction is more likely to occur. Among them, hypothyroidism and Hashimoto's thyroiditis are more common than hyperthyroidism and Graves' disease [ 20 ] . In addition, a survey of 607 adult patients with CSU who were continuously referred showed that 10.1% of the patients were known to have thyroid disease [ 21 ] . Based on these findings, our study employed a two-sample MR design to investigate the causal relationship between hypothyroidism and urticaria. The study provides robust genetic evidence suggesting that hypothyroidism is a risk factor for an increased incidence of urticaria. Although the mechanism of hypothyroidism and CSU is not fully understood, many literature supports a generally accepted view: CSU is associated with elevated levels of IgG AAbs [22] . These autoantibodies have been shown in vitro to stimulate the degranulation of eosinophils and skin mast cells in the blood, particularly by stimulating the release of complement and Complement Component 5a (C5a) to enhance the activation of eosinophils [ 23 – 24 ] . In addition, studies have shown that high levels of IgE antibodies in the blood of chronic urticaria patients be a new pathogenesis, in which thyroid peroxidase (TPO) IgE sensitize mast cells and induce degranulation after exposure to specific circulating antigens [25–28] . Mast cells play an important role in immune and inflammatory diseases, containing a large amount of histamine, hormones, proteases, and other mediators. Studies have shown that mast cells interact with thyroid function, especially in storing thyroid hormones and expressing thyroid-stimulating hormone receptors (TSHRs) [29] . There is pharmacological evidence showing an increase in thyroid hormone metabolites such as 3,5-diiodothyronine (3,5-T2), 3-iodothyrogenamine (T1AM), and 3-iodothyroacetic acid (TA1) in inflammatory diseases involving mast cells [ 30 ] . These compounds increase in inflammatory diseases and produce neuroprotective effects and regulate itch and pain sensitivity through mechanisms such as mediating histamine release.This explain the association between hypothyroidism and urticaria observed in our current study. Additionally, we conducted research on hypothyroidism and urticaria, utilizing harmonized SNPs data to investigate their potential disease-driving gene features and functional networks. Through enrichment analysis, we confirmed core modules and signaling pathways closely associated with both conditions. These core genes play crucial roles in regulating immune cell activation, cytokine production, and exhibit significant enrichment features in modulating immune responses, inflammatory reactions, and immune-related disease processes.Additionally, pivotal genes were identified: PTPN22 , CTLA4 , CD247 , among others. PTPN22 is one of the potential susceptibility genes for autoimmune diseases, encoding Lyp protein. Specific variations in PTPN22 have been associated with various autoimmune diseases such as type 1 diabetes, rheumatoid arthritis, systemic lupus erythematosus, etc [ 31 ] . Furthermore, PTPN22 regulates anti-thyroid peroxidase IgG and anti-thyroglobulin IgG [ 32 ] . Studies have shown that allelic variations in PTPN22 are also associated with increased susceptibility to CSU in Iranian populations and Chinese Han adults [ 33 – 34 ] . CTLA4 is one of the susceptibility genes for multiple autoimmune diseases, playing a crucial role in immune regulation by inhibiting the activation of T cells on antigen-presenting cells. Numerous studies have shown an association between the CTLA-4 gene and autoimmune hypothyroidism [ 35 – 37 ] .Our study results further demonstrate a correlation between hypothyroidism and urticaria, suggesting that investigating the potential impact of CTLA-4 on urticaria could be a future research direction. However, it should be noted that not all patients with hypothyroidism will develop urticaria, nor will all urticaria patients have thyroid dysfunction. This phenomenon be caused by various factors such as Individual Differences, severity of the disease, and treatment options. Therefore, in clinical practice, it is necessary to check the thyroid status of patients with urticaria in order to determine prevention and treatment methods. Although our study provides valuable insights, some limitations need to be taken into account. Our study only involved individuals of European descent, which limits the applicability of the research results to other populations and reduces bias caused by racial and regional differences. Although our study supports causality through genetic evidence, other studies, including functional studies, are needed to further reveal potential mechanisms. Nevertheless, our study provides important leads for understanding the relationship between hypothyroidism and urticaria. 5. Conclusion The study provides genetic evidence to support the causal relationship between hypothyroidism and urticaria. The analysis results provide further molecular biological support for the possible immune regulation abnormalities between hypothyroidism and urticaria. These findings not only help to deepen the understanding of the pathogenesis of these two diseases, but also provide important leads for the future development of therapeutic strategies for these diseases. By revealing the core gene and protein interaction network, we better understand the association between hypothyroidism and urticaria and potential therapeutic targets, providing a useful information foundation for personalized medicine and precision treatment. Declarations Data availability statement The original contributions presented in the study are included in the article/Supplementary Material. Further inquiries can be directed to the corresponding authors. Acknowledgements We authors are grateful to all the participants of this work. Appreciating for the tools provided by website:www.xiantao.love. Author contributions Qianqian Chen and Shifeng Qiu wrote the main manuscript text. Ting Yang is responsible for drawing and analysis.Min Xiao review and modify the manuscript.All authors reviewed the manuscript. Funding This work was supported by the Sichuan Provincial Administration of Traditional Chinese Medicine (2023MS470)and Chengdu University of Traditional Chinese Medicine “Apricot Grove Scholars” Discipline Talent Research Enhancement Program(QJRC2021007). References Radonjic-Hoesli S, Hofmeier KS, Micaletto S, Schmid-Grendelmeier P, Bircher A, Simon D. Urticaria and Angioedema: an Update on Classification and Pathogenesis. Clin Rev Allergy Immunol. 2018;54(1):88-101. Gimenez-Arnau AM, Maibach H. Contact Urticaria. Immunol Allergy Clin North Am. 2021;41(3):467-480. Bagnasco M, Minciullo PL, Saraceno GS, Gangemi S, Benvenga S. Urticaria and thyroid autoimmunity. Thyroid. 2011;21(4):401-410. 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Genetic Polymorphism of PTPN22 in Autoimmune Diseases: A Comprehensive Review. Medicina (Kaunas) . 2022;58(8):1034. Published 2022 Aug 2. Jacobson EM, Tomer Y. The CD40, CTLA-4, thyroglobulin, TSH receptor, and PTPN22 gene quintet and its contribution to thyroid autoimmunity: back to the future. J Autoimmun . 2007;28(2-3):85-98. Sadr M, Khalili N, Mohebbi B, Mosharmovahed B, Afradi P, Rezaei N. Association of PTPN22 single nucleotide polymorphisms with chronic spontaneous urticaria. Allergol Immunopathol (Madr) . 2021;49(2):40-45. Published 2021 Mar 1. Zhang L, Qiu L, Wu J, et al. GWAS of Chronic Spontaneous Urticaria Reveals Genetic Overlap with Autoimmune Diseases, Not Atopic Diseases. J Invest Dermatol . 2023;143(1):67-77.e15. Patel H, Mansuri MS, Singh M, Begum R, Shastri M, Misra A. Association of Cytotoxic T-Lymphocyte Antigen 4 (CTLA4) and Thyroglobulin (TG) Genetic Variants with Autoimmune Hypothyroidism. PLoS One . 2016;11(3):e0149441. Published 2016 Mar 10. 1 Ueda H, Howson JM, Esposito L, et al. Association of the T-cell regulatory gene CTLA4 with susceptibility to autoimmune disease. Nature . 2003;423(6939):506-511. Vaidya B, Pearce S. The emerging role of the CTLA-4 gene in autoimmune endocrinopathies. Eur J Endocrinol . 2004;150(5):619-626. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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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-4252621","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":299046308,"identity":"8c4b27ea-5cad-48fe-9cbb-1e366b7a6465","order_by":0,"name":"Qianqian Chen","email":"","orcid":"","institution":"The 925th Hospital Of The Joint Logistics Support Force.Guiyang","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Qianqian","middleName":"","lastName":"Chen","suffix":""},{"id":299046309,"identity":"4c2931b2-4b2a-4ba5-9a5b-0e2ff4bdc609","order_by":1,"name":"Ting Yang","email":"","orcid":"","institution":"The Traditional Chinese Medicine Hospital Of Longquanyi","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ting","middleName":"","lastName":"Yang","suffix":""},{"id":299046310,"identity":"fbb8e824-d4ad-433e-b1eb-5b0f349d3f99","order_by":2,"name":"Shifeng Qiu","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA1ElEQVRIiWNgGAWjYBACNvaGxAcSP2x4+NmbDxCnhY/nwGMDy540GcmeYwnEaZGTSHwmUcF22MbgRo4BkQ5jSE6QuMGTxsNw5szHG28Y7OR0GwhqOZZgOMPChoexvXez5RyGZGOzA4S0MPYkJEsAbWHmObtNmofhQOI2glqY+T8c/sN2mIdNIucZkVrYGBIbJIBaeCRy2IjUwsOQzCDZk8YjwXPM2HKOARF+kZ//IP0HMCrt7Y83P7zxpsJOjqAWFCDBQ2TUIGshVccoGAWjYBSMCAAAuHk/xlF5xPMAAAAASUVORK5CYII=","orcid":"","institution":"Chongzhou People's Hospital","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Shifeng","middleName":"","lastName":"Qiu","suffix":""},{"id":299046311,"identity":"6b9b8c64-15e3-4c6d-9a12-0da6291c444c","order_by":3,"name":"Min Xiao","email":"","orcid":"","institution":"Hospital of Chengdu University of Traditional Chinese Medicine","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Min","middleName":"","lastName":"Xiao","suffix":""}],"badges":[],"createdAt":"2024-04-11 13:05:00","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4252621/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4252621/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":56122436,"identity":"743ed0d7-fdd0-4f5a-a189-1a48dcee9d4d","added_by":"auto","created_at":"2024-05-08 20:32:23","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":64993,"visible":true,"origin":"","legend":"\u003cp\u003eMR's three major hypotheses\u003c/p\u003e\n\u003cp\u003eMR: Mendelian randomization\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4252621/v1/eed0d9468e19403d36ec7ca3.png"},{"id":56122437,"identity":"f7246ab7-1419-4a96-8042-8c610714dab5","added_by":"auto","created_at":"2024-05-08 20:32:24","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":141600,"visible":true,"origin":"","legend":"\u003cp\u003eHypothyroidism significantly associated with risk of urticaria.(a)Two-sample MR assessed the risk of hypothyroidism and urticaria.(b)The scatterplot of hypothyroidism against urticaria.(c)Leave-one-out sensitivity analysis plot for hypothyroidism versus urticaria.\u003c/p\u003e\n\u003cp\u003eMR: Mendelian randomization\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4252621/v1/275c1bf67856111df45838aa.png"},{"id":56122438,"identity":"e82f667a-44a3-4354-8495-3d127ab76d21","added_by":"auto","created_at":"2024-05-08 20:32:24","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":254304,"visible":true,"origin":"","legend":"\u003cp\u003eFunctional assessment of causal SNPs.(a)GO enrichment analysis of SNP-associated genes.(b)KEGG pathway analysis of SNP-associated genes.\u003c/p\u003e\n\u003cp\u003eSNP:single-nucleotide polymorphism;GO: Gene Ontology, KEGG：Kyoto Encyclopedia of Genes and Genomes.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-4252621/v1/395d9f079c46d5d511db0726.png"},{"id":56122439,"identity":"1168484b-9f53-4744-aaa9-bf34957dce48","added_by":"auto","created_at":"2024-05-08 20:32:24","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":235696,"visible":true,"origin":"","legend":"\u003cp\u003ePPI Network Construction.(a)PPI network diagram for SNPs. (b)The processed PPI network diagram for SNPs.SNP:single-nucleotide polymorphism\u003c/p\u003e\n\u003cp\u003ePPI:Protein–protein interaction\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4252621/v1/e63159bd67a82ef3e7818ce0.png"},{"id":59389895,"identity":"52ca212a-3224-498f-9e53-6a64c257bb14","added_by":"auto","created_at":"2024-07-01 07:48:23","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1058199,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4252621/v1/da2dbeb0-a055-4673-a3ca-e41c449b2c7e.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Using Mendelian randomization and bioinformatics methods to explore the causal effect of hypothyroidism on urticaria","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eUrticaria is a common disease characterized by localized edema reactions caused by dilation and increased permeability of small blood vessels in the skin and mucous membranes. Typical symptoms include pale red patches and itching on the skin, which usually subside on their own in a short period of time\u003csup\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]\u003c/sup\u003e. Currently, more than 70% of urticaria patients cannot find the exact cause, but immune responses induced by genetic, physical, endocrine metabolism, and environmental factors are considered important steps in its pathogenesis\u003csup\u003e[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eUrticaria occurs in one out of every five people worldwide. The disease is easy to diagnose, but due to its persistence, recurrence, and often developing into chronic spontaneous skin allergic reactions, as well as various pathogenic factors, there are certain difficulties in treatment, which easily burden economic and social life. A common cause of urticaria is autoimmune diseases. Current research shows that the proportion of thyroid dysfunction in people with chronic urticaria is significantly increased\u003csup\u003e[\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]\u003c/sup\u003e. Some studies have shown that patients with chronic urticaria have high affinity receptors for Immunoglobulin E (IgE) FcepsilonRI, anti-IgE, and anti-thyroid antibodies. Early treatment with thyroid drugs may help alleviate chronic spontaneous urticaria (CSU) in patients with both autoimmune thyroid disease and CSU\u003csup\u003e[\u003cspan additionalcitationids=\"CR5\" citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003c/sup\u003e.The relationship between urticaria and hypothyroidism is worth studying.\u003c/p\u003e \u003cp\u003eMendelian Randomization (MR) is an effective causal inference analysis method that deduces the causal relationship between exposure and outcome based on genetic variation, and largely avoids confounding bias in traditional epidemiological studies\u003csup\u003e[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]\u003c/sup\u003e. Currently, the relationship between urticaria and hypothyroidism remains unclear. This study employs MR analysis to explore potential causal relationships between the two from a genetic perspective, aiming to discover new therapeutic strategies and preventive measures.\u003c/p\u003e"},{"header":"2. Methodology","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n \u003ch2\u003e2.1 Research Design\u003c/h2\u003e\n \u003cp\u003eTwo-sample MR was used to study the causal relationship between hypothyroidism and urticaria. MR is an effective method to derive the causal relationship between exposure and outcome using genetic variation as an instrumental variable (SNPs). The instrumental variable must satisfy three core assumptions: ①there is a strong correlation between SNPs and exposure factors;②SNPs only affect outcomes through exposure factors (exclusivity hypothesis);③SNPs and confounding factors affecting the \u0026quot;exposure-outcome\u0026quot; relationship are independent of each other\u003csup\u003e[\u003cspan class=\"CitationRef\"\u003e9\u003c/span\u003e]\u003c/sup\u003e.The analysis workflow in this bidirectional study is shown in Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n \u003ch2\u003e2.2 Data source\u003c/h2\u003e\n \u003cp\u003eThe datasets used in this MR analysis are all from publicly available IEU OpenGwas data and are all from European populations, reducing potential biases caused by significant differences in genetic material among populations. Summary statistical data on GWAS exposures (hypothyroidism, ebi-a-GCST90018862) and outcomes (urticaria, fin-b-L12_URTICARIA) were obtained through \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://gwas.mrcieu.ac.uk/access\u003c/span\u003e\u003c/span\u003e. These summary data have obtained informed consent and ethical approval, so no additional ethical approval or informed consent is required for this study.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\n \u003ch2\u003e2.3 Instrumental variable selection\u003c/h2\u003e\n \u003cp\u003eTo satisfy the correlation hypothesis in MR, we set the genome-wide significance threshold to p\u0026thinsp;\u0026lt;\u0026thinsp;5 \u0026times; 10\u0026thinsp;\u0026minus;\u0026thinsp;8 to identify instrumental variables significantly correlated with exposure. To avoid linkage disequilibrium, two parameters kb\u0026thinsp;=\u0026thinsp;10,000 and r2\u0026thinsp;=\u0026thinsp;0.01 were used for clustered SNPs. When checking the exposure and outcome Statistical Data, palindrome sequences and incompatible SNPs were removed; at the same time, SNPs related to exposure that could not be matched in the GWAS results statistical data were also excluded. Finally, F statistic was used for SNPs correlation assessment (F\u0026thinsp;\u0026gt;\u0026thinsp;10) to avoid bias caused by weak instrumental variables\u003csup\u003e[\u003cspan class=\"CitationRef\"\u003e10\u003c/span\u003e\u0026ndash;\u003cspan class=\"CitationRef\"\u003e11\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n \u003ch2\u003e2.4 Statistical analysis\u003c/h2\u003e\n \u003cp\u003eMR analysis methods are performed using the \u0026quot;TwoSampleMR\u0026quot; package in the R version of the 4.3.1. (IVW) method is the primary causal effect estimation in MR research and has strong causal relationship detection ability\u003csup\u003e[\u003cspan class=\"CitationRef\"\u003e12\u003c/span\u003e]\u003c/sup\u003e. MR-Egger method and WME method test the pleiotropy of the results. In order to evaluate the stability of the results and whether there is bias, we use MR-egger intercept to test whether multiple IVs have horizontal pleiotropy, whether the evaluation results are robust, and whether the conclusions are reliable\u003csup\u003e[\u003cspan class=\"CitationRef\"\u003e13\u003c/span\u003e]\u003c/sup\u003e.By observing the values of Cochran\u0026apos;s Q test and MR-PRESSO, evaluate whether there is potential bias in the evaluation results. When P\u0026thinsp;\u0026gt;\u0026thinsp;0.05, there is no heterogeneity in the data, and when there is heterogeneity, random effects IVW is used. The \u0026quot;leave-one-out\u0026quot; method removes SNPs one by one and performs MR analysis to evaluate whether there is a certain instrumental variable that seriously affects the outcome variable. In addition, MR Steiger tests the causal direction of verification.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n \u003ch2\u003e\u003cstrong\u003e2.5. Enrichment analysis of\u003c/strong\u003e SNP-associated \u003cstrong\u003egenes and PPI network construction\u003c/strong\u003e\u003c/h2\u003e\n \u003cp\u003eBy integrating SNPs from exposure and result tables, ensure their directionality and effect size are consistent. Use the core solo polymorphism database (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ncbi.nlm.nih.gov/snp/\u003c/span\u003e\u003c/span\u003e) of the US National Human Genome Research Institute to label the corresponding gene name for each rsID. GO and KEGG pathway enrichment analysis was performed using the clusterProfiler package, and the R package ggplot2 [3.3.6] was used for visualization. The threshold screening satisfied FDR (qvalue)\u0026thinsp;\u0026lt;\u0026thinsp;0.25, and the corrected p-value\u0026thinsp;\u0026lt;\u0026thinsp;0.05. The SNP-associated genes were imported into the String online tool (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://string-db.org/\u003c/span\u003e\u003c/span\u003e), and the Confidence Level was set to moderate Confidence Level (\u0026gt;\u0026thinsp;0.4) to obtain the PPI network diagram. The PPI network was visualized using Cytoscape 3.10.1 software\u003csup\u003e[\u003cspan class=\"CitationRef\"\u003e14\u003c/span\u003e\u0026ndash;\u003cspan class=\"CitationRef\"\u003e15\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"3. Results","content":"\u003cp\u003e\u003cstrong\u003e3.1 Hypothyroidism is significantly associated with an increased risk of urticaria\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWhen conducting a two-sample MR analysis of the association between hypothyroidism and urticaria, the selected genetic SNPs were selected based on their check point significance (p \u0026lt; 5 \u0026times; 10 ^ -8). SNPs with intermediate allele frequencies and palindromes, such as rs2921053, were excluded. The correlation of SNPs was evaluated by F statistics (F \u0026gt; 10) (supplementary table 1). After screening, 85 SNPs met the criteria for inclusion in the analysis. Different methods were used for Mendelian randomization analysis, and the results showed that the SNPs of hypothyroidism significantly increased the risk of urticaria, as shown in Figure 2a: IVW method (OR = 1.11, 95% CI: 1.06-1.17, p = 2.78 x 10 ^ (-5)); MR Egger (OR = 1.13, 95% CI: 1.01-1.26, p = 3.38 x 10 ^ (-2)); WME median (OR = 1.09, 95% CI: 1.01-1.18, p = 3.16 x 10^(-2)). Sensitivity analysis results are listed in Table 1. IVW analysis (p = 0.35) and MR-Egger analysis (p = 0.32) were performed in Cochran\u0026apos;s Q test, and no significant heterogeneity was found between IVs. MR-Egger intercept (p = 0.82) and MR-PRESSO (p = 0.35) analysis showed no potential confounding factors and horizontal pleiotropy. Leave-one-out analysis confirmed the robustness and reliability of MR analysis, excluding any significant impact of SNP on the overall results (Figures 2b and 2c).\u003c/p\u003e\n\u003cp\u003eTable 1 Sensitivity analysis of the MR study on the associations between hypothyroidism and the risk of urticaria.\u003c/p\u003e\n\u003cp\u003eMR:Mendelian randomization\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"677\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.699115044247787%\"\u003e\n \u003cp\u003e\u003cstrong\u003eExposures\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.32448377581121%\"\u003e\n \u003cp\u003e\u003cstrong\u003eOutcome\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"23.746312684365783%\" colspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eHeterogeneity test\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"48.08259587020649%\" colspan=\"6\"\u003e\n \u003cp\u003e\u003cstrong\u003eHorizontal pleiotropy test\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.14749262536873156%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.725258493353028%\" rowspan=\"3\"\u003e\n \u003cp\u003e\u003cstrong\u003eHypothyroiddism\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.3397341211226%\" rowspan=\"3\"\u003e\n \u003cp\u003e\u003cstrong\u003eUrticaria\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.282127031019204%\"\u003e\n \u003cp\u003e\u003cstrong\u003emethod\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.942392909896602%\" colspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eQ\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.646971935007386%\"\u003e\n \u003cp\u003e\u003cstrong\u003eQ_df\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.567208271787297%\"\u003e\n \u003cp\u003e\u003cstrong\u003eQ_pval\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.805022156573116%\"\u003e\n \u003cp\u003e\u003cstrong\u003emethod\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.862629246676514%\"\u003e\n \u003cp\u003e\u003cstrong\u003ese\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.828655834564254%\" colspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003epval\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"24.024640657084188%\"\u003e\n \u003cp\u003e\u003cstrong\u003eInverse variance weighted\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.650924024640657%\" colspan=\"2\"\u003e\n \u003cp\u003e88.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.240246406570842%\"\u003e\n \u003cp\u003e84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.90965092402464%\"\u003e\n \u003cp\u003e0.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"21.971252566735114%\"\u003e\n \u003cp\u003e\u003cstrong\u003eegger_intercept\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.320328542094456%\"\u003e\n \u003cp\u003e6.01\u0026times;10^(-3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.882956878850102%\" colspan=\"2\"\u003e\n \u003cp\u003e0.82\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"24.024640657084188%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMR Egger\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.650924024640657%\" colspan=\"2\"\u003e\n \u003cp\u003e88.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.240246406570842%\"\u003e\n \u003cp\u003e83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.90965092402464%\"\u003e\n \u003cp\u003e0.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"21.971252566735114%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMR-PRESSO\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.320328542094456%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.882956878850102%\" colspan=\"2\"\u003e\n \u003cp\u003e0.35\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003e3.2 GO and KEGG enrichment analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBy harmonizing exposure and outcome data, we obtained SNPs with consistent effect alleles and effect amounts, and searched for the genes corresponding to each rsID number (Supplementary Table 2). GO analysis showed that core genes play important roles in regulating immune cell activation, intercellular adhesion, cell proliferation, and cytokine production. These genes are mainly enriched in positive regulation of leukocyte activation, positive regulation of lymphocyte activation, positive regulation of cell activation, regulation of leukocyte cell-cell adhesion, regulation of T cell activation, leukocyte cel-cell adhesion, regulation of cell-cell adhesion, mononuclear cell proliferation, ymphocyte proliferation, leukocyte proliferation, positive regulation of cytokine production. (Figure 3a, Supplementary Data Table 3) KEGG analysis showed that core genes are mainly enriched in Th1 and Th2 cell differentiation, Th17 cell differentiation, Influenza A, Influtory bowel disease, T cell receptor signaling pathway, Hematopoietic cell lineage, Autoimmune thyroid disease. (Figure 3b, Supplementary Data Table 4)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.3 Construction of PPI network\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eUsing the STRING database to construct a protein-protein interaction (PPI) network and exclude isolated node genes. In this network, the depth of node color reflects the strength of gene association(Figure4). The analysis results show that \u003cem\u003ePTPN22\u003c/em\u003e,\u003cem\u003e\u0026nbsp;CTLA4\u003c/em\u003e, \u003cem\u003eCD247\u003c/em\u003e, \u003cem\u003eTBX21\u003c/em\u003e, \u003cem\u003eSH2B3\u003c/em\u003e, \u003cem\u003eIL2RA\u003c/em\u003e, \u003cem\u003eTYK2\u003c/em\u003e, \u003cem\u003eSTAT4\u003c/em\u003e, and \u003cem\u003eTLR3\u003c/em\u003e play important roles in the association mechanism between hypothyroidism and urticaria.\u003c/p\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eUrticaria is a common skin disease characterized by pruritic erythema and localized skin lesions, closely related to angioedema, and be divided into acute and chronic types. Among them, chronic urticaria be related to autoimmune diseases such as hypothyroidism\u003csup\u003e[16\u0026ndash;17]\u003c/sup\u003e. Studies have shown that most CSU patients have elevated levels of thyroid peroxidase antibody(TPO-Ab) and thyroglobulin antibody (TG-Ab), and CSU patients with hypothyroidism are more likely to be older women with specific biochemical markers\u003csup\u003e[\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e19\u003c/span\u003e]\u003c/sup\u003e. In addition, A systematic review study showed that Immunoglobulin G (IgG) levels of anti-thyroid autoantibodies (AAbs) and thyroid peroxidase (TPO) are often elevated in CSU patients, especially in adult patients, and thyroid dysfunction is more likely to occur. Among them, hypothyroidism and Hashimoto's thyroiditis are more common than hyperthyroidism and Graves' disease\u003csup\u003e[\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e20\u003c/span\u003e]\u003c/sup\u003e. In addition, a survey of 607 adult patients with CSU who were continuously referred showed that 10.1% of the patients were known to have thyroid disease\u003csup\u003e[\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e21\u003c/span\u003e]\u003c/sup\u003e. Based on these findings, our study employed a two-sample MR design to investigate the causal relationship between hypothyroidism and urticaria. The study provides robust genetic evidence suggesting that hypothyroidism is a risk factor for an increased incidence of urticaria.\u003c/p\u003e \u003cp\u003eAlthough the mechanism of hypothyroidism and CSU is not fully understood, many literature supports a generally accepted view: CSU is associated with elevated levels of IgG AAbs\u003csup\u003e[22]\u003c/sup\u003e. These autoantibodies have been shown in vitro to stimulate the degranulation of eosinophils and skin mast cells in the blood, particularly by stimulating the release of complement and Complement Component 5a (C5a) to enhance the activation of eosinophils\u003csup\u003e[\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e24\u003c/span\u003e]\u003c/sup\u003e. In addition, studies have shown that high levels of IgE antibodies in the blood of chronic urticaria patients be a new pathogenesis, in which thyroid peroxidase (TPO) IgE sensitize mast cells and induce degranulation after exposure to specific circulating antigens\u003csup\u003e[25\u0026ndash;28]\u003c/sup\u003e. Mast cells play an important role in immune and inflammatory diseases, containing a large amount of histamine, hormones, proteases, and other mediators. Studies have shown that mast cells interact with thyroid function, especially in storing thyroid hormones and expressing thyroid-stimulating hormone receptors (TSHRs)\u003csup\u003e[29]\u003c/sup\u003e. There is pharmacological evidence showing an increase in thyroid hormone metabolites such as 3,5-diiodothyronine (3,5-T2), 3-iodothyrogenamine (T1AM), and 3-iodothyroacetic acid (TA1) in inflammatory diseases involving mast cells\u003csup\u003e[\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e30\u003c/span\u003e]\u003c/sup\u003e. These compounds increase in inflammatory diseases and produce neuroprotective effects and regulate itch and pain sensitivity through mechanisms such as mediating histamine release.This explain the association between hypothyroidism and urticaria observed in our current study. Additionally, we conducted research on hypothyroidism and urticaria, utilizing harmonized SNPs data to investigate their potential disease-driving gene features and functional networks. Through enrichment analysis, we confirmed core modules and signaling pathways closely associated with both conditions. These core genes play crucial roles in regulating immune cell activation, cytokine production, and exhibit significant enrichment features in modulating immune responses, inflammatory reactions, and immune-related disease processes.Additionally, pivotal genes were identified: \u003cem\u003ePTPN22\u003c/em\u003e, \u003cem\u003eCTLA4\u003c/em\u003e, \u003cem\u003eCD247\u003c/em\u003e, among others. \u003cem\u003ePTPN22\u003c/em\u003e is one of the potential susceptibility genes for autoimmune diseases, encoding Lyp protein. Specific variations in \u003cem\u003ePTPN22\u003c/em\u003e have been associated with various autoimmune diseases such as type 1 diabetes, rheumatoid arthritis, systemic lupus erythematosus, etc\u003csup\u003e[\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e31\u003c/span\u003e]\u003c/sup\u003e. Furthermore, \u003cem\u003ePTPN22\u003c/em\u003e regulates anti-thyroid peroxidase IgG and anti-thyroglobulin IgG\u003csup\u003e[\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e32\u003c/span\u003e]\u003c/sup\u003e. Studies have shown that allelic variations in \u003cem\u003ePTPN22\u003c/em\u003e are also associated with increased susceptibility to CSU in Iranian populations and Chinese Han adults\u003csup\u003e[\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e34\u003c/span\u003e]\u003c/sup\u003e.\u003cem\u003eCTLA4\u003c/em\u003e is one of the susceptibility genes for multiple autoimmune diseases, playing a crucial role in immune regulation by inhibiting the activation of T cells on antigen-presenting cells. Numerous studies have shown an association between the \u003cem\u003eCTLA-4\u003c/em\u003e gene and autoimmune hypothyroidism\u003csup\u003e[\u003cspan additionalcitationids=\"CR36\" citationid=\"CR32\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e37\u003c/span\u003e]\u003c/sup\u003e.Our study results further demonstrate a correlation between hypothyroidism and urticaria, suggesting that investigating the potential impact of \u003cem\u003eCTLA-4\u003c/em\u003e on urticaria could be a future research direction.\u003c/p\u003e \u003cp\u003eHowever, it should be noted that not all patients with hypothyroidism will develop urticaria, nor will all urticaria patients have thyroid dysfunction. This phenomenon be caused by various factors such as Individual Differences, severity of the disease, and treatment options. Therefore, in clinical practice, it is necessary to check the thyroid status of patients with urticaria in order to determine prevention and treatment methods.\u003c/p\u003e \u003cp\u003eAlthough our study provides valuable insights, some limitations need to be taken into account. Our study only involved individuals of European descent, which limits the applicability of the research results to other populations and reduces bias caused by racial and regional differences. Although our study supports causality through genetic evidence, other studies, including functional studies, are needed to further reveal potential mechanisms. Nevertheless, our study provides important leads for understanding the relationship between hypothyroidism and urticaria.\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eThe study provides genetic evidence to support the causal relationship between hypothyroidism and urticaria. The analysis results provide further molecular biological support for the possible immune regulation abnormalities between hypothyroidism and urticaria. These findings not only help to deepen the understanding of the pathogenesis of these two diseases, but also provide important leads for the future development of therapeutic strategies for these diseases. By revealing the core gene and protein interaction network, we better understand the association between hypothyroidism and urticaria and potential therapeutic targets, providing a useful information foundation for personalized medicine and precision treatment.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eData availability statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe original contributions presented in the study are included in the article/Supplementary Material. Further inquiries can be directed to the corresponding authors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe authors are grateful to all the participants of this work.\u0026nbsp;Appreciating for the tools provided by website:www.xiantao.love.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eQianqian Chen and Shifeng Qiu wrote the main manuscript text. Ting Yang is responsible for drawing and analysis.Min Xiao review and modify the manuscript.All authors reviewed the manuscript.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by the Sichuan Provincial Administration of Traditional Chinese Medicine (2023MS470)and Chengdu University of Traditional Chinese Medicine \u0026ldquo;Apricot Grove Scholars\u0026rdquo; Discipline Talent Research Enhancement Program(QJRC2021007).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eRadonjic-Hoesli S, Hofmeier KS, Micaletto S, Schmid-Grendelmeier P, Bircher A, Simon D. Urticaria and Angioedema: an Update on Classification and Pathogenesis. Clin Rev Allergy Immunol. 2018;54(1):88-101.\u003c/li\u003e\n \u003cli\u003eGimenez-Arnau AM, Maibach H. Contact Urticaria. 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Association of Cytotoxic T-Lymphocyte Antigen 4 (CTLA4) and Thyroglobulin (TG) Genetic Variants with Autoimmune Hypothyroidism. \u003cem\u003ePLoS One\u003c/em\u003e. 2016;11(3):e0149441. Published 2016 Mar 10. 1\u003c/li\u003e\n \u003cli\u003eUeda H, Howson JM, Esposito L, et al. Association of the T-cell regulatory gene CTLA4 with susceptibility to autoimmune disease. \u003cem\u003eNature\u003c/em\u003e. 2003;423(6939):506-511.\u003c/li\u003e\n \u003cli\u003eVaidya B, Pearce S. The emerging role of the CTLA-4 gene in autoimmune endocrinopathies. \u003cem\u003eEur J Endocrinol\u003c/em\u003e. 2004;150(5):619-626.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"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":"Mendelian randomization study, SNPs, hypothyroidism, urticaria","lastPublishedDoi":"10.21203/rs.3.rs-4252621/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4252621/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eObservational studies connect hypothyroidism with urticaria, but causality is unclear. This study explores genetic variations in hypothyroidism and their potential influence on urticaria risk.We collected statistical data from open genome-wide association studies in the Integrative Epidemiology Unit (IEU OpenGwas) project, analyzed genetic data from patients with hypothyroidism and urticaria to study their causal relationship using various methods such as the inverse variance weighting (IVW) method, MR-Egger method, weighted median estimator (WME) method, and single-nucleotide polymorphism (SNP) analysis through gene ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, and protein-protein interaction (PPI) network evaluation.Our research identified SNPs that significantly elevate urticaria risk in hypothyroidism. GO analysis highlighted crucial genes regulating immune cell function, adhesion, growth, and cytokine production. KEGG analysis demonstrated enrichment of these genes in immune response and autoimmune pathways. PPI network analysis pinpointed key genes involved in these processes. The study provides strong genetic evidence that there is a causal relationship between hypothyroidism and an increased incidence of urticaria. This finding is expected to help develop more effective treatment strategies for urticaria patients.\u003c/p\u003e","manuscriptTitle":"Using Mendelian randomization and bioinformatics methods to explore the causal effect of hypothyroidism on urticaria","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-05-08 20:32:18","doi":"10.21203/rs.3.rs-4252621/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":"4c6cb9c1-7863-4be6-9ceb-9e612e140f99","owner":[],"postedDate":"May 8th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":31547195,"name":"Biological sciences/Computational biology and bioinformatics"},{"id":31547196,"name":"Health sciences/Medical research/Genetics research"},{"id":31547197,"name":"Health sciences/Medical research/Epidemiology"}],"tags":[],"updatedAt":"2024-07-01T07:40:16+00:00","versionOfRecord":[],"versionCreatedAt":"2024-05-08 20:32:18","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4252621","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4252621","identity":"rs-4252621","version":["v1"]},"buildId":"uwybb5PU2iWlRI8EIam5Y","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}