Uncovering a role for METTL13 in malignant transformation of human hematopoietic stem cells and in the progression of pediatric leukemia

Uncovering a role for METTL13 in malignant transformation of human hematopoietic stem cells and in the progression of pediatric leukemia | 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 Uncovering a role for METTL13 in malignant transformation of human hematopoietic stem cells and in the progression of pediatric leukemia Frida Holm, Sabina Enlund, Chae-Eun Lim, Isabella Hoang, Sonali Joshi, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7652169/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract Post-transcriptional RNA modifications, such as N6-methyladenosine (m6A) methylation and adenosine to inosine (A-to-I) editing, are critical regulators of hematopoietic stem cell (HSC) self-renewal and differentiation, yet their precise contributions to malignant transformation are not fully elucidated. In this study, we uncovered the epitranscriptomic landscape caused by knockdown of genes from the methyltransferase (METTL)-family in hematopoietic stem and progenitor cells (HSPCs). We identified both converging and distinct roles of METTL3 and METTL14, known members of the m6A writer complex, as well as orphan gene METTL13. Notably, METTL13 was uniquely upregulated by adenosine deaminase acting on RNA 1 (ADAR1) overexpression, while other METTL genes were downregulated. Knockdown of METTL13 altered the expression of multiple genes involved in oncogenic development in HSPCs. Furthermore, METTL13 was associated with a high-risk profile in pediatric T-cell acute lymphoblastic leukemia (T-ALL), and functional studies confirmed that METTL13 is required for T-ALL cell proliferation and survival both in vitro and in vivo. Collectively, our results indicate a previously unrecognized, oncogenic role for METTL13 in pre-leukemic transformation and T-ALL pathogenesis. Biological sciences/Stem cells/Cancer stem cells Health sciences/Diseases/Haematological diseases/Haematological cancer/Leukaemia/Acute lymphocytic leukaemia Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Full Text Additional Declarations (Not answered) Supplementary Files Supplementalfigure1.png Supplemental figure 1 Supplementaltable5.xls Supplemental table 5 Supplementalfigure6.png Supplemental figure 6 Supplementalfigure7.png Supplemental figure 7 Supplementaltable3.xls Supplemental table 3 Supplementaltable1.xls Supplemental table 1 Supplementalfigure5.png Supplemental figure 5 Supplementalfigure3.png Supplemental figure 3 Supplementaltable4.xls Supplemental table 4 Supplementalfigure4.png Supplemental figure 4 Supplementalfigure2.png Supplemental figure 2 Supplementaltable2.xls Supplemental table 2 uncroppedwesternblots.png Uncropped western blots Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: revise 29 Oct, 2025 Review # 2 received at journal 26 Oct, 2025 Review # 1 received at journal 20 Oct, 2025 Reviewer # 2 agreed at journal 12 Oct, 2025 Reviewer # 1 agreed at journal 10 Oct, 2025 Reviewers invited by journal 09 Oct, 2025 Submission checks completed at journal 26 Sep, 2025 First submitted to journal 25 Sep, 2025 Unknown event 19 Sep, 2025 Editor assigned by journal 18 Sep, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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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-7652169","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":527192582,"identity":"4be7a8ba-bef7-4e6f-b341-f4f73cf149b7","order_by":0,"name":"Frida Holm","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAwElEQVRIiWNgGAWjYBAC9gbGBgkGhgMMfAzMhxkSiNHCcwCqhY2BLZlYLQwMUC08xsQ5jEf6cOONHwx35NjEznw2eMBQJ0dYC19is2UPwzNjNunczQkJDIcJW2XPw9gmwcNwOLENqOVAAsOBxAaCtgC1SP4Ba8l5DNRSV0+UFmmILTnMQIcxJxD2Cw9js7WMwWGgX9KMDRIMDhsSYQv7w5tvKg7L8UsnP5b8UVEnT9AWCDDAYIyCUTAKRsEooAgAABsTNLra0Bf1AAAAAElFTkSuQmCC","orcid":"","institution":"Karolinska Institutet","correspondingAuthor":true,"prefix":"","firstName":"Frida","middleName":"","lastName":"Holm","suffix":""},{"id":527192583,"identity":"ea41fca2-264a-4963-8bd9-38d432568122","order_by":1,"name":"Sabina Enlund","email":"","orcid":"","institution":"Karolinska Institutet","correspondingAuthor":false,"prefix":"","firstName":"Sabina","middleName":"","lastName":"Enlund","suffix":""},{"id":527192584,"identity":"3104f1a3-65bb-47fb-92e3-eb43c12b5923","order_by":2,"name":"Chae-Eun Lim","email":"","orcid":"","institution":"University 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1","display":"","copyAsset":false,"role":"figure","size":220147,"visible":true,"origin":"","legend":"\u003cp\u003eADAR1 overexpression suppressed members of the m6A complex. A) Heat map of the top 500 differentially expressed genes in ADAR1 overexpressed human CD34+ HSPCs (ADAR1 OE, n=3) compared to the backbone control (pCDH, n=3). Created in Qlucore Omics Explorer, significance was calculated using unpaired two-tailed t-test, p\u0026lt;0.05. B) Distribution of differentially expressed genes in ADAR1 overexpressed HSPCs compared to the pCDH control. Significance was calculated using unpaired two-tailed t-test, p\u0026lt;0.05. C) Volcano plot of dysregulated genes from the METTL-family following ADAR1 overexpression. Significance was calculated usin unpaired two-tailed t-test, results are displayed as L2FC (ADAR OE/pCDH) and negative log10 p-value. D) Expression of genes from the m6A writer complex (METTL3, METTL14 and WTAP) and from the METTL-family (METTL13) in ADAR1 overexpressed cells compared to compared to the pCDH control. Significance was calculated using unpaired two-tailed t-test, results are displayed as TPM, mean ± SEM. E) Significantly differentially expressed m6A erasers FTO and ALKBH5 and readers HNRNPC and YTHDF1 in ADAR1 overexpressed cells compared to compared to the pCDH control. Significance was calculated using unpaired two-tailed t-test, results are displayed as TPM, mean ± SEM.\u003c/p\u003e","description":"","filename":"Figure1ADAROE.png","url":"https://assets-eu.researchsquare.com/files/rs-7652169/v1/8825fd9870fbe0a973c5771e.png"},{"id":94207506,"identity":"bd95e473-63b0-4a30-ba5b-33bd0eb18ceb","added_by":"auto","created_at":"2025-10-23 14:57:56","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":420194,"visible":true,"origin":"","legend":"\u003cp\u003eOverlapping and distinct gene regulation by METTL3, METTL9, METTL13 and METTL14 in HSPCs. A) PCA plot of METTL3 (shMETTL3, n=4), METTL5 (shMETTL5, n=2), METTL9 (shMETTL9, n=3), METTL13 (shMETTL13, n=4) and METTL14 (shMETTL14, n=3) knockdown in human CD34+ HSPCs compared to the backbone control (shCTRL, n=5). Created in Qlucore Omics Explorer. B) Heatmap of shMETTL3, shMETTL5, shMETTL9, shMETTL13 and shMETTL14 in HSPCs compared to shCTRL. Created in Qlucore Omics Explorer, significance was calculated using multi-group ANOVA, q\u0026lt;0.1, SD\u0026lt;0.05. C) Venn diagram of differentially expressed genes in shMETTL3, shMETTL9, shMETTL13 and shMETTL14 in HSPCs compared to shCTRL. Significance was calculated using unpaired twotailed t-test, p\u0026lt;0.05, in each shMETTL compared to shCTRL. D) Dysregulated genes (TP53, MDM2, CDKN1A, CASP3, DFFB and c-MYC) in shMETTL3, shMETTL9, shMETTL13 and shMETTL14 compared to shCTRL. Significance was calculated using ordinary one-way ANOVA with multiple comparisons compared to shCTRL, as well as Dunnett correction, results are displayed as TPM, mean ± SEM. E) Differentially expressed genes from the ADAR family in shMETTL3, shMETTL13 and shMETTL14 compared to shCTRL. Significance was calculated using unpaired two-tailed t-test, results are displayed as TPM, mean ± SEM.\u003c/p\u003e","description":"","filename":"Figure2METTLKDinHSPCs.png","url":"https://assets-eu.researchsquare.com/files/rs-7652169/v1/2360b01855aad3b02fb07de6.png"},{"id":94207511,"identity":"d74e0de0-d699-4c36-8089-d968ad5d4a99","added_by":"auto","created_at":"2025-10-23 14:57:56","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":916020,"visible":true,"origin":"","legend":"\u003cp\u003eMETTL3, METTL13 and METTL14 converged in regulating immune signaling, with many distinct pathways altered uniquely by METTL13. A) Distribution of significantly enriched pathways in each shMETTL compared to shCTRL generated through GSEA. Created in GSEA and MSigDB, FDR q\u0026lt;0.1, including only proteincoding genes, using three different gene set libraries (Wiki Pathways, Reactome and KEGG). Groups are based on which pathways are uniquely altered by shMETTL3 (n=4), shMETTL9 (n=3), shMETTL13 (n=4) or shMETTL14 (n=3), as well pathways altered in several conditions; the m6A writer complex (METTL3 and METTL14) with and without METTL13, as well as all other combined conditions, compared to shCTRL (n=5) in human CD34+ HSPCs. B) Network plot of GSEA Wiki Pathways in shMETTL3, shMETTL13 and shMETTL14 compared to shCTRL (FDR q\u0026lt;0.1). C) Top 10 significant GSEA Reactome pathways in shMETTL3, shMETTL9, shMETTL13 and shMETTL14 compared to shCTRL (FDR q\u0026lt;0.1). Results are displayed as normalized enrichment score (NES). D) Dysregulated genes involved in inflammatory signaling (TNF, IL1A, CXCL8, IL6, IL11 and IFNB1) in shMETTL3, shMETTL9, shMETTL13 and shMETTL14 compared to shCTRL. Significance was calculated using ordinary one-way ANOVA with multiple comparisons compared to shCTRL, as well as Dunnett correction, results are displayed as TPM, mean ± SEM. E) GSEA of shMETTL3, shMETTL9, shMETTL13 and shMETTL14 compared to shCTRL using KEGG legacy pathways (FDR q\u0026lt;0.1) Results are displayed as NES and FDR q-value.\u003c/p\u003e","description":"","filename":"Figure3METTLKDGSEA.png","url":"https://assets-eu.researchsquare.com/files/rs-7652169/v1/b911236527918b7371c51339.png"},{"id":94207513,"identity":"de99e86d-6d05-49ef-a642-f9bd0456bd92","added_by":"auto","created_at":"2025-10-23 14:57:56","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":941654,"visible":true,"origin":"","legend":"\u003cp\u003eMETTL13 knockdown caused dysregulation of pathways involved in malignant transformation. A) Volcano plot of differentially expressed genes following METTL13 knockdown (shMETTL13, n=4) compared to the control (shCTRL, n=5) in human CD34+ HSPCs. Significance was calculated using unpaired two-tailed t-test, results are displayed as L2FC and negative log10 pvalue (p \u0026lt;0.05). B) ORA of the top enriched KEGG pathways in shMETTL13 compared to shCTRL. Created in R, with packages clusterprofiler and enrichPlot, statistics was set to: q.value \u0026lt;0.1, L2FC cutoff = 1, p-adjust method = BH. C) Network plot of the KEGG gene set pathways in cancer in shMETTL13 compared to shCTRL, nodes are colored by L2FC. D) ORA of disease ontologies focused on hematological malignances. Created in R, packages with clusterprofiler, DOSE and enrichplot, statistics were set to: q.value ≤ 0.1, L2FC cutoff = 1, p-adjust method = BH.\u003c/p\u003e","description":"","filename":"Figure4METTLKDincancer.png","url":"https://assets-eu.researchsquare.com/files/rs-7652169/v1/5c6b178a866cf3e349e73dc9.png"},{"id":94207515,"identity":"83c71c41-17de-4c47-a451-7c0c1018c2fe","added_by":"auto","created_at":"2025-10-23 14:57:56","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":418749,"visible":true,"origin":"","legend":"\u003cp\u003eUpregulation of METTL13 associated with a high-T-ALL A) Dysregulated genes (TAL1, LEF1, PHF6 and PTEN) following METTL3 (n=4), METTL9, (n=3) METTL13 (n=4) and METTL14 (n=3) knockdown in human CD34+ HSPCs compared to the control (shCTRL, n=5). Significance was calculated using ordinary one-way ANOVA with multiple comparisons compared to shCTRL, as well as Dunnett correction, results are displayed as TPM, mean ± SEM. B) Uniquely dysregulated genes (FBXW7 and FOXO3) in shMETTL13 (not affected by METTL3, METTL9 or METTL14 knockdown) compared to shCTRL. Significance was calculated using unpaired two-tailed t-test, results are displayed as TPM, mean ± SEM. C) Table of ALL patient characteristics and subgroups used for RNA-sequencing analysis (publicly available by the TARGET Initiative). Samples were divided into T-ALL (n=190) or B-ALL (n=78), diagnosis or relapse, as well as different high-risk factors (CNS infiltration, KMT2A-r and WBC). D) Expression levels of METTL3, METTL5, METTL9, METTL13 and METTL14 in T-ALL (n=162) and B-ALL (n=39) diagnosis samples (publicly available by the TARGET Initiative) compared to normal HSPCs (CD34+ CB, n=5) obtained through RNA-sequencing. Significance was calculated using multiple unpaired t-test, results are displayed as TPM, mean ± SEM. E) Expression levels of METTL3, METTL5, METTL9, METTL13 and METTL14 in T-ALL patient samples (publicly available by the TARGET Initiative) generated through RNAsequencing, displayed as TPM, mean ± SEM. T-ALL samples were grouped by risk stratification, into standard risk (CNS negative, non KMT2A-r and low WBC, n=70), and high risk (CNSinfiltrated, KMT2A-r or high WBC, n=87). Significance was calculated using multiple unpaired ttest. F) Expression levels of METTL3, METTL5, METTL9, METTL13 and METTL14 in T-ALL patient samples (publicly available by the TARGET Initiative) generated through RNAsequencing, displayed as TPM, mean ± SEM. T-ALL samples were grouped by disease stage, into diagnosis (standard risk, n=87) and relapse samples (BM relapse, n=5). Significance was calculated using multiple unpaired t-test. G) Survival probability in B-ALL patients samples (publicly available by the TARGET Initiative) by METTL13 expression levels (top and bottom 15%, n=6 in each group). Significance was calculated using Log-rank (Mantel-Cox) test. H) Correlation of METTL13 and TP53 as well as METTL13 and NOTCH1 in T-ALL patient samples, colored by the disease stage (diagnosis = blue, n=162 and relapse = red, n=18), results are displayed as TPM. Correlation was calculated using Pearson correlation coefficients with a two-tailed with 95% confidence interval. I) Correlation of METTL13 and TP53 in B-ALL patient samples, colored by disease stage (diagnosis = blue (n=39), relapse = red (n=39)), results are displayed as TPM. Correlation was calculated using Pearson correlation coefficients with a two-tailed with 95% confidence interval.\u003c/p\u003e","description":"","filename":"Figure5METTLsinALL.png","url":"https://assets-eu.researchsquare.com/files/rs-7652169/v1/9df88661df10eb45cb8a50a5.png"},{"id":94208222,"identity":"ab528a26-6493-473c-96b6-b1700d0dc3ee","added_by":"auto","created_at":"2025-10-23 15:05:57","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":371794,"visible":true,"origin":"","legend":"\u003cp\u003eLoss of METTL13 impaired T-ALL cell proliferation and survival. A) Western blot image showing the expression level of METTL13 and beta-actin in normal PBMCs and T-ALL cell lines (SUP-T1, Jurkat, MOLT4, CEM and CUTTL1). B) Validation of METTL13 knockdown (shMETTL13) in T-ALL cell lines Jurkat (n=3), MOLT4 (n=3) and SUP-T1 (n=3) compared to the control (shCTRL, n=3 for each cell line) through RNAsequencing. Significance was calculated using unpaired two-tailed t-test, results are displayed as TPM, mean ± SEM. C) Total number of viable cells following METTL13 knockdown in T-ALL cell lines MOLT4 (n=3), Jurkat (n=3) and SUP-T1 (n=3) from day 4 to day 14 post transduction compared to the control (n=3 for each cell line). Significance was calculated using two-way ANOVA. D) Cell viability (in percent) following METTL13 knockdown in T-ALL cell lines MOLT4 (n=3), Jurkat (n=3) and SUP-T1 (n=3) from day 4 to day 14 post transduction compared to the control (n=3 for each cell line). Significance was calculated using two-way ANOVA). E) Representative flow cytometry showing human EGFP+CD45+ leukemia engraftment in NSGSGM3 mice transplanted with SUP-T1 cells. F) Engraftments of human EGFP+CD45+ were quantified by flow cytometry in bone marrow (BM) and spleen (SP) of SUP-T1 (blue) and MOLT4 (orange) transplanted mice (n = 10-17 mice per condition).\u003c/p\u003e","description":"","filename":"Figure6METTL13KDinTALL.png","url":"https://assets-eu.researchsquare.com/files/rs-7652169/v1/9611434cff44122c662cda94.png"},{"id":94207520,"identity":"5c57b947-569f-41a7-81dd-3589ed4a8ba2","added_by":"auto","created_at":"2025-10-23 14:57:57","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":473034,"visible":true,"origin":"","legend":"\u003cp\u003eKnockdown of METTL13 promoted pathways inducing apoptosis and suppressing DNA synthesis in T-ALL cells. A) PCA plot of T-ALL cell lines as biological replicates following METTL13 knockdown (shMETTL13, n= 3) compared to the control (shCTRL, n=3). Significance was calculated using unpaired two-tailed t-test, p\u0026lt;0.05. B) Heat map of the top 400 differentially expressed genes looking at T-ALL cell lines as biological replicates in shMETTL13 (n=3) compared to the shCTRL (n=3). Significance was calculated using unpaired two-tailed t-test, p\u0026lt;0.05. C) Pie chart of the percentage of significantly expressed genes (p\u0026lt;0.05) in T-ALL cell lines as biological replicates (n=3) in shMETTL13 (n=3) compared to shCTRL (n=3). Significance was calculated using unpaired two-tailed t-test, p\u0026lt;0.05. D) Pie chart of the distribution of upregulated versus downregulated significantly expressed genes in T-ALL cell lines as biological replicates in shMETTL13 (n=3) compared to the shCTRL (n=3). Significance was calculated using unpaired two-tailed t-test, p\u0026lt;0.05. E) Volcano plot of significantly expressed genes (p\u0026lt;0.05) in T-ALL cell lines as biological replicates in shMETTL13 (n=3) compared to the shCTRL (n=3). Only protein-coding genes were included in this plot. Significance was calculated using unpaired two-tailed t-test, results are displayed as L2FC and negative log10 p-value (p \u0026lt;0.05). F) Dysregulated genes (MDM2, NRAS, ERBB3 and BTG1) in shMETTL13 (n=3) compared to shCTRL (n=3 Significance was calculated using unpaired two-tailed t-test, p\u0026lt;0.05, results are displayed as TPM, mean ± SEM. G) GSEA of some of the top significantly enriched Reactome pathways (Created in GSEA and MSigDB, nominal p-value \u0026lt;0.05) in shMETTL13 (n=3) compared to shCTRL (n=3) in T-ALL cells, using only protein-coding genes, displayed as activated (positive NES) or suppressed (negative NES).\u003c/p\u003e","description":"","filename":"Figure7METTL13KDinTALLRNAseq.png","url":"https://assets-eu.researchsquare.com/files/rs-7652169/v1/a60cfd8108ddbef5d8412bca.png"},{"id":94209609,"identity":"0137e52c-5c02-4eee-8432-9d3977a6ff41","added_by":"auto","created_at":"2025-10-23 15:22:05","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3662367,"visible":true,"origin":"","legend":"Article 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leukemia","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"cell-death-and-disease","isNatureJournal":false,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"cddis","sideBox":"Learn more about [Cell Death \u0026 Disease](http://www.nature.com/cddis/)","snPcode":"41419","submissionUrl":"https://mts-cddis.nature.com/cgi-bin/main.plex","title":"Cell Death \u0026 Disease","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-7652169/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7652169/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Post-transcriptional RNA modifications, such as N6-methyladenosine (m6A) methylation and adenosine to inosine (A-to-I) editing, are critical regulators of hematopoietic stem cell (HSC) self-renewal and differentiation, yet their precise contributions to malignant transformation are not fully elucidated. In this study, we uncovered the epitranscriptomic landscape caused by knockdown of genes from the methyltransferase (METTL)-family in hematopoietic stem and progenitor cells (HSPCs). We identified both converging and distinct roles of METTL3 and METTL14, known members of the m6A writer complex, as well as orphan gene METTL13. Notably, METTL13 was uniquely upregulated by adenosine deaminase acting on RNA 1 (ADAR1) overexpression, while other METTL genes were downregulated. Knockdown of METTL13 altered the expression of multiple genes involved in oncogenic development in HSPCs. Furthermore, METTL13 was associated with a high-risk profile in pediatric T-cell acute lymphoblastic leukemia (T-ALL), and functional studies confirmed that METTL13 is required for T-ALL cell proliferation and survival both in vitro and in vivo. Collectively, our results indicate a previously unrecognized, oncogenic role for METTL13 in pre-leukemic transformation and T-ALL pathogenesis.","manuscriptTitle":"Uncovering a role for METTL13 in malignant transformation of human hematopoietic stem cells and in the progression of pediatric leukemia","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-23 14:57:51","doi":"10.21203/rs.3.rs-7652169/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"revise","date":"2025-10-29T10:07:56+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"This content is not available.","date":"2025-10-26T17:09:25+00:00","index":2,"fulltext":"This content is not available."},{"type":"editorInvitedReview","content":"This content is not available.","date":"2025-10-20T12:49:55+00:00","index":1,"fulltext":"This content is not available."},{"type":"reviewerAgreed","content":"This content is not available.","date":"2025-10-12T04:59:02+00:00","index":2,"fulltext":"This content is not available."},{"type":"reviewerAgreed","content":"This content is not available.","date":"2025-10-10T05:37:41+00:00","index":1,"fulltext":"This content is not available."},{"type":"reviewersInvited","content":"","date":"2025-10-09T14:55:43+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-09-26T10:35:44+00:00","index":"","fulltext":""},{"type":"submitted","content":"Cell Death \u0026 Disease","date":"2025-09-25T19:56:51+00:00","index":"","fulltext":""},{"type":"checksFailed","content":"","date":"2025-09-19T12:02:44+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-18T18:07:09+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"cell-death-and-disease","isNatureJournal":false,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"cddis","sideBox":"Learn more about [Cell Death \u0026 Disease](http://www.nature.com/cddis/)","snPcode":"41419","submissionUrl":"https://mts-cddis.nature.com/cgi-bin/main.plex","title":"Cell Death \u0026 Disease","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b2a260d7-caee-4849-9e68-51124b262791","owner":[],"postedDate":"October 23rd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":56039468,"name":"Biological sciences/Stem cells/Cancer stem cells"},{"id":56039469,"name":"Health sciences/Diseases/Haematological diseases/Haematological cancer/Leukaemia/Acute lymphocytic leukaemia"}],"tags":[],"updatedAt":"2026-04-10T08:37:30+00:00","versionOfRecord":[],"versionCreatedAt":"2025-10-23 14:57:51","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7652169","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7652169","identity":"rs-7652169","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}