FLI1 and GATA1 govern TLN1 transcription: new insights into FLI1-related platelet disorders.

IF 8.2 1区 医学 Q1 HEMATOLOGY
Haematologica Pub Date : 2025-07-01 Epub Date: 2025-01-02 DOI:10.3324/haematol.2024.286372
Elisa Gabinaud, Laurent Hannouche, Mathilde Veneziano-Broccia, Johannes Van Agthoven, Justine Suffit, Julien Maurizio, Delphine Potier, Dominique Payet-Bornet, Delphine Bastelica, Elisa Andersen, Manal Ibrahim-Kosta, Timothée Bigot, Céline Falaise, Anne Vincenot, Pierre-Emmanuel Morange, Paul Saultier, Marie-Christine Alessi, Marjorie Poggi, Hemostasis Unit Of Lille
{"title":"FLI1 and GATA1 govern <i>TLN1</i> transcription: new insights into FLI1-related platelet disorders.","authors":"Elisa Gabinaud, Laurent Hannouche, Mathilde Veneziano-Broccia, Johannes Van Agthoven, Justine Suffit, Julien Maurizio, Delphine Potier, Dominique Payet-Bornet, Delphine Bastelica, Elisa Andersen, Manal Ibrahim-Kosta, Timothée Bigot, Céline Falaise, Anne Vincenot, Pierre-Emmanuel Morange, Paul Saultier, Marie-Christine Alessi, Marjorie Poggi, Hemostasis Unit Of Lille","doi":"10.3324/haematol.2024.286372","DOIUrl":null,"url":null,"abstract":"<p><p>Germline variants of FLI1, essential for megakaryopoiesis, are linked to bleeding disorders, platelet aggregation defects and mild thrombocytopenia. However, the mechanisms behind these abnormalities remain unclear. This study aims to elucidate the impact of FLI1 variants on human megakaryocytes and platelets. We focused on four FLI1 variants, two of which are novel: p.G307R and p.R340C. We assessed the impact of FLI1 variants on megakaryopoiesis using single-cell RNA sequencing, and defects were confirmed in patients' platelets and cell lines. Results showed variants p.R337Q, p.K345E and p.R340C exhibited faulty nuclear localization and defective transcriptional activity in vitro and variants p.K345E and p.G307R affected protein stability. A total of 626 genes were differentially expressed in patient megakaryocytes, including genes associated with the platelet activation pathway. TLN1 was among the most down-regulated genes, with an 88% reduction in talin-1 protein levels in FLI1 patients' platelets. Analysis of chromatin immunoprecipitation sequencing data revealed FLI1-binding regions in the TLN1 gene. Luciferase reporter gene assays revealed the functional role of an intronic binding region in cooperation with GATA1. FLI1 variants were linked to reduced cooperative transcriptional activity. These findings reveal novel mechanisms underlying the pathogenicity of FLI1 variants. Defective cooperation between FLI1 variants and GATA1 may play a role in talin-1 deficiency in FLI1 patients' platelets, thus contributing to platelet dysfunction. Moreover, talin-1 could serve as a biomarker for classifying the pathogenicity of FLI1 variants.</p>","PeriodicalId":12964,"journal":{"name":"Haematologica","volume":" ","pages":"1584-1595"},"PeriodicalIF":8.2000,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12208161/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Haematologica","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3324/haematol.2024.286372","RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/1/2 0:00:00","PubModel":"Epub","JCR":"Q1","JCRName":"HEMATOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

Germline variants of FLI1, essential for megakaryopoiesis, are linked to bleeding disorders, platelet aggregation defects and mild thrombocytopenia. However, the mechanisms behind these abnormalities remain unclear. This study aims to elucidate the impact of FLI1 variants on human megakaryocytes and platelets. We focused on four FLI1 variants, two of which are novel: p.G307R and p.R340C. We assessed the impact of FLI1 variants on megakaryopoiesis using single-cell RNA sequencing, and defects were confirmed in patients' platelets and cell lines. Results showed variants p.R337Q, p.K345E and p.R340C exhibited faulty nuclear localization and defective transcriptional activity in vitro and variants p.K345E and p.G307R affected protein stability. A total of 626 genes were differentially expressed in patient megakaryocytes, including genes associated with the platelet activation pathway. TLN1 was among the most down-regulated genes, with an 88% reduction in talin-1 protein levels in FLI1 patients' platelets. Analysis of chromatin immunoprecipitation sequencing data revealed FLI1-binding regions in the TLN1 gene. Luciferase reporter gene assays revealed the functional role of an intronic binding region in cooperation with GATA1. FLI1 variants were linked to reduced cooperative transcriptional activity. These findings reveal novel mechanisms underlying the pathogenicity of FLI1 variants. Defective cooperation between FLI1 variants and GATA1 may play a role in talin-1 deficiency in FLI1 patients' platelets, thus contributing to platelet dysfunction. Moreover, talin-1 could serve as a biomarker for classifying the pathogenicity of FLI1 variants.

FLI1和GATA1调控TLN1转录:对FLI1相关血小板疾病的新见解
对巨核生成至关重要的FLI1的种系变异与出血性疾病、血小板聚集缺陷和轻度血小板减少症有关。然而,这些异常背后的机制仍不清楚。本研究旨在阐明FLI1变异对人类巨核细胞和血小板的影响。我们重点研究了四个FLI1变体,其中两个是新的(p.G307R和p.R340C)。我们使用单细胞RNA测序评估了FLI1变异对巨核生成的影响,并在患者血小板和细胞系中证实了缺陷。结果表明,变异p.R337Q、p.K345E和p.R340C在体外表现出核定位缺陷和转录活性缺陷,变异p.K345E和p.G307R影响蛋白稳定性。共有626个基因在患者巨核细胞中差异表达,包括与血小板激活途径相关的基因。TLN1是最下调的基因之一,FLI1患者血小板中talin-1蛋白水平降低88%。染色质免疫沉淀测序数据分析显示在TLN1基因中存在fli1结合区。荧光素酶报告基因分析揭示了与GATA1合作的内含子结合区域的功能作用。FLI1变异与协同转录活性降低有关。这些发现揭示了FLI1变异致病性的新机制。FLI1变异体与GATA1之间的合作缺陷可能在FLI1患者血小板talin-1缺乏中起作用,从而导致血小板功能障碍。此外,talin-1可以作为分类FLI1变异致病性的生物标志物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Haematologica
Haematologica 医学-血液学
CiteScore
14.10
自引率
2.00%
发文量
349
审稿时长
3-6 weeks
期刊介绍: Haematologica is a journal that publishes articles within the broad field of hematology. It reports on novel findings in basic, clinical, and translational research. Scope: The scope of the journal includes reporting novel research results that: Have a significant impact on understanding normal hematology or the development of hematological diseases. Are likely to bring important changes to the diagnosis or treatment of hematological diseases.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信