Targeting splicing in myelodysplasia through GSK-3

通过 GSK-3 靶向骨髓增生异常中的剪接

• 批准号: 10677505
• 负责人: PETER S KLEIN
• 金额: $ 10万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-16 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10677505
• 关键词: Address; Alternative Splicing; Apoptosis; CASP8 gene; CASP9 gene; Cell Death; Cell Line; Cells; Clinical; Data; Disease; Drug usage; Dysmyelopoietic Syndromes; Flow Cytometry; Future; Glycogen Synthase Kinase 3; Goals; Hematopoietic; Human; K-562; Measures; Messenger RNA; Mutation; Neoplasms; Pathogenicity; Patients; Protein Family; RNA Splicing; Recurrence; Role; SRSF2 gene; Signal Transduction; Signaling Protein; Somatic Mutation; System; Testing; Therapeutic; Western Blotting; experimental study; gain of function mutation; in vivo; inhibitor; mutant; novel; novel strategies; novel therapeutic intervention; side effect; small molecule; therapeutic target

(PLEASE KEEP IN WORD, DO NOT PDF) Recurrent mutations in the splicing factors SRSF2 and SF3B1 are common in myelodysplasia (MDS). These gain of function mutations are mutually exclusive and invariably heterozygous, indicating that cells with splicing factor mutations require some wild-type splicing activity to survive. Further perturbation of the splicing apparatus is lethal, creating a therapeutic vulnerability for splicing factor mutant neoplasms. Thus, small molecule splicing inhibitors are currently being explored to treat splicing factor mutant MDS, but they have had intolerable side effects and limited efficacy so far, indicating that new approaches to inhibit splicing are needed. We found that the signaling protein glycogen synthase kinase-3 (GSK-3) phosphorylates multiple splicing factors and regulates the alternative splicing of a broad range of mRNAs. Inhibition of GSK-3 disrupts splicing and promotes cell death selectively in hematopoietic cell lines with heterozygous mutations in SRSF2 or SF3B1. Our long-term goals are to define the role of GSK-3 as a novel, global regulator of splicing and to test whether GSK-3 inhibitors are selectively lethal in vivo for MDS cells with splicing factor mutations. Our preliminary data support this hypothesis in hematopoietic cell lines, but it is important to establish these findings in primary human cells from patients with MDS and CMML. We will express SRSF2wt or the pathogenic SRSF2P95H mutation in primary cells from patients with MDS to ask whether expression of the pathogenic mutation confers sensitivity to GSK-3 inhibition in an otherwise isogenic context. These experiments will provide critical preliminary data and lay the groundwork for future experiments to investigate the role of GSK-3 as a global regulator of splicing and as a potential therapeutic target in splicing factor mutant MDS.

（请以 WORD 形式保存，请勿以 PDF 形式保存） 剪接因子 SRSF2 和 SF3B1 的反复突变在骨髓增生异常中很常见 （MDS）。这些功能获得突变是相互排斥的并且总是杂合的， 表明具有剪接因子突变的细胞需要一些野生型剪接活性才能 存活。拼接装置的进一步扰动是致命的，造成治疗脆弱性 用于剪接因子突变肿瘤。因此，目前正在研究小分子剪接抑制剂 探索治疗剪接因子突变体MDS，但它们具有难以忍受的副作用 迄今为止效果有限，表明需要新的方法来抑制剪接。我们发现 信号蛋白糖原合酶激酶 3 (GSK-3) 磷酸化多重剪接 因素并调节多种 mRNA 的选择性剪接。 GSK-3 的抑制 在杂合造血细胞系中选择性地破坏剪接并促进细胞死亡 SRSF2 或 SF3B1 突变。我们的长期目标是将 GSK-3 的作用定义为 新颖的全局剪接调节剂并测试 GSK-3 抑制剂是否具有选择性 对于具有剪接因子突变的 MDS 细胞具有体内致死性。我们的初步数据支持这一点 造血细胞系中的假设，但重要的是在原代细胞中建立这些发现 来自 MDS 和 CMML 患者的人类细胞。我们将表达SRSF2wt或致病性 MDS 患者原代细胞中 SRSF2P95H 突变，询问是否表达 致病突变赋予在其他同基因背景下对 GSK-3 抑制的敏感性。 这些实验将提供关键的初步数据，并为未来奠定基础 实验研究 GSK-3 作为剪接全局调节剂的作用以及作为潜在的 剪接因子突变MDS的治疗靶点。