Role of DDX41 in HSC development and MDS/AML

DDX41 在 HSC 发育和 MDS/AML 中的作用

基本信息

批准号：
10216402
负责人：
SUSAN R ROSS
金额：
$ 23.99万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-16 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10216402
关键词：
AML/MDS Acute Myelocytic Leukemia Adult Affect Alleles Biogenesis Biological Models Cell Proliferation DNA Defect Development Disease Dysmyelopoietic Syndromes Embryo Engineering Enterobacteria phage P1 Cre recombinase Equilibrium Erythro Familial disease Family Gene Expression Genes Growth Hematologic Neoplasms Hematopoietic Hematopoietic stem cells Interferons Knock-out Knockout Mice Late-Onset Disorder LoxP-flanked allele Modeling Mus Mutate Mutation Myelogenous Myeloid Cells Natural Immunity Nuclear Nucleic Acids Nude Mice Organelles Patients Play Protein Isoforms Proteins RNA RNA Helicase RNA Splicing Retroviridae Ribosomes Role Spliced Genes Stem Cell Development Survival Rate Testing Therapeutic Time Tissues Translations Tumor Suppressor Genes Tumor-Suppressor Gene Inactivation Work aspartylglutamate conditional knockout glutamylalanine helicase hematopoietic stem cell differentiation knock-down mRNA Precursor member mutant pathogen protein function protein protein interaction response retroviral transduction sensor transcriptome sequencing tumor growth

项目摘要

Dead-box helicase 41 (DDX41) is a known sensor of nucleic acids that contributes to the interferon response to retroviruses. DDX41 belongs to a family of RNA helicases, with distinct DEAD/H box (Asp-Glu-Ala-Asp/His) domains, whose members have been implicated in translation, ribosome biogenesis, nuclear-cytoplasmic transport, organelle gene expression and pre-mRNA splicing. DDX41 was also recently identified as a tumor suppressor gene in familial and sporadic myelodysplastic syndrome/acute myeloid leukemia (MDS/AML), as well as other hematological malignancies. In MDS/AML, DDX41 is thought to interact with spliceosomal components and alter splicing, resulting in the inactivation of tumor suppressor genes or alterations in the balance of gene isoforms, although whether this occurs through protein-RNA, protein-DNA or protein-protein interactions is not known. We recently developed mice with a floxed allele of DDX41, which will allow us for the first time to study DDX41's role in hematopoietic development and transformation. DDX41 germline knockouts die prior to E10 in development, but mice heterozygous for the knockout are completely viable and will be used to determine whether these mice develop myelodysplasias. We have also generated conditional knockouts that delete the gene in hematopoietic stem cells (HSC). The HSCs will be used to study the role of DDX41 in hematopoietic lineage differentiation. RNA Seq analysis on the knockout HSCs will be performed, to determine if loss of DDX41 results in altered gene expression or splicing defects. The HSCs will be transduced with wild type and mutant DDX41; in particular, we will engineer patient-derived mutations as well those in the DEAD and helicase domains, to begin to determine how this protein functions in development and transformation. We propose to use these mice to study the role of DDX41 in HSC development, as well as to determine the mechanism by which it regulates transformation of myeloid cells. These studies are likely to further our understanding of AML as well as to generate a model system for testing potential therapeutics.

死盒解旋酶 41 (DDX41) 是一种已知的核酸传感器，有助于对逆转录病毒的干扰素反应。 DDX41 属于 RNA 解旋酶家族，具有独特的功能 DEAD/H 盒 (Asp-Glu-Ala-Asp/His) 结构域，其成员涉及翻译、核糖体生物合成、核质运输、细胞器基因表达和前mRNA剪接。 DDX41最近也被鉴定为家族性肿瘤抑制基因和散发性骨髓增生异常综合征/急性髓系白血病（MDS/AML），以及其他血液系统恶性肿瘤。在 MDS/AML 中，DDX41 被认为与剪接体相互作用成分并改变剪接，导致肿瘤抑制基因失活或基因亚型平衡的改变，尽管这是否通过蛋白质-RNA 发生，蛋白质-DNA 或蛋白质-蛋白质相互作用尚不清楚。我们最近开发了具有 DDX41的floxed等位基因，这将使我们能够首次研究DDX41在造血中的作用发展与转型。 DDX41 种系敲除在 E10 开发之前就已死亡，但是基因敲除杂合小鼠完全存活，将用于确定是否这些小鼠出现骨髓增生异常。我们还生成了删除的条件淘汰赛造血干细胞（HSC）中的基因。 HSC 将用于研究 DDX41 的作用在造血谱系分化中。对敲除 HSC 进行 RNA 测序分析进行，以确定 DDX41 的丢失是否会导致基因表达改变或剪接缺陷。 HSC 将用野生型和突变型 DDX41 转导；特别是，我们将设计患者衍生的突变以及 DEAD 和解旋酶结构域中的突变，开始确定这种蛋白质如何在发育和转化中发挥作用。我们建议使用这些小鼠研究 DDX41 在 HSC 发育中的作用，并确定其机制它调节骨髓细胞的转化。这些研究可能会加深我们的理解 AML 以及生成用于测试潜在疗法的模型系统。