Role of DDX41 in HSC development and MDS/AML

DDX41 在 HSC 发育和 MDS/AML 中的作用

• 批准号: 10373097
• 负责人: SUSAN R ROSS
• 金额: $ 19.99万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 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/10373097
• 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

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，其有助于在细胞内的表达。 干扰素对逆转录病毒的反应。DDX41属于RNA解旋酶家族，具有不同的 DEAD/H box（Asp-Glu-Ala-Asp/His）结构域，其成员与 翻译、核糖体生物合成、核质转运、细胞器基因表达和 前mRNA剪接。DDX41也是最近发现的家族性肿瘤抑制基因。 和散发性骨髓增生异常综合征/急性髓性白血病（MDS/AML），以及其他 血液恶性肿瘤在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-蛋白质相互作用是未知的。我们最近培育了一种 DDX41的floxed等位基因，这将使我们首次研究DDX41在造血中的作用， 发展和转型。DDX41种系敲除在发育过程中在E10之前死亡，但 敲除的杂合子小鼠是完全存活的，并将用于确定是否 这些小鼠发展成骨髓增生异常。我们还生成了条件敲除， 造血干细胞（HSC）中的基因。HSC将被用于研究DDX41的作用。 造血谱系分化。将对敲除HSC进行RNA Seq分析。 以确定DDX41的缺失是否导致基因表达改变或剪接缺陷。 HSC将用野生型和突变体DDX41转导;特别地，我们将工程化 患者来源的突变以及DEAD和解旋酶结构域中的突变，以开始确定 这种蛋白质是如何在发育和转化中发挥作用的我们打算用这些老鼠 研究DDX41在HSC发育中的作用，以及确定其机制， 它调节骨髓细胞的转化。这些研究很可能会加深我们对 以及生成用于测试潜在疗法的模型系统。