Cellular Interactions with Thrombospondin

细胞与血小板反应蛋白的相互作用

• 批准号: 10702998
• 负责人: david d roberts
• 金额: $ 146.71万
• 依托单位: DIVISION OF CLINICAL SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10702998
• 关键词: 7-methylguanosine; Acute; Alkylation; Angiogenesis Inhibition; Anthracycline; Autophagocytosis; Blood Platelets; Blood Pressure; Blood flow; Bone Marrow; CD47 gene; CD8-Positive T-Lymphocytes; Cell Surface Receptors; Cell Survival; Cells; Code; Complex; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; DNA Repair; Data; Eukaryotic Initiation Factor-4E; Extracellular Matrix; Family suidae; Genes; Glycoproteins; Growth; Hemostatic function; Ionizing radiation; Irradiated tumor; Ischemia; KDR gene; Knockout Mice; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Messenger RNA; Metabolism; MicroRNAs; Mus; Myocardium; NOS3 gene; Natural Killer Cells; Nitric Oxide; Non-Malignant; Nuclear; Nuclear Export; Null Lymphocytes; Play; Proteins; RNA; Radiation Injuries; Rattus; Receptor Cell; Receptor Signaling; Recovery; Regulation; Reperfusion Injury; Reporting; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Smooth Muscle Myocytes; Soluble Guanylate Cyclase; Stromal Cells; T-Lymphocyte; Therapeutic; Thrombospondin 1; Thrombospondins; Tissue Survival; Tissues; Transgenic Mice; Tumor Immunity; UBQLN1 gene; Untranslated RNA; Vascular Smooth Muscle; adaptive immune response; angiogenesis; antagonist; anti-tumor immune response; base; c-myc Genes; cancer cell; cancer immunotherapy; cancer stem cell; cell type; cytotoxic; exportin 1 protein; extracellular vesicles; gene discovery; immune checkpoint; inhibitor; ischemic injury; leptomycin B; macrophage; malignant breast neoplasm; mouse model; neoplastic cell; novel; radiation resistance; receptor; response; self-renewal; soft tissue; stem; stem cells; tissue culture; tissue stress; trafficking; transcription factor; transcriptomics; treatment response; tumor; tumor progression; tumor xenograft; vesicular release

CD47 is a marker of self and a signaling receptor for thrombospondin-1 that is also a component of extracellular vesicles (EVs) released by various cell types. Previous studies identified CD47-dependent functional effects of T cell EVs on target cells, mediated by delivery of their RNA contents, and enrichment of specific subsets of coding and noncoding RNAs in CD47+ EVs. Mass spectrometry was employed here to identify potential mechanisms by which CD47 regulates the trafficking of specific RNAs to EVs. Specific interactions of CD47 and its cytoplasmic adapter ubiquilin-1 with components of the exportin-1/Ran nuclear export complex were identified and confirmed by coimmunoprecipitation. Exportin-1 is known to regulate nuclear to cytoplasmic trafficking of 5'-7-methylguanosine (m7G)-modified microRNAs and mRNAs that interact with its cargo protein EIF4E. Interaction with CD47 was inhibited following alkylation of exportin-1 at Cys528 by its covalent inhibitor leptomycin B. Leptomycin B increased levels of m7G-modified RNAs, and their association with exportin-1 in EVs released from wild type but not CD47-deficient cells. In addition to perturbing nuclear to cytoplasmic transport, transcriptomic analyses of EVs released by wild type and CD47-deficient Jurkat T cells revealed a global CD47-dependent enrichment of m7G-modified microRNAs and mRNAs in EVs released by CD47-deficient cells. Correspondingly, decreasing CD47 expression in wild type cells or treatment with thrombospondin-1 enhanced levels of specific m7G-modified RNAs released in EVs, and re-expressing CD47 in CD47-deficient T cells decreased their levels. Therefore, CD47 signaling limits the trafficking of m7G-modified RNAs to EVs through physical interactions with the exportin-1/Ran transport complex.

CD47是自我的标记，也是血小板传播1的信号受体，它也是各种细胞类型释放的细胞外囊泡（EV）的组成部分。先前的研究确定了T细胞EVS对靶细胞的CD47依赖性功能效应，通过递送其RNA含量介导，以及在CD47+ EV中富集编码和非编码RNA的特定子集。这里采用了质谱法来确定CD47调节特定RNA向电动汽车的运输的潜在机制。通过共免疫沉淀确定并确认了CD47及其细胞质适配器泛素泛素-1与Exportin-1/RAN核出口复合物的成分的特异性相互作用。已知Exportin-1会调节与5'-7-甲基鸟苷（M7G）改性的microRNA和MRNA相互作用的核质量运输，这些核蛋白（M7G）与其货物蛋白EIF4E相互作用。在CYS528的Exportin-1烷基化在CYS528处的烷基化抑制剂瘦霉素B.钩霉素B的相互作用增加了M7G修饰的RNA水平，并且它们与野生型但不是CD47缺乏细胞的EVS中的EVS中的EVS中的相关性。除了扰动核对细胞质转运外，野生型和CD47缺乏的Jurkat T细胞对电动汽车的转录组分析表明，CD47缺乏细胞释放的M7G修饰的microRNA和MRNA的全球CD47依赖性富集M7G修饰的microRNA和mRNA。相应地，野生型细胞中的CD47表达降低或用血小板传播1的处理增强了EV中释放的特定M7G修饰的RNA水平，并在CD47缺陷型T细胞中重新表达CD47降低了其水平。 因此，CD47信号传导通过与Exportin-1/RAN运输复合物的物理相互作用将M7G修饰的RNA的运输限制为电动汽车。