Control of ribosomal function in cells of the hematopoietic system

造血系统细胞核糖体功能的控制

• 批准号: 10509989
• 负责人: Andrew Levine
• 金额: $ 11.11万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10509989
• 关键词: Affect; Area; Award; Biochemical; Biological Assay; Biology; Bone Marrow; Bone marrow failure; Cell physiology; Cells; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Craniofacial Abnormalities; Data Science; Defect; Development; Development Plans; Diagnostic; Disease; Doctor of Philosophy; Elements; Environment; FMRP; Foundations; Functional disorder; Future; Genes; Genetic Translation; Goals; Grant; Hematopoiesis; Hematopoietic; Hematopoietic System; Housekeeping; Human; Immune; Immunologist; Immunology; In Vitro; Infection; Inflammation; Inflammatory; Inflammatory Response; Innate Immune System; Lipopolysaccharides; Macrophage Activation; Mediating; Mentors; Messenger RNA; Methods; Modification; Molecular; Mouse Protein; Mutagenesis; Mutation; Nature; Nutrient; Pathologist; Phase; Physicians; Poly I-C; Process; Production; Protein Biosynthesis; Proteins; Proteomics; Protocols documentation; RNA; RNA Binding; RNA Sequences; Regulation; Regulatory Element; Research; Research Ethics; Research Personnel; Ribosomal Interaction; Ribosomal Proteins; Ribosomal RNA; Ribosomes; Role; Scientist; Signal Pathway; Signal Transduction; Specificity; Stimulus; Structure; Testing; Tissues; Training; Transcript; Translating; Translation Initiation; Translational Regulation; Translations; United States National Institutes of Health; Work; career development; cell type; cytokine; experimental study; extracellular; immune activation; immune function; immunoregulation; interest; macrophage; monocyte; neutrophil; novel; programs; proteostasis; response; ribosome profiling

PROJECT SUMMARY/ABSTRACT Translation is a dynamic process with a high degree of specificity conferred by multiple factors including mRNA transcript sequence, composition of the translation machinery, and cellular milieu. A striking demonstration of this specificity is the tissue-specific (e.g. craniofacial anomalies, bone marrow failure) rather than global nature of the developmental defects seen in human ribosomopathies caused by congenital mutations in ribosomal factors. Moreover, ribosomopathy-driven bone marrow failure is characterized by selective, rather than global, defects in mRNA translation. Thus, even the composition of the ribosome can affect mRNA translational efficiency in a tissue-specific manner. These observations point to a particularly nuanced regulation of translation in the hematopoietic system; elucidating the mechanisms of this regulation is therefore essential for understanding hematopoietic cell development, function, and dysfunction in disease states. As such, my long-term research goal is to elucidate mechanisms of translational control in hematolymphoid cells during normal immune function and in disease states, focusing on ribosome-associated proteins (RAPs)—a poorly studied class of proteins which physically interact with the core ribosomal proteins to tune the translational efficiency of specific mRNAs. My central hypothesis is that, in immune cells poised for rapid responses to infection, inflammatory stimuli may induce dynamic interactions of RAPs with ribosomes, where they may effect rapid changes in the translational efficiency of specific mRNAs with important immunoregulatory consequences. In previous work, I have defined the repertoire of ribosome-bound RAPs in in vitro bone marrow-derived macrophages. In Aim 1 of this proposal, I will use global, unbiased proteomics to temporally profile RAP-ribosome interactions in activated macrophages following TLR stimulation. In Aim 2, I will use CRISPR gene editing, ribosome profiling, and cytokine secretion assays to 1) identify mRNA targets that are specifically regulated by two RAPs I have already identified, Rnf213 and Helz2, as well as other RAPs identified in Aim 1, and 2) determine the function of these RAPs in the macrophage inflammatory response. Having completed MD-PhD training and clinical training as a clinical pathologist and hematopathologist, I am applying for the K38 Award to support my goal of becoming an independent physician investigator. UCSF’s exceptional training environment, especially in the areas of immunology and proteostasis, make it an ideal place for me to pursue my efforts. Critical elements of my career development plan include leveraging the expertise of my primary mentor Dr. Davide Ruggero, a pioneer in the field of ribosome biology, and my co-mentors Dr. Jason Cyster, an expert immunologist, and Dr. Karthik Ganapathi, an expert diagnostic hematopathologist; guidance by physician-scientist advisors; coursework in data science and research ethics; and other professional development activities. All together, I wholeheartedly believe this career development plan will provide a strong foundation on which to ultimately build an independent research program.

项目摘要/摘要 翻译是一个动态过程，具有高度特异性，包括mrna 转录序列，翻译机械的组成和细胞环境。 该特异性是组织规范（例如颅面异常，骨髓衰竭），而不是全球性质 核糖体先天性突变引起的人类核糖体病中发现的发育缺陷 此外，核糖瘤驱动的骨髓衰竭的特征是选择性而不是全球 因此，mRNA翻译中的缺陷。 以组织特异性的效率。 在造血系统中的翻译； 了解疾病状态下的造血细胞发育，功能和功能障碍。 因此，我的长期研究目标是阐明翻译控制机制 正常免疫功能和疾病状态下的血压细胞，重点是核糖体相关 蛋白质（RAP） - 一种较差的蛋白质类别，物质与核心核糖体蛋白相互作用 调整了特定mRNA的翻译功效。 对感染的快速反应，炎症刺激可能引起Rapos与核糖的动态相互作用， 其中效果快速改变了特定mRNA的转化效率 免疫调节奉献。 体外骨髓来源的巨噬细胞。 在TLR刺激后，临时的巨噬细胞中的Rap-ribosis相互作用，I AIM 2。 将使用CRISPR基因编辑，核糖体分析和细胞因子分泌测定1）确定mRNA靶标 由我确定Allyady的两个说唱，RNF213和HELZ2以及其他说唱 在AIM 1和2）确定说唱在巨噬细胞炎症反应中的功能。 完成了MD-PHD培训并担任临床病理学家和血管病理学家的粘贴培训后 我正在申请K38奖，以支持我的独立调查员的目标 杰出的培训环境，尤其是在免疫学和蛋白质的领域，使其成为理想的地方 我追求我的职业发展计划的关键要素包括利用专业知识 我的主要导师戴维德·鲁格罗（Davide Ruggero）博士是核糖体生物学领域的先驱，以及我的联合主管博士 专家免疫学家Jason Cyster和专家诊断血管病医生Karthik Ganapathi博士； 医师科学顾问的指导； 整个开发活动，我全心全意地相信Thiser开发计划 最终建立独立研究计划的基础。