The role of Rpl5 haploinsufficiency in hematopoietic stem/progenitor cell function in Diamond Blackfan anemia

Rpl5 单倍体不足在 Diamond Blackfan 贫血造血干/祖细胞功能中的作用

• 批准号: 10368461
• 负责人: Sharon A Singh
• 金额: $ 17.11万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10368461
• 关键词: Affect; Age; Age-Months; Anemia; Animal Model; Aplastic Anemia; Basic Science; Biological Assay; Birth; Blood Transfusion; Bone Marrow; CFU-E; Cell Count; Cell Culture Techniques; Cell Cycle; Cell Cycle Regulation; Cells; Cellular Stress; Cellular biology; Characteristics; Child; Childhood; Chronic; Clinical; Collaborations; Complex; Data; Defect; Development; Diamond-Blackfan anemia; Disease; Disease remission; Dysmyelopoietic Syndromes; Embryo; Environment; Erythrocytes; Erythropoiesis; Etiology; Evaluation; Failure; Functional disorder; Funding; GATA1 gene; Gene Proteins; Genetic; Genetic Transcription; Germ-Line Mutation; Goals; Health; Hematopoiesis; Hematopoietic stem cells; Heme; Hemoglobin; Human; Hydrops Fetalis; In Vitro; Inbred Mouse; Individual; Inherited; Innate Immune System; Junior Physician; Laboratory Research; Lead; Longevity; Macrocytic Anemia; Mediator of activation protein; Mentors; Mentorship; Michigan; Modeling; Mus; Mutant Strains Mice; Mutation; Neonatal; Newborn Infant; Normal Cell; Oncologist; Pathogenesis; Pathway interactions; Patients; Pediatric Hematologist; Pediatric Hematology; Penetrance; Persons; Phenotype; Physicians; Play; Pronormoblasts; Protein Biosynthesis; Protein Deficiency; Proteins; Reactive Oxygen Species; Research; Resolution; Resources; Ribosomal Proteins; Ribosomes; Role; Scientist; Severities; Spontaneous Remission; Stress; Testing; Toxic effect; Training; Transfusion; Translations; Transplantation; Universities; Work; base; bone marrow failure syndrome; burden of illness; career; critical developmental period; critical period; design; erythroid differentiation; exhaustion; experimental study; hematopoietic cell transplantation; immune activation; in utero; in vivo; mortality; mouse model; mutant; novel; novel therapeutics; patient registry; prevent; stem; stem cell biology; stem cell function; stem cells

PROJECT SUMMARY/ABSTRACT Anemia is an extraordinarily common condition that affects approximately 1 in 3 people around the globe. Erythropoiesis is exquisitely sensitive to both endogenous and exogenous stress, which contributes to this massive global disease burden. Genetic defects in ribosomes that lead to severe anemia are associated with rare inherited bone marrow failure syndromes as well as more commonly acquired myelodysplastic syndrome. The pathway from ribosomal protein haploinsufficiency to defective erythropoiesis in Diamond Blackfan anemia (DBA) is complex and likely involves multiple factors (e.g. diminished GATA1 translation, heme toxicity and/or nucleolar stress). Patients with DBA can undergo spontaneous remission such that chronic transfusions are not required. This phenomenon is poorly understood using current patient registries and animal models. Understanding normal pathways that lead to the resolution or improvement of anemia is critical in order to advance therapies for DBA and other causes of anemia. The applicant is a pediatric hematologist/oncologist who actively treats patients with all types of genetic and acquired anemias and her long term goal is to become a physician-scientist with expertise in hematopoiesis. To achieve this goal, the candidate has proposed the following training plan: 1) To develop expertise in isolation and analysis of hematopoietic stem/progenitor cells; 2) To further develop expertise employing mouse models and human hematopoietic stem cell culture to study disease-specific mechanisms; 3) To develop an independently funded basic research laboratory; 4) To develop into a successful physician-scientist focused on mentorship and collaboration. The candidate has assembled a mentorship team with expertise in hematopoiesis and mouse models (Drs. Engel and Li). The environment at the University of Michigan is highly conducive to the training of successful junior physician-scientists with exceptional resources, educational and collaborative opportunities. The scientific proposal centers on the hypothesis that DBA remission, even in these inbred mice, is based on a threshold effect on hematopoietic stem/progenitor cells that responds to both exogenous and endogenous modifiers. The rationale for this hypothesis is based on observations in our unique DBA mouse model where all neonatal Rpl5 (ribosomal protein gene) mutant mice are born with anemia, which contributes to early mortality in some mice while others survive and maintain normal hemoglobin characteristics throughout their lifespan. Aim 1 will determine if there is an intrinsic defect in stem/progenitor cells and/or the bone marrow microenvironment in mutant mice by examining transplantation of hematopoietic cells from different developmental periods. Aim 2 will test whether cellular stress alters the niche and/or hematopoietic stem/progenitor cell function. Aim 3 will examine whether cell cycle dysregulation is responsible for defective hematopoiesis during developmentally vulnerable periods or during cellular stress. Completion of training under the guidance of senior mentors will enable the applicant to develop into an independent physician-scientist with greater expertise in hematopoiesis.

项目摘要/摘要 贫血是一种非常普遍的疾病，影响了全球三分之一的人。 红细胞生成对内源性和外源性压力非常敏感，这有助于此 巨大的全球疾病负担。核糖体中导致严重贫血的遗传缺陷与 罕见的遗传性骨髓衰竭综合征以及更常见的骨髓增生综合征。 从核糖体蛋白单倍弥弥漫到有缺陷的钻石黑芬贫血的促红细胞生成的途径 （DBA）很复杂，可能涉及多种因素（例如，GATA1翻译减少，血红素毒性和/或 核仁应力）。 DBA患者可以自发缓解，因此慢性输血是 不需要。使用当前的患者登记和动物模型，这种现象对这种现象的理解很少。 了解导致贫血分辨率或改善的正常途径对于为了 DBA和其他贫血原因的预先疗法。申请人是儿科血液学家/肿瘤学家 他们积极治疗各种遗传和获得性贫血的患者，她的长期目标是成为 具有造血专业知识的医师科学家。为了实现这一目标，候选人提出了 遵循培训计划：1）发展造血干/祖细胞的孤立和分析方面的专业知识； 2）使用小鼠模型和人类造血干细胞培养进一步发展专业知识来研究 疾病特异性机制； 3）开发一个独立的基础研究实验室； 4）开发 成为一个成功的医师科学家，专注于指导和协作。候选人组装了 指导团队具有造血和鼠标模型（恩格尔博士和李）的专业知识。环境在 密歇根大学非常有利于培训成功的初级医师科学家 卓越的资源，教育和协作机会。科学提议集中在 假设DBA缓解即使在这些近交小鼠中，也基于对造血的阈值作用 对外源和内源性修饰剂反应的茎/祖细胞。理由 假设基于我们独特的DBA小鼠模型中的观察结果，其中所有新生儿RPL5（核糖体） 蛋白质基因）突变小鼠出生于贫血，这在某些小鼠中导致早期死亡，而另一些小鼠则有助于 在整个生命周期中生存并保持正常的血红蛋白特征。 AIM 1将确定是否在那里 是茎/祖细胞和/或突变小鼠中的骨髓微环境的固有缺陷 检查来自不同发育时期的造血细胞的移植。 AIM 2将测试是否 细胞应力改变利基和/或造血茎/祖细胞功能。 AIM 3将检查是否 细胞周期失调负责发育脆弱时期的造血缺陷 或在细胞应激期间。在高级导师的指导下完成培训将使申请人能够 发展成独立的医师科学家，在造血方面具有更高的专业知识。