Establishing CUX1 as a determinant of Hematopoietic Stem cell heterogeneity

建立 CUX1 作为造血干细胞异质性的决定因素

• 批准号: 10707891
• 负责人: Tanner Clark Martinez
• 金额: $ 5.27万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2027-09-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10707891
• 关键词: Address; Binding; Bioinformatics; Biological; Biology; Blood Platelets; C-terminal; Catastrophic Illness; Cations; Cell Compartmentation; Cell Cycle; Cell Line; Cell surface; Cells; Childhood; Chromatin; Chromatin Remodeling Factor; Chromosomes; Clinical; Communication; Cues; Cytoplasm; Data; Deposition; Development; Developmental Biology; Diagnosis; Disease; Dose; Dysplasia; Enhancers; Equilibrium; Erythropoiesis; Etiology; Exhibits; Fluorescence; Gene Expression; Generations; Genes; Genetic Transcription; Genomics; Goals; Hematologic Neoplasms; Hematopoiesis; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic Stem Cell heterogeneity; Hematopoietic stem cells; Heterogeneity; Homologous Gene; Human; Immunophenotyping; K562 Cells; Link; Longevity; Lymphoid; Lymphopoiesis; Maps; Mentors; Methods; Modeling; Molecular; Multipotent Stem Cells; Mus; Mutation; Myelogenous; Myelopoiesis; Myeloproliferative disease; Organism; Outcome; Patients; Performance; Phenotype; Physicians; Play; Population; Property; Proteins; Recording of previous events; Recurrence; Regulation; Reporter; Reporting; Reproducibility; Role; Scientist; Sorting; System; Tissues; Training; Transplantation; Tumor Suppressor Proteins; Undifferentiated; Variant; body system; cell behavior; dosage; epigenomics; extracellular; gene repression; high risk; homeodomain; improved outcome; insight; knock-down; knowledgebase; molecular scale; mouse model; novel therapeutics; prospective; recruit; self-renewal; skills; stem cell biology; stem cells; transcription factor; transcriptome

Project Summary/Abstract Hematopoietic stem cells (HSCs) exhibit heterogeneity with respect to repopulating capacity, lineage bias, and cell cycle participation. Teleologically, heterogeneity is the result of stem cells making fundamental decisions regarding the population of the tissue. Differences in stem cell behavior have been linked to the expression of cell surface and cytoplasmic markers without mechanistic explanation. HSC heterogeneity has not previously been attributed to a transcription factor. We propose the observed heterogeneity in HSCs can be explained by the actions of a dose-dependent, homeodomain-containing transcription factor, CUX1. Three lines of evidence suggest CUX1 to be a putative orchestrator of HSC heterogeneity: it recurrently acts in a dose-dependent manner across developmental systems, its recurrent loss in high-risk hematopoietic disease, and its role in chromatin regulation. I report the generation of a CUX1mCherry reporter mouse to study the role of CUX1 in hematopoiesis. The Cux1mCherry mouse is the result of an in-frame, C-terminal mCherry tag at the endogenous CUX1 locus. The addition of the tag creates no aberrant hematopoietic phenotype, suggesting this is a suitable model for the proposed studies. The immunophenotypic long-term HSC (LT-HSC) compartment has among the greatest variances in CUX1 expression. Across CUX1 protein levels in the LT-HSC compartment, we report several correlations to strongly suggest that the observed variation in CUX1 protein results in meaningful differences in stem cell behavior. For example, CUX1Bright HSCs are more likely to be cycling than CUX1Int and CUX1Dim HSCs at steady state. Thus, our studies suggest that CUX1 is playing a dose-dependent role in murine hematopoietic stem and progenitor cells (HSPCs). This proposal aims to (i) establish the role of CUX1 in lineage bias and repopulating capacity and (ii) determine the mechanism by which CUX1 exerts a dose- dependent role. Accomplishing the proposed studies will illuminate an important paradigm in developmental biology: how a small pool of stem cells balance self-renewal and differentiation to give rise to all the mature cells in a tissue. An etiological understanding of stem cell behavior will provide new insights into the development of new therapies for the many diseases that arise in HSCs. The project I propose here is accompanied by a training plan developed by me and my mentors that delineates four goals I will need to accomplish to propel me towards becoming a successful independent physician-scientist. These four goals include gaining expertise in hematopoiesis, gaining expertise in bioinformatics, developing proficiency in scientific communication, and integrating the scientific and clinical aspects of my training. Realizing these goals will give me the skills that I need to be a physician-scientist well- equipped to address meaningful biological questions related to the catastrophic illnesses of childhood.

项目摘要/摘要 造血干细胞（HSC）表现出相对于重新流动能力，谱系偏见的异质性 和细胞周期参与。从图上，异质性是干细胞产生基本的结果 关于组织种群的决定。干细胞行为的差异已与 没有机械解释的细胞表面和细胞质标记的表达。 HSC异质性具有 以前不是归因于转录因子。我们提出了HSC中观察到的异质性可以 可以用剂量依赖性的，同源域的转录因子Cux1的作用来解释。三 证据线表明CUX1是HSC异质性的推定编排：它反复作用于 跨发育系统的剂量依赖性方式，其在高风险造血疾病中的复发损失， 及其在染色质调节中的作用。 我报告了CUX1MCHERRY记者小鼠的产生，以研究CUX1在造血中的作用。这 Cux1-Mcherry小鼠是内源性CUX1基因座处的框架内C末端MCHERRY标签的结果。这 标签的添加不会产生异常的造血表型，这表明这是适合该表型的模型 拟议的研究。免疫表型长期HSC（LT-HSC）室具有最大的 CUX1表达中的方差。在LT-HSC室中的CUX1蛋白水平上，我们报告了几个 与强烈的相关性表明，观察到的Cux1蛋白的变化导致有意义的差异 干细胞行为。例如，Cux1bright HSC比Cux1int和Cux1Dim更有可能骑自行车 HSC处于稳定状态。因此，我们的研究表明，Cux1在鼠中起剂量依赖性作用 造血干和祖细胞（HSPC）。该建议旨在（i）确定Cux1在 谱系偏差和重新载能能力，（ii）确定Cux1施加剂量的机制 依赖的角色。完成拟议的研究将阐明发展中的重要范式 生物学：一小部分干细胞如何平衡自我更新和分化以产生所有成熟的 细胞中的细胞。对干细胞行为的病因理解将为您提供新的见解 开发HSC中许多疾病的新疗法。 我在这里提出的项目伴随着我和我的导师制定的培训计划 描述我需要实现的四个目标，以推动我成为成功的独立 医师科学家。这四个目标包括获得造血方面的专业知识，获得专业知识 生物信息学，发展科学交流的水平以及整合科学和临床 我的培训方面。意识到这些目标将为我提供成为医师科学家的技能 能够解决与儿童灾难性疾病有关的有意义的生物学问题。