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.

项目概要/摘要 造血干细胞 (HSC) 在再生能力、谱系偏倚、 和细胞周期参与。从目的论来看，异质性是干细胞做出根本性作用的结果 关于组织群体的决定。干细胞行为的差异与 细胞表面和细胞质标记物的表达，没有机制解释。 HSC 异质性 之前并未被归因于转录因子。我们认为 HSC 中观察到的异质性可以 可以通过剂量依赖性、含有同源结构域的转录因子 CUX1 的作用来解释。三 一系列证据表明 CUX1 被认为是 HSC 异质性的协调者：它经常以一种 整个发育系统的剂量依赖性方式，其在高风险造血疾病中的反复损失， 及其在染色质调控中的作用。 我报道了 CUX1mCherry 报告小鼠的产生，以研究 CUX1 在造血中的作用。这 Cux1mCherry 小鼠是内源性 CUX1 基因座上框内 C 端 mCherry 标签的结果。这 添加标签不会产生异常的造血表型，表明这是一个合适的模型 拟议的研究。免疫表型长期 HSC (LT-HSC) 区室具有最大的免疫表型 CUX1 表达的差异。在 LT-HSC 区室中的 CUX1 蛋白水平上，我们报告了几个 相关性强烈表明观察到的 CUX1 蛋白变化会导致有意义的差异 干细胞行为。例如，CUX1Bright HSC 比 CUX1Int 和 CUX1Dim 更有可能循环 HSC 处于稳定状态。因此，我们的研究表明 CUX1 在小鼠体内发挥剂量依赖性作用 造血干细胞和祖细胞（HSPC）。该提案旨在 (i) 确立 CUX1 在以下方面的作用： 谱系偏倚和重新增殖能力，以及 (ii) 确定 CUX1 发挥剂量作用的机制 依赖角色。完成拟议的研究将阐明发展中的一个重要范式 生物学：一小群干细胞如何平衡自我更新和分化以产生所有成熟的细胞 组织中的细胞。对干细胞行为的病因学理解将为我们提供新的见解 开发针对 HSC 中出现的许多疾病的新疗法。 我在这里提出的项目附有我和我的导师制定的培训计划，该计划 描绘了我需要实现的四个目标，以推动我成为一名成功的独立人士 医生科学家。这四个目标包括获得造血的专业知识、获得 生物信息学，培养科学交流的能力，并将科学与临床相结合 我的训练的各个方面。实现这些目标将使我具备成为一名医生科学家所需的技能—— 有能力解决与儿童灾难性疾病相关的有意义的生物学问题。