Clarifying the Origins of Blood Stem Cell Heterogeneity by Single-Cell Epigenetic State Profiling

通过单细胞表观遗传状态分析阐明血液干细胞异质性的起源

• 批准号: 10701145
• 负责人: Hao Yuan Kueh
• 金额: $ 9.94万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-23 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10701145
• 关键词: Biological Assay; Blood; Blood Cells; Bone Marrow Transplantation; Cell Differentiation process; Cell Lineage; Cell division; Cells; Clonal Hematopoietic Stem Cell; DNA biosynthesis; Decision Making; Development; Disease; Environment; Epigenetic Process; Evaluation; Future; Gene Activation; Generations; Genes; Genetic Transcription; Goals; Health; Hematological Disease; Hematopoiesis; Hematopoietic Stem Cell heterogeneity; Hematopoietic stem cells; Heritability; Heterogeneity; Histones; Human; Image; Immune; Individual; Malignant Neoplasms; Measures; Methodology; Methods; Microscopy; Physiology; Polymerase; Post-Translational Protein Processing; Regulator Genes; Reporting; Research; Research Personnel; Shapes; Somatotype; Specific qualifier value; Testing; Text; Time; Validation; Work; base; cell type; epigenetic regulation; genomic locus; hematopoietic stem cell self-renewal; histone modification; imaging modality; premature; regenerative therapy; self-renewal; stem cell biology; stem cell population; stem cells; tool

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Evaluating the extent and dynamics of epigenetic heterogeneity in blood stem cells at the single-cell level Proper control of hematopoietic stem cell (HSC) self-renewal and differentiation is essential for all humans, not only during healthy conditions, but also for the treatment of various hematological disorders through bone marrow transplantations. Understanding the mechanisms of hematopoiesis is therefore vital for understanding healthy physiology and for creating efficient and rational approaches for blood cell regeneration therapies. Intriguingly, prior research has reported substantial heterogeneity in the distributions of final cell fates for the descendants of individual HSCs, even for clonally related cells in identical environments. These differences in lineage potential could result from differences in epigenetic state, however, the study of epigenetic states at the level of individual stem cells has been hindered by limitations in available methodology. To overcome these limitations, we have developed a new microscopy-based assay called SCEPTRE (Single-Cell Evaluation of Post-TRanslational Epigenetic encoding) that can sensitively resolve epigenetic states at single gene loci in single cells (Woodworth et al. 2021). In Aim 1 we will use SCEPTRE to study histone post-translational modifications in HSCs at key gene loci governing lymphomyeloid differentiation. In Aim 2, toward the goal of analyzing epigenetic state dynamics, we will develop a live imaging assay for tracking HSC clones over multiple cell generations that we will then combine with SCEPTRE to probe epigenetic heritance at the clonal level. If successful, this work will open the door for future studies that clarify the origins and consequences of heterogeneity in HSC biology.

在此处输入文本，这是您应用程序的新摘要信息。本节必须不超过30行文本。 评估单细胞水平血干细胞表观遗传异质性的程度和动力学 适当控制造血干细胞（HSC）自我更新和分化对于所有人类都至关重要，不仅在健康状况下，而且对于通过骨髓移植治疗各种血液学疾病。因此，了解造血的机制对于理解健康的生理学以及为血细胞再生疗法的有效和合理的方法至关重要。有趣的是，先前的研究报道了单个HSC后代的最终细胞命运分布中的实质性异质性，即使是在相同环境中具有克隆相关细胞的分布。这些谱系潜力的这些差异可能是由于表观遗传状态的差异而导致的，但是，对单个干细胞水平的表观遗传态的研究受到了可用方法的限制的阻碍。为了克服这些局限性，我们开发了一种新的基于显微镜的测定法，称为权杖（翻译后表观遗传编码的单细胞评估），该测定方法可以灵敏地解决单个细胞中单基因基因座的表观遗传态（Woodworth等人，Woodworth等，2021年）。在AIM 1中，我们将使用权杖研究在关键基因座位上淋巴细胞分化的HSC中翻译后修饰。在AIM 2中，为了分析表观遗传状态动力学，我们将开发一种实时成像测定法，用于在多个细胞世代跟踪HSC克隆，然后我们将与权杖结合以在克隆水平上探测表观遗传遗传。如果成功，这项工作将为将来的研究打开大门，以阐明HSC生物学中异质性的起源和后果。