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，这是您的应用程序的新摘要信息。此部分不得超过30行文本。 在单细胞水平上评估造血干细胞表观遗传异质性的程度和动态 造血干细胞（HSC）自我更新和分化的适当控制对所有人都是必不可少的，不仅在健康状况期间，而且对于通过骨髓移植治疗各种血液疾病也是如此。因此，了解造血机制对于理解健康生理学和创造有效合理的血细胞再生疗法方法至关重要。有趣的是，先前的研究已经报道了个体HSC后代的最终细胞命运分布的实质性异质性，即使是相同环境中的克隆相关细胞。这些谱系潜力的差异可能是由于表观遗传状态的差异造成的，然而，在个体干细胞水平上的表观遗传状态的研究受到现有方法的限制。为了克服这些局限性，我们开发了一种新的基于显微镜的检测方法，称为SCEPTRE（单细胞翻译后表观遗传编码评估），可以灵敏地解析单细胞中单基因位点的表观遗传状态（Woodworth等人。2021）。在目的1中，我们将使用SCEPTRE研究组蛋白的翻译后修饰在HSC的关键基因位点控制淋巴髓样分化。在目标2中，为了分析表观遗传状态动力学的目标，我们将开发一种活的成像测定，用于跟踪多代细胞中的HSC克隆，然后我们将联合收割机与SCEPTRE结合，以在克隆水平上探测表观遗传。如果成功，这项工作将为未来的研究打开大门，阐明HSC生物学异质性的起源和后果。