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

在此输入文本，它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血液干细胞表观遗传异质性的程度和动态 适当地控制造血干细胞(HSC)的自我更新和分化对所有人类来说都是至关重要的，不仅在健康条件下，而且在通过骨髓移植治疗各种血液病方面也是如此。因此，了解造血机制对于了解健康的生理学和为血细胞再生治疗创造有效和合理的方法至关重要。有趣的是，先前的研究已经报道了单个造血干细胞后代的最终细胞命运的显著异质性，即使是在相同环境中的克隆相关细胞。这些谱系潜力的差异可能是表观遗传状态的差异造成的，然而，在单个干细胞水平上的表观遗传状态的研究一直受到现有方法学的限制。为了克服这些限制，我们开发了一种新的基于显微镜的分析方法，称为Sceptre(翻译后表观遗传编码的单细胞评估)，它可以敏感地解决单细胞中单个基因座位的表观遗传状态(Woodworth等人)。2021年)。在目标1中，我们将使用Sceptre来研究组蛋白在HSCs中的翻译后修饰，这些修饰位于控制淋巴髓样分化的关键基因位点。在目标2中，为了分析表观遗传状态动力学的目标，我们将开发一种实时成像分析方法，用于跟踪多代细胞中的HSC克隆，然后我们将与Sceptre结合，在克隆水平上探索表观遗传。如果成功，这项工作将为未来澄清HSC生物学中异质性的起源和后果的研究打开大门。

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.