Generation of a temporal, spatial, and molecular map of in situ hematopoiesis

生成原位造血的时间、空间和分子图

• 批准号: 10415468
• 负责人: Fernando Camargo
• 金额: $ 100万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10415468
• 关键词: Advanced Development; Aging; Anatomy; Architecture; Behavior; Biological; Biological Assay; Blood; Bone Marrow; Bone Marrow Cells; Catalogs; Cell Transplantation; Cells; Clone Cells; Collection; Development; Disease; Event; Flow Cytometry; Fluorescent in Situ Hybridization; Generations; Goals; Gold; Growth; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic System; Hematopoietic stem cells; Image; In Situ; Individual; Inflammation; Knowledge; Label; Lasers; Location; Maps; Marrow; Measurement; Mediating; Microscope; Microscopy; Molecular; Molecular Analysis; Mus; Optics; Outcome; Process; Production; Reagent; Reporter; Resolution; Resources; Role; Seminal; Sister; System; Techniques; Technology; Text; Textbooks; Time; Tissues; Transcriptional Regulation; Visualization software; Work; aged; base; blood formation; clinically relevant; daughter cell; hematopoietic stem cell differentiation; hematopoietic stem cell niche; image guided; imaging scientist; insight; interdisciplinary approach; mouse model; multiple omics; neglect; novel; novel strategies; progenitor; single molecule; stem cell biology; stem cells; success; tool; transcriptome; transcriptomics; two-photon

(PLEASE KEEP IN WORD, DO NOT PDF) Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. The hematopoietic system has been, arguably, the best well studied tissue from a developmental and stem cell biology perspective. Historically, the study of hematopoiesis has taken advantage of flow cytometry and cell transplantation approaches as the gold standard assays to examine functional behaviors. While these studies have provided an elegant textbook view of hematopoiesis, we argue here that our knowledge of how early blood formation works is largely incomplete. This is predominantly due to the limitations of utilizing traditional assays and readouts that disrupt the normal marrow tissue archicture and neglect the anatomical localization of cells. Thus, one key biological component missing in many studies in the field is that of spatial organization. Basic questions such as when and where hematopoietic fate commitment occurs, what the exact cellular intermediates are between a stem cell and mature daughter cells, and what the identity and role of the cellular microenvironement (niche) is in the hematopoietic process remain largely unsettled. Here, we propose an interdisciplinary approach based on state-of-the-art live microscopy manipulations in combination with novel hematopoietic fate reporters to enable a comprehensive study of hematopoiesis entirely in situ. Using novel two photon-guided collection of stem cell clones directly from the bone marrow, and longitudinal live-imaging analysis of stem cell progenies, we aim to generate a spatial, temporal and molecular map of hematopoiesis at single cell resolution. Complementary proximity-based photolabelling approaches will be taken to catalogue the cellular composition of the niche in the steady-sate, diseased, and aged settings. By moving away from tissue disruptive approaches to a system that relies on observation, manipulation, and analysis of single hematopoietic cells within the bone marrow, we hope to obtain unique insights into the cellular organization of blood production.

（请保持言语，不要PDF） 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。 可以说，造血系统是从发育和干细胞生物学的角度进行的最好的组织。从历史上看，造血的研究利用了流式细胞术和细胞移植方法作为检查功能行为的金标准测定法。尽管这些研究提供了造血的优雅教科书观点，但我们在这里认为我们对早期血液形成的起作用的了解在很大程度上是不完整的。这主要是由于利用传统测定和读数的局限性破坏了正常的骨髓组织档案并忽略了细胞的解剖学定位。因此，该领域许多研究中缺少一个关键的生物学成分是空间组织。基本问题，例如造血命运的何时何地发生，确切的细胞中间体在干细胞和成熟的子细胞之间是什么，以及细胞微环境（NIHEE）在造血过程中的认同和作用在很大程度上保持不确定。在这里，我们提出了一种基于最先进的实时显微镜操作的跨学科方法，并结合新型的造血命运记者，以实现对造血的全面研究。使用新型的两个光子引导的干细胞克隆收集，直接从骨髓中，以及对干细胞后代的纵向生物成像分析，我们旨在在单细胞分辨率下生成造血的空间，时间和分子图。将采用基于互补的接近光标记方法来对稳态，患病和老化的环境中的细胞组成分类。通过从组织破坏性方法转移到依赖于对骨髓中单个造血细胞的观察，操纵和分析的系统，我们希望获得对血液产生细胞组织的独特见解。