Intravital analysis of hematopoietic stem cells in their bone marrow niche - Resubmission - 1

骨髓微环境中造血干细胞的活体分析 - 重新提交 - 1

• 批准号: 10676859
• 负责人: David R Fooksman
• 金额: $ 64.51万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-20 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10676859
• 关键词: Address; Adult; Affect; Animals; Behavior; Blood; Blood Vessels; Bone Marrow; Bone Marrow Transplantation; CXCR4 Receptors; CXCR4 gene; Calcium; Calcium Signaling; Cellular Morphology; Clinic; Collaborations; Complex; Computer Analysis; Computer Models; Development; Enterobacteria phage P1 Cre recombinase; Environment; Genetic; Growth Factor; Hematopoiesis; Hematopoietic; Hematopoietic Stem Cell Mobilization; Hematopoietic stem cells; Hour; Immunity; Impairment; Inflammation; Inflammatory; Integrins; Interferons; Knowledge; Label; Mediator; Molecular; Morphology; Mouse Strains; Mus; Natural regeneration; Nature; Pathologic; Play; Process; Reporter; Role; Signal Transduction; Stem Cell Factor; Stem cell transplant; Stimulus; Stromal Cell-Derived Factor 1; Stromal Cells; System; Tamoxifen; Testing; Time; Transgenes; Transplantation; Visualization; antagonist; behavioral study; cell behavior; cell motility; chemokine; clinically relevant; genetic analysis; genetic manipulation; hematopoietic stem cell niche; human old age (65+); in vivo; inducible Cre; insight; intravital imaging; intravital microscopy; multiphoton imaging; multipotent cell; novel; peripheral blood; pharmacologic; ratiometric; reconstitution; response; sensor; stem cell biology; stem cell function; stem cell growth; stem cell homeostasis

ABSTRACT Hematopoietic stem cells (HSC) in the adult mammalian bone marrow (BM) are rare multipotent cells that can regenerate and sustain multilineage hematopoiesis upon transplantation. Given the rarity and limited availability of HSC in the BM, strategies to "mobilize" HSC from the BM to the peripheral blood show great promise to enhance the efficiency of hematopoietic reconstitution. On the other hand, the reconstitution capacity of HSC can be impaired in old age or following inflammatory insults. HSC are localized in a unique functional BM compartment called the niche. Its key component is a network of perivascular stromal cells expressing the HSC growth factor stem cell factor (SCF or Kit ligand) and chemokine CXCL12, whose receptor CXCR4 helps retain HSC in the BM. However, the morphology and dynamics of endogenous live HSC and their interactions with the BM niche components have not been well characterized. We have developed a system for specific fluorescent labeling of adult murine HSC, and adapted it for intravital imaging of HSC within the BM. Preliminary studies revealed an unexpectedly dynamic morphology and complex motility of HSC in the steady state. Furthermore, we were able to visualize interactions between live HSC and the stromal cells. We will apply this system to systematically study the behavior of live HSC and their interaction with the niche in live animals. The project involves a collaboration between labs with the expertise in genetic manipulation and analysis of HSC (Reizis), intravital microscopy of the BM (Fooksman) and computational analysis and modeling of cell behavior (Krichevsky). In Aim 1, we will visualize the behavior of live HSC in the BM in the steady state and during inflammation. In Aim 2, we will characterize the interaction of HSC with their BM environment, both in normal conditions and after mobilization from the BM. In Aim 3, we will analyze the molecular basis of HSC dynamics and correlate it with HSC function in supporting hematopoiesis. The proposed studies would address major gaps in our knowledge of basic HSC biology, and provide novel insights into the clinically relevant process of HSC mobilization.

抽象的 成年哺乳动物骨髓（BM）中的造血干细胞（HSC）是罕见的多能细胞 移植后可以再生并维持多素造血。鉴于稀有性和有限 HSC在BM中的可用性，将HSC从BM到外周血“动员” HSC的策略很棒 承诺提高造血重建的效率。另一方面，重组 HSC的能力可能会在老年或炎症性损害后受损。 HSC本地化为独特 功能性BM室称为利基市场。它的关键成分是血管周间基细胞网络 表达HSC生长因子干细胞因子（SCF或KIT配体）和趋化因子CXCL12 受体CXCR4有助于将HSC保留在BM中。但是，内生的形态和动力学 HSC及其与BM利基组件的相互作用尚未得到很好的特征。我们有 开发了一种用于成年鼠HSC的特定荧光标记的系统，并将其改编成浸润 BM内HSC的成像。初步研究揭示了一种意外的动态形态和 HSC在稳定状态下的复杂运动。此外，我们能够可视化 活HSC和基质细胞。我们将应用该系统系统地研究实时HSC的行为 以及他们与活动物中利基市场的互动。该项目涉及实验室与 HSC（REIZIS）的遗传操作和分析的专业知识，BM的浸润显微镜 （Fooksman）以及细胞行为的计算分析和建模（Krichevsky）。在AIM 1中，我们将 在稳态和炎症期间可视化活体力体中实时HSC的行为。在AIM 2中，我们将 在正常条件下和之后，都表征了HSC与其BM环境的相互作用 来自BM的动员。在AIM 3中，我们将分析HSC动力学的分子基础并将其关联 HSC功能在支持造血。拟议的研究将解决我们的主要差距 基本HSC生物学知识，并提供有关HSC临床相关过程的新见解 动员。

项目成果

Clonal barcoding of endogenous adult hematopoietic stem cells reveals a spectrum of lineage contributions.

• DOI: 10.1073/pnas.2317929121
• 发表时间: 2024-01-23
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Feng, Jue;Jang, Geunhyo;Esteva, Eduardo;Adams, Nicholas M.;Jin, Hua;Reizis, Boris
• 通讯作者: Reizis, Boris