Imaging the formation of an hematopoietic niche

造血生态位形成的成像

• 批准号: 10808347
• 负责人: STEPHEN Francis DINARDO
• 金额: $ 16.25万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-22 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10808347
• 关键词: Address; Adopted; Applications Grants; Architecture; Biological; Cells; Cis Tests; Communication; DNA Binding Domain; DNA Sequencing Facility; Data; Defect; Development; Dorsal; Drosophila genus; Embryo; Event; Gene Expression Profile; Genes; Health; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Homeostasis; Image; Imaging Device; Immune; Innate Immune System; Insecta; Investigation; Label; Letters; Macrophage; Maintenance; Mesoderm; Mesoderm Cell; Morphogenesis; Organ; Pattern; Population; Positioning Attribute; Process; Production; Property; RNA; Reagent; Regenerative Medicine; Regulation; Resolution; Role; Sampling; Series; Signal Transduction; Source; Specific qualifier value; Specificity; Testing; Testis; Time; Tissues; Transactivation; Transgenic Organisms; Visceral; Visualization; Work; antimicrobial; candidate identification; cell behavior; daughter cell; experimental study; follow-up; improved; in vivo; insight; interest; lymph nodes; migration; next generation sequencing; novel marker; optogenetics; progenitor; prospective; selective expression; single-cell RNA sequencing; spatial relationship; stem cell function; stem cell niche; stem cells; success; tool; virtual

Stem cells are necessary for tissue homeostasis, and are often localized to specialized niches that control their function. In this manner, niches control virtually all aspects of stem cell dynamics, properties essential to tissue maintenance. Recent work has shown that precise cellular architecture is important in order for a niche to communicate with fidelity to the stem cells it controls. A major issue is that the field does not fully understand how niches are initially formed in a tissue, nor the key cell biological steps that cause a group of cells to create an effective niche, nor how that organization impacts stem cell regulation. Our lab recently made significant advances on such questions working on the testis niche. In particular we used live-imaging to define the dynamic steps in Drosophila gonadal niche assembly. These observations led directly to a series of experiments revealing a mechanistic understanding of the assembly of this niche. Insect hematopoiesis closely parallels our innate immune system, using several conserved factors important for specifying macrophage-like and anti-microbial-producing cells. The progenitors for these immune cells are controlled by a niche called the Posterior Signaling Center (PSC). Work of others using fixed embryos and end-point analysis showed that the PSC is derived from a cell cluster that must migrate to take up its proper position and begin functional as a niche. How the pro-niche cells navigate to the correct position and assemble into a functional niche is unknown. We propose here to develop tools to address these questions. We will build tools to lineage-label the PSC at an early-enough stage to visualize its construction in vivo. This includes the construction of transgenic lines that should confer spatial and temporal optogenetic control for labeling and real-time visualization. These same tools would enable follow-up experiments to expore the mechanisms underlying niche assembly. We will additionally profile the transcriptional landscape of PSC cells in order to identify new markers aiding investigation of PSC morphogenesis. Collectively, our approaches should provide us with the reagents and preliminary results to support a substantive, longer-term grant proposal addressing underlying mechanisms directly.

干细胞对于组织稳态是必需的，并且通常位于控制其自身功能的专门生态位中。 功能。通过这种方式，生态位几乎控制干细胞动力学的所有方面，以及组织所必需的特性 维护。最近的研究表明，精确的细胞结构对于利基市场的发展非常重要。 与其控制的干细胞保持忠实的沟通。一个主要问题是该领域没有完全理解 组织中最初如何形成生态位，也不是导致一组细胞产生的关键细胞生物学步骤 一个有效的利基市场，以及该组织如何影响干细胞调节。我们的实验室最近取得了重大进展 关于睾丸生态位的此类问题的进展。特别是，我们使用实时成像来定义 果蝇性腺生态位组装的动态步骤。这些观察结果直接导致了一系列 实验揭示了对这个生态位组装的机械理解。 昆虫的造血作用与我们的先天免疫系统密切相关，利用了几个重要的保守因素 用于指定巨噬细胞样细胞和抗微生物产生细胞。这些免疫细胞的祖细胞是 由一个称为后信号中心（PSC）的利基控制。其他人使用固定胚胎的工作和 终点分析表明，PSC 源自细胞簇，该细胞簇必须迁移才能占据其适当的位置。 定位并开始作为利基市场发挥作用。亲利基细胞如何导航到正确的位置并组装 进入一个功能性利基市场是未知的。我们在此建议开发工具来解决这些问题。 我们将在早期阶段构建工具来对 PSC 进行谱系标记，以可视化其在体内的构建。这 包括构建应赋予空间和时间光遗传学控制的转基因系 标签和实时可视化。这些相同的工具将使后续实验能够揭示 利基组装的机制。我们还将进一步分析 PSC 细胞的转录景观 以确定有助于 PSC 形态发生研究的新标记。 总的来说，我们的方法应该为我们提供试剂和初步结果来支持 直接涉及基本机制的实质性、长期拨款提案。