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，通常将其定位于控制其的专业生态平衡。 功能。以这种方式，壁ches几乎控制干细胞动力学的所有方面，对组织必不可少的特性 维护。最近的工作表明，精确的蜂窝结构对于利基市场很重要 用它控制的干细胞以保真度进行通信。一个主要问题是该领域不完全理解 壁ches最初是如何在组织中形成的，也不是导致一组细胞创建的关键细胞生物学步骤 一个有效的利基市场，也不是该组织如何影响干细胞调节。我们的实验室最近变得很重要 在睾丸利基上工作的此类问题的进展。特别是我们使用现场模仿来定义 果蝇促性腺藻的动态步骤。这些观察直接导致了一系列 实验揭示了对这种利基组装的机械理解。 昆虫造血与我们的先天免疫系统紧密相似，使用了几个重要因素 用于指定巨噬细胞样和抗菌产生的细胞。这些免疫细胞的祖细胞是 由一个名为后信号中心（PSC）的利基控制。使用固定胚胎和他人的工作 端点分析表明，PSC源自必须迁移以占据其适当的细胞簇 位置并开始充当利基市场。亲甲基细胞如何导航到正确的位置并组装 进入功能性利基是未知的。我们在这里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，以使其在体内形象化其构造。这 包括构造转基因线，应赋予空间和时间光遗传控制的转基因线 标签和实时可视化。这些相同的工具将使后续实验实现 利基组装的基础机制。我们还将介绍PSC细胞的转录格局 为了确定有助于PSC形态发生的新标记。 总的来说，我们的方法应该为我们提供试剂和初步结果，以支持 直接解决潜在机制的实质性长期赠款建议。