High-dimensional characterization of phagosome composition, control and phagocytic receptor redundancy

吞噬体组成、控制和吞噬受体冗余的高维表征

• 批准号: 10447138
• 负责人: Bryan David Bryson
• 金额: $ 42万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-15 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10447138
• 关键词: Address; Apoptotic; Biochemistry; Biological Process; Biology; Cell membrane; Cellular biology; Complex; Coupled; Dendritic Cells; Development; Engineering; Event; Fibroblasts; Goals; Homeostasis; Immune; Immunology; Ions; Knowledge; Label; Ligands; Metals; Microbiology; Modeling; Molecular; Organelles; Phagocytes; Phagocytosis; Phagosomes; Positioning Attribute; Protein Engineering; Research; Series; Shapes; Signal Transduction; Stereotyping; System; Systems Biology; Tissues; cell type; enzyme activity; experimental study; extracellular; high dimensionality; insight; lens; macrophage; neutrophil; novel; pathogen; programs; receptor; trafficking

Project Summary The phagosome is a dynamically formed organelle that is generated upon phagocyte encounter with cargo. Phagocytic receptors and other extracellular receptors engage with cargo-derived ligands prior to the formation of the phagocytic cup at the cell membrane and subsequent phagocytosis. Following phagosome formation, a dynamic series of steps proceed involving organelle trafficking and fusion. Ultimately, these collective molecular events influence and shape phagosome function which is often characterized through the lens of phagosome biochemistry (pH, metal ion abundance, oxidative radicals, and enzyme activity). While many of the stereotyped features of phagosome maturation and biochemistry have been studied, there has been relatively fewer studies that take an integrated systems-level view from signaling to phagosome biochemistry. Furthermore, while the field has defined several features of general phagocytosis, phagosome biology is incredibly complex. Several distinct cell types can perform phagocytosis ranging from professional phagocytes (ex: macrophages, neutrophils, dendritic cells) to non-professional phagocytes (ex: fibroblasts). Adding another layer of complexity, phagocytes engulf a diverse array of cargo ranging from pathogens to apoptotic bodies. Combined with the temporal maturation of the phagosome, these three axes construct a complex landscape for phagosome biology. In depth study of this landscape has not been performed limiting our fundamental understanding of molecular control of this organelle. Here, we propose a research program centered around the question: “how is control of phagosome biology achieved?” To address these questions, my research program integrates approaches in genetics, protein engineering, systems biology, immunology, and microbiology. We seek to address three knowledge gaps in our program initially. (1) Is there crosstalk in signaling among receptors (phagocytic and soluble ligand) during phagocytosis? (2) How do cargo and phagocyte identity instruct phagosome composition? (3) What are the molecular circuits that control phagosome biochemistry? Over the next five years, we will develop a strategy to examine higher-order interactions in phagocytosis signaling. Furthermore, we will engineer specific cargo capable of performing proximity labeling in the phagosome. Lastly, we will define the molecular circuits that control phagosome biochemistry. These questions are inextricably coupled, and our program operates in a highly collaborative manner. Supporting our experimental systems is a strong quantitative modeling and analytical framework equipped to derive novel insights from high-throughput experiments and propose new experimental directions. Together, these strengths position us in a unique manner to address longstanding questions in phagosome biology.

项目摘要 吞噬体是一种动态形成的细胞器，在吞噬细胞相遇时生成 货物。吞噬受体和其他细胞外受体与货物衍生的配体互动 在细胞膜和随后的吞噬作用上形成吞噬杯。跟随吞噬体 形成，一系列动态的步骤，涉及细胞器贩运和融合。最终，这些 集体分子事件会影响和形状吞噬体功能，通常通过 吞噬体生物化学的镜头（pH，金属离子丰度，氧化自由基和酶活性）。 虽然吞噬体成熟和生物化学的许多定型特征已经研究了，但 从信号到吞噬体的集成系统级视图的研究相对较少。 生物化学。此外，尽管该领域定义了一般吞噬作用的几个特征，但 生物学非常复杂。几种不同的细胞类型可以执行吞噬作用，从专业 吞噬细胞（例如：巨噬细胞，中性粒细胞，树突状细胞）到非专业吞噬细胞（例如：成纤维细胞）。 增加了另一层复杂性，吞噬吞噬了一​​系列从病原体到病原体的货物阵列 凋亡的身体。与吞噬体的暂时成熟相结合，这三个轴构造了A 吞噬体生物学的复杂景观。在深入研究该景观的情况下，尚未进行限制我们的 对该细胞器的分子控制的基本理解。 在这里，我们提出了一个研究计划，围绕一个问题：“吞噬体的控制如何 生物学实现了吗？”为了解决这些问题，我的研究计划整合了遗传学的方法 工程，系统生物学，免疫学和微生物学。我们试图解决我们的三个知识差距 程序最初。 （1）在接收器（吞噬和固体配体）之间是否存在串扰 吞噬作用？ （2）货物和吞噬细胞身份如何指导吞噬体组成？ （3）什么是 控制吞噬体生物化学的分子回路？在接下来的五年中，我们将制定一项策略 检查吞噬作用信号传导中的高阶相互作用。此外，我们将设计特定的货物 能够在吞噬体中执行接近性标记。最后，我们将定义分子电路 控制吞噬体生物化学。这些问题是密不可分的，我们的计划在一个高度运行 协作方式。支持我们的实验系统是强大的定量建模和分析性的 配备了从高通量实验和提案中获得新见解的框架 方向。这些优势共同将我们定位为独特的方式，以解决长期存在的问题 吞噬体生物学。