Programming of Resident Macrophages by the Brain Environment Following Transplantation

移植后大脑环境对常驻巨噬细胞的编程

• 批准号: 10790219
• 负责人: Frederick Bennett
• 金额: $ 47.54万
• 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: 2023-09-16 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10790219
• 关键词: ATAC-seq; Adopted; Adoption; Atlases; Automobile Driving; Back; Biological; Blood; Brain; Brain Diseases; Cells; Cellular biology; Complex; Data; Development; Disease; Engraftment; Environment; Epigenetic Process; Exposure to; Fingerprint; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Transcription; Goals; Growth; Harvest; Homeostasis; Immune; In Vitro; Infiltration; Injections; Knowledge; Kupffer Cells; Link; Liver; Macrophage; Maps; Measures; Methods; Microglia; Modeling; Molecular; Morphology; Neuroglia; Pathogenesis; Pathway interactions; Peripheral; Process; RNA; Regulation; Resolution; Rest; Role; Sex Differences; Shapes; Signal Transduction; Specific qualifier value; Testing; Time; Transplantation; brain parenchyma; cell type; chemotherapy; data resource; epigenomics; genetic signature; improved; in vivo; in vivo Model; monocyte; nervous system disorder; neurotransmission; new therapeutic target; prevent; programs; receptor; response; sex; single cell analysis; single cell sequencing; single-cell RNA sequencing; therapy development; transcription factor; transcriptomics

PROJECT SUMMARY A striking feature of microglia, the brain's resident macrophages, is their ability to adapt in response to changes in the brain environment. Microglial state changes occur in development and nearly all diseases, often linked to harmful or helpful functions. A better understanding of the regulatory mechanisms underlying microglia state change will therefore improve understanding of brain diseases, and uncover new therapeutic targets. Dozens of disease reactive states have been identified, but little is known about how microglia transition between them. Even microglial “homeostasis” is a state actively maintained by brain environmental signals, and lost in a culture dish. We are experts in the isolation and manipulation of microglia, and created a unique model for intracranial transplantation of microglia and other macrophages following genetic microglia depletion. After transplantation, macrophages engraft the brain and over 14 days undergo dramatic changes in gene expression. In preliminary data, we harvested transplanted macrophages at several timepoints, and by single cell RNA sequencing (scRNAseq), measured the progressive acquisition of microglial identity over time. With this highly controlled in vivo model, we will generate a comprehensive fingerprint of how transplanted microglia are programmed by the brain environment, and use it to identify the genes, pathways, regulatory networks likely to be responsible. In aim 1A, we will capture the environmental programming of cultured microglia after transplant, using paired single cell RNA/ATACseq to identify intermediate states, and to predict the external signals, transcription factors, receptors, pathways and networks responsible. In aim 1B, we will compare transplantation with all combinations of donor and host sex, in order to determine its role in microglia identity specification. Finally, since blood infiltrating macrophages can resemble microglia but remain a distinct cell type, aim 2 will measure their programming after transplantation, to better understand why they cannot become microglia. In summary, completion of these aims will fill knowledge gaps about the regulation of microglial state and identity, create a new foundational data resource, and substantiate an R01 proposal to causally test identity regulators uncovered here.

项目摘要 小胶质细胞的一个惊人特征，大脑的居民巨噬细胞是它们适应变化的能力 在大脑环境中。小胶质状态的变化发生在开发中，几乎所有疾病通常与 有害或有用的功能。更好地了解小胶质细胞状态的调节机制 因此，变化将改善对脑部疾病的理解，并发现新的治疗靶标。数十个 已经确定了疾病反应性状态，但对它们之间的小胶质细胞过渡几乎不知道。 即使是小胶质的“体内平衡”，也是一个由大脑环境信号积极维护的国家，在文化中迷失了 盘子。我们是小胶质细胞隔离和操纵的专家，并为颅内创建了独特的模型 遗传小胶质细胞耗竭后，小胶质细胞和其他巨噬细胞的移植。移植后， 巨噬细胞植入大脑，超过14天的基因表达发生了巨大变化。在初步 数据，我们在几个时间点收集了移植的巨噬细胞，并通过单细胞RNA测序 （SCRNASEQ）测量了随着时间的推移的逐步逐步获得小胶质身份。有了这个高度控制的 在体内模型中，我们将生成有关移植小胶质细胞的全面指纹 由大脑环境编程，并使用它来识别基因，途径，监管网络 可能负责。在AIM 1A中，我们将捕获培养的小胶质细胞的环境编程 移植，使用成对的单细胞RNA/atacseq识别中间状态，并预测外部 负责的信号，转录因子，接收器，路径和网络。在AIM 1B中，我们将比较 与捐赠者和宿主性别的所有组合一起移植，以确定其在小胶质细胞身份中的作用 规格。最后，由于血液浸润的巨噬细胞可能类似于小胶质细胞，但仍然是独特的细胞类型，所以 AIM 2将在移植后衡量其编程，以更好地理解为什么它们不能成为 小胶质细胞。总之，这些目标的完成将填补有关小胶质状态调节的知识空白 和身份，创建一个新的基础数据资源，并证实R01的建议来检测身份 监管机构在这里发现。