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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要小胶质细胞（大脑中常驻的巨噬细胞）的一个显着特征是它们能够适应变化在大脑环境中，小胶质细胞状态的变化发生在发育和几乎所有疾病中，通常与这些疾病有关。更好地了解小胶质细胞状态的调节机制。因此，改变将增进对脑部疾病的了解，并发现新的治疗靶点。疾病反应状态已经被确定，但关于小胶质细胞如何在它们之间转变却知之甚少。甚至小胶质细胞的“稳态”也是一种由大脑环境信号主动维持的状态，并且在文化中丢失了我们是小胶质细胞分离和操作方面的专家，并创建了独特的颅内模型。遗传性小胶质细胞耗竭后移植小胶质细胞和其他巨噬细胞。巨噬细胞植入大脑后，在 14 天的时间里，基因表达发生了巨大的变化。根据数据，我们在几个时间点收获了移植的巨噬细胞，并通过单细胞 RNA 测序 (scRNAseq)，测量了小胶质细胞身份随时间的逐渐获得。在体内模型中，我们将生成移植小胶质细胞如何进行的全面指纹由大脑环境编程，并用它来识别基因、通路、调节网络在目标 1A 中，我们将捕获培养的小胶质细胞的环境编程。移植，使用配对的单细胞 RNA/ATACseq 来识别中间状态，并预测外部在目标 1B 中，我们将比较信号、转录因子、受体、通路和网络。移植供体和宿主性别的所有组合，以确定其在小胶质细胞身份中的作用最后，由于血液浸润巨噬细胞可能类似于小胶质细胞，但仍然是一种独特的细胞类型，目标2将在移植后测量他们的编程，以更好地理解为什么他们不能成为总之，这些目标的完成将填补有关小胶质细胞状态调节的知识空白。和身份，创建新的基础数据资源，并证实 R01 提案以因果测试身份监管机构在这里揭晓。