Creation of new tools to study human microglia using blood cells

创建利用血细胞研究人类小胶质细胞的新工具

• 批准号: 10378989
• 负责人: Frederick Bennett
• 金额: $ 5.83万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2021-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10378989
• 关键词: Alzheimer' s Disease; Basic Science; Biology; Blood; Blood Cells; Bone Marrow; Brain; CSF1 gene; Cells; Data; Development; Development Plans; Disease; Fostering; Goals; Hematopoietic; Human; Immune; K-Series Research Career Programs; Knock-out; Mental Depression; Mental Health; Mental disorders; Mentors; Microglia; Mood Disorders; Mus; Neurobiology; Patients; Pattern; Psychiatry; Research; Research Personnel; Resources; Rest; Role; Schizophrenia; Scientist; Specimen; System; Training; Translating; Translational Research; Transplantation; Universities; Work; autism spectrum disorder; base; brain tissue; career; career development; cell type; design; humanized mouse; improved; in vivo; receptor; tool; transcriptome; transcriptome sequencing; transcriptomics

Contact PD/PI: BENNETT, FREDERICK PROJECT SUMMARY/ABSTRACT Microglia, the brain's resident immune cells, are essential for normal brain development and function, and likely involved in mental illnesses such as depression and schizophrenia. Because it is nearly impossible to obtain suitable human brain specimens, we are severely restricted from studying microglial function in these diseases, especially in vivo. This Mentored Career Development Award, by a clinician-scientist in the Departments of Psychiatry and Neurobiology at Stanford University, mentored by Dr. Ben Barres, will make the study of human microglia in mental illness a reality through related basic and translational research goals. The basic research goal is to advance our understanding of microglial biology by comparing the transcriptomes of microglia and blood-derived “Microglia Like Cells” (MLCs) that enter the brain during disease. The translational goal is to use these results to generate human microglia from blood cells, allowing study of microglia without human brain tissue. This proposal outlines a 5 year mentored career development plan that harnesses the abundant educational and scientific resources at Stanford to accomplish the research aims described below while providing the candidate with needed training to function as an independent investigator. Preliminary data show that when mice lacking microglia due to knock-out of the CSF1 receptor (CSF1R) are transplanted with wild type (WT) bone marrow (BM), donor-derived cells fill the brain in a near-identical pattern to WT microglia, and express nearly all specific markers of microglial fate. These engrafted MLCs can be purified based on expression of Tmem119, a highly specific mouse and human microglial marker. RNAseq data shows that MLCs are highly similar to WT microglia. The proposed research will expand on these intriguing findings to better understand differences between MLCs and microglia, and create a reliable system for studying bone marrow-derived MLCs in vivo. Aim 1 will determine how closely MLCs resemble WT microglia, and in what ways they are different. Aim 2 will identify the specific hematopoietic cell types capable of generating MLCs. Aim 3 will translate these findings for use with human blood cells, by generating RNAseq profiles for highly pure resting human microglia, and comparing them to MLCs generated by transplantation of human BM into immunodeficient humanized mice. This work will advance our understanding of core differences between microglia and MLCs, identify the transcriptomic signature of pure human microglia, and create a tractable system to study microglia using patient-derived hematopoietic cells, while fostering an independent research career centered around the role of microglia in mental health. Page 6 Project Summary/Abstract

联系PD/PI：班尼特，FREDERICK 项目总结/摘要 小胶质细胞是大脑的常驻免疫细胞，对大脑的正常发育和功能至关重要， 与抑郁症和精神分裂症等精神疾病有关。因为几乎不可能得到 合适的人脑标本，我们从研究这些小胶质细胞的功能受到严重限制， 疾病，尤其是在体内。这个指导职业发展奖，由一位临床医生，科学家在 由本·巴雷斯博士指导的斯坦福大学精神病学和神经生物学系将使 通过相关的基础和转化研究目标，人类小胶质细胞在精神疾病中的研究成为现实。的 基本的研究目标是通过比较小胶质细胞的转录组来促进我们对小胶质细胞生物学的理解。 小胶质细胞和血液来源的“小胶质细胞样细胞”（MLCs）在疾病期间进入大脑。平移 我们的目标是利用这些结果从血细胞中产生人类小胶质细胞，允许研究小胶质细胞，而不需要 人脑组织该提案概述了一个5年的指导职业发展计划，利用 斯坦福大学拥有丰富的教育和科学资源，以实现下述研究目标 同时为候选人提供必要的培训，使其能够作为独立调查员开展工作。 初步数据显示，当由于CSF 1受体（CSF 1 R）敲除而缺乏小胶质细胞的小鼠， 移植野生型（WT）骨髓（BM）后，供体来源的细胞以几乎相同的模式填充大脑 转化为WT小胶质细胞，并表达几乎所有小胶质细胞命运的特异性标志物。这些嫁接的MLC可以是 基于高度特异性的小鼠和人小胶质细胞标志物Tmem 119的表达纯化。RNAseq 数据显示MLC与WT小胶质细胞高度相似。拟议的研究将扩大这些 有趣的发现，以更好地了解MLCs和小胶质细胞之间的差异，并创建一个可靠的系统， 用于体内研究骨髓来源的MLCs。目标1将确定MLC与WT的相似程度 小胶质细胞，以及它们在哪些方面不同。目的2将确定特定的造血细胞类型， 生成MLC。Aim 3将通过生成RNAseq， 高纯度静息人小胶质细胞的特征，并将它们与通过移植 人BM到免疫缺陷人源化小鼠中。这项工作将促进我们对核心的理解 小胶质细胞和MLC之间的差异，鉴定纯人类小胶质细胞的转录组学特征，以及 创建一个易于处理的系统，使用患者来源的造血细胞研究小胶质细胞，同时培养一个 独立的研究生涯围绕小胶质细胞在心理健康中的作用。 第6页 项目总结/摘要