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：BENNETT、FREDERICK 项目概要/摘要 小胶质细胞是大脑的常驻免疫细胞，对于正常的大脑发育和功能至关重要，并且可能 涉及抑郁症和精神分裂症等精神疾病。因为几乎不可能获得 合适的人脑标本，我们在这些样本中研究小胶质细胞功能受到严格限制 疾病，尤其是体内疾病。该指导职业发展奖由一位临床医生科学家颁发 斯坦福大学精神病学和神经生物学系在本·巴雷斯 (Ben Barres) 博士的指导下，将 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。这 基础研究目标是通过比较小胶质细胞的转录组来增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”（MLC）在疾病期间进入大脑。翻译的 目标是利用这些结果从血细胞中产生人类小胶质细胞，从而无需对小胶质细胞进行研究即可 人类脑组织。该提案概述了一个为期 5 年的指导性职业发展计划，该计划利用 斯坦福大学丰富的教育和科学资源可实现下述研究目标 同时为候选人提供作为独立调查员所需的培训。 初步数据显示，当因 CSF1 受体 (CSF1R) 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 WT小胶质细胞，并表达小胶质细胞命运的几乎所有特定标记。这些植入的 MLC 可以是 基于 Tmem119 的表达进行纯化，Tmem119 是一种高度特异性的小鼠和人类小胶质细胞标记物。 RNA测序 数据显示 MLC 与 WT 小胶质细胞高度相似。拟议的研究将扩展这些 有趣的发现可以更好地理解 MLC 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 MLC。目标 1 将确定 MLC 与 WT 的相似程度 小胶质细胞，以及它们有何不同。目标 2 将确定能够产生作用的特定造血细胞类型 生成MLC。目标 3 将通过生成 RNAseq 将这些发现转化为用于人类血细胞 高纯度静息人小胶质细胞的概况，并将其与移植产生的 MLC 进行比较 将人骨髓转化为免疫缺陷人源化小鼠。这项工作将增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来研究小胶质细胞，同时培养 独立研究生涯围绕小胶质细胞在心理健康中的作用展开。 第6页 项目概要/摘要