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 项目摘要/摘要 小胶质细胞是大脑的居民免疫核素，对于正常的大脑发育和功能至关重要，可能 参与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，我们严重限制了研究中的小胶质功能 疾病，尤其是体内。这个指导的职业发展奖，由临床科学家 由Ben Barres博士指导的斯坦福大学精神病学和神经生物学系将成为 研究人类小胶质细胞在精神疾病中的研究通过相关的基本和翻译的研究目标是现实。这 基础研究目标是通过比较的转录组来提高我们对小胶质生物学的理解 在疾病期间进入大脑的小胶质细胞和血液衍生的“像细胞相似的小胶质细胞”（MLC）。翻译 目的是使用这些结果从血细胞中产生人类小胶质细胞，从而可以研究小胶质细胞 人脑组织。该提案概述了一项为期5年的职业发展计划，该计划利用 斯坦福大学的理查德教育和科学资源完成以下所述的研究目的 在为候选人提供所需的培训以作为独立研究人员的同时。 初步数据表明，当由于CSF1受体（CSF1R）敲除小鼠时， 用野生型（WT）骨髓（BM）移植，供体衍生的细胞以几乎相同的模式填充大脑 至WT小胶质细胞，并表达小胶质命运的几乎所有特定标记。这些植入的MLC可以是 根据TMEM119的表达（一种高度特异性的小鼠和人类小胶质细胞标记）纯化。 rnaseq 数据表明，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MLC和小胶质细胞之间的差异，并创建一个可靠的系统 用于研究体内骨髓衍生的MLC。 AIM 1将确定MLC类似于WT 小胶质细胞，以及它们不同的方式。 AIM 2将确定特定的造血细胞类型 生成MLC。 AIM 3将通过产生RNASEQ来翻译这些发现以与人类血细胞一起使用 高度纯净的人类小胶质细胞的轮廓，并将其与通过移植产生的MLC进行比例 人BM进入免疫缺陷的人性化小鼠。这项工作将提高我们对核心的理解 小胶质细胞和MLC之间的差异，确定纯人类小胶质细胞的转录组特征，以及 创建一个可使用患者衍生的造血细胞研究小胶质细胞的可拖动系统，同时促进 独立的研究职业围绕小胶质细胞在心理健康中的作用。 第6页 项目摘要/摘要