Unlocking microglia targeting for neurotherapeutics

解锁神经治疗药物的小胶质细胞靶向

基本信息

批准号：
10696509
负责人：
Mariko L. Bennett
金额：
$ 44.5万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
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：
2023-09-20 至 2028-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10696509
关键词：
Acceleration Affect Anti-Retroviral Agents Award Benchmarking Bone Marrow Brain CRISPR screen Cell Therapy Cells Central Nervous System Child Care Childhood Chimera organism Clinical Clustered Regularly Interspaced Short Palindromic Repeats Custom Engineered Gene Engineering Ensure Future Gene Expression Genes Genetic Genetic Engineering Genome Goals Growth HIV Herpes encephalitis Immune Impaired cognition In Vitro Infection Injury Institution Interferons Knock-out Knockout Mice Lentivirus Libraries Light Link Macrophage Mediating Mediator Mentors Mentorship Methods Microglia Molecular Mus Neuroglia Neuroimmune Neurologic Neurologist Neurology Neurons Nucleic Acids Pathogenesis Pathway interactions Patients Pediatric Hospitals Pharmaceutical Preparations Philadelphia Point Mutation Postdoctoral Fellow Predisposition Production Recombinant adeno-associated virus (rAAV)Regulation Research Resistance Resources Role Signal Transduction Spinal Cord Stimulator of Interferon Genes Students Surveys Testing Tissues Training Transgenic Mice Transplantation Tropism Validation Vesicular stomatitis Indiana virus Viral Viral Physiology Virus Diseases Vision Work antiviral immunity brain cell design frontier gain of function gene therapy in vivo inhibitor insight lentivirally transduced leukodystrophy multidisciplinary nervous system disorder neuroimmunologic disease novel strategies novel therapeutics preservation prevent professor programs receptor residence response sensor single-cell RNA sequencing targeted treatment therapeutic target therapy development tool transcriptome transcriptomics transduction efficiency

项目摘要

PROJECT SUMMARY/ABSTRACT Microglia are the long-lived resident macrophages of the brain and spinal cord, which constantly survey the microenvironment to respond rapidly to injury and infection. Unlike neurons or other glia, microglia are transplantable, making them ideal candidates for targeted therapies to treat neurological diseases. A major barrier to creating and refining such treatments is an inability to genetically engineer microglia with lentiviruses. This proposal aims to identify and overcome microglia restriction factors to enable gene editing with lentiviruses and gain insights into microglia-mediated antiviral immunity. Aim 1 will test the hypothesis that cGAS/STING, a known anti-viral pathway, underlies microglial viral restriction using inhibitors and transgenic mouse microglia in vitro, with in vivo validation using a new microglia-specific Cre mouse. In Aim 2, I will perform an unbiased microglia-focused CRISPR knockout screen using newly created AAVs with microglia tropism to discover new anti-viral pathways that prevent lentiviral infection. Aim 2 demonstrates the first CRISPR screen in primary microglia. This Early Independence application offers a vision for my independent research program. It is based on a platform of microglia-based discovery tools I created over the last decade, and driven by my work as a pediatric neurologist who cares for children with neuroimmunological conditions including leukodystrophies and interferonopathies. It will provide new approaches to genetic targeting of microglia for neurotherapeutics, direct testing of microglia antiviral immunity, and will influence future directions related to the neurological sequelae of HIV latency and the pathogenesis of neuroimmunological disease.

项目概要/摘要小胶质细胞是大脑和脊髓中长寿命的常驻巨噬细胞，它不断地调查微环境以对伤害和感染做出快速反应。与神经元或其他神经元不同神经胶质细胞、小胶质细胞是可移植的，使它们成为靶向治疗的理想候选者神经系统疾病。创建和完善此类治疗方法的一个主要障碍是无法用慢病毒对小胶质细胞进行基因改造。该提案旨在确定并克服小胶质细胞限制因子使慢病毒基因编辑成为可能并深入了解小胶质细胞介导的抗病毒免疫。目标 1 将检验以下假设：cGAS/STING，a 已知的抗病毒途径，是使用抑制剂和转基因限制小胶质细胞病毒的基础小鼠小胶质细胞体外实验，并使用新型小胶质细胞特异性 Cre 小鼠进行体内验证。瞄准 2，我将使用新创建的以小胶质细胞为中心的 CRISPR 敲除筛选具有小胶质细胞趋向性的 AAV 可以发现预防慢病毒感染的新抗病毒途径。目标 2 展示了原代小胶质细胞中的首次 CRISPR 筛选。早期的独立申请为我的独立研究计划提供了一个愿景。它是基于一个平台我在过去十年中创建了基于小胶质细胞的发现工具，并受到我作为一名科学家的工作的推动儿科神经科医生，照顾患有神经免疫疾病的儿童，包括脑白质营养不良和干扰素病。它将为基因靶向提供新方法小胶质细胞用于神经治疗，直接检测小胶质细胞抗病毒免疫力，并将影响与艾滋病毒潜伏期的神经系统后遗症和发病机制相关的未来方向神经免疫疾病。