Amyloid Beta CAR Macrophages: a cell engineering strategy to clear pathogenic proteins

淀粉样蛋白 Beta CAR 巨噬细胞：清除致病蛋白的细胞工程策略

基本信息

批准号：
10562093
负责人：
Frederick Bennett
金额：
$ 71.08万
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：
2023-01-21 至 2027-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10562093
关键词：
Abeta clearance Acute Address Affect Alzheimer&apos s Disease Alzheimer&apos s disease risk Alzheimer&apos s disease therapy Amyloid Amyloid beta-Protein Antigen Receptors Behavior Behavioral Assay Behavioral Symptoms Binding Biochemical Biochemistry Biology Brain CSF1R gene Cell Therapy Cells Cellular immunotherapy Chronic Collaborations Data Engineering Engraftment Ensure Epitopes Foundations Genetic Gills Gliosis Goals Histology Hyperactive behavior Immune Immune system Immunotherapy Impairment Inflammasome Inflammatory Inflammatory Response Injections Intravenous Knowledge Lead Macrophage Measures Methods Microglia Modeling Monoclonal Antibodies Mus Nerve Degeneration Passive Immunization Pathogenesis Pathogenicity Pathology Patients Phagocytes Phagocytosis Principal Investigator Proteins Research Resource Sharing Role Signal Transduction Specimen TREM2 gene Testing Therapeutic Tissues Transplantation abeta oligomer amyloid pathology beta amyloid pathology brain cell cellular engineering chemotherapy chimeric antigen receptor cytokine design gain of function human disease immunoregulation improved in vivo individualized medicine knowledge base loss of function mouse model neuroinflammation neuropathology prevent programs response targeted treatment tool transcriptomics uptake

Abeta clearance Acute Address Affect Alzheimer&apos s Disease Alzheimer&apos s disease risk Alzheimer&apos s disease therapy Amyloid Amyloid beta-Protein Antigen Receptors Behavior Behavioral Assay Behavioral Symptoms Binding Biochemical Biochemistry Biology Brain CSF1R gene Cell Therapy Cells Cellular immunotherapy Chronic Collaborations Data Engineering Engraftment Ensure Epitopes Foundations Genetic Gills Gliosis Goals Histology Hyperactive behavior Immune Immune system Immunotherapy Impairment Inflammasome Inflammatory Inflammatory Response Injections Intravenous Knowledge Lead Macrophage Measures Methods Microglia Modeling Monoclonal Antibodies Mus Nerve Degeneration Passive Immunization Pathogenesis Pathogenicity Pathology Patients Phagocytes Phagocytosis Principal Investigator Proteins Research Resource Sharing Role Signal Transduction Specimen TREM2 gene Testing Therapeutic Tissues Transplantation abeta oligomer amyloid pathology beta amyloid pathology brain cell cellular engineering chemotherapy chimeric antigen receptor cytokine design gain of function human disease immunoregulation improved in vivo individualized medicine knowledge base loss of function mouse model neuroinflammation neuropathology prevent programs response targeted treatment tool transcriptomics uptake

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项目摘要

Phagocytosis by microglia, the brain's resident macrophages, is central to Alzheimer Disease (AD) risk and pathogenesis. Current β-amyloid (Aβ) clearing immunotherapies use monoclonal antibodies to indirectly elicit phagocytosis, in an attempt to reverse AD manifestations. This form of passive immunization induces microglial reactivity and neuroinflammation, two features abundant in AD neuropathologic specimens. Due to a dearth of tools, prior mechanistic study of microglia-Aβ interactions used loss of function strategies such as Trem2 KO or microglial depletion, leaving uncertain whether microglial Aβ phagocytosis itself is sufficient to alter neuropathology. To address this knowledge gap, we clearance engraftment test engineered macrophages to achieve specific amyloid with antigen receptors ( AβCAR) , and developed methods for efficient macrophage in the brain. This dual PI proposal unites experts in immune cell therapies and microglial biology to the overarching hypothesis that Aβ-chimeric AβCAR-engineered macrophages effectively clear amyloid in vivo and that associated neuroinflammation is prevented by manipulation of the inflammasome. In preliminary data we successfully engineered AβCAR engulf into determine engraft neuropathology and AβCAR exposure NLRP3 on transplant determine in Aim inflammasome close to expressing macrophages, found that they specifically amyloid chemotherapy-dependent and - independent methods to transplant macrophages the mouse brain at unprecedented efficiency, ensuring the feasibility of this proposal. In Aim 1 we will how active cell-based Aβ targeting affects amyloid pathology in murine models. To do so, we will AβCAR macrophages into the 5xFAD model of Aβ pathology and measure their effects on and behavior. Aim 1 will clarify the controversial relationship between microglial Aβ phagocytosis pathology using a cell engineering strategy with high t herapeutic potential. Where Aim 1 focuses on how macrophages affect the brain Aβ, Aim 2 addresses how brain Aβ in turn affects macrophages. Amyloid induces microglial state changes and neuroinflammation, with strong mechanistic data implicating inflammasome signaling as a major driver. Aim 2 will therefore elucidate the effects of AβCAR targeting microglial reactivity, neuroinflammation, and their degree of rescue by NLRP3 eficiency. To do so, we will AβCAR macrophages engineered from Nlrp3 KO or Nlrp3 WT donor mice into the 5xFAD model and a) the degree to which CAR-augmented Aβ uptake increases macrophage inflammatory profiles and turn neuroinflammation b) whether this is rescued by Nlrp3 loss, and c) the consequences on Aβ pathology. 2 will improve our understanding of how immunotherapies lead to neuroinflammation and determine whether inhibition is a strategy to enhance cell therapies for neurodegeneration. Together, these aims will knowledge gaps about the role of microglial phagocytosis in AD treatment and create a cell-based strategy specifically target proteins in the CNS, using principles our multi-PI team has already tested in human disease. , and developed d

小胶质细胞吞噬作用，大脑的居民巨噬细胞是阿尔茨海默氏病（AD）风险和发病。当前的β-淀粉样蛋白（Aβ）清除免疫疗法使用单克隆抗体间接引起吞噬作用，试图逆转AD表现。这种被动免疫刺激诱导小胶质细胞反应性和神经炎症，在AD神经病理标本中丰富的两个特征。由于死亡工具，小胶质细胞-Aβ相互作用的先前机械研究使用了功能策略的丧失，例如TREM2 KO或小胶质部署，尚不确定小胶质细胞Aβ吞噬作用本身是否足以改变神经病理学。为了解决这个知识差距，我们清除植入测试工程巨噬细胞以实现特定的淀粉样蛋白抗原受体（AβCAR），并开发了有效巨噬细胞的方法在大脑中。该双PI提案单位免疫细胞疗法和小胶质生物学专家总体假设 Aβ-晶体 AβCAR工程巨噬细胞有效地清除体内淀粉样蛋白并且通过操纵炎症体可以阻止相关的神经炎症。在初步数据我们成功地设计了AβCAR 吞噬进入决定灌输神经病理学和 AβCAR 接触 nlrp3 在移植决定在目的炎症关闭到表达巨噬细胞，发现他们专门淀粉样蛋白化学疗法依赖性和 - 独立的移植巨噬细胞方法小鼠大脑以前所未有的效率，确保了该提议的可行性。在目标1中，我们将在鼠模型中，活性细胞的Aβ靶向如何影响淀粉样病理。为此，我们会 AβCAR巨噬细胞进入Aβ病理的5xFAD模型，并测量其对和行为。 AIM 1将阐明小胶质Aβ吞噬作用之间有争议的关系使用具有较高T肌潜力的细胞工程策略的病理学。 AIM 1的重点是如何巨噬细胞影响大脑Aβ，AIM 2解决了大脑Aβ又如何影响巨噬细胞。淀粉样蛋白诱导小胶质状态变化和神经炎症，具有强大的机械数据暗示炎症的信号作为主要驱动力。因此，AIM 2将阐明AβCAR靶向的影响小胶质反应性，神经炎症及其通过NLRP3效率挽救程度。为此，我们会 AβCAR巨噬细胞从NLRP3 KO或NLRP3 WT供体小鼠设计为5xFAD模型，并 a）汽车增强的Aβ摄取的程度增加了巨噬细胞的炎症特征和转动神经炎症b）是否是通过NLRP3损失挽救的，c）对Aβ病理的后果。 2将提高我们对免疫疗法如何导致神经炎症的理解，并确定是否是否抑制是一种增强神经退行性细胞疗法的策略。这些目标将在一起关于小胶质细胞增多症在AD治疗中的作用的知识差距，并创建基于细胞的策略特别是在中枢神经系统中靶向蛋白质，使用我们的多PI团队已经在人类疾病中进行了测试。，开发 d