Elucidating and harnessing the molecular mechanisms of protective clearance in endogenous and engineered phagocytes

阐明和利用内源性和工程化吞噬细胞保护性清除的分子机制

• 批准号: 10729935
• 负责人: Adam Patrick Williamson
• 金额: $ 41.86万
• 依托单位: BRYN MAWR COLLEGE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-08 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10729935
• 关键词: Anti-Inflammatory Agents; Antibodies; Antigen-Antibody Complex; Antigens; Apoptotic; Autoantibodies; Autophagocytosis; Behavior; Binding; Biological; Biological Assay; Biological Models; Birth; Brain; CRISPR/Cas technology; Cell Culture Techniques; Cell model; Cell physiology; Cells; Central Nervous System; Central Nervous System Diseases; Childhood; Chloride Channels; Chlorides; Chronic; Collaborations; Collection; Data; Deposition; Development; Disease; Drosophila genus; Elements; Engineering; Excision; Extracellular Domain; Family; Functional disorder; Genes; Genetic; Goals; Homeostasis; Human; IgG Receptors; Immune; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Ingestion; Inherited; Knowledge; Learning; Ligands; Link; Lysosomes; Macrophage; Maps; Measures; Mediating; Membrane; Mendelian disorder; Methods; Modeling; Molecular; Mus; Mutate; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neuronal Ceroid-Lipofuscinosis; Orthologous Gene; Pathology; Phagocytes; Phagocytosis; Play; Process; Protein Dynamics; Proteins; Recovery; Retina; Retinal Degeneration; Rod Outer Segments; Role; Signal Transduction; Spielmeyer-Vogt Disease; Structure of retinal pigment epithelium; Synapses; System; Testing; Texas; Therapeutic; Time; Tissues; Transplantation; Tumor Antigens; Work; axon injury; cancer cell; design; extracellular; in vivo; mutant; novel; particle; programs; protein function; receptor; reconstitution; retinal rods; success; transplant model

Project Summary/Abstract Protective clearance describes the process of the removal of membrane-intact cells or parts of cells without induction of pro-inflammatory responses; it is a central mode of normal development and homeostasis across tissues and phyla. Executing this process in a regulated and anti-inflammatory fashion requires exquisite orchestration of a collection of activities including receptor-mediated phagocytosis, lysosome formation, and intracellular degradation. Protective clearance plays a particularly important role in maintaining function and homeostasis in the central nervous system (CNS). The lysosomal storage disorders are a broad family of diseases characterized by dysregulated protective clearance in the CNS. Batten disease is a class of 13 fatal neurodegenerative lysosomal storage disorders that usually appear in childhood and comprise the most common inherited pediatric neurodegenerative disease worldwide. The pathology of Batten disease is linked to synaptic dysfunction and auto antibody deposition in the CNS. All genetically mapped forms of the disease are monogenic, caused by mutations in one of 13 ceroid lipofuscinosis (cln) genes. Despite the success mapping the cln genes, the cell biological mechanisms governing the CLN proteins in space and time remain an open problem. Further understanding of the fundamental mechanisms underlying CLN protein function may identify new avenues to treat Batten disease. The goal of this proposal is to elucidate the molecular and cellular mechanisms underlying protective clearance in endogenous phagocytes and engineer the process in for therapy by programming phagocytes to eliminate auto antigen-antibody complexes in the CNS in an anti-inflammatory manner. This project will use three powerful model systems comprised of living phagocytes and defined targets to define the molecular and cellular mechanisms underlying protective clearance. In Aim 1, we will use a simplified cell model of protective clearance to explore a connection we recently discovered between a CLN protein and a conserved phagocyte receptor. In Aim 2, we will use a novel model of endogenous protective clearance in the retina to systematically define the functions of each CLN protein during protective clearance. In Aim 3, we will use our expertise in immune cell programming to engineer phagocytes that eliminate antigen-antibody complexes from the CNS via protective clearance and test these molecules in vivo in an advanced mouse CNS macrophage transplant model. Completion of these aims will clarify molecular mechanisms underlying protective clearance, define how Batten disease mutations dysregulate the process, and investigate the therapeutic potential of synthetic receptors to eliminate antigen-antibody complexes from the CNS in an anti-inflammatory manner.

项目摘要/摘要 保护性清除是指去除膜完好的细胞或部分细胞的过程。 在没有诱导促炎反应的情况下；它是正常发育和 组织和门之间的动态平衡。在受监管和抗炎的情况下执行此过程 时尚需要精心编排一系列活动，包括受体介导的活动 吞噬、溶酶体形成和细胞内降解。保护间隙起到了特别的作用 在维持中枢神经系统(CNS)的功能和动态平衡方面发挥重要作用。这个 溶酶体储存障碍是一大类疾病，其特征是保护功能失调。 在中枢神经系统的通关。巴顿病是一种致命性的13种神经变性溶酶体存储 通常出现在儿童时期的疾病，构成最常见的遗传性儿科疾病 世界各地的神经退行性疾病。巴顿病的病理与突触功能障碍有关 自身抗体沉积在中枢神经系统。这种疾病的所有基因图谱形式都是单基因的， 由13种蜡样脂褐素沉积症(CLN)基因中的一种突变引起。尽管成功地绘制了CLN 基因，调控CLN蛋白的细胞生物学机制在空间和时间上仍然是一个开放的 有问题。进一步了解CLN蛋白功能的基本机制可能会 寻找治疗巴顿病的新途径。这项提议的目标是阐明分子和 内源性吞噬细胞保护性清除的细胞机制和工程 通过编程吞噬细胞来清除体内的自身抗原抗体复合体来进行治疗 中枢神经系统的抗炎作用。该项目将使用三个强大的模型系统，包括 活的吞噬细胞和明确的靶点，以确定潜在的分子和细胞机制 保护净空。在目标1中，我们将使用保护性清除的简化细胞模型来探索 我们最近发现了CLN蛋白和一个保守的吞噬细胞受体之间的联系。在目标2中， 我们将使用一个新的视网膜内源性保护性清除模型来系统地定义 保护清除过程中各CLN蛋白的功能。在目标3中，我们将利用我们在免疫方面的专业知识 设计吞噬细胞的细胞编程，通过 在晚期小鼠中枢神经系统巨噬细胞移植中保护性清除和体内测试这些分子 模特。这些目标的完成将澄清保护清除的分子机制， 明确巴顿病突变是如何扰乱这一过程的，并研究 合成受体在抗炎中消除中枢神经系统中的抗原-抗体复合体 举止。