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

项目概要/摘要 保护性清除描述了去除膜完整细胞或部分细胞的过程 不诱导促炎反应;它是正常发育的中心模式， 跨组织和门的内稳态。执行这一过程中，在一个调节和抗炎 时尚需要一系列活动的精心编排，包括受体介导的 吞噬作用、溶酶体形成和细胞内降解。保护性间隙尤其重要 在维持中枢神经系统（CNS）的功能和体内平衡中起重要作用。的 溶酶体贮积症是一个广泛的疾病家族，其特征在于失调的保护性免疫缺陷。 CNS中的清除率。Batten病是一类13种致命的神经退行性溶酶体贮积症 通常出现在儿童期的疾病，包括最常见的遗传性儿科疾病， 神经退行性疾病巴顿病的病理学与突触功能障碍有关 以及自身抗体在CNS中的沉积。所有的基因图谱形式的疾病都是单基因的， 由13种蜡样质脂褐质沉积症（cln）基因中的一种突变引起。尽管成功绘制了cln 基因，在空间和时间上控制CLN蛋白的细胞生物学机制仍然是开放的。 问题.进一步了解CLN蛋白功能的基本机制， 寻找治疗巴滕氏病的新途径。这项提案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， 一种通过编程吞噬细胞以消除细胞中的自身抗原-抗体复合物来进行治疗的方法， 中枢神经系统以抗炎的方式。该项目将使用三个强大的模型系统， 活的吞噬细胞和确定的目标，以确定潜在的分子和细胞机制 保护间隙在目标1中，我们将使用简化的保护性清除细胞模型来探索 我们最近发现了CLN蛋白和保守的吞噬细胞受体之间的联系。在目标2中， 我们将使用视网膜内源性保护性清除的新模型来系统地定义 每种CLN蛋白在保护性清除过程中的功能。在目标3中，我们将利用我们在免疫方面的专业知识， 细胞编程以工程化吞噬细胞，其通过以下途径从CNS消除抗原-抗体复合物： 保护性清除，并在晚期小鼠CNS巨噬细胞移植物中体内测试这些分子 模型这些目标的完成将阐明保护性清除的分子机制， 定义Batten病突变如何失调这一过程，并研究 合成受体，以消除抗原抗体复合物从中枢神经系统的抗炎 方式

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.