Macrophage driven, profibrotic adrenergic nerve remodeling in SSc-ILD

SSc-ILD 中巨噬细胞驱动的促纤维化肾上腺素神经重塑

• 批准号: 10579990
• 负责人: Erica L Herzog
• 金额: $ 60.77万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-05 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10579990
• 关键词: Ablation; Adrenergic Agents; Adrenergic Receptor; Affect; American; Animal Model; Autoimmune Diseases; Axon; Biology; Bone Marrow; Cells; Cessation of life; Chemical Sympathectomy; Chimera organism; Cicatrix; Clinical; Coculture Techniques; Cutaneous; Cytometry; DCC gene; Dependence; Dermal; Diagnosis; Disease; Disease Progression; Event; FDA approved; Fibroblasts; Fibrosis; Fostering; Functional disorder; Human; Image; Immune; Inflammation; Injury; Interstitial Lung Diseases; Knockout Mice; Laminin; Link; Lung; Lung diseases; Macrophage; Modeling; Mus; NTN1 gene; Natural Immunity; Nerve; Nerve Regeneration; Neurosciences; Norepinephrine; Organ; Patients; Peripheral; Pharmaceutical Preparations; Process; Proteins; Pulmonary Fibrosis; Raynaud Disease; Receptor Activation; Receptor Inhibition; Research Personnel; Role; Sampling; Scleroderma; Signal Transduction; Slice; Sympathetic Ganglia; System; Testing; Therapeutic; Tissues; Visceral; afferent nerve; cost; fibrotic lung; improved outcome; in vivo; injury and repair; insight; interest; lung injury; microCT; migration; mouse model; nerve supply; neuronal guidance; pharmacologic; postnatal; receptor; repaired; side effect; single cell sequencing; translational study

Scleroderma is a difficult to treat and poorly understood autoimmune condition characterized by dermal and visceral fibrosis, sympathetic overactivity in the form of Raynaud’s phenomenon, and the accumulation of scar- promoting innate immune cells such as macrophages. Despite the well accepted clinical relationship between these entities, a mechanistic link has been neither proposed nor shown. Because we have a longstanding interest in the role of neuronal guidance proteins (NGPs) such as Netrin-1 in SSc-ILD, we decided to study whether Netrin-1 could connect these processes. Netrin-1 is a secreted, laminin-like protein that through attractive and repulsive interactions with dependence receptors regulates critical events in injury, inflammation, remodeling, and repair. In the developing CNS, NTN-1 guides the migration of axons, and in the periphery NTN- 1 guides both postnatal adrenergic innervation and sensory nerve regeneration via interactions with its attractive receptor deleted in colorectal carcinoma (DCC). However, despite growing interest in nerve associated mechanisms of injury and repair in peripheral organs, essentially nothing is known about how NTN-1 interacts with adrenergic innervation in the fibrotic lung and, while nerve-derived signals are increasingly recognized as regulators of macrophage function, the process by which macrophages might reciprocally maintain and remodel adrenergic nerves in the healthy and diseased lung is obscure. Given SSc’s known association with sympathetic dysfunction, better understanding of how adrenergic nerves are remodeled and maintained in SSc-ILD could allow fundamental insights into the neuroscience of fibrosis. In this application we use mouse modeling and SSc-ILD tissues to demonstrate aberrant adrenergic innervation, augmented Noradrenaline (NA), and NA- responsive fibroblasts in the fibrotic lung. These factors interact to drive fibrosis, which can be inhibited via chemical sympathectomy, pharmacologic inhibition of alpha1 adrenergic receptors, or specific deletion of NTN- 1 from macrophages. These findings support the hypothesis that macrophage derived signals such as Netrin-1 foster profibrotic adrenergic nerve remodeling in the lung and will be explored in three aims. This application unites world class investigators in lung biology, scleroderma, innate immunity, adrenergic innervation, Netrin-1, and neuroscience to conduct an integrated project using two animal models, cell specific knockout mice, shielded bone marrow chimeras, pharmacologic receptor activation and inhibition, ex vivo study of human cells and tissues, precision cut lung slices, single cell sequencing approaches, and microCT. Aim 1 will determine the mechanism through which adrenergic nerve derived NA drives pulmonary fibrosis. Aim 2 will determine the mechanism(s) through which Netrin-1+ macrophages regulates pulmonary fibrosis. Aim 3 will define how macrophage derived Netrin-1 controls post-injury adrenergic nerve remodeling. These studies have the potential to produce paradigm shifting results that will change the way we view fibrosis and potentially develop new treatments for SSc-ILD.

硬皮病是一种难以治疗且知之甚少的自身免疫性疾病，其特征在于皮肤和 内脏纤维化，雷诺现象形式的交感神经过度活跃，以及疤痕的积累， 促进先天免疫细胞如巨噬细胞。尽管广泛接受的临床关系， 在这些实体中，既没有提出也没有显示出机械联系。因为我们长期以来 由于对神经元引导蛋白（NGP）如Netrin-1在SSc-ILD中的作用感兴趣，我们决定研究 Netrin-1是否可以连接这些进程。Netrin-1是一种分泌的层粘连蛋白样蛋白， 与依赖性受体的吸引和排斥相互作用调节损伤，炎症， 重塑和修复。在发育中的CNS中，NTN-1引导轴突的迁移，而在外周中，NTN-1引导轴突的迁移。 1通过与其吸引力的相互作用指导出生后肾上腺素能神经支配和感觉神经再生。 受体缺失的大肠癌（DCC）。然而，尽管对神经相关的兴趣越来越大， 在外周器官的损伤和修复机制，基本上是什么都不知道NTN-1如何相互作用 纤维化肺中的肾上腺素能神经支配，而神经源性信号越来越被认为是 巨噬细胞功能的调节剂，巨噬细胞可能维持和重塑的过程 健康和患病肺中的肾上腺素能神经是模糊的。考虑到SSc与同情心 功能障碍，更好地了解SSc-ILD中肾上腺素能神经是如何重塑和维持的， 让我们对纤维化的神经科学有了基本的了解。在这个应用程序中，我们使用鼠标建模， SSc-ILD组织显示肾上腺素能神经支配异常、去甲肾上腺素（NA）增强和NA- 纤维化肺中的反应性成纤维细胞。这些因素相互作用导致纤维化，可以通过以下方式抑制纤维化 化学交感神经切除术，α 1肾上腺素能受体的药理学抑制，或NTN- 1巨噬细胞。这些发现支持巨噬细胞衍生的信号如Netrin-1 促进肺中的促纤维化肾上腺素能神经重塑，并将在三个目标中进行探索。本申请 联合世界一流的研究人员在肺生物学，硬皮病，先天免疫，肾上腺素能神经支配，Netrin-1， 和神经科学进行一个综合项目，使用两种动物模型，细胞特异性敲除小鼠，屏蔽 骨髓嵌合体、药理学受体激活和抑制、人类细胞的离体研究和 组织、精确切割肺切片、单细胞测序方法和microCT。目标1将决定 肾上腺素能神经源性NA驱动肺纤维化的机制。目标2将决定 Netrin-1+巨噬细胞调节肺纤维化的机制。目标3将定义如何 巨噬细胞衍生的Netrin-1控制损伤后肾上腺素能神经重塑。这些研究有可能 产生范式转变的结果，这将改变我们看待纤维化的方式，并可能开发新的 治疗SSc-ILD。