ATG13: A new player in ME/CFS

ATG13：ME/CFS 的新玩家

• 批准号: 10666899
• 负责人: AVIK ROY
• 金额: $ 22.5万
• 依托单位: SIMMARON RESEARCH INC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10666899
• 关键词: Ablation; Acute; Adopted; Afferent Neurons; Age; Agonist; Anxiety; Apoptosis; Attention; Attentional deficit; Autonomic Dysfunction; Autophagocytosis; Blood Pressure; Brain; Brain Stem; Calcium; Cardiopulmonary; Cells; Chronic; Chronic Disease; Chronic Fatigue Syndrome; Clinical; Cognitive deficits; Computer software; Data; Demyelinations; Development; Disease; Disease model; Dizziness; Drops; Energy Metabolism; Exercise; Exercise Test; Exertion; Exhibits; FRAP1 gene; Fatigue; Female; Genes; Growth; Hand Strength; Heart Rate; Homeostasis; Impaired cognition; Impairment; Inflammation; Inflammatory; Investigation; Knock-out; Knockout Mice; Malaise; Measurement; Measures; Mental Depression; Metabolic; Mitochondria; Molecular; Molecular Analysis; Monitor; Mononuclear; Mouse Strains; Mus; Muscle; Muscle Fatigue; Muscle Weakness; Mutation; Myalgia; Neurocognitive Deficit; Pain; Pathogenesis; Pathology; Pathway interactions; Patients; Peripheral; Peripheral Nervous System; Pharmaceutical Preparations; Play; Production; Proteins; Protocols documentation; RANTES; Regimen; Role; Running; Spinal Cord; Spinal Ganglia; Spleen; Stress; Symptoms; System; Tamoxifen; Testing; Th1 Cells; Tissues; anaerobic glycolysis; brain cell; chronic inflammatory disease; knock-down; lymph nodes; motor impairment; mouse model; neural; novel; orthostatic intolerance; treadmill

Project summary: Myalgic encephalomyelitis and chronic fatigue syndrome (ME/CFS) is a multisystem chronic illness characterized by extreme fatigue, muscle weakness, muscle pain, dizziness, a cognitive deficit of attention, and depression. Despite intense investigation, the molecular mechanism of this disease is still unknown. Our recent finding suggests that the functional inactivation of ATG13, an essential protein of cellular autophagy, contributes to the pathogenesis of ME/CFS. To further characterize the role of ATG13 in the pathogenesis of ME/CFS, a reliable disease model is required that exhibits some of the cardinal disease symptoms such as post-exertional malaise (PEM) and orthostatic intolerance (OI). PEM is characterized by severe muscle fatigue and pains immediately after exercise, whereas OI is characterized by the sudden drop in blood pressure in an upright condition. Our preliminary results suggest that MHY1485, an agonist of mammalian target of rapamycin (mTOR), inactivates ATG13-dependent autophagy and induces severe fatigue and PEM in 3-4 weeks old female mice. Therefore, to establish the direct role of Atg13 in ME/CFS pathogenesis, our overall hypothesis is that atg13 depletion will display a spontaneous PEM pathology. In aim1, we will generate a mouse strain with muscle-specific knock-out of the atg13 gene (atg13∆muscle). A 20-minute acute treadmill running protocol and 2-days CPET tests would be adopted in these mice followed by simultaneous measurement of M- wave amplitude through EMG recording, decreased mobility recorded by Stoelting ANY-maze tracking software, muscle pain, stress, and neurocognitive impairment of attention (Aim 1.1). Molecular analysis of mitochondrial impairment in energy metabolism, calcium homeostasis, apoptosis, and anaerobic glycolysis would be evaluated in these mice (Aim 1.2). Under aim2, we would see if neural mutation of atg13 in myelinating cells of the brain (atg13∆brain) or ablation of atg13 in sensory neurons (atg13∆PNS) contributes to the pathogenesis of ME/CFS such as autonomic dysfunction, the cognitive deficit of attention, anxiety, and pain. Briefly, atg13∆brain and atg13∆PNS mice would be generated and then analyzed for chronic fatigue, followed by a 2-Days CPET test. After another 3-4 weeks, muscle fatigue, pain, stress, and neurocognitive impairment (aim 2.1) would be evaluated. Since chronic inflammation is one of the cardinal symptoms of ME/CFS, we would assess peripheral inflammation of CD4 Th1 cell activation in the spleen- and lymph node-derived mononuclear cells (Aim 2.2). Central inflammation will be assessed by monitoring the activation of NF-κB, iNOS, and RANTES production in the brain and spinal cord (Aim 2.2). If successful, our current proposal identifies the first mouse model to study ME/CFS and ME/CFS- associated PEM.

项目概要：肌痛性脑脊髓炎和慢性疲劳综合征（ME/CFS）是一个多系统疾病 以极度疲劳、肌肉无力、肌肉疼痛、头晕、认知缺陷为特征的慢性疾病 注意力、抑郁症。尽管进行了大量研究，但该疾病的分子机制仍不清楚 未知。我们最近的发现表明，细胞必需蛋白 ATG13 的功能失活 自噬有助于 ME/CFS 的发病机制。为了进一步表征 ATG13 在 ME/CFS 的发病机制，需要一个可靠的疾病模型来表现一些主要疾病 诸如劳累后不适 (PEM) 和直立不耐受 (OI) 等症状。质子交换膜的特点是 运动后立即出现严重的肌肉疲劳和疼痛，而 OI 的特点是运动强度突然下降 直立状态下的血压。我们的初步结果表明，MHY1485，哺乳动物的激动剂 雷帕霉素 (mTOR) 的靶点，使 ATG13 依赖性自噬失活，并在体内诱导严重疲劳和 PEM 3-4周大的雌性小鼠。因此，为了确定 Atg13 在 ME/CFS 发病机制中的直接作用，我们的总体 假设 atg13 耗尽将表现出自发的 PEM 病理。在 Target1 中，我们将生成一个 肌肉特异性敲除 atg13 基因 (atg13Δmuscle) 的小鼠品系。 20分钟跑步机急性跑 将在这些小鼠中采用方案和 2 天的 CPET 测试，然后同时测量 M- 通过 EMG 记录的波振幅，通过 Stoelting ANY-maze 跟踪软件记录的移动性下降， 肌肉疼痛、压力和注意力的神经认知障碍（目标 1.1）。线粒体的分子分析 将评估能量代谢、钙稳态、细胞凋亡和无氧糖酵解的损害 在这些小鼠中（目标 1.2）。在目标2下，我们将了解大脑髓鞘细胞中atg13的神经突变是否 (atg13Δbrain) 或感觉神经元中 atg13 的消融 (atg13ΔPNS) 有助于 ME/CFS 的发病机制，例如 如自主神经功能障碍、注意力、焦虑和疼痛的认知缺陷。简而言之，atg13Δbrain 和 atg13ΔPNS 将生成小鼠，然后分析其慢性疲劳情况，然后进行 2 天的 CPET 测试。又一个 3-4 周后，将评估肌肉疲劳、疼痛、压力和神经认知障碍（目标 2.1）。自从 慢性炎症是 ME/CFS 的主要症状之一，我们将评估周围炎症 脾脏和淋巴结来源的单核细胞中 CD4 Th1 细胞的激活（目标 2.2）。中枢炎症 将通过监测大脑和脊髓中 NF-κB、iNOS 和 RANTES 生成的激活情况进行评估 绳（目标 2.2）。如果成功，我们目前的提案将确定第一个研究 ME/CFS 的小鼠模型以及 ME/CFS- 相关的PEM。