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.

肌痛性脑脊髓炎和慢性疲劳综合征（ME/CFS）是一个多系统 慢性疾病的特点是极度疲劳，肌肉无力，肌肉疼痛，头晕，认知缺陷， 注意力和抑郁症尽管进行了深入的研究，但这种疾病的分子机制仍然是 未知我们最近的发现表明，细胞内必需的蛋白质ATG 13的功能失活， 自噬是ME/CFS的发病机制。为了进一步表征ATG 13在细胞凋亡中的作用， ME/CFS的发病机制，需要一个可靠的疾病模型，表现出一些主要疾病 运动后不适（PEM）和直立不耐受（OI）等症状。PEM的特征在于： 严重的肌肉疲劳和疼痛后立即运动，而OI的特点是突然下降， 直立状态下的血压。我们的初步结果表明，MHY 1485，哺乳动物的激动剂， 雷帕霉素（mTOR）的靶点，灭活ATG 13依赖性自噬，并诱导严重的疲劳和PEM， 3-4几周大的雌性老鼠因此，为了确定Atg 13在ME/CFS发病机制中的直接作用，我们的总体研究结果表明， 假设atg 13耗竭将显示自发PEM病理。在aim 1中，我们将生成一个 肌肉特异性敲除atg 13基因的小鼠品系（atg 13肌肉）。一个20分钟的急性跑步机运行 方法：采用常规方法和2天CPET试验，同时测定小鼠的M- 通过EMG记录的波幅，通过Stoelting ANY迷宫跟踪软件记录的移动性下降， 肌肉疼痛、压力和注意力神经认知障碍（目标1.1）。线粒体的分子分析 评估能量代谢、钙稳态、细胞凋亡和无氧糖酵解的损伤 在这些小鼠中（目标1.2）。在aim 2下，我们将观察大脑髓鞘细胞中atg 13的神经突变是否 （atg 13脑）或感觉神经元中atg 13的消融（atg 13脑PNS）有助于ME/CFS的发病机制， 如自主神经功能障碍，注意力的认知缺陷，焦虑和疼痛。简单地说，atg 13 β-brain和atg 13 β-PNS 产生小鼠，然后分析慢性疲劳，接着进行2天CPET试验。个又一个 3-4周，将评估肌肉疲劳、疼痛、压力和神经认知障碍（目标2.1）。以来 慢性炎症是ME/CFS的主要症状之一，我们将评估外周炎症， 脾和淋巴结来源的单核细胞中的CD 4 Th 1细胞活化（目标2.2）。中枢性炎症 将通过监测脑和脊髓中NF-κB、iNOS和RANTES产生的活化来评估 软线（目标2.2）。如果成功，我们目前的提议确定了第一个研究ME/CFS和ME/CFS的小鼠模型- 相关PEM