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.

项目摘要：肌痛性脑脊髓炎和慢性疲劳综合征(ME/CFS)是一个多系统的疾病 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病 注意力和忧郁。尽管进行了大量的研究，但该病的分子机制仍未得到证实。 未知。我们最近的发现表明，细胞内必需的蛋白质ATG13的功能失活 自噬，参与了ME/CFS的发病机制。为了进一步表征ATG13在生物信息学中的作用 ME/CFS的发病机制，需要一个可靠的疾病模型来显示一些主要疾病 运动后不适(PEM)和立位不耐受(OI)等症状。PEM的特点是 运动后立即出现严重的肌肉疲劳和疼痛，而OI的特征是突然下降 血压处于正常状态。我们的初步结果表明，哺乳动物的激动剂MHY1485 雷帕霉素靶标(MTOR)，抑制ATG13依赖的自噬并诱导严重疲劳和PEM 3-4周龄雌性小鼠。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的耗竭将表现为自发的PEM病理。在aim1中，我们将生成一个 肌肉特异敲除atg13基因的小鼠品系(atg13∆肌肉)。在跑步机上急速跑20分钟 这些小鼠将采用常规和为期2天的CPET试验，然后同时测量M- 通过肌电记录的波幅，通过Stoelting任意迷宫跟踪软件记录的移动性降低， 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方面的损害。 在这些小鼠中(目标1.2)。在AIM2下，我们将看到大脑髓鞘细胞中atg13的神经突变 (atg13∆脑)或消融感觉神经元中的atg13(atg13∆pns)在ME/CFs的发病机制中起重要作用。 作为自主神经功能障碍，注意力、焦虑和疼痛的认知缺陷。简单地说，atg13∆Brain和atg13∆PNS 将产生小鼠，然后分析慢性疲劳，然后进行为期2天的CPET测试。一个接一个 3-4周，评估肌肉疲劳、疼痛、应激和神经认知功能障碍(Aim 2.1)。自.以来 慢性炎症是ME/CFS的主要症状之一，我们将评估 脾和淋巴来源的单个核细胞中的CD4Th1细胞活化(目标2.2)。中枢性炎症 将通过监测脑和脊髓中核因子-κB、诱导型一氧化氮合酶和RANTES产物的激活来进行评估 CORD(目标2.2)。如果成功，我们目前的建议确定了第一个研究ME/CFS和ME/CFS的小鼠模型- 关联的PEM。