Exosomal-mediated pyroptosis in ventilation-associated preterm brain injury

通气相关早产性脑损伤中外泌体介导的细胞焦亡

• 批准号: 10039670
• 负责人: Augusto F Santos Schmidt
• 金额: $ 15.55万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-15 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10039670
• 关键词: Address; Adoptive Transfer; Advisory Committees; Age-Months; Animals; Area; Blood - brain barrier anatomy; Blood Circulation; Brain; Brain Injuries; Bronchopulmonary Dysplasia; CASP1 gene; Caspase; Cell Death; Cell membrane; Cells; Cessation of life; Clinical; Collaborations; Complement; Data; Development; Dyes; Enzyme-Linked Immunosorbent Assay; Face; Fellowship; Foundations; Genetic; Goals; Hyperoxia; Immunohistochemistry; Infant; Inflammation; Inflammatory; Injury; Intervention; Knock-out; Knockout Mice; Knowledge; Label; Laboratories; Life; Link; Low Birth Weight Infant; Lung; Lytic; Mechanical ventilation; Mechanics; Mediating; Medical Students; Mentors; Modeling; Morbidity - disease rate; Mus; Neonatal; Neonatal Brain Injury; Neonatology; Neurodevelopmental Impairment; Neurodevelopmental Problem; Neurons; Newborn Infant; Outcome; Paralysed; Pathogenesis; Pathway interactions; Pediatric Hospitals; Pediatrics; Pharmacology; Physicians; Placebos; Play; Population; Postdoctoral Fellow; Premature Infant; Preterm brain injury; Principal Investigator; Process; Randomized; Rattus; Research; Research Proposals; Residencies; Risk; Rodent; Role; Savings; Scientist; Serum; Survival Rate; Techniques; Testing; Therapeutic; Therapeutic Intervention; Tidal Volume; Time; Training; Training Programs; Universities; Water; Western Blotting; brain cell; career; career development; cytokine; dalton; doctoral student; ex vivo imaging; exosome; genetic approach; high risk infant; improved; inhibitor/antagonist; insight; interest; interleukin-1beta-converting enzyme inhibitor; loss of function; lung injury; mouse model; myelination; neonatal lung injury; neurobehavioral; neurobehavioral test; neuroinflammation; neuron loss; neurosurgery; new therapeutic target; novel; preterm newborn; prevent; professor; pulmonary function; respiratory; skills; student mentoring; ventilation; vesicular release

ABSTRACT This proposal describes a five-year training program and research proposal for the career development of Dr. Augusto Schmidt as a physician-scientist in Neonatology with expertise in neonatal brain injury. Dr. Schmidt completed his Pediatrics residency and Neonatology fellowship at Cincinnati Children’s Hospital and is currently Assistant Professor of Pediatrics in the Division of Neonatology at the University of Miami (UM). Dr. Schmidt is interested in determining the mechanistic relationship between mechanical ventilation and brain injury in preterm newborns, and is particularly interested in how mechanical ventilation causes the lung to release exosomes, which may induce brain cellular death and injury in preterm newborns. Dr. Schmidt will be mentored by Dr. Dalton Dietrich and Dr. Shu Wu with the support of a scientific advisory committee. Dr. Dietrich, a world-renowned expert in brain injury, will mentor Dr. Schmidt’s scientific progress and career development. Dr. Dietrich is the Director of the Miami Project to Cure Paralysis and Distinguished Chair of Neurosurgery at UM and has been committed to training scientists throughout his career having served as the principal investigator of a T32 Training Program, thesis advisor of 9 PhD students, and mentored over 30 postdoctoral fellows. Dr. Wu is a neonatologist and expert in neonatal lung injury and will complement Dr. Dietrich as a co-mentor. Dr. Wu has successfully mentored more than 20 clinical and postdoctoral fellows and medical students. The advisory committee will support Dr. Dietrich and Dr. Wu in mentoring Dr. Schmidt and will consist of Dr. Roberta Brambilla, an expert in neuroinflammation, Dr. Eduardo Bancalari, a neonatologist and expert in neonatal chronic lung disease and Dr. Alan Jobe, a neonatologist and accomplished physician-scientist, who has previously mentored Dr. Schmidt. In this proposal, Dr. Schmidt will test the hypothesis that mechanical ventilation causes the lung to release exosomes containing caspase-1 and that these exosomes cross the blood-brain barrier to activate Gasdermin D (GSDMD) in the brain, inducing cellular death and brain injury. Dr. Schmidt will use a survival model of neonatal rodent ventilation to test the following aims: (1) to determine the mechanistic function of GSDMD in inducing neuronal cell death in ventilation-associated brain injury (VABI); (2) to determine the mechanistic function of pulmonary-derived exosomal caspase-1 in activating neuronal GSDMD in VABI. Dr. Schmidt will use a combination of pharmacological and genetic loss-of function mice models for GSDMD and caspase-1 to address these aims. Brain injury and cellular death will be determined by a combination of techniques including immunohistochemistry, western blotting, ELISA, and real time-PCR. Long-term outcomes will be assessed by neurobehavioral testing. These pharmacological and genetic approaches will delineate the role of the exosomal caspase-1 / neuronal GSDMD pathway in the development of VABI. This research will provide insight into new pathways for therapeutic intervention to prevent or treat brain injury in ventilated preterm newborns and provide the foundation for Dr. Schmidt’s research career focused on neonatal brain injury.

抽象的 本提案描述了博士职业发展的五年培养计划和研究建议。 奥古斯托·施密特 (Augusto Schmidt) 是一名新生儿科医师科学家，拥有新生儿脑损伤方面的专业知识。 在辛辛那提儿童医院完成了儿科住院医师培训和新生儿科研究金，目前正在 施密特博士是迈阿密大学 (UM) 新生儿科的儿科助理教授。 对确定早产儿机械通气与脑损伤之间的机制关系感兴趣 新生儿，并且对机械通气如何导致肺部释放外泌体特别感兴趣， 施密特博士将接受道尔顿博士的指导，这可能会导致早产儿的脑细胞死亡和损伤。 Dietrich 和 Shu Wu 博士在世界著名的 Dietrich 博士的科学顾问委员会的支持下。 脑损伤专家，将指导施密特博士的科学进步和职业发展。 迈阿密大学瘫痪治疗项目主任兼神经外科杰出主席 作为 T32 培训项目的首席研究员，他在整个职业生涯中致力于培训科学家 吴博士是一名新生儿科医生，指导了 9 名博士生，并指导了 30 多名博士后。 作为新生儿肺损伤方面的专家，吴博士将成功补充 Dietrich 博士的指导。 顾问委员会将指导 20 多名临床和博士后研究员以及医学生。 支持 Dietrich 博士和 Wu 博士指导 Schmidt 博士，其中包括 Roberta Brambilla 博士，他是以下领域的专家 神经炎症，新生儿学家和新生儿慢性肺病专家 Eduardo Bancalari 博士和 艾伦·乔布 (Alan Jobe) 是一位新生儿学家，也是一位卓有成就的医师科学家，曾指导过施密特博士。 根据这项提议，施密特博士将测试机械通气导致肺部释放的假设 含有 caspase-1 的外泌体，这些外泌体可以穿过血脑屏障激活 Gasdermin 施密特博士将使用新生儿的生存模型。 啮齿类动物通气试验目的如下：（1）确定GSDMD诱导通气的机制功能 (2) 确定通气相关性脑损伤（VABI）中神经细胞死亡的机制功能； Schmidt 博士将使用肺源性外泌体 caspase-1 激活 VABI 中的神经元 GSDMD。 GSDMD 和 caspase-1 的药理学和遗传功能丧失小鼠模型相结合，以解决 这些目标将通过多种技术来确定，包括： 将通过免疫组织化学、蛋白质印迹、ELISA 和实时 PCR 来评估长期结果。 这些药理学和遗传学方法将描述外泌体的作用。 caspase-1/神经元 GSDMD 通路在 VABI 发展中的作用将提供新的见解。 预防或治疗通气早产儿脑损伤的干预治疗途径，并提供 施密特博士的研究生涯奠定了新生儿脑损伤的基础。