Exosomal-mediated pyroptosis in ventilation-associated preterm brain injury

摘要 这份提案描述了一项为期五年的培训计划和研究方案，以促进博士的职业发展。 奥古斯托·施密特是新生儿学的内科科学家，擅长新生儿脑损伤。施密特博士 在辛辛那提儿童医院完成儿科住院医师和新生儿科研究员学位，目前 迈阿密大学(UM)新生儿科儿科学助理教授。施密特医生是 有兴趣确定机械通气与早产儿脑损伤的机制关系 他特别感兴趣的是机械通风如何导致肺释放外周体， 这可能导致早产儿脑细胞死亡和损伤。施密特医生将由道尔顿医生指导 迪特里希和舒芜博士在一个科学顾问委员会的支持下进行了研究。迪特里希博士，世界著名的 脑损伤专家，将指导施密特博士的科学进步和职业发展。迪特里希博士是 迈阿密瘫痪治疗项目主任和密歇根大学神经外科杰出主席 在他的整个职业生涯中致力于培训科学家，曾担任T32培训的首席研究员 项目，9名博士生的论文导师，30多名博士后研究员。吴医生是一名新生儿科医生 他是新生儿肺损伤方面的专家，并将作为共同导师补充Dietrich博士。吴博士已经成功地 指导了20多名临床和博士后研究员和医学生。咨询委员会将 支持迪特里希博士和吴博士指导施密特博士，并将由罗伯塔·布拉比拉博士组成，他是 新生儿专家、新生儿慢性肺部疾病专家爱德华多·班卡拉里博士说，他是一名神经炎专家。 艾伦·乔布是一位新生儿科医生，也是一位颇有成就的内科科学家，此前曾指导过施密特博士。在……里面 在这项提议中，施密特博士将检验机械通风导致肺释放的假设 含有caspase-1的外切体以及这些外切体穿过血脑屏障激活Gasdermin D(GSDMD)在脑内，导致细胞死亡和脑损伤。施密特博士将使用新生儿存活模型 目的：(1)测定GSDMD在诱导大鼠肺损伤中的作用。 呼吸机相关性脑损伤(VABI)的神经细胞死亡；(2)确定VABI的机制 肺源性胞外caspase-1在激活VABI神经元GSDMD中的作用施密特博士将使用一种 联合应用GSDMD和caspase-1的药理学和遗传性功能丧失小鼠模型 这些目标。脑损伤和细胞死亡将通过组合技术来确定，包括 免疫组织化学、免疫印迹、酶联免疫吸附试验和实时荧光定量聚合酶链式反应。将通过以下方式评估长期结果 神经行为测试。这些药理学和遗传学方法将描述外体的作用。 Caspase-1/神经元GSDMD通路在VABI发生发展中的作用这项研究将为新的 治疗干预预防或治疗呼吸机早产儿脑损伤的途径和提供 施密特博士研究生涯的基础是新生儿脑损伤。

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.