Exosomal-mediated pyroptosis in ventilation-associated preterm brain injury

摘要 本建议书描述了一个为期五年的培训计划和研究建议，为博士的职业发展。 奥古斯托施密特作为一个医生，科学家在新生儿脑损伤的专业知识。施密特博士 在辛辛那提儿童医院完成了他的儿科住院医师和新生儿奖学金，目前 迈阿密大学（UM）新生儿科学助理教授。施密特博士是 感兴趣的是确定机械通气和早产儿脑损伤之间的机械关系， 新生儿，特别感兴趣的是机械通气如何导致肺释放外泌体， 这可能导致早产新生儿的脑细胞死亡和损伤。施密特博士将由道尔顿博士指导 迪特里希和吴舒博士在一个科学顾问委员会的支持下。迪特里希博士，世界著名的 他是脑损伤方面的专家，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UM神经外科杰出主席， 在他的职业生涯中，他一直致力于培训科学家，曾担任T32培训的首席研究员。 计划，9名博士生的论文导师，并指导了30多名博士后研究员。吴医生是一位妇科医生 他是新生儿肺损伤方面的专家，将作为迪特里希博士的共同导师。吴博士成功地 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Dietrich博士和Wu博士指导施密特博士，并将由Roberta Brambilla博士组成，她是 Eduardo Bancalari博士是一位儿科医生和新生儿慢性肺病专家， 艾伦·乔布，一位神经学家和有成就的物理学家，他曾指导过施密特博士。在 施密特博士将检验机械通气导致肺释放的假设 外泌体含有半胱天冬酶-1，这些外泌体穿过血脑屏障激活Gasdermin D（GSDMD）在大脑中，诱导细胞死亡和脑损伤。施密特医生将使用新生儿存活模型 实验目的：（1）确定GSDMD诱导大鼠肺动脉高压的机制 通气相关脑损伤（VABI）中的神经元细胞死亡;（2）确定 肺源性外泌体caspase-1在VABI中激活神经元GSDMD中的作用施密特医生会用 GSDMD和半胱天冬酶-1的药理学和遗传功能丧失小鼠模型的组合， 这些目标。脑损伤和细胞死亡将通过以下技术的组合来确定， 免疫组化、蛋白质印迹、ELISA和真实的时间PCR。将通过以下方式评估长期结局： 神经行为测试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发生中的作用这项研究将提供新的见解 预防或治疗机械通气早产儿脑损伤的治疗干预途径， 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.