Exosomal Gasdermin D Mediated Lung to Brain Crosstalk in Preterm Brain Injury

摘要 我衷心感谢NIH给我这个机会申请这份行政副刊，以延续 美国国立卫生研究院研究项目资助首次获得者的生物医学和行为研究 面临重大人生事件的奖项(非OD-23-032)。我的研究目前得到了R01拨款的支持 来自NHLBI(1RO1HL156803-01A1)，我于2020年8月获得。这笔赠款的主要目标是 确定循环细胞外小泡(EVS)介导肺到脑的串扰和 它们在早产儿脑损伤和神经发育障碍(NDI)中的作用。电动汽车扮演着至关重要的角色 在器官间沟通中的作用。上睑下垂是一种新描述的炎性细胞死亡形式，仅 受Gasdermin D(GSDMD)调节，GSDMD是一种炎症体激活的膜“造孔”蛋白。激活 GSDMD的发生依赖于caspase-1的激活，caspase-1是通过组装一个炎症体复合体来实现的 与凋亡相关的斑点样蛋白含有一张卡片(ASC)。我们的研究表明，早产 患有慢性肺部疾病--支气管肺发育不良(BPD)的婴儿表达增加。 1周龄循环电动汽车中的ASC和GSDMD。重要的是，这些电动汽车的采用转移到 正常新生小鼠的血液循环导致肺部炎症和脑发育受损。这些 这些发现导致我们提出了一个新的模型，将BPD、EV-ASC、EV-GSDMD和脑损伤联系起来。中环 该模型的假设是含有ASC和GSDMD的EV从肺释放到 BPD发展阶段的循环和脑神经元对这些EVS的摄取导致神经细胞 上睑下垂和NDI。我们在家长拨款中提出了三个具体目标来检验这一假设。在……里面 这份行政副刊，我们将集中研究工作的具体目标1，以进一步 确定EV-ASC和EV-GSDMD的来源，它们是否来自肺泡上皮细胞 肺泡巨噬细胞(AM)。在具体目标2中，我们将开发新的方法来分离AEC来源的 EVS和AM来源EVS及其对脑损伤和NDI影响的过继转移实验 在新生的老鼠身上。

ABSTRACT I sincerely thank NIH for the opportunity to apply for this Administrative Supplement for Continuity of Biomedical and Behavioral Research Among First-Time Recipients of NIH Research Project Grant Awards Facing Critical Life Events (NOT-OD-23-032). My research is currently supported by an R01 grant from NHLBI (1RO1HL156803-01A1), which I obtained in August 2020. The major goal of this grant is to determine the mechanisms by which circulating extracellular vesicles (EVs) mediate lung-to-brain crosstalk and their contribution to brain injury and neurodevelopmental impairment (NDI) in preterm infants. EVs play a crucial role in inter-organ communications. Pyroptosis is a newly described form of inflammatory cell death that is solely regulated by gasdermin D (GSDMD), an inflammasome-activated membrane “pore-forming” protein. Activation of GSDMD depends on the activation of caspase-1, which is achieved by assembling an inflammasome complex with the apoptosis-associated speck-like protein containing a card (ASC). Our studies demonstrated that preterm infants who develop bronchopulmonary dysplasia (BPD), a chronic lung disease, have increased expression of ASC and GSDMD in their circulating EVs at 1 week of age. Importantly, the adoptive transfer of these EVs into the circulation of normal newborn mice induced lung inflammation and impaired brain development. These findings led us to propose a novel model that links BPD, EV-ASC, EV-GSDMD, and brain injury. The central hypothesis of this model is that ASC-containing and GSDMD-containing EVs are released from the lung into the circulation during the evolving stage of BPD, and brain neuronal uptake of these EVs results in neural cell pyroptosis and NDI. We proposed three Specific Aims in the parent grant to test this hypothesis. In this Administrative Supplement, we will concentrate our research efforts on Specific Aim 1 to further characterize the sources of EV-ASC and EV-GSDMD, whether they are derived from alveolar epithelial cells (AEC) or alveolar macrophages (AM). In Specific Aim 2, we will develop new methods to isolate AEC-derived EVs and AM-derived EVs and determine their effects on brain injury and NDI by adoptive transfer experiments in newborn mice.