Pulmonary pathophysiology sub-phenotypes of pneumonia

肺炎会导致不良后果，包括住院、重症监护和死亡。主机响应为 关键，免疫活动和细胞功能障碍是病理生理的责任，包括但不限于 微生物过度生长，肺泡泛滥，隔膜破坏。许多不同的生物途径可以 导致感染肺部的不良后果。更好地了解免疫学和细胞学 需要在严重感染的肺部内发生活动，以区分疾病的亚型和 促进宿主导向疗法的开发和应用。在拟议的研究中，我们将利用 通过快速尸检收集的大量死后人类肺炎肺样本以确定肺炎 根据他们的肺部病理生物学进行亚型分析。我们将整合半定量的组织病理学 (由委员会认证的病理学家评分)，定量多重荧光免疫组织化学(mfIHC；to 对这些肺内的免疫细胞进行计数和定位)，以及单核RNA测序(SnRNA-seq；to 扩大和加深这些肺的细胞和分子分辨率)以区分不同的肺 肺炎期间的病理生物学。此外，我们还将检查多个电流和紧急实验 重症肺炎的模型，以确定这些肺的哪些元素(如果有)重现 病理生物学。我们建议对严重肺炎聚集成不同类型的中心假设进行检验。 肺病理生物学亚型，通过追求以下特定目标：目的1)测试人类 死亡肺炎分离成肺病理亚型，用快速尸检标本进行分析 通过组织病理学评分和mFIHC定量。目的2)验证和阐明肺炎亚单位 使用与SnRNA-seq匹配的独立队列显微镜来定义细胞类型和表型 感染人肺组织中细胞特异性基因的表达。目标3)测试现有的和新兴的 重症肺炎的实验模型概括了人类肺脏的一些和哪些元素 病理生物学。这项拟议的研究将对从肺和血液样本中产生新的发现具有重要意义 在老年人肺炎病例中，肺炎是一个特别高风险的群体。拟议的研究将 提高对肺炎异质性的分辨率，这是当前紧迫的研究重点。结果将 揭示肺炎肺内测量的亚表型或选定特征是否不同 肺炎球菌、流感和SARS-CoV-2引起的肺炎。确定肺炎的亚型 肺中不同的分子和细胞变化将改善宿主定向的发育和使用 治疗或预防肺炎的方法。

Pneumonia causes adverse outcomes including hospitalization, intensive care, and death. Host response is pivotal, with immune activities and cell dysfunctions responsible for pathophysiologies including but not limited to excessive microbial growth, alveolar flooding, and septal destruction. Many diverse biological pathways can lead to adverse outcomes within the infected lungs. A better understanding of the immunological and cellular activities occurring inside severely infected lungs is needed, to distinguish sub-phenotypes of disease and improve the development and application of host-directed therapies. In the proposed studies, we will leverage large sets of post-mortem human pneumonic lung samples collected via rapid autopsy for defining pneumonia sub-phenotypes based on their pulmonary pathobiology. We will integrate semi-quantitative histopathology (scored by board-certified pathologists), quantitative multiplex fluorescent immunohistochemistry (mfIHC; to enumerate and localize immune cells within these lungs), and single nuclei RNA sequencing (snRNA-seq; to broaden and deepen cell and molecular resolution of these lungs) in order to differentiate distinct lung pathobiologies during pneumonia. In addition, we will examine multiple current and emergent experimental models of severe pneumonia to determine which if any recapitulate elements of these pulmonary pathobiologies. We propose to test the central hypothesis that severe pneumonias cluster into distinct lung pathobiology sub-phenotypes, by pursuing the following specific aims: Aim 1) To test whether humans dying with pneumonia segregate into lung pathology sub-phenotypes, using rapid autopsy samples analyzed via histopathologic scoring and quantitative mfIHC. Aim 2) To validate and elucidate pneumonia sub- phenotypes using an independent cohort with microscopy matched to snRNA-seq for defining cell-types and cell-specific gene expression in infected human lungs. Aim 3) To test whether established and emerging experimental models of severe pneumonia recapitulate some and which elements of human pulmonary pathobiology. The proposed studies will be significant for generating discoveries from lung and blood samples of pneumonia cases in elderly subjects, an especially high risk group for pneumonia. Proposed studies will increase resolution into the heterogeneity of pneumonia, a currently pressing research priority. Results will reveal whether sub-phenotypes or select features measured within the pneumonic lungs differ between pneumonias caused by pneumococcus, influenza, and SARS-CoV-2. Defining sub-types of pneumonia with distinct molecular and cellular changes in the lungs will improve the development and use of host-directed approaches for treating or preventing pneumonia.