Spatial analysis of Alcoholic Hepatitis Liver Tissue

酒精性肝炎肝组织的空间分析

• 批准号: 10261590
• 负责人: Jon Jacobs
• 金额: $ 24.38万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-10 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10261590
• 关键词: Address; Alcoholic Hepatitis; Alcoholic Liver Diseases; Anti-Inflammatory Agents; Apoptotic; Area; Biopsy; Cells; Cellular Morphology; Cessation of life; Cirrhosis; Collection; Coupling; Diagnosis; Disease; Disease Progression; Event; Fibrosis; Functional disorder; Goals; Heavy Drinking; Hepatocyte; Heterogeneity; Histologic; Histology; Hospitals; Infiltration; Inflammatory Infiltrate; Inflammatory Response; Link; Liver; Lobular; Mammalian Cell; Maps; Mass Spectrum Analysis; Molecular; Onset of illness; Pathology; Pathway interactions; Patients; Preparation; Protein Analysis; Proteins; Proteome; Public Health; Resolution; Sampling; Severities; Spectrum Analysis; Steroids; Surface; Technology; Tissues; Treatment Efficacy; United States; Variant; Work; analytical method; base; cohort; interest; laser capture microdissection; liver biopsy; mortality; nanoDroplet; patient response; problem drinker

SUMMARY Excessive alcohol consumption is a major global public health issue, with alcoholic liver disease (ALD) being a leading cause of liver-related death in the United States. Alcoholic hepatitis (AH) is a particularly serious form of ALD with high short term mortality rates, ~35% after one month. Although anti-inflammatory steroid treatments and other general therapies exist, patient response to such therapies varies widely. There is a need to understand both the mechanisms of AH disease onset and progression as well as identify markers of treatment efficacy. To this end, we aim to apply more precise analytical methods to move beyond gross tissue analytics and towards more cell specific and spatial protein abundances. Specifically, by more precisely analyzing the spectrum of AH liver tissue, we will gain a better understanding of how hepatocytes progress to apoptotic ballooning and the effect of these cells on adjacent hepatocytes particularly in moderate or confounded AH cases. We have developed a breakthrough technology, termed nanoPOTS (Nanodroplet Preparation in One pot for Trace Samples) which currently enables effective analysis of <10 mammalian cells with extensive proteome coverage. Coupling nanoPOTS with laser capture microdissection (LCM) and downstream high throughput and high sensitivity mass spectrometry analysis platforms now enables the technical capability to capture global protein abundances at the micro scale level. Our objective is to apply nanoPOTS to spatially map with high resolution heterogenous AH biopsy tissues, guided by histologically defined markers of AH. We hypothesize that through the analysis of precise intra-liver protein abundance variations, we will gain greater resolution of protein level mechanisms in the control and dysregulation of the liver in AH. The overall specific aims include 1) Correlate histological events associated with AH pathology, i.e., cell ballooning, steatosis, infiltration, with their overlapping spatially resolved protein abundances utilizing the LCM-nanoPOTS MS platform. 2) Identify spatial protein abundance variations across AH, Alcoholic cirrhotic, and normal liver tissues.

概括 过度饮酒是全球主要的公共卫生问题，酒精性肝病（ALD）是 美国与肝有关的主要原因。酒精性肝炎（AH）是一种特别严重的形式 短期死亡率高的ALD在一个月后约为35％。虽然抗炎类固醇治疗 还有其他一般疗法，患者对这种疗法的反应差异很大。有必要 了解AH疾病发作和进展的机制，并确定治疗的标志 功效。为此，我们旨在应用更精确的分析方法以超越总体组织分析 并朝着更多细胞特异性和空间蛋白丰度。具体而言，通过更精确地分析 AH肝组织的光谱，我们将更好地了解肝细胞如何发展为凋亡 气球和这些细胞对相邻的肝细胞的影响，特别是在中度或混杂的AH中 案例。我们已经开发了一项突破性技术，称为纳米（纳米圆形机） 对于痕量样品），目前可以有效分析具有广泛蛋白质组的<10个哺乳动物细胞 覆盖范围。与激光捕获微分解（LCM）和下游高吞吐量耦合纳米机耦合 高灵敏度质谱分析平台现在使技术能力能够捕获全球 蛋白质丰度在微尺度水平上。我们的目的是将纳米机应用于高空间映射 分辨率异性AH活检组织，由组织学定义的AH的标记引导。我们假设 通过分析精确的肝内蛋白丰度变化，我们将获得更大的分辨率 肝脏对AH中肝脏的控制和失调的蛋白质水平机制。总体具体目的包括 1）与AH病理学相关的组织学事件，即细胞气囊，脂肪变性，浸润与其 利用LCM-Nananopots MS平台的空间分辨蛋白质丰度重叠。 2）识别空间 AH，酒精性肝硬化和正常肝组织的蛋白质丰度变化。