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.

摘要 过量饮酒是一个主要的全球公共卫生问题，酒精性肝病(ALD)是一种 在美国，与肝脏相关的死亡的主要原因。酒精性肝炎(AH)是一种特别严重的 ALD近期病死率高，1个月后病死率约为35%。虽然抗炎类固醇治疗 和其他普通疗法一样，患者对这些疗法的反应差异很大。有必要 了解AH病的发病和发展机制，并找出治疗的标志物 功效。为此，我们的目标是应用更精确的分析方法，超越大体组织分析 以及更多的细胞特异性和空间蛋白质丰度。具体地说，通过更准确地分析 ，我们将更好地了解肝细胞是如何进展到凋亡的 气球膨胀及其对邻近肝细胞的影响，尤其是在中度或混浊的急性肝炎中 案子。我们开发了一项突破性技术，称为纳米POTS(一锅制备纳米液滴 对于痕量样本)，它目前能够有效地分析具有广泛蛋白质组的哺乳动物细胞 覆盖范围。将纳米POTS与激光捕获显微切割(LCM)和下游高通量和 高灵敏度质谱分析平台现在使技术能力能够捕获全球 在微观水平上的蛋白质丰度。我们的目标是将纳米点应用于高分辨率的空间映射 在组织学确定的AH标志物的引导下，分辨异种AH活检组织。我们假设 通过对肝内蛋白质丰度变化的精确分析，我们将获得更高的分辨率 急性肝炎肝脏调控和失调的蛋白质水平机制。总体具体目标包括 1)与AH病理相关的组织学事件，即细胞膨胀、脂肪变性、浸润性与其 利用LCM-NanPOTS MS平台重叠空间解析的蛋白质丰度。2)识别空间 蛋白丰度在酒精性肝炎、酒精性肝硬变和正常肝组织中的变化。