NanoPOTS Proteomic Characterization of Alcoholic Hepatitis liver tissue
EMSL Project ID
61018
Abstract
Mass spectrometry (MS) has been used extensively for the analysis of protein abundances in diverse sample types; however, limitations on sample handling and sensitivity have prohibited extension to extremely small tissue/cell collections. We have developed a breakthrough technology, termed nanoPOTS (Nanodroplet Preparation in One pot for Trace Samples) which currently enables effective analysis <10 mammalian cells with extensive proteome coverage. Coupling nanoPOTS with laser capture microdissection (LCM) and downstream high throughput and high sensitivity MS 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 features of AH. We will work with Drs. Ramon Bataller at U. of Pittsburgh and Tim Morgan at the Long Beach VA hospital to link histologically defined AH pathologies to spatially identified protein molecular mechanisms. We hypothesize that through the analysis of precise intra-liver protein abundance variations, we will gain greater resolution and hence greater mechanistic understanding of the inflammatory response control and dysregulation of the liver in AH. The overall specific aims include:
Specific Aim 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. Using a limited number of AH liver biopsies, we plan to capture and analyze hundreds of LCM isolations to map histologically defined AH disease conditions. The goal will be to determine to what extent histological results spatially correlate with protein abundance findings. Results will also help determine the overall surface area resolution required for SA2 analyses.
Specific Aim 2- Identify spatial protein abundance variations across AH, Alcoholic cirrhotic (AC), and normal liver tissues. Expanding the LCM-nanoPOTS MS platform analysis into larger patient cohorts, we will analyze liver tissue/biopsies utilizing the determined optimal spatial resolution found in SA1, to capture protein abundances of normal liver cellular morphology as well as variations in both AH and AC liver tissues. The goal will be to provide exceptional spatial and quantitative resolution of key AH discriminatory proteins and related network/pathways beyond those previously described via histology.
Project Details
Start Date
2023-11-13
End Date
N/A
Status
Active
Released Data Link
Team
Principal Investigator