ANALYSIS OF INTRAHEPATIC MACROPHAGE PROFILES FOR PREDICTING RISK OF FIBROSIS DEVELOPMENT IN PATIENTS WITH DIFFERENT TYPES OF CHRONIC LIVER DISEASE - PROJECT SUMMARY/ABSTRACT Cirrhosis and hepatocellular carcinoma are increasing health and economic burdens. Non-alcoholic fatty liver disease (NAFLD/NASH), alcohol-associated liver disease (AALD), chronic hepatitis (CHC), and autoimmune hepatitis (AIH) are common etiologies. Unfortunately, many patients do not adhere to recommended life style modifications, thus, we need better techniques for predicting risk of fibrosis progression and personalizing therapies prior to development of poor outcomes. Intrahepatic macrophages (Macs), liver sinusoidal endothelial cells (LSECs), and stellate cells (HSCs) can greatly influence the composition of the hepatic microenvironment and development of fibrosis. Therapies targeting these initiators of fibrosis are being investigated in phase II-III clinical trials; however, the underlying hepatic microenvironment and patient variability in these cells and expression of these targets is not being considered prior to treatment. We use cutting-edge spectral imaging microscopy combined with NanoString technology to evaluate these cells and associated pro-fibrotic gene expression profiles in the same patient's liver biopsy at the time of initial diagnosis. From our liver tissue biobank, we identified 225 biopsies with different chronic liver diseases (NASH, AALD, CHC, and AIH) that were collected at the time of diagnosis from patients that had adequate follow-up either with a repeat biopsy or by liver replacement (for those that later developed cirrhosis). The majority showed no progression of hepatic fibrosis over time (n = 150) while a portion rapidly developed cirrhosis (n = 75). We use the above platforms to assess differences in these patients' hepatic microenvironments in their initial liver biopsies. We propose to test the hypothesis that patients with definable pro-fibrotic variations in their hepatic microenvironment early in the course of disease predicts their propensity to develop fibrosis. Preliminary data showed that initial liver biopsies from patients with a predisposition to rapidly develop cirrhosis have increased profibrotic macrophages (e.g., Mac387+ and CD163+, respectively), enhanced cellular interactions of Mac-LSEC-HSCs, increased expression of therapy-related targets (e.g., CCR2 and galectin 3) and increased pro-inflammatory/pro-fibrotic gene expression profiles (e.g., CCL2, TNF, and TGF-beta). Imaging and molecular bioinformatics will be used for data analyses. For Aim 1, we will use three panels to phenotype intrahepatic Macs and examine their interactions with LSECs and HSCs, and will assess differences in expression of pro-fibrotic therapy-related targets. For Aim 2, we will analyze over 200 Mac-LSEC-HSC-related and pro-fibrotic genes in the other half of the biopsy from Aim 1. The proposed approach will lay the groundwork for our long-term objective: personalization of targeted therapies (e.g., cenicriviroc or obeticholic acid), similar to the manner in which the response to immunotherapy is predicted by staining of tissue in patients with cancer. In this retrospective longitudinal study, we will determine which platform (Spectral imaging-Aim1 vs. NanoString-Aim 2) is the most performant for determining potential targets of fibrosis progression and most cost efficient for clinical implementation in the future.
