High through-put screening to repurpose drugs for lupus therapies
We propose to implement a highly effective screening pipeline to repurpose drugs for lupus therapeutics. The purpose of this project is to assess the therapeutic potentials of existing drugs in mouse models of lupus and in T and B cells obtained from the peripheral blood of lupus patients. Based on results that we have obtained in a mouse model of lupus, and recent progress showing that cellular metabolism controls immune effector functions, we propose a pilot study that will focus on three compounds targeting immunometabolism – dichloroacetate, metformin, and rapamycin. The pipeline is designed to be scalable and to serve as a template for systematic and efficient screening of other classes of compounds in mouse models and PBLs from patients. This project will be divided into two linked proposals. Dr. Derry Roopenian, at the Jackson Laboratory will be the Principal Investigator of proposal 1, and I will be the Principal Investigator of proposal 2.
Proposal 1 will be performed at the Jackson Laboratory. It will be focused on performing mouse treatments, assessing disease progression, and collecting tissue samples that will be sent to Project 2.
Proposal 2 will be performed at the University of Florida and at Stanford University. It will have 3 aims: 1. To determine treatment efficacy on renal pathology and autoantibody production. Mouse kidneys received from Project 1 will be processed and evaluated for immune complex deposition and renal pathology. Autoantibodies in the serum of these mice will be screened with protein arrays. 2. To characterize the metabolic profile of T and B cells from treated mice and controls. The direct effect of metabolism inhibitors will be evaluated on glucose metabolism and oxidative phosphorylation (OXPHOS) of CD4+ T cells and B cells isolated from treated and control mice in Project 1. Metabolism parameters will be correlated with treatment outcomes.3. To assess the efficacy of the treatments tested in mice on T and B cells obtained from lupus patients and healthy controls. The metabolic inhibitors tested in Project 1 will be applied to CD4+ T cells and B cells isolated from the PBL of patients and controls. Responses to antigen-receptor dependent and independent stimulations will be evaluated by flow cytometry and metabolic parameter analysis.