P.192 High-throughput screening of antifibrotic and antiadipogenic drugs using human FAP cells

Abstract

Fibroadipogenic precursor cells (FAPs) are the main cells involved in the expansion of fibro-fatty tissue in muscles of muscular dystrophy patients. FAPs can differentiate into fibroblast or adipocytes leading to the increase of fibrotic and adipogenic tissue. Growing studies have shown that FAP regulation in muscular dystrophies is altered, contributing to muscle degeneration and also reducing the effective delivery of potential drugs into the muscle tissue. There are different pathways governing this process, however, the precise mechanisms underlying FAP differentiation in muscular dystrophies remains not fully understood. Many drugs can modulate signaling pathways involved in that differentiation, suggesting that modulation of FAPs with pharmacological agonists or antagonists may provide a potential therapy to slow down the progression of muscular dystrophies. This suggestion raises an attractive route for drug discovery for exploring chemical modulators of FAP regulation. Our principal aim is to obtain a high throughput method that allow us to screen large libraries of drugs to find therapeutical approaches able to modulate FAPs differentiation. Herein, we provide an application to measure the rate of differentiation of FAPs into fibroblast or adipocytes after drug treatment. The method is based on human FAPs differentiated in vitro and the differentiation rate is quantified by the detection of specific proteins of fibroblast and adipocytes. This protocol has allowed us to set-up a drug screening using batteries of a total of 526 drugs. A final group of 8 drugs were selected to have a reduced fibrotic and adipogenic differentiation in human FAPs in in vitro experiments. The high throughput screening is a powerful tool for drug discovery that may accelerate the screening of FAP modulators to identify drugs that effectively reduce fibrofatty expansion. Therapies that modify the differentiation of FAPs could influence disease's progression in muscular dystrophies.