The Role of Tryptophan Metabolism in Pulmonary Fibrosis

University Of Michigan At Ann Arbor, Ann Arbor MI

Investigators

Abstract

Project Summary / Abstract Idiopathic Pulmonary Fibrosis (IPF) is a progressive fibrotic disease of the lungs that is more common in aging and men, with 1 in 200 adults over age of 70 diagnosed. Patients have an estimated 2-5 years mean survival time following diagnosis, and only lung transplant is considered curative. Understanding the causes and immunological environment of the fibrotic lung could provide new targets for therapeutic or prophylactic treatment. Tryptophan metabolism is associated with increased inflammation in the lungs of IPF patients, as well as in our bleomycin-induced murine lung fibrosis model, producing several metabolites, such as kynurenine (kyn) that are recognized by the Aryl-hydrocarbon receptor (AHR). AHR has been well-characterized as affecting the immune system through several mechanisms both pro- and anti-inflammatory. Preliminary data from our lab show that inhibiting one of the early enzymes in the kynurenine pathway (KP) of tryptophan metabolism, TDO, results in significant protection from fibrosis in our murine bleomycin (blm) model. Data from human IPF patients support that TDO is uniquely expressed in alveolar fibroblasts of IPF patients, but not healthy controls. Furthermore, AHR is mostly highly expressed in dendritic cells (DCs) which play a critical role in fibrosis. We hypothesize that KP metabolites from fibroblasts are activating AHR in DCs to stimulate pro-inflammatory signals which in turn drive pulmonary fibrosis. The long term goal of this proposal is to understand how tryptophan metabolism impacts lung fibrosis in order to identify novel therapies for IPF. The objectives of this project are to determine how TDO inhibition affects downstream metabolites, and whether those metabolites are signaling to relevant immune cells which are important for fibrosis, specifically DCs, in a pro-inflammatory way. The specific aims of my proposal are: 1) to determine the in vivo effects of TDO inhibition on the kyn pathway and dendritic cells, and 2) to evaluate the mechanism of fibroblast-dendritic cell cross talk through ex vivo co-culture and conditioned media experiments. Under the first aim, I will perform metabolomics on healthy mice and on mice with blm-induced fibrosis with or without treatment with a TDO inhibitor in order to understand how fibrosis alters the metabolome of the lung and to establish the effect of TDO inhibition on downstream immunomodulatory metabolites. I will also perform RNAseq on primary DCs from the same conditions to determine how their maturation and pro-inflammatory pathways are altered, as well as evaluate their functional capacity to polarize naïve T-cells. Under the second aim, I will test primary fibroblasts, DCs, and T-cells of both our murine blm model and human IPF patients to determine the specific mechanism of anti-fibrotic activity by both directly treating cells with TDO-inhibitors and Tryptophan metabolites of interest, as well as using conditioned media experiments to evaluate signaling.