Investigation of REVERBalpha as a therapeutic target in pulmonary fibrosis

Idiopathic pulmonary fibrosis (IPF) is a lung disease that kills 5000 people a year in the UK with few effective treatments. Myofibroblasts are a key cell type in IPF, and I have discovered a new way to regulate them. Myofibroblasts are not normally found in healthy tissue but occur in response to injury. This development/ differentiation is partly under the control of key messengers secreted by epithelial cells or macrophages.
The importance of myofibroblasts in IPF is shown by the pathological accumulation of these cells observed in biopsies from patients. Therefore an attractive treatment approach would be to inhibit the development of myofibroblasts, and their function potentially stopping disease progression and permit healing. I have discovered that an important cell regulator protein, REVERBalpha, can prevent the formation of myofibroblasts from normal, healthy lung cells. I now want to take this exciting development forward, to determine if targeting REVERBalpha is a viable treatment option for patients, potentially providing a life saving treatment.
REVERBalpha belongs to a family of proteins called nuclear hormone receptors. REVERBalpha is a master regulator of the body's capacity to handle fats from the diet, targeting them for storage, or utilisation. In my previous MRC sponsored research I developed novel compounds that alter the function of REVERBalpha. I showed that these compounds can regulate the function of inflammatory cells from the human lung. I have now extended these findings and shown that these compounds stop the development of myofibroblasts in the lab.
My proposed research will investigate the function of REVERBalpha in lung fibrosis. Initially I will examine how loss of REVERBalpha affects development of lung fibrosis in models of human fibrotic lung disease. I will then test how my novel "drug-like" small molecules can prevent fibrosis development. I have developed new resources to permit rapid progress in this area, including a new antibody which permits precise measurement of REVERBalpha protein levels in patients with lung fibrosis, comparing against normal, healthy lung tissue.
I have also discovered that REVERBalpha can alter how cells manage their energy requirements, leading to distinct changes in a number of small molecules, including some types of fat, together termed metabolites. This is very relevant for IPF as I have also shown that some of these pulmonary metabolites can alter the differentiation of myofibroblasts. I therefore plan to sample metabolites in both healthy and diseased lungs in the same patient; this will allow me to identify metabolic changes caused by IPF. I then have the option of investigating candidate metabolites with the aim of discovering which are regulated by REVERBalpha and play a causative role in disease progression. Finally, I will apply new technology to investigate which genes in the lung cells are targets for REVERBalpha including how the regulation is managed and what the consequences are for the cell. This will identify new pathways and additional drug targets for further studies beyond the current fellowship.
I have brought together an international collaboration of researchers to address this novel hypothesis, which provides added benefit by offering me a superb research training experience. Though the research will be mainly conducted in Manchester, researchers, both basic scientists and clinical investigators, from other parts of the U.K. as well as Toronto will also be involved.
In summary, I will be exploring a potential treatment option for an increasingly common and fatal lung disease for which no effective treatments currently exist.

Pulmonary fibrosis (IPF) kills 5,000 people a year in the U.K. I have discovered that REVERBalpha prevents myofibroblast differentiation, critical for fibrosis. I propose that REVERBalpha inhibits pulmonary fibrosis by inhibiting myofibroblast differentiation.
Aim1: The role of REVERBalpha in pulmonary fibrosis development
I will repress the exprssion of REVERBalpha, in two distinct fibrosis models. I will investigate how REVERBalpha affects fibrosis development and resolution. Based on these studies I will proceed to target REVERBalpha expression in specific lung cell types, to identify which cells are mediating the fibrotic effects. In parallel studies I will test the therapeutic effects of novel REVERBalpha ligands in a lung fibrosis model. Finally I will compare REVERBalpha expression between human fibrotic explants and healthy lung explants.
Aim2: Examine the pulmonary fibrotic pathological changes in the BAL metabolome
I have developed a metabolomics approach capable of detecting differences between fibrotic and control human lungs sampled in-vivo. I will apply this discovery platform to identify metabolites that predict fibrosis development, and progression in the murine studies above. I will develop this approach further, analysing additional human samples, and applying a bioinformatics approach to find metabolites for functional analysis below.
Aim3: Analysis of the metabolomic and transcriptional changes through which REVERBalpha alters the pulmonary fibrotic phenotype
I will use ChIP-SEQ and RNA-SEQ to identify the REVERBalpha cistrome, and the network of REVERBalpha effects, in the cell-types identified above. This powerful approach, coupled with targeted validation using siRNA, and pharmacological inhibition, will discover new control points for fibrosis regulation. In addition I will identify the function of candidate metabolites identified from aim 2 using in vitro cell culture models.

The research questions posed within this proposal are of significant interest to ACADEMIC GROUPINGS in Biological and Biomedical Sciences. The academic community will benefit from elucidation of novel mechanisms that control fibrosis. Fibrosis is a promising field for research, since there are few effective treatments; the pathobiology is still poorly understood including cellular communication between different cell types. This fellowship will contribute to answering these important questions and it is expected that as mechanisms of fibrosis share some common pathways it will have impact not only IPF but other fibrotic diseases
Fibrosis is a consequence of an ageing population and therefore it is placing an increasing burden on the health of the population. As such, research findings will impact greatly on the HEALTH CARE COMMUNITY. We will disseminate findings by publishing primary papers and reviews in high impact journals, and presenting work at national and international meetings. We anticipate that the proposed work will produce 1-2 high-quality primary research papers.
All findings will be of high interest to the GENERAL PUBLIC due to the prevalence of lung disease, and the relative lack of effective intervention, particularly in fibrotic disease. In addition, REVERBalpha, is a clock protein controlling circadian variation. This raises questions for the 24hr lifestyles in our modern society especially in regard to exposure industrial air pollution. At its most basic, the work will engage sections of the populous who wish to learn about their health and human physiology. Research findings will be delivered to the general public through public engagement activities (e.g. brain awareness week, annual science open days at the UoM, Café Scientifique presentations), through the local transplant charity New Start and mass media. For example, several of our recent papers have been reported widely in national and international newspapers, on local radio, and on the internet following press releases issued by the University of Manchester and BBSRC.
The proposed research is of interest to PHARMACEUTICAL COMPANIES due to direct implications for human metabolic disease. Pharmaceutical industry investment into circadian biology is rapidly growing due to the fact that circadian dysfunction has been linked to sleep disorders, mental health disorders, cancer, inflammation, and aging. In the context of the MRC's mission to "build partnerships to enhance take-up and impact, thereby contributing to the economic competitiveness of the United Kingdom", our laboratories have taken a major lead within the extensive community of researchers at the University of Manchester by developing significant interactions and links with GSK and developing a joint ChronoBiology Programme.
Industrial interest is evidenced by the substantive contributions of GSK, provision of chemical biology assets, compound synthesis, and expertise (Stuart Farrow). The Faculty of Medical and Human Sciences at Manchester has taken a strong proactive role in developing links with major pharmaceutical companies, enhancing public communication of science, as well as identification and development of commercialisation opportunities. There are dedicated members of staff employed within the Faculty to assist in these areas. In addition since Stuart Farrow is a key collaborator on the project, results will be rapidly disseminated within pharma ensuring rapid development of key results into clinical trials of compounds. The timescale for these developments are impossible to predict with any degree of certainty however I would expect translation within 10-20 years.
In addition to my training during the fellowship I have also requested funding for a research associate. This will provide training for them in chronobiology as well as systems level biology with a view to helping the research associate prepare their own fellowship application in the future.