This WP aims to characterize the potential therapeutic usefulness of novel modulators of the TGF-? pro-fibrotic signalling pathways to prevent and/or regress MIF in HF. To this end we will focus on studying some novel modulators of the pro-fibrotic master regulator TGF-?, providing further evidence on the mechanisms of actions of those potential modulators and the experimental validation of new anti-fibrotic strategies. Given the broad range of effects of TGF-? and its important role in tissue homeostasis, long term direct inhibition of TGF-? may have detrimental effects, accelerating atherosclerotic plaque formation and interfering with the immune system. This may help to explain why the available anti-TGF-? agents aimed to treat MIF associated with cardiovascular disease have failed so far. Therefore, the search for novel, safe and efficient anti-fibrotic strategies based on the blockade of TGF-?-related fibrogenic mechanisms is necessary.
Here we aim to validate the therapeutic prospects of non-structural matrix proteins in different models of HF, to better understand the biological role by which these matricellular proteins regulate intra- and inter-cellular functions. These targets have been chosen based upon screens in vitro and validation studies in vivo in hypertensive and ischemic heart disease. We will study how Thrombospondin-2 (TSP-2), Osteonectin (SPARC), Osteoglycin (OGN) and the cartilage intermediated layer protein (CILP-1) could be used as novel therapeutic targets against adverse cardiac fibrosis, leading to cardiac dilatation and systolic dysfunction after acute myocardial infarction (MI) and pressure overload. The main sub-studies here are the cross-talk with TGF-? pathway (e.g. SPARC acting as a TGF-? co-factor) (close interaction with WP1), miRNA modulation (such as miR-29 and its down-regulation of SPARC expression), and the cross-talk between inflammatory cells and fibroblasts -where inflammatory cells drive the fibroblasts to improve the collagen quality in the infarct scar – protecting against systolic heart failure.
WP3 aims to apply discovery research tools to clarify the mechanistic role of novel miRNA targets in myocardial interstitial fibrosis (MIF) and to develop both diagnostic and therapeutic miRNA-based strategies of cardiac fibrosis. First, miRNAs will be identified that activate pro-fibrotic cytokine secretion in MIF by functional high-throughput miRNA library screens. Then, effects of pro-fibrotic stimuli (TGF-beta, WP1; non structural matrix proteins, WP2; mineralocorticoid receptor activation and its targets; WP4) on identified miRNAs will be tested in vitro in fibroblast-miRNA-fluorescence-tag reporter cell lines and small molecular compound libraries will be tested for miRNA inhibiting capacity. The role in MIF of selected miRNA candidates will be then validated in vivo in small animals using genetic and pharmacologic silencing techniques (antagomirs). Based on their efficacy to block fibroblast activation and pathologic cytokine secretion in vivo, enrichment in cardiac fibroblasts and genetic sequence conservation amongst species, the most promising anti-fibrotic miRNA candidate will subsequently be tested in a large pig model of myocardial infarction-induced pathological cardiac remodelling (WP6). The fibrosis-inducing miRNAs will be also tested in biopsy material and body fluids of small-sized patient samples of cardiac fibrosis and will be validated in WP7 in larger cohorts.
WP4 aims to 1) identify the mechanisms whereby NGAL, CT-1 and Gal-3 on one hand and aldo-mineralocorticoid (MR) modulated miRNAs on the other hand, are biotargets of Aldo-induced MIF 2) develop and validate novel molecules aimed to decrease expression/activity of these targets 3) explore and validate their relevance as novel therapeutic targets reducing MIF, especially when aldo/MR is involved. We will take advantage of a series of dedicated experimental models in which the level of these targets are changed as well as small molecules designed to interfere with NGAL/CT-1/Gal-3 activity or expression and with miRNAs expression. These molecules could be used as novel therapeutic agents in Aldo-induced MIF. The most promising molecules will then be optimized in terms of activity and ADMET profile then tested in a large animal model with myocardial fibrosis related to myocardial infarct (Explored in WP6). Moreover, selected biomarkers related to these targets may serve to identify and stratify distinct mechanistically defined "fibrogenetic' patient profiles with the aim of improving the targeting of anti-fibrotic interventions (Explored in WP5). This will maximize response rates in phenotypically defined potential responders and avoid treating unnecessarily potential non-responders. This personalized medicine outcome is typically another major impact that clinicians should anticipate from therapeutic targets research.
WP5 : Identification and "in silico" and "in vitro" screening for therapeutic small molecule therapeutic candidates, targeted to Myocardial Interstitial Fibrosis.
The main objective of this WP5 is to identify molecules (hits) that could be further used in the design of therapeutics for myocardial interstitial fibrosis treatment (MIF). This WP will be closely related to the rest of WPs. WP1-4 in accordance with GP and INNO, will suggest targets that will be tested in order to screen putative small molecules that would have therapeutic interest in MIF treatment. By means of chemo informatics approach, lists of molecules will be provided to be "in vitro" screened in this WP6 in order to identify hits that would prevent MIF or block its progression.
The objective is to prove the beneficial effect of antagomir and aptamers in small and large animals, regarding the read-out fibrosis characterization :
a) Haemodynamic consequences;
b) In vivo imaging,
d) Tissue histology, and
e) Molecular biology.
INSERM 1b will perform small animal experiments, while MUW will serve as a core facility for large animal model of postmyocardial infarction treatment. Emerging drugs from WP1-4 will be tested in the small and large animals (pig) for translational research. This WP will address the feasibility, safety and efficacy of the selected substances using clinical settings and clinical cameras under similar conditions as used in humans. The standardized human methods (e.g. coronary angiography, left ventricular angiography, echocardiography, PET-MRI) enable the direct comparison between substances and controls, to evaluate ventricular remodelling and extent of fibrosis quantitatively. The gene array analysis of the fibrotic per infarct areas and the remodelled part of the non-ischemic myocardial tissue will give information on the direct effect of the selected drugs at molecular level.
Characterization of circulating and tissue biomarkers pathophysiologically related to the therapeutic targets identified in WP 1 to 4, involved in MIF for a better detection of fibrogenesis in the animals (pigs) from WP5 as well as in patients at risk of or with HF and that may also subsequently serve for the evaluation of the effectiveness of MIF targeted therapies on the transition to and the progression of HF.
- To set up an effective management framework for the FIBROTARGETS consortium, this will ensure the progress of the project towards its planned objectives;
- To act as the interface between the FIBROTARGETS consortium and the European Commission;
- To ensure that all actions are performed correctly and within the rules and regulations established by the European Commission and the Consortium Agreement, including financial and legal management, and to ensure that the received funds are correctly distributed and accounted for;
- To ensure the work and tasks are performed on time, within budget and to the highest quality and create an early warning system.
- To keep each partner, including the Commission, fully informed about the project status, scientific issues, the work planning (adjustments) and all other issues which are important and relevant to partners in order to obtain maximum transparency and achieve synergy of the cooperation; to ensure that all partners are informed of all important and impacting information that can influence the outcome of the project.
- Promote integration and exchanges within and outside the FIBROTARGETS consortium
- Promote integration and exchanges within and outside the FIBROTARGETS consortium
- Ensure effective communication and public dissemination of project data to the research community, main stakeholders and general public
- Train and educate scientists in approaches relevant to the project
- Manage intellectual property issues arising from the project