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Development of microRNA Medicines for Cardiac Metabolic Diseases; A Targeted European Exchange Programme

Final Report Summary - CARDIOMIR (Development of microRNA Medicines for Cardiac Metabolic Diseases;A Targeted European Exchange Programme)

The CardiomiR project has identified novel therapeutic targets based upon mRNA and non-coding RNA mechanisms, by using in vivo models of metabolic and inflammatory heart failure. Their biological role is being studied by using the screening in vitro technology of Cenix Bio GmbH, which will be further validated in the same mouse/rat models for their therapeutic prospects. Since the last report, the major progress consists of the in vitro screen done at Cenix by Robin Verjans and Wouter Derks (attachment).

Partner Cenix Bio GmbH in Germany has seconded one ESR (Fellow 2) from CENIX to UM. She was introduced to the HF models available at UM and has learned to isolate primary cardiomyocytes from neonatal rats. Two ESRs (Fellows 7 and 8, Robin Verjans and Wouter Derks, now back in Maastricht since February 2015, from UM have been introduced to the Cenix screening platform. They have been able to finalize the screening assays for primary rat and human cardiomyoctes, fibroblasts and macrophages (working at Cenix till the end of February 2015, after the breakdown in January). In addition to the previous report in December 2014, the CardiomiR consortium now has a complete report of the successful screening of over 200 inflammatory/metabolic microRNAs in these different cell types. These will form the basis for further validation of selected targets in animal experiments. We have thereby identified novel microRNA candidates within this screening at CENIX, and preclinical validation of these compounds is currently being performed in animal experiments at UM (Robin Verjans and Wouter Derks, but being funded by UM).

The University of Liège (ULg), contributed to the project by performing and analyzing the initial mRNA and microRNA screens necessary to identify the first therapeutic targets. ULg is now conducting research on algorithms for cell/tissue image analysis based on machine learning algorithms (combining the extraction of random sub-windows and the use of ensemble of randomized trees). These algorithms are implemented within CYTOMINE, a rich web-based application for storage, visualization, collaborative annotation and automatic analyses of large-scale images (

University of Maastricht (UM), the Netherlands, has also participated in the mRNA and miRNA screens for inflammatory and metabolic heart failure. They have performed in vivo screens in different heart failure models, including the metabolic risk induced heart failure (ZSF1 rats, attachment), in viral myocarditis in mice, in cardiac transplantation induced inflammatory heart failure and in Angiontensin-II-induced cardiac inflammation, fibrosis and hypertrophy. UM has learned from the expertise of Liège in relation to bio-informatical analysis of RNA arrays in mouse and human cardiac failure samples.

As described in the initial project proposal the expected final results are the identification of potential therapeutic targets, their biological role, for the future animal validation of their therapeutic prospects using the LNA-technology. The expertise of UM allowed Cenix to perform the necessary in vitro screens for studying the biological mechanisms by which the therapeutic targets identified in the initial in vivo arrays and screens, could affect cardiac inflammation, fibrosis or hypertrophy. UM, by learning of the screening technology of Cenix, has learned in regard to for future therapeutic validation, how the screening-determined mechanisms could explain the therapeutic effects of these targets. Finally, ULg, has learned from Cenix whether their imaging technology could be implemented for this screening technology.

The key aims that have been obtained during the CardiomiR are:
• Study the biological role of the therapeutic targets identified in the in vivo and human
screening/validation rounds, by
• Exchanging the expertise of in vitro screening from CENIX towards UM (screens are finalized).
• Training the imaging/bio-informatical analysis tools of ULg for CENIX application (been used to analyze the screen).

• Study and validate the in vivo therapeutic prospects of these therapeutic targets, pending on
their biological role in fibrosis, inflammation and/or hypertrophy.
• Exiqon has provided the expertise for LNA-technology for inhibition of microRNAs
• UM has performed and will perform animal experiments to validate these targets.

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