Identification of druggable non-coding RNAs (ncRNAs) and novel therapeutic entities in the cardiopulmonary system
The role of small non-coding microRNAs (miRs) in ischemic cardiac injury has been studied by our group (Fiedler et al., Circulation, 2011). Pro-apoptotic function of miR-24 was shown to impact on endothelial cell biology and reduced capillary density in a mouse model of myocardial infarction (MI). Therapeutic miR-24 antagonism counteracted loss of vascular structures and highlights novel therapeutic potential after MI. In addition miR-24 has been shown recently to alter smooth muscle cell phenotype contributing to vascular density as well (Fiedler et al., ARS, 2013). Besides vascular miRs, other non-coding RNA (e.g. lncRNA, Fiedler et al., JACC, 2015, circRNA) is also of interest to identify novel miRs and small molecules regulating fibroblast biology which is part of our research. Conclusively, miR-based therapeutic strategies are of great importance for basic translational research and novel non-coding RNA key players are needed to be identified.
Staff:
- Jan Fiedler (PhD), Group Leader
- Kristina Sonnenschein (MD), Postdoc
- Jan Weusthoff (cand. med.), Strucmed
- Angelika Pfanne, Technician
- Annette Just, Technician
Methodology:
A) In vitro
- global miR profiling (Real-time PCR) with TaqMan-based arrays (ABI cards)
- single miR validation TaqMan Real-time PCR
- miR target and promoter validation: luciferase reporter gene assays
- Ago2 RNA immunoprecipitation for targetome analysis
- functional studies: vascular phenotyping
- FACS assays: apoptosis, reactive oxygen species, cell cycle analysis
- Western blotting and proteome profiling
- lentiviral cloning and viral transduction
- miR or gene modulation via specific precursors/viruses or antagonists/siRNAs/shRNAs
- antigen or ncRNA detection in cell culture or tissue samples
- CRISPR/Cas9 genome editing
B) In vivo, Ex vivo
Studies are performed in collaboration with the research groups of “Cardiovascular Phenotyping and Translational Strategies” and "Ex vivo technologies". Our main focus is on cardiac remodelling after MI and cardiac ischemia/reperfusion (IR) injury. Here, we try to interfere with novel miR therapeutics and further translational research also aims for transfer in larger animals. In addition, assays for peripheral angiogenesis testing are used (e.g. angioreactors).
Projects:
1. NcRNAs in the vasculature
NcRNAs are investigated in endothelial and smooth muscle cells to understand downstream signalling.
2. MiRs / Non-codings and fibroblast biology
Novel fibrotic miRs are investigated to decipher downstream molecular mechanisms.
3. MiRs / Non-codings and cardiac injury
Overall role of certain miRs during cardiac IR injury is validated in different in vitro and in vivo settings.