Project's code: PN-III-P1-1.1-TE-2019-1893
Project's title: MIR-210 genomic locus at the overlap between hypoxia signalling and inflammatory networks
Contract number: TE186/07.01.2021
Coordinator: Dr. Mihai Bogdan Preda
Budget: 431.900 Lei
Period: 07.01.2021 – 31.12.2022
Accumulating evidence suggest that hypoxia and inflammation are intimately linked on many levels with a significant crosstalk between the transcription factors that are classically understood to respond to each one of them. The molecular machinery that regulates the activity of genes in response to hypoxia is subject to many regulatory feedbacks, mediated by transcription factors, metabolites, and the more recently appreciated, yet insufficiently understood, non-coding RNAs. To date, the best characterized hypoxia responsive non-coding RNA is miR210, which has diverse functions that cover almost every aspect of cellular hypoxic response.
An integrated inspection of mouse and human genomic locus shows multiple long-noncoding RNA transcripts and a promoter/enhancer regulatory element, suggesting that miR210 locus functions as a coordinated unit with the neighbouring genetic elements. Furthermore, the consequences of genetically disrupting this locus are not equivalent to a conventional inactivation of the mature miR210.
The goal of this project is to determine cell-specific cis- and trans- effects of miR210 locus during hypoxia and inflammation. The specific objectives of this project are: (i) Dissection of miR210 genomic locus in immune cells (T cells, B cells, and monocytes/ macrophages) in two transgenic mouse models for miR210; (ii) Evaluate the impact of hypoxia and inflammation on the inactivation of miR210 genomic locus after CRISPR-mediated experimental dissection.
Dissecting RNA biology of non-coding RNAs is generally a difficult and laborious process, however to get these objectives, we will work under the direct supervision of recognized experts in the hypoxia signalling field, and will use powerful and modern tools for deciphering genome organization (CRISPR-mediated genome editing, RNA-seq, 3C assay), with the goal of shedding the light into the biology of a genomic region long considered important for the immune and inflammatory response.
Mircea Ivan, MD, PhD, (Associate Professor of Microbiology & Immunology at Indiana University School of Medicine, USA)
Fabio Martelli, PhD (Molecular Cardiology Laboratory, I.R.C.C.S. Policlinico San Donato, Milan, Italy)
Sergiu-Bogdan Catrina, MD, PhD (Department of Molecular Medicine and Surgery, Karolinska Institutet, Sweden)
Dissemination of results
Presentations at international scientific conferences:
1. M.B. Preda. CRISPR-based disruption of miR-210 genomic locus affects cell growth and proliferation. CardioRNA Live Reboot. EU-CardioRNA COST ACTION online meeting. 5-7 july 2021 (Oral presentation)
2. Preda M.B. Current research activity and results. Prezentare orala, IRCCS Ospedale San Raffaele, Millano, 30 august 2021 (Oral presentation).
3. Preda M.B. Local microenvironment activates in vivo apoptosis of mesenchymal stromal cells after transplantation. 38th Annual scientific session of the Romanian Society for Cell Biology. 4-6 nov, 2021 (Oral presentation).
4. Preda M.B., Neculachi C.A., Fenyo I.M., Vacaru A.M., Publik M.A., Simionescu M. and Burlacu A. Local microenvironment triggers in vivo apoptosis of mesenchymal stromal cells after transplantation. Nature Conferences “Innovative Tissue Therapies from Bench to Bedside”. November 16-18, 2021 (Poster).