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Battling monocyte misbehavior by targeting non-coding RNAs as novel intervention in systemic sclerosis

Thursday 26 March 2015

Battling monocyte misbehavior by targeting non-coding RNAs as novel intervention in systemic sclerosis
Dr. Marzia Rossato / Prof. Tim Radstake

Systemic Sclerosis (SSc) is a disfiguring autoimmune disease with high mortality and morbidity, characterized by progressive and uncontrollable fibrosis of the skin and internal organs. Genetic association studies demonstrated that SSc susceptibility is linked to immune-related genes, thus pointing at immune system as the driving force of SSc development. However the molecular mechanisms underlying immune system alteration in SSc, and their implications in the development of fibrosis characterizing SSc patients, are still obscure.
Mononuclear cell infiltration, consisting of monocytes-macrophages and T cells, has been detected in SSc fibrotic lesions as in the skin and lungs. In addition, the number of peripheral blood monocytes and the proportions of monocyte subsets are altered in SSc patients. Our recent transcriptome data demonstrated that multiple genes are dysregulated in SSc monocytes, including pro-fibrotic and type I interferon-dependent genes. In parallel, we observed that several non-coding transcripts such as long-non-coding RNA, antisense-RNA and enhancer RNAs are either upregulated or reduced in monocytes isolated from patients.
Non-coding RNAs (ncRNAs) constitute a recently discovered class of RNAs longer than 200nt without coding potential, that can regulate gene expression either at the transcriptional or post-transcriptional level. ncRNAs have the ability to form networks of ribonucleoproteins (RNP), chromatin regulators and transcription factors, and then target their enzymatic activities to appropriate locations in the genome, thus influencing gene expression. Underscoring the importance of lncRNAs’ regulatory roles is their involvement in both physiological processes and in the aetiology of diseases, such as cancers. Very recently, lncRNAs have been implicated in the regulation of inflammatory mediator release (including IFNs and IL-6) by human monocytes.

Hypothesis
At the light of the aforementioned results, we hypothesize that the aberrant expression of ncRNAs underlie the monocyte dysregulation observed in SSc patients. Being cell-type specific and potentially targetable by using specific silencing-RNAs, they also represent potential therapeutic targets to restore the aberrant phenotype and activation profile of SSc monocytes.

The verify the above-mentioned hypothesis, the following objectives will be addressed during the internship:
1) Validation of altered ncRNAs identified by RNAseq, in an additional cohort of healthy and SSc monocytes.
2) Identification of stimuli that can modulate the expression of ncRNAs in monocytes.
3) Deployment of RNA-silencing approaches to inhibit the expression of ncRNAs and identify their function.

During the internship, the student will be directly supervised from the scientific point of view by Dr. Rossato (post-doc, head of epigenetic unit in Prof. Radstake group) and on the lab-practice by Maarten van der Kroef (PhD student). The student will take advantage by a young and enthusiast group focusing on the emerging field of epigenetic in autoimmunity, with a long-standing technical and clinical expertise.
The project involves the implementation of delicate molecular and cellular biology techniques. The maximal accuracy and commitment to the project are therefore required and only highly motivated students will be considered.

Techniques: isolation of monocytes from peripheral blood, culture of monocyte-derived dendritic cells, Isolation of RNA, Real-Time quantitative PCR, cell transfection, western-blot, flow cytometry, Chromatin-IP.

Duration: 6 or 9 months

Contact: Dr. Rossato, M.Rossato-2@umcutrecht.nl;
Prof. Radstake, T.R.D.J.Radstake@umcutrecht.nl