The proposed technology is based on the endogenous mechanisms regulating cellular homeostasis and, in particular, on mechanisms that control cell proliferation and apoptosis.
From a molecular point of view, the methodological approach is based on the design of RNA suitable for gene silencing, the so-called small interfering RNAs/siRNAs.
Once delivered to the cells as such or as shRNA using suitable vectors (also inducible), these siRNA are able to induce the alternative translation of the specific mRNA target.
We set up the technology using as prototype target a gene (ENO1/MBP-1) that, in response to cellular stress, generates a protein variant that negatively regulates cell growth, thus exerting a potential “pharmacological activity”.
The technology has been validated in vitro using breast carcinoma and neuroblastoma cell lines.
We generated a synthetic siRNA that induces the alternative translation of endogenous mRNA target by an AUG codon downstream of the canonical initiation site, thus switching on the cell response to stress; this induces the expression of a protein variant (MBP-1) and, consequently, the downregulation of cell proliferation and stimulation of the apoptotic process
Several genes encoding regulatory proteins such as protooncogenes, transcription factors, protein kinases and growth factors carry diverse translation initiation sites; the mRNA alternative translation IRES-mediated or “cap-indipendent” is cell-type specific, and depends on the stage of differentiation and/or of cellular growth. .
The proposed technology is synthetic siRNA-mediated and takes advantage of the cell endogenous mechanisms of “RNA interference”, which are active in all cell types, thus it has the advantage of a general use with possible applications in the therapeutic field.
Therapeutic siRNA-based approaches have already been applied to various diseases and clinical trials are currently underway but, to date, it is not documented in the scientific literature the usage we proposed. Numerous studies have allowed to establish the best conditions to delivery siRNA, to avoid off-target immune responses and/or toxic effects, already making feasible the development of a preclinical model for the proposed technology.
The technology developed enables the patent filing. This step would allow to value the results obtained so far.