Stem cells for drug screening targeting splices – SEDUCE

The SEDUCE proposal aims at establishing an innovative approach to identify new therapeutic molecules for alternate-splicing related diseases and to develop new screening tools for drug discovery approaches driven by academia and industry.
Pre-mRNA splicing is a fundamental process in mammalian gene expression and alternative RNA splicing plays a considerable role in generating protein diversity. During this process, particular exons of a gene will be included within or excluded from the final maturated mRNA, and the resulting transcripts generate diverse protein isoforms. Alternative splicing has been proposed as one of the major mechanisms contributing to protein diversity. Increasing evidence has shown that the disruption of alternative splicing negatively impacts health and contributes to human diseases, including cancer, diabetes, and neuromuscular diseases. The recent realization that up to 50% of genetic diseases involve splicing mutations has driven the development of several therapeutic approaches to correct aberrant splicing. Among these, the identification of small molecules capable to modulate splicing has been accelerated in the last decade with the technical development of large-scale cell-based screens. However, up to date, the success rate for identifying a splicing modulator that reaches the market is extremely low. Several parameters can explain this failure 1). the use of reporter constructs that contain only a restricted part of the gene of interest 2). the use of transformed cells instead of disease-relevant cell types. Therefore, one of the most important challenges for the future development of small molecule modulator will depend upon how well the specificity of the effects can be optimized.
Altogether, these bottlenecks largely block the deployment of drug discovery campaigns and therefore abrogate the development of new medicines curing alternate splicing related diseases early on in the drug development process.
In this context, SEDUCE consortium proposes to combine the use of disease-specific human stem cells differentiated into relevant cell types with high throughput RT-qPCR screening to identify new therapeutics for two distinct alternative splicing related neuromuscular diseases: spinal muscular atrophy and myotonic dystrophy type 1. Overall, our proposal has several objectives (1) identify and optimize new splice modulators for SMA and DM1, (2) decipher their mechanisms of action (3) validate their action in vivo and (4) convert these new molecules into marketable products. Ultimately, our aim is to deliver a meaningful technology that will accelerate the development of therapeutics for the growing list of diseases in which the process of pre-mRNA splicing is altered while ensuring its direct availability to academic lab or pharmaceuticals companies for screening campaigns.
To achieve these goals, the consortium has secured all the necessary expertise : Partner 1 (Cécile Martinat, I-Stem) was one of the first laboratory developing new cellular models based on disease-specific human pluripotent stem cells such as DM1 and SMA. Partner 2 (Eric Perret, Evotec) brings an industrial expertise in the development of successful RT-qPCR HTS based, in the access and optimization of chemical librairies and the expertise of hit optimization. Partner 3 and Partner 4 (Denis Furling, Institute of Myology and Nicolas Charlet, IGBMC) are leading experts in deciphering molecular mechanisms involved in normal and pathological alternate splicing as well as in testing splicing and phenotype correction in vivo in mouse models. Our proposal offers a genuinely innovative opportunity to push beyond the limitations of current models and promises to open up major new “assay development space” by increasing our understanding of the regulation of alternate splicing, identifying disease specific splicing modulators and offering a platform that can be applied to a wide range of research areas.