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2018-09-12 - Colloque/Présentation - poster - Anglais - page(s)

Neaga Ioan-Ovidiu, Baroni Alexandra, Hambye Stéphanie , Bodoki Ede, Blankert Bertrand , Oprean Radu, "Affinity capillary electrophoresis: an useful tool for screening drug candidates in  myotonic dystrophy type 1" in DA-PBA 2018 : 11th International symposium on Drug Analysis - 29th International Symposium on Pharmaceutical and Biomedical Analysis, Leuven, Belgium, 2018

  • Codes CREF : Analyse et contrôle pharmaceutique (DI3450), Chimie analytique (DI1314), Sciences pharmaceutiques (DI3400)
  • Unités de recherche UMONS : Analyse pharmaceutique (M130)
  • Instituts UMONS : Institut des Sciences et Technologies de la Santé (Santé)

Abstract(s) :

(Anglais) Myotonic dystrophy type 1 is an autosomal dominantly inherited degenerative disease, being the most prevalent forms of muscular dystrophy in adults. The disease presents a broad range of signs and symptoms, including, but not limited to: progressive or loss of muscle mass, cataract, hypersomnia, fatigue, heart conduction abnormalities, and respiratory problems [1]. The genetic mechanism of the disease is related to the presence of the cytosine-thymine-guanine (CTG) triplet repeat on the site of the DMPK (dystrophia myotonica protein kinase) gene. The CTG triplet is then transcribed into (CUG)n RNA repeats, which in turn is able to bind splicing factors from which MBLN1 and CUGBP1. The deficiency in these factors will result in RNA mis-splicing and protein synthesis defects. Since at the present there is no available treatment for the disease, several groups focused their study on finding molecules that could disrupt the MBNL-1/CUG complex or bind to DNA CTG repeat sequence and preventing its transcription [2,3]. In this study we evaluate affinity capillary electrophoresis as a technique for screening potential drug candidates in myotonic dystrophy type 1. A small library of compounds was tested, containing among others two previously proven ligands (pentamidine and neomycin) and two novel ones, a pentamidine analogue (EBAB) with potential lower toxicity and a polycarbonate guanidine polymer. As nucleic acid probes, we used DNA and RNA molecules with lengths corresponding with those present in both physiological (14 CUG/CTG) and pathological states (95 CUG/CTG). The affinity and efficiency of the tested ligands to bind the DNA/RNA probes were quantified in terms of binding constants and stoichiometry. For all the experiments, fused silica capillary coated dynamically with polyethylene oxide (200k) was used. The coating was performed in order to prevent interactions between the capillary wall and the different biomolecules tested and also to reduce the electroosmotic flow. From the tested ligands, the polycarbonate guanidine polymer showed interesting behavior. While pentamidine presented similar affinity for both RNA chains (Kb = 11205 M-1 for 14 CUG and Kb = 15985 M-1 for 95 CUG), the guanidine carbonate polymer seems to bind mostly to the longer 95 CUG RNA chains (Kb = 212521 M-1) The developed ACE method proved to be efficient and robust, allowing the testing of a ligand in a few hours, with minimum costs of reagents. References [1] Turner C, Hilton-Jones D. The myotonic dystrophies: diagnosis and management. J. Neurol. Neurosurg. Psychiatry. 2010;81:358–67. [2] Warf MB, Nakamori M, Matthys CM, Thornton CA, Berglund JA. Pentamidine reverses the splicing defects associated with myotonic dystrophy - supporting information. Proc. Natl. Acad. Sci. 2009;106:18551–6. [3] Coonrod LA, Nakamori M, Wang W, Carrell S, Hilton CL, Bodner MJ, et al. Reducing Levels of Toxic RNA with Small Molecules. ACS Chem. Biol. 2013;8:2528–37.