Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology
Session: 2
Room: Exhibit Hall A
Rachel Owusu-Takyi - University at Albany
Myotonic Dystrophy Type 1 (DM1) is a neurodegenerative disease characterized by muscle weakness/wasting, cataracts, myotonia, cardiac abnormalities, and respiratory insufficiency and affects many different tissues in the body. DM1 has a high prevalence rate and affects at least 1 individual in every 8000 people worldwide. This disease is caused by a CTG repeat expansion in the 3’ UTR of the DMPK (dystrophia myotonica protein kinase) gene. Transcription from the DMPK gene with expanded CTG repeats, results in RNA transcripts containing CUG repeats that sequester RNA binding proteins such as the muscleblind-like (MBNL1) family of RNA binding proteins. The MBNL proteins are splicing regulatory proteins and their sequestration to r(CUG)exp RNA cause the deregulation in splicing of their hundreds of different target genes. These genes include the auto-regulation of MBNL1 and MBNL2 and many other genes such as NUMA1, SYNE1, INSR, and CLASP1 genes. The transcript of expanded CUG repeats i.e. r(CUG)exp also undergoes repeat-associated non-AUG translation (RAN) translation that results in the formation of toxic dipeptide repeat protein aggregates. Although there are currently no treatments available to patients with DM1, many different approaches are under development. A recent study in the lab revealed colchicine selectively reduced the production of expanded repeat RNA at nanomolar concentrations. In a separate study, the lab found that furamidine rescued DM1 splicing defects through multiple mechanisms in different DM1 models. In developing potential therapeutic strategies for DM1, we are testing the activity of combination treatments in DM1 models. I will present my research efforts on testing the combined activity of furamidine and colchicine in a DM1 HeLa cell model and DM1 patient-derived myoblast cell line. The experimental approaches include using cell culture, quantitative reverse-transcriptase polymerase chain reaction and splicing assays to assess the activities of colchicine and furamidine in the DM1 cell models. My current preliminary data has shown that colchicine partially blocked transcription of the expanded CUG repeats as expected, but unexpectedly furamidine was not active in the DM1 HeLa cell model and reduced colchicine’s activity. Further experiments will be performed in DM1 patient-derived myoblast cells with these molecules and more broadly we will test different combinations of small molecules.
Funder Acknowledgement(s): LSAMP; CSTEP
Faculty Advisor: Andrew Berglund, aberglund@albany.edu
Role: My research project is based on Myotonic Dystrophy DM. Myotonic Dystrophy is an autosomal dominant disease which is caused by repeat expansion of noncoding CTG/CCTG in resulting in a toxic RNA. DM is a microsatellite disease including cardiac abnormalities, respiratory insufficiency, cataracts, muscle weakness, etc. Main factors causing these symptoms are MBNL protein sequestration, aberrant splicing and RAN translation. My role in this project was to use a combination of small molecules such as Furamidine and Colchicine to target toxic RNA to avoid the symptoms. This research was done to analyze if Colchicine and Furamidine will be effective in reducing repeat expansion and splicing effects. My research has contributed in concluding that, small molecules such as Colchicine was effective than Furamidine in rescuing abnormal splicing and reducing RNA level.