TITLE: Assay to Screen for Small Molecules that Suppress CUG RNA-induced Neurotoxicity
CASE NUMBER: C14213
INVENTION NOVELTY
A neuronal-like cell model that may be used as a powerful tool to screen for small molecule compounds that can suppress JPH3 CUG RNA-mediated cytotoxicity.
VALUE PROPOSITION
Huntington’s disease-like 2 (HDL2) is a devastating autosomal dominant neurodegenerative diseases that is caused by a CTG repeat expansion in exon 2A of junctophilin 3 (JHP3) gene . JHU researchers have successfully created a neuronal-like cell model that may be used as a powerful tool to screen for small molecule compounds that can suppress JPH3 CUG RNA-mediated cytotoxicity. Advantages are:
1. It can be used to screen for compounds that can reduce neurotoxicity of JPH3 RNA
2. Expression and cytotoxicity of non-translatable JPH3 transcripts with the normal or expanded JPH3 CUG repeat is induced by doxycycline
3. The cells specifically model RNA toxicity, in the absence of any toxic protein expression from the inserts
TECHNICAL DETAILS
The cell model can be used to both perform mechanistic studies relevant to the role of RNA in neurodegeneration.
Identify novel tool compounds or drugs that can ameliorate RNA-induced neurotoxicity.
They have engineered published SK-N-MC cell line to stably and inducibly express either normal or expanded JPH3 fragment.
1: A junctophilin 3 (JPH3) fragment with 14 or 55 CUG repeats was cloned into an episomal vector and stable cell lines were established. Cell line generation and QC. Both constructs were inserted into SK-N-MC cells already engineered to express rtTA. Using a 96-well plate format, with ~104 cells/well, and a 72-hour culture, we selected clonal cell lines expressing non-translatable JPH3 transcripts with either 14 or 55 CUG repeats.
DISEASE INDICATION
Huntington Diseases
ASSOCIATED PUBLICATIONS
1. Ann Neurol, 2007. 61(3): p. 272-82.
2. Ann Neurol, 2012. 71(2): p. 245-57. 3.
3. Neuron, 2011. 70(3): p. 427-40.
4. Hum Mol Genet, 2011. 20(11): p. 2161-70
