Abstract Details

Title ATXN2 Is Regulated by a Promoter Associated Antisense Long Noncoding RNA (lncRNA)

Topic Movement Disorders

Presentation(s) P05 - (-)

Poster/Presentation
Number 030

Objective

Background BACKGROUND: SCA2 is an autosomal dominant inherited disorder caused by CAG repeat expansion in the ATXN2 gene. The ATXN2 gene protein product ataxin-2 is characterized by gains of function upon ATXN2 mutation, caused by expansion of the CAG-encoded polyglutamine tract in ataxin-2. In seeking ways to develop therapeutics for SCA2 that lower ATXN2 expression we discovered a long non-coding RNA encoded by a portion of the ATXN2 antisense strand corresponding to the ATXN2 upstream sequence and 5'-UTR. The upstream region of the ATXN2-lncRNA includes a CTG repeat (antisense of the ATXN2 CAG repeat).

Design/Methods DESIGN/METHODS: ATXN2-lncRNA expression in ATXN2-BAC mouse cerebellum was determined by reverse-transcription PCR (rtPCR). The effects of ATXN2-lncRNA expression on ATXN2 was studied in transfected HEK293 or SH-SY5Y cells by luciferase assays and western blotting.

Results RESULTS: The ATXN2 antisense lncRNA is a spliced transcript with two exons of 56 and 429 bp separated by a 274 bp intron. The spliced 485 bp human ATXN2-lncRNA was present in one online database, HEK293, SH-SY5Y, and cerebellum of ATXN2-BAC mice but not in wildtype littermates, by rtPCR. ATXN2-lncRNA-luciferase constructs with CTG58 or CTG100 had 30-40% reduced expression compared to CTG22 (normal length). CMV-ATXN2-lncRNA expression reduced endogenous ATXN2 by 60% on western blots. ATXN2-lncRNA driven by its own promoter reduced ATXN2-luciferase expression by 30-50%; CTG length in the ATXN2-lncRNA upstream sequence did not significantly correlate with the magnitude of ATXN2 inhibition.

Conclusions CONCLUSIONS: Increased expression of the promoter associated antisense ATXN2-lncRNA was associated with reduced ATXN2 expression. We believe this is due to a sequence-specific interaction between the ATXN2-lncRNA and the ATXN2 promoter. This study supports ATXN2-lncRNA as a therapeutic target for the treatment of SCA2.

Authors/Disclosures
Daniel R. Scoles, PhD, FAAN (University of Utah) PRESENTER	The institution of Dr. Scoles has received research support from NIH.
Aaron L. Boster, MD (Ohiohealth)	Dr. Boster has received personal compensation in the range of $50,000-$99,999 for serving as a Consultant for sanofi. Dr. Boster has received personal compensation in the range of $50,000-$99,999 for serving as a Consultant for roche. Dr. Boster has received personal compensation in the range of $5,000-$9,999 for serving as a Consultant for novartis. Dr. Boster has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Medtronic. Dr. Boster has received personal compensation in the range of $50,000-$99,999 for serving as a Consultant for Serono. The institution of Dr. Boster has received research support from Sanofi. The institution of Dr. Boster has received research support from Roche.
	No disclosure on file
	No disclosure on file
Stefan M. Pulst, MD, FAAN (University of Utah)	Dr. Pulst has received personal compensation in the range of $500-$4,999 for serving as a Consultant for venrock. Dr. Pulst has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Arrowhead. Dr. Pulst has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Leverna. Dr. Pulst has received personal compensation in the range of $10,000-$49,999 for serving as an Editor, Associate Editor, or Editorial Advisory Board Member for AAN. Dr. Pulst has received personal compensation in the range of $500-$4,999 for serving as an Expert Witness for Leninthal LLC. The institution of Dr. Pulst has received research support from NINDS. Dr. Pulst has received intellectual property interests from a discovery or technology relating to health care.

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