Human Molecular Genetics Advance Access originally published online on April 17, 2009
Human Molecular Genetics 2009 18(13):2462-2471; doi:10.1093/hmg/ddp184
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Activating the synthesis of progerin, the mutant prelamin A in Hutchinson–Gilford progeria syndrome, with antisense oligonucleotides
1 Department of Medicine 2 Department of Human Genetics, David Geffen School of Medicine, University of California, 695 Charles E. Young Drive South, 4524 Gonda Building, Los Angeles, CA 90095, USA 3 Department of Antisense Research, ISIS Pharmaceuticals, Carlsbad, CA 92008, USA 4 Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, KY 40536, USA
* To whom correspondence should be addressed. Tel: +1 3102674380; Fax: +1 3102672722; Email: lfong{at}mednet.ucla.edu or sgyoung{at}mednet.ucla.edu
Received March 14, 2009; Accepted April 14, 2009
Hutchinson–Gilford progeria syndrome (HGPS) is caused by point mutations that increase utilization of an alternate splice donor site in exon 11 of LMNA (the gene encoding lamin C and prelamin A). The alternate splicing reduces transcripts for wild-type prelamin A and increases transcripts for a truncated prelamin A (progerin). Here, we show that antisense oligonucleotides (ASOs) against exon 11 sequences downstream from the exon 11 splice donor site promote alternate splicing in both wild-type and HGPS fibroblasts, increasing the synthesis of progerin. Indeed, wild-type fibroblasts transfected with these ASOs exhibit progerin levels similar to (or greater than) those in fibroblasts from HGPS patients. This progerin was farnesylated, as judged by metabolic labeling studies. The synthesis of progerin in wild-type fibroblasts was accompanied by the same nuclear shape and gene-expression perturbations observed in HGPS fibroblasts. An ASO corresponding to the 5' portion of intron 11 also promoted alternate splicing. In contrast, an ASO against exon 11 sequences 5' to the alternate splice site reduced alternate splicing in HGPS cells and modestly lowered progerin levels. Thus, different ASOs can be used to increase or decrease ‘HGPS splicing’. ASOs represent a new and powerful tool for recreating HGPS pathophysiology in wild-type cells.