Antisense oligonucleotides correct the familial dysautonomia splicing defect in IKBKAP transgenic mice

作者：Sinha, Rahul; Kim, Young Jin; Nomakuchi, Tomoki; Sahashi, Kentaro; Hua, Yimin; Rigo, Frank; Bennett, C. Frank; Krainer, Adrian R.^*

来源：Nucleic Acids Research, 2018, 46(10): 4833-4844.

DOI：10.1093/nar/gky249

• TRANSFER-RNA MODIFICATION
• MOUSE MODEL
• ELONGATOR COMPLEX
• GENE
• RESTORATION
• EXPRESSION
• PROTEIN
• KINETIN
• RESCUE
• BRAIN

Familial dysautonomia (FD) is a rare inherited neurodegenerative disorder caused by a point mutation in the IKBKAP gene that results in defective splicing of its pre-mRNA. The mutation weakens the 5' splice site of exon 20, causing this exon to be skipped, thereby introducing a premature termination codon. Though detailed FD pathogenesis mechanisms are not yet clear, correcting the splicing defect in the relevant tissue(s), thus restoring normal expression levels of the full-length IKAP protein, could be therapeutic. Splice-switching antisense oligonucleotides (ASOs) can be effective targeted therapeutics for neurodegenerative diseases, such as nusinersen (Spinraza), an approved drug for spinal muscular atrophy. Using a two-step screen with ASOs targeting IKBKAP exon 20 or the adjoining intronic regions, we identified a lead ASO that fully restored exon 20 splicing in FD patient fibroblasts. We also characterized the corresponding cis-acting regulatory sequences that control exon 20 splicing. When administered into a transgenic FD mouse model, the lead ASO promoted expression of full-length human IK-BKAP mRNA and IKAP protein levels in several tissues tested, including the central nervous system. These findings provide insights into the mechanisms of IKBKAP exon 20 recognition, and pre-clinical proof of concept for an ASO-based targeted therapy for FD.

• 出版日期2018-6-1
• 单位苏州大学