DEVELOPMENTAL-CELL-D-19-00590R1 Misra et al

Published: 14-01-2020| Version 1 | DOI: 10.17632/csmd7kz9r8.1
Chaitali Misra


Myotonic dystrophy type 1 (DM1) is a dominantly inherited neuromuscular disease caused by a CTG repeat expansion in 3’-UTR of DMPK gene. DM1 affects multiple tissues, but cardiac dysfunctions are the second leading cause of death. The best characterized pathogenic mechanism of DM1 is toxic gain-of-function of expanded CUG repeat (CUGexp) RNA that accumulates to form ribonuclear foci causing sequestration of MBNL and overexpression of CELF1 family of splicing factors. However, loss of MBNL or gain of CELF1 activity does not explain the cardiac phenotypes observed in DM1. Here we report that steady-state protein levels of RBFOX2, a critical splicing regulator, are drastically upregulated in DM1 heart tissue. This is accompanied by aberrant skipping of a muscle-specific 43bp exon in RBFOX2 transcript, resulting in selective up-regulation of the non-muscle RBFOX2 splice isoform in adult cardiomyocytes. We demonstrate that expression of CUGexp RNA in DM1 affects RBFOX2 in two distinct ways: reduced expression of a subset of microRNAs causes de-repression of RBFOX2 protein production; and CELF1 overexpression promotes skipping of the muscle-specific exon of RBFOX2. Remarkably, tet-inducible overexpression of the non-muscle RBFOX2 isoform, or CRISPR/Cas9 mediated deletion of the muscle-specific RBFOX2 exon in the mouse heart results in prolonged PR and QR intervals, slower conduction velocity and spontaneous cardiac arrhythmias that mirror human DM1 pathology. Integration of RBFOX2-CLIP with RNA-seq data from cardiomyocytes of mice expressing the non-muscle RBFOX2 isoform identified a core network of mRNA splicing defects in genes encoding sodium, potassium, and calcium ion channels that are similarly misspliced in hearts of DM1 patients. These aberrantly spliced ion channels alter their rate of ion diffusion and electrophysiological properties thereby induce cardiac arrhythmias in DM1 heart. Thus, we have uncovered a crucial role for RBFOX2 isoform switching in DM1 cardiac pathogenesis


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