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| Status |
Public on Nov 27, 2024 |
| Title |
Regulation of sarcomere formation and function in the healthy heart requires a titin intronic enhancer |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Heterozygous truncating variants in the sarcomere protein titin (TTN) are the most common genetic cause of heart failure. To understand mechanisms that regulate abundant cardiomyocyte TTN expression we characterized highly conserved intron 1 sequences that exhibited dynamic changes in chromatin accessibility during differentiation of human cardiomyocytes from induced pluripotent stem cells (hiPSC-CMs). Homozygous deletion of these sequences in mice caused embryonic lethality, while heterozygous mice demonstrated allele-specific reduction in Ttn expression. A 296 bp fragment of this element, denoted E1, was sufficient to drive the expression of a reporter gene in hiPSC-CMs. Deletion of E1 downregulated TTN expression, impaired sarcomerogenesis, and decreased contractility in hiPSC-CMs. Site-directed mutagenesis of predicted NKX2-5- and MEF2-binding sites within E1 abolished its transcriptional activity. Embryonic mice expressing E1 reporter gene constructs validated in vivo cardiac-specific activity of E1 and the requirement for NKX2-5 and MEF2 binding sequences. Moreover, isogenic hiPSC-CMs containing a rare E1 variant in the predicted MEF2 binding motif that was identified in a patient with unexplained DCM showed reduced TTN expression. Together, these discoveries define an essential, functional enhancer that regulates TTN expression. Manipulation of this element may advance therapeutic strategies to treat DCM caused by TTN haploinsufficiency.
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| Overall design |
RNAseq of WT or mice carrying heterozygous E0 deletion. We generated E0del/+ mice in a hybrid genetic background by crossing C57BL/6 E0del/+ mice and 129SvEv WT mice. We performed RNAseq of transcripts from left ventricle (LV) and skeletal muscle where Ttn is highly expressed.
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| Contributor(s) |
Kim S, Kim Y, Wakimoto H, Gorham JM |
| Citation missing |
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| |
| Submission date |
Nov 22, 2024 |
| Last update date |
Nov 27, 2024 |
| Contact name |
Seong Won Kim |
| E-mail(s) |
[email protected]
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| Organization name |
Harvard Medical School
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| Department |
Genetics
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| Street address |
77 Ave Louis Pasteur
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| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02215 |
| Country |
USA |
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| Platforms (1) |
| GPL24247 |
Illumina NovaSeq 6000 (Mus musculus) |
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| Samples (10)
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| Relations |
| BioProject |
PRJNA1189576 |
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