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| Status |
Public on Jul 03, 2024 |
| Title |
Innate immune activation modulates contractile and electrical dysfunction in engineered heart tissue models fordesmoplakin(DSP)cardiomyopathy. |
| Organisms |
Homo sapiens; Rattus norvegicus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Desmosomes are specialized cell-cell junctions critically important in the heart, where their disruption promotes arrhythmogenic cardiomyopathy (ACM). Individuals carrying truncating variants in the desmosomal gene desmoplakin (DSP) often show a prominent inflammatory component in the absence of known antigenic stimuli, termed “hot phase” arrythmia. To assess the role of innate immune activation in DSP-cardiomyopathy, we generated engineered heart tissues (EHTs) from two patients with heterozygous DSP truncation variants (DSPtv, p.E1597X and p.R1951X). We also generated EHTs with homozygous DSP deletion (DSP-/-) for modeling. DSP-/- EHTs revealed impaired force production and reduced conduction velocity compared to isogenic controls, features consistent with ACM. RNA sequencing and protein characterization demonstrated a dramatic increase in innate immune activation in DSP-/- EHTs. Similarly, patient-derived DSPtv EHTs show enhanced sensitivity to inflammatory stimuli seen as reduced contractility and greater cytokine release. Inhibition of NFκB signaling improved baseline force production and mechanical response to strain in DSPtv EHTs, indicating that anti-inflammatory modulation may improve mechanical function in DSP deficient hearts. Furthermore, genomic correction of p.R1951X using adenine base editing reduced inflammatory biomarkers in cultured EHTs. Taken together, these data identify that DSPtv promote innate immune activation, creating an arrhythmogenic substrate in the concealed phase of ACM.
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| Overall design |
To investigate the gene-specific disease processes of desomsomal proteins in ACM, we performed bulk RNA-seq on neonatal rat ventricular myocytes (NRVMs) treated with siRNA to knockdown expression of Desmoplakin (Dsp), Plakoglobin (Jup) and Plakophilin-2 (Pkp2). We then performed differential expression and gene ontology analysis.
We then generated engineered heart tissues (EHTs) from human induced pluripotent stem cells (hiPSC) that contained homozygous deletion of DSP (DSP-/-) and performed bulk RNA-Seq alongside isogenic controls (DSP+/+) to identify differential gene expression in an alternate genetic model for severe ACM. We then performed differential expression and gene ontology analysis as well as motif enrochment analysis among differentially expressed genes.
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| Contributor(s) |
Selgrade D, Dubash A |
| Citation(s) |
38768074 |
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| Submission date |
Nov 17, 2023 |
| Last update date |
Jul 03, 2024 |
| Contact name |
Daniel Selgrade |
| E-mail(s) |
dselgrade@gmail.com
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| Organization name |
Northwestern University
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| Lab |
McNally Lab
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| Street address |
303 E. Superior Street
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| City |
Chicago, |
| State/province |
IL |
| ZIP/Postal code |
60611 |
| Country |
USA |
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| Platforms (2) |
| GPL22396 |
Illumina HiSeq 4000 (Rattus norvegicus) |
| GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
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| Samples (30)
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| GSM7907201 |
NRVMs, si-RNA Pkp2 biol rep1 |
| GSM7907202 |
NRVMs, si-RNA Pkp2 biol rep2 |
| GSM7907203 |
NRVMs, si-RNA Pkp2 biol rep3 |
| GSM7907204 |
NRVMs, si-RNA Jup biol rep1 |
| GSM7907205 |
NRVMs, si-RNA Jup biol rep2 |
| GSM7907206 |
NRVMs, si-RNA Jup biol rep3 |
| GSM7907207 |
NRVMs, si-RNA Dsp biol rep1 |
| GSM7907208 |
NRVMs, si-RNA Dsp biol rep2 |
| GSM7907209 |
NRVMs, si-RNA Dsp biol rep3 |
| GSM7907210 |
NRVMs, si-RNA scramble biol rep4 |
| GSM7907211 |
NRVMs, si-RNA scramble biol rep5 |
| GSM7907212 |
NRVMs, si-RNA scramble biol rep6 |
| GSM7907213 |
NRVMs, si-RNA Pkp2 biol rep4 |
| GSM7907214 |
NRVMs, si-RNA Pkp2 biol rep5 |
| GSM7907215 |
NRVMs, si-RNA Pkp2 biol rep6 |
| GSM7907216 |
NRVMs, si-RNA Jup biol rep4 |
| GSM7907217 |
NRVMs, si-RNA Jup biol rep5 |
| GSM7907218 |
NRVMs, si-RNA Jup biol rep6 |
| GSM7907219 |
NRVMs, si-RNA Dsp biol rep4 |
| GSM7907220 |
NRVMs, si-RNA Dsp biol rep5 |
| GSM7907221 |
NRVMs, si-RNA Dsp biol rep6 |
| GSM7907222 |
EHTs, DSP-/- biol rep1 |
| GSM7907223 |
EHTs, DSP-/- biol rep2 |
| GSM7907224 |
EHTs, DSP-/- biol rep3 |
| GSM7907225 |
EHTs, DSP+/+ biol rep1 |
| GSM7907226 |
EHTs, DSP+/+ biol rep2 |
| GSM7907227 |
EHTs, DSP+/+ biol rep3 |
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| Relations |
| BioProject |
PRJNA1041887 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE248187_DSP_Differential_Expression_Table.txt.gz |
1.5 Mb |
(ftp)(http) |
TXT |
| GSE248187_DSP_EHTs_Differential_Expression_Table.txt.gz |
1.5 Mb |
(ftp)(http) |
TXT |
| GSE248187_JUP_Differential_Expression_Table.txt.gz |
1.0 Mb |
(ftp)(http) |
TXT |
| GSE248187_PKP2_Differential_Expression_Table.txt.gz |
1.0 Mb |
(ftp)(http) |
TXT |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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