TY  - JOUR
A1  - Eberl, Hanna
A1  - Rebs, Sabine
A1  - Hoppe, Stefanie
A1  - Sedaghat-Hamedani, Farbod
A1  - Kayvanpour, Elham
A1  - Meder, Benjamin
A1  - Streckfuss-Bömeke, Katrin
T1  - Generation of an RBM20-mutation-associated left-ventricular non-compaction cardiomyopathy iPSC line (UMGi255-A) into a DCM genetic background to investigate monogenetic cardiomyopathies
JF  - Stem Cell Research
N2  - RBM20 mutations account for 3 % of genetic cardiomypathies and manifest with high penetrance and arrhythmogenic effects. Numerous mutations in the conserved RS domain have been described as causing dilated cardiomyopathy (DCM), whereas a particular mutation (p.R634L) drives development of a different cardiac phenotype: left-ventricular non-compaction cardiomyopathy. We generated a mutation-induced pluripotent stem cell (iPSC) line in which the RBM20-LVNC mutation p.R634L was introduced into a DCM patient line with rescued RBM20-p.R634W mutation. These DCM-634L-iPSC can be differentiated into functional cardiomyocytes to test whether this RBM20 mutation induces development of the LVNC phenotype within the genetic context of a DCM patient.
KW  - cell biology
KW  - developmental biology
KW  - general medicine
KW  - RBM20 mutations
KW  - DCM genetic background
KW  - monogenetic cardiomyopathies
Y1  - 2024
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-350565
SN  - 1873-5061
VL  - 74
ER  - 
TY  - JOUR
A1  - Sedaghat-Hamedani, Farbod
A1  - Rebs, Sabine
A1  - Kayvanpour, Elham
A1  - Zhu, Chenchen
A1  - Amr, Ali
A1  - Müller, Marion
A1  - Haas, Jan
A1  - Wu, Jingyan
A1  - Steinmetz, Lars M.
A1  - Ehlermann, Philipp
A1  - Streckfuss-Bömeke, Katrin
A1  - Frey, Norbert
A1  - Meder, Benjamin
T1  - Genotype complements the phenotype: identification of the pathogenicity of an LMNA splice variant by nanopore long-read sequencing in a large DCM family
JF  - International Journal of Molecular Sciences
N2  - Dilated cardiomyopathy (DCM) is a common cause of heart failure (HF) and is of familial origin in 20–40% of cases. Genetic testing by next-generation sequencing (NGS) has yielded a definite diagnosis in many cases; however, some remain elusive. In this study, we used a combination of NGS, human-induced pluripotent-stem-cell-derived cardiomyocytes (iPSC-CMs) and nanopore long-read sequencing to identify the causal variant in a multi-generational pedigree of DCM. A four-generation family with familial DCM was investigated. Next-generation sequencing (NGS) was performed on 22 family members. Skin biopsies from two affected family members were used to generate iPSCs, which were then differentiated into iPSC-CMs. Short-read RNA sequencing was used for the evaluation of the target gene expression, and long-read RNA nanopore sequencing was used to evaluate the relevance of the splice variants. The pedigree suggested a highly penetrant, autosomal dominant mode of inheritance. The phenotype of the family was suggestive of laminopathy, but previous genetic testing using both Sanger and panel sequencing only yielded conflicting evidence for LMNA p.R644C (rs142000963), which was not fully segregated. By re-sequencing four additional affected family members, further non-coding LMNA variants could be detected: rs149339264, rs199686967, rs201379016, and rs794728589. To explore the roles of these variants, iPSC-CMs were generated. RNA sequencing showed the LMNA expression levels to be significantly lower in the iPSC-CMs of the LMNA variant carriers. We demonstrated a dysregulated sarcomeric structure and altered calcium homeostasis in the iPSC-CMs of the LMNA variant carriers. Using targeted nanopore long-read sequencing, we revealed the biological significance of the variant c.356+1G>A, which generates a novel 5′ splice site in exon 1 of the cardiac isomer of LMNA, causing a nonsense mRNA product with almost complete RNA decay and haploinsufficiency. Using novel molecular analysis and nanopore technology, we demonstrated the pathogenesis of the rs794728589 (c.356+1G>A) splice variant in LMNA. This study highlights the importance of precise diagnostics in the clinical management and workup of cardiomyopathies.
KW  - familial DCM
KW  - laminopathy
KW  - long-read sequencing
KW  - nanopore
KW  - induced pluripotent stem cell cardiomyocytes
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-290415
SN  - 1422-0067
VL  - 23
IS  - 20
ER  -