SNTA1-deficient human cardiomyocytes demonstrate hypertrophic phenotype and calcium handling disorder
Data files
Oct 14, 2024 version files 5.78 GB
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H9CM_30da_1.fq.gz
964.34 MB
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H9CM_30db_1.fq.gz
952.50 MB
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H9CM_30dc_1.fq.gz
968.31 MB
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README.md
2.76 KB
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README.txt
274 B
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SNTA1_30da_1.fq.gz
969.09 MB
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SNTA1_30db_1.fq.gz
957.48 MB
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SNTA1_30dc_1.fq.gz
965.51 MB
Abstract
Background: α-1-syntrophin (SNTA1), a protein encoded by SNTA1, is highly expressed in human cardiomyocytes. Mutations in SNTA1 are associated with arrhythmia and cardiomyopathy. Previous research on SNTA1 has been based on non-human cardiomyocytes. This study was designed to identify the phenotype ofSNTA1-deficiency using human cardiomyocytes.
Methods: SNTA1 was knocked out in the H9 embryonic stem cell line using the CRISPR-Cas9 system. H9SNTA1KO cells were then induced to differentiate into cardiomyocytes using small molecule inhibitors. The phenotypic discrepancies associated with SNTA1-deficient cardiomyocytes were investigated.
Results: SNTA1 was truncated at the 149th amino acid position of PH1 domain by a stop codon (TGA) using the CRISPR-Cas9 system. SNTA1-deficiency did not affect the pluripotency of H9SNTA1KO, and they retain their in vitro ability to differentiate into cardiomyocytes. However, H9SNTA1KO derived cardiomyocytes exhibited hypertrophic phenotype, lower cardiac contractility, weak calcium transient intensity, and lower level of calcium in the sarcoplasmic reticulum. Early treatment of SNTA1-deficient cardiomyocytes with ranolazine improved the calcium transient intensity and cardiac contractility.
Conclusion: SNTA1-deficient cardiomyocytes can be used to research the etiology, pathogenesis, and potential therapies for myocardial diseases. The SNTA1-deficient cardiomyocyte model suggests that the maintenance of cardiac calcium homeostasis is a key target in the treatment of myocardial-related diseases.
https://doi.org/10.5061/dryad.3xsj3txj7
Description of the data and file structure
Animal cardiomyocytes are widely used in the cardiovascular diseases research, but there are obvious differences in structure and function between animal and human cardiomyocytes. As usual, it is very difficult to obtain human cardiomyocytes. Myocardial cells derived from pluripotent stem cells can solve the problem of human cardiomyocytes supply. Human embryonic stem cells were used to induce the cardiomyocytes. Using the CRISPR/Cas9 technique, SNTA1 was knocked out in embryonic stem cell, and then the SNTA1 knockout human embryonic stem cells were differentiated into SNTA1-deficient cardiomyocytes. RNA-seq data were collected from the control and SNTA1-deficient cardiomyocytes. We obtained the gene expression profile of human cardiomyocytes, which provides potentially useful transcriptional data for researchers studying human cardiomyocytes, particularly inherited heart diseases.
- The process of myocardial differentiation followed the instructions from the CardioEasy kit (Cellapy, China).
- Cardiomyocytes were derived from the human embryonic stem cell line H9.
- Data were collected on the 30th day after myocardial differentiation.
For the file name "H9CM_30da":
"H9CM" represents cardiomyocytes derived from H9 embryonic stem cells,
"30d" indicates that the cardiomyocytes are 30 days old, and
"a" refers to group A.
For the file name "H9CM_30db":
"H9CM" represents cardiomyocytes derived from H9 embryonic stem cells,
"30d" indicates that the cardiomyocytes are 30 days old, and
"b" refers to group B.
For the file name "H9CM_30dc":
"H9CM" represents cardiomyocytes derived from H9 embryonic stem cells,
"30d" indicates that the cardiomyocytes are 30 days old, and
"c" refers to group C.
The files "H9CM_30da", "H9CM_30db", and "H9CM_30dc" represent the three replicates for the control group.
For the file name "SNTA1_30da":
"SNTA1" represents cardiomyocytes derived from H9SNTA1KO embryonic stem cells,
"30d" indicates that the cardiomyocytes are 30 days old, and
"a" refers to group A.
For the file name "SNTA1_30db":
"SNTA1" represents cardiomyocytes derived from H9SNTA1KO embryonic stem cells,
"30d" indicates that the cardiomyocytes are 30 days old, and
"b" refers to group B.
For the file name "SNTA1_30dc":
"SNTA1" represents cardiomyocytes derived from H9SNTA1KO embryonic stem cells,
"30d" indicates that the cardiomyocytes are 30 days old, and
"c" refers to group C.
The files "SNTA1_30da", "SNTA1_30db", and "SNTA1_30dc" represent the three replicates for the experimental group.
Cardiomyocytes were derived from human embryonic stem cell line H9. The data were obtained on the 30th day after Myocardial differentiation.
1. Establish knockout embryonic stem cell.
2. The process of myocardial differentiation.
3. Total RNA was extracted from the cells.
4.mRNA Library Construction.
5. RNA-seq data analysis and Statistics.
- Dong, Tao; Zhang, Siyao; Chang, Yun et al. (2021). The establishment of a homozygous SNTA1 knockout human embryonic stem cell line (WAe009-A-50) using the CRISPR/Cas9 system. Stem Cell Research. https://doi.org/10.1016/j.scr.2021.102196
- Dong, Tao; Zhao, Yan; Jin, Hai-Feng et al. (2022). SNTA1-deficient human cardiomyocytes demonstrate hypertrophic phenotype and calcium handling disorder. Stem Cell Research & Therapy. https://doi.org/10.1186/s13287-022-02955-4