Related Experiment Video
Updated: Jul 4, 2025
A Doxorubicin-Induced Murine Model of Dilated Cardiomyopathy In Vivo
Published on: May 16, 2020
LncRNA CHKB-DT Downregulation Enhances Dilated Cardiomyopathy Through ALDH2
Xiang Nie1, Jiahui Fan1, Beibei Dai1,2
1Division of Cardiology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College (X.N., J.F., B.D., Z.W., H.L., C.C., D.W.W.), Huazhong University of Science and Technology, Wuhan, China.
Insights
The long noncoding RNA CHKB-DT is downregulated in dilated cardiomyopathy (DCM). Restoring CHKB-DT improves cardiac function and energy metabolism, suggesting it as a potential therapeutic target for DCM.
Area of Science:
- Cardiovascular Biology
- Molecular Genetics
- Noncoding RNA Research
Background:
- Dilated cardiomyopathy (DCM) is associated with altered cardiac long noncoding RNA (lncRNA) profiles.
- The lncRNA CHKB-DT (choline kinase beta divergent transcript) is notably downregulated in human DCM hearts.
Purpose of the Study:
- To elucidate the functional role of CHKB-DT in the pathogenesis of DCM.
- To investigate CHKB-DT as a potential therapeutic target for DCM.
Main Methods:
- Quantitative reverse transcription-polymerase chain reaction and in situ hybridization for lncRNA expression analysis.
- CRISPR/Cas9-generated CHKB-DT knockout mouse models and adeno-associated virus-mediated gene delivery.
- Assessment of cardiomyocyte contractility, mitochondrial function (Seahorse XF assay), and ATP production.
- Mechanistic studies including quantitative proteomics, ribosome profiling, and RNA-protein interaction assays (e.g., RNA pull-down, luciferase assay).
Main Results:
- CHKB-DT downregulation was confirmed in DCM patients and heart failure mouse models.
- CHKB-DT heterozygous knockout in cardiomyocytes led to cardiac dilation, dysfunction, and reduced contractility.
- Mitochondrial dysfunction, decreased ATP production, and impaired cardiac energy metabolism were observed in CHKB-DT knockout models.
- CHKB-DT directly targets ALDH2 mRNA, influencing its stability and downstream 4-HNE production. Restoring ALDH2 ameliorated cardiac defects in knockout mice.
Conclusions:
- CHKB-DT is significantly downregulated in DCM and plays a crucial role in maintaining cardiac function and energy metabolism.
- CHKB-DT acts as an energy metabolism-associated lncRNA and represents a promising therapeutic target for DCM.
Background:
Human cardiac long noncoding RNA (lncRNA) profiles in patients with dilated cardiomyopathy (DCM) were previously analyzed, and the long noncoding RNA CHKB (choline kinase beta) divergent transcript (CHKB-DT) levels were found to be mostly downregulated in the heart. In this study, the function of CHKB-DT in DCM was determined.
Methods:
Long noncoding RNA expression levels in the human heart tissues were measured via quantitative reverse transcription-polymerase chain reaction and in situ hybridization assays. A CHKB-DT heterozygous or homozygous knockout mouse model was generated using the clustered regularly interspaced palindromic repeat (CRISPR)/CRISPR-associated protein 9 system, and the adeno-associated virus with a cardiac-specific promoter was used to deliver the RNA in vivo. Sarcomere shortening was performed to assess the primary cardiomyocyte contractility. The Seahorse XF cell mitochondrial stress test was performed to determine the energy metabolism and ATP production. Furthermore, the underlying mechanisms were explored using quantitative proteomics, ribosome profiling, RNA antisense purification assays, mass spectrometry, RNA pull-down, luciferase assay, RNA-fluorescence in situ hybridization, and Western blotting.
Results:
CHKB-DT levels were remarkably decreased in patients with DCM and mice with transverse aortic constriction-induced heart failure. Heterozygous knockout of CHKB-DT in cardiomyocytes caused cardiac dilation and dysfunction and reduced the contractility of primary cardiomyocytes. Moreover, CHKB-DT heterozygous knockout impaired mitochondrial function and decreased ATP production as well as cardiac energy metabolism. Mechanistically, ALDH2 (aldehyde dehydrogenase 2) was a direct target of CHKB-DT. CHKB-DT physically interacted with the mRNA of ALDH2 and fused in sarcoma (FUS) through the GGUG motif. CHKB-DT knockdown aggravated ALDH2 mRNA degradation and 4-HNE (4-hydroxy-2-nonenal) production, whereas overexpression of CHKB-DT reversed these molecular changes. Furthermore, restoring ALDH2 expression in CHKB-DT+/- mice alleviated cardiac dilation and dysfunction.
Conclusions:
CHKB-DT is significantly downregulated in DCM. CHKB-DT acts as an energy metabolism-associated long noncoding RNA and represents a promising therapeutic target against DCM.
More Related Videos
05:58Mouse Electroacupuncture Fixation Device Fabrication for Electroacupuncture Pretreatment in Diabetic Cardiomyopathy Mouse Model
Published on: April 18, 2025
09:41Visualization of Cell Cycle Variations and Determination of Nucleation in Postnatal Cardiomyocytes
Published on: February 24, 2017
