Targeting RyR2 with a phosphorylation site-specific nanobody reverses dysfunction of failing cardiomyocytes in rats
| Autor: | Yongji Yang, Shi Hu, Ying Tang, Chuqi Wang, Yafeng Shen, Changhai Lei, Jizhou Liang, Wenyan Fu, Tian Li, Xuting Ye, Fangxing Lin, Xiaoyan Fan, Shuowu Liu, Min Ding |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Male Genetic Vectors chemistry.chemical_element Gene delivery Calcium Biochemistry Ryanodine receptor 2 Calcium in biology Contractility Rats Sprague-Dawley 03 medical and health sciences 0302 clinical medicine In vivo Genetics Animals Myocytes Cardiac Phosphorylation Molecular Biology Heart Failure Gene Transfer Techniques Ryanodine Receptor Calcium Release Channel Dependovirus Single-Domain Antibodies Cell biology Rats 030104 developmental biology chemistry Animals Newborn cardiovascular system 030217 neurology & neurosurgery Intracellular Biotechnology |
| Zdroj: | FASEB journal : official publication of the Federation of American Societies for Experimental Biology. 33(6) |
| ISSN: | 1530-6860 |
| Popis: | Chronic PKA phosphorylation of ryanodine receptor 2 (RyR2) has been shown to increase diastolic sarcoplasmic reticulum (SR) Ca2+ leakage and lead to cardiac dysfunction. We hypothesize that intracellular gene delivery of an RyR2-targeting phosphorylation site-specific nanobody could preserve the contractility of the failing myocardium. In the present study, we acquired RyR2-specific nanobodies from a phage display library that were variable domains of Camelidae heavy chain-only antibodies. One of the nanobodies, AR185, inhibited RyR2 phosphorylation in vitro and was chosen for further investigation. We investigated the potential of adeno-associated virus (AAV)9-mediated cardiac expression of AR185 to combat postischemic heart failure (HF). AAV gene delivery elevated the intracellular expression of the AR185 protein in a rat model of ischemic HF, and this treatment normalized the systolic and diastolic dysfunction of the failing myocardium in vivo by reversing myocardial Ca2+ handling. Furthermore, AR185 gene transfer to failing cardiomyocytes reduced the frequency of SR calcium leaks, thereby restoring the attenuated intracellular calcium transients and SR calcium load. Moreover, AR185 gene transfer inhibited the PKA-mediated phosphorylation of RyR2 in failing cardiomyocytes. Our results provide preclinical experimental evidence that the cardiac expression of RyR2 nanobodies with AAV9 vectors is a promising therapeutic strategy for HF.-Li, T., Shen, Y., Lin, F., Fu, W., Liu, S., Wang, C., Liang, J., Fan, X., Ye, X., Tang, Y., Ding, M., Yang, Y., Lei, C., Hu, S. Targeting RyR2 with a phosphorylation site-specific nanobody reverses dysfunction of failing cardiomyocytes in rats. |
| Databáze: | OpenAIRE |
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