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腺相关病毒介导的微小RNA在大鼠心力衰竭模型中的递送方案。

Protocol for adeno-associated virus-mediated miRNA delivery in a rat heart failure model.

作者信息

Mushtaq Iqra, Kao Yu-Hsun, Chen Yi-Jen

机构信息

International Ph.D. Program in Medicine, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan.

International Ph.D. Program in Medicine, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan; Graduate Institute of Clinical Medicine, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan; Department of Medical Education and Research, Wan Fang Hospital, Taipei Medical University, Taipei 11031, Taiwan.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103498. doi: 10.1016/j.xpro.2024.103498. Epub 2024 Dec 12.

DOI:10.1016/j.xpro.2024.103498
PMID:39671280
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11699408/
Abstract

Delivering microRNA (miRNA) to treat cardiac disease is a significant challenge, and selecting an efficient delivery method to target the affected organ is critical for therapeutic success. Here, we present a protocol for delivering miRNA to the heart of rats with heart failure using adeno-associated virus (AAV9) coupled with a hydrodynamic transfusion strategy. We describe steps for inducing heart failure, echocardiography, and AAV9 application. Further, we provide a comprehensive procedure to detect miRNA expression in cardiac tissues using stem-loop RT-qPCR. For complete details on the use and execution of this protocol, please refer to Mushtaq et al..

摘要

递送微小RNA（miRNA）来治疗心脏病是一项重大挑战，选择一种有效的递送方法以靶向受影响器官对于治疗成功至关重要。在此，我们展示了一种使用腺相关病毒（AAV9）并结合流体动力学输血策略将miRNA递送至心力衰竭大鼠心脏的方案。我们描述了诱导心力衰竭、超声心动图检查和应用AAV9的步骤。此外，我们提供了一种使用茎环逆转录定量聚合酶链反应（stem-loop RT-qPCR）检测心脏组织中miRNA表达 的全面程序。有关此方案的使用和执行的完整详细信息，请参考穆斯塔克等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f5f/11699408/81177b22409c/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f5f/11699408/a244eab94591/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f5f/11699408/9e3f43996181/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f5f/11699408/62e1e391e921/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f5f/11699408/81177b22409c/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f5f/11699408/a244eab94591/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f5f/11699408/9e3f43996181/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f5f/11699408/62e1e391e921/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f5f/11699408/81177b22409c/gr3.jpg

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本文引用的文献

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iScience. 2024 May 23;27(6):110084. doi: 10.1016/j.isci.2024.110084. eCollection 2024 Jun 21.
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Challenges in scaling up AAV-based gene therapy manufacturing.大规模生产基于腺相关病毒的基因治疗的挑战。
Trends Biotechnol. 2023 Oct;41(10):1268-1281. doi: 10.1016/j.tibtech.2023.04.002. Epub 2023 Apr 30.
3
MicroRNAs: diagnostic, prognostic and therapeutic role in heart failure-a review.
微小 RNA：心力衰竭的诊断、预后和治疗作用——综述。
ESC Heart Fail. 2023 Apr;10(2):753-761. doi: 10.1002/ehf2.14153. Epub 2022 Nov 8.
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The microRNA Expression Profiling in Heart Failure: A Systematic Review and Meta-Analysis.心力衰竭中的微小RNA表达谱：一项系统评价与荟萃分析
Front Cardiovasc Med. 2022 Jun 15;9:856358. doi: 10.3389/fcvm.2022.856358. eCollection 2022.
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Quantitative analysis of genome packaging in recombinant AAV vectors by charge detection mass spectrometry.通过电荷检测质谱法对重组腺相关病毒（AAV）载体中的基因组包装进行定量分析。
Mol Ther Methods Clin Dev. 2021 Aug 26;23:87-97. doi: 10.1016/j.omtm.2021.08.002. eCollection 2021 Dec 10.
6
Adeno-associated virus-mediated delivery of anti-miR-199a tough decoys attenuates cardiac hypertrophy by targeting .腺相关病毒介导的抗miR-199a强力诱饵的递送通过靶向……减轻心肌肥大。
Mol Ther Nucleic Acids. 2020 Nov 17;23:406-417. doi: 10.1016/j.omtn.2020.11.007. eCollection 2021 Mar 5.
7
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Lab Anim. 2019 Apr;53(2):190-201. doi: 10.1177/0023677218786982. Epub 2018 Aug 8.
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Emerging Issues in AAV-Mediated Gene Therapy.腺相关病毒介导的基因治疗中的新问题
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