期刊
BIOMATERIALS
卷 275, 期 -, 页码 -出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.biomaterials.2021.121000
关键词
Extracellular vesicles; CD47; MSC; miRNA-21; Intravenous injection; Myocardial ischemia reperfusion; Biodistribution
资金
- Programs of Shanghai Science and Technology Committee Foundation [18411950200]
- Postgraduate Research & Practice Innovation Program of Jiangsu Province [KYCX18_1462]
- National Natural Science Foundation of China [81870358, 81601539, 81700392]
- Key Projects of Science and Technology of Jiangsu Province [BE2016607, BE2019602]
- Funds for Jiangsu Provincial Key Medical Discipline [ZDXKB2016013]
- Nanjing Medical Science and technique Development Foundation [QRX17057]
- Municipal Medical Science Technology Development Foundation of Nanjing [YKK19063]
- Programs of the Science Foundation in Nanjing [ZKX17011]
The study demonstrated that enhancing the biodistribution of CD47-EVs in the heart could potentially serve as a therapeutic approach to alleviate myocardial ischemia-reperfusion injury.
Mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) with anti-apoptotic and anti-inflammatory properties have been intensively studied. However, rapid clearance by the mononuclear phagocyte system remains a huge barrier for the delivery of extracellular vesicle contents into target organs and restricts its wider application, particularly in the heart. CD47 is a transmembrane protein that enables cancer cells to evade clearance by macrophages through CD47(-) signal regulatory protein alpha binding, which initiates a don't eat me signal. This study aimed to explore the biodistribution and delivery efficiency of EVs carrying the membrane protein CD47 and specific anti-apoptotic miRNAs. EVs were isolated from MSCs overexpressing CD47 (CD47EVs) and identified. Fluorescence-labeled EVs were injected through the tail vein and tracked using fluorescence imaging. In silico analysis was performed to determine miRNA profiles in MSCs and in a heart-derived H9c2 cardiomyoblast cell line under hypoxia vs. normoxia conditions. Electro CD47-EV was constructed by encapsulating purified CD47-EV with miR-21a via electroporation. The effect of miR21-EVs on the pro-apoptotic gene encoding phosphatase and tensin homolog (PTEN) was evaluated by dual-luciferase assay, qPCR, and western blotting. Exogenous miR21 distribution, PTEN protein level, blood vessel density, anti-apoptotic effect by TdTmediated dUTP nick-end labeling staining, and macrophage and leukocyte infiltration in the myocardium were assessed by immunofluorescence staining. Cardiac functional recovery during the early stage and recovery period was evaluated using echocardiography. The results showed that CD47-EVs were still detectable in the plasma 120 min after the tail vein injection, compared to the detection time of less than 30 min observed with the unmodified EVs. More strikingly, CD47-EVs preferentially accumulated in the heart in the ischemia-reperfusion (I/R) + CD47-EV group [heart total fluorescence radiance (x 10(5) Photons/sec/cm(2)/sr) 51.62 +/- 11.30 v.s. 10.08 +/- 3.15 in the I/R + unmodified EVs group] 8 h post-injection. Exogenous miR-21 is efficiently internalized into cardiomyocytes, inhibits apoptosis, alleviates inflammation, and improves cardiac function. In conclusion, electro CD47-EVs efficiently improve biodistribution in the heart, shedding new light on the application of a two-step EV delivery method (CD47 genetic modification followed by therapeutic content electrotransfection) as a potential therapeutic tool for myocardial I/R injury that may benefit patients in the future.
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