Downregulating the P2X3 receptor in the carotid body to reduce blood pressure via acoustic gene delivery in canines
Downregulating the P2X3 receptor in the carotid body to reduce blood pressure via acoustic gene delivery in canines
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通过声学基因传递下调颈动脉体中的 P2X3 受体以降低犬科动物的血压
DOI：
10.1016/j.trsl.2020.06.005
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发表时间：
2021
影响因子：
7.8
通讯作者：
Huang Jing
中科院分区：
文献类型：
--
作者：
Xue Qian;Wang Ruiyu;Wang Liang;Xiong Bo;Li Lingjiao;Qian Jun;Hao Lan;Wang Zhigang;Liu Dichuan;Deng Changming;Rong Shunkang;Yao Yuanqing;Jiang Yonghong;Zhu Que;Huang Jing
颈动脉体（CB）中的嘌呤能P2 X3受体被认为是治疗高血压的新靶点，但缺乏靶向调节P2 X3受体表达的方法。在这里，我们探讨了通过局部低强度聚焦超声（LIFU）介导的基因递送来降低血压的CBs中靶向P2 X3受体下调的可行性。将32只昆明种犬随机分为治疗组（n = 14）、阴性对照组（n = 10）、LIFU +阳离子微泡组（n = 4）和单纯LIFU组（n = 4）。注射载有质粒的阳离子微泡，并用基于LIFU的换能器照射双侧CB。流式细胞仪检测结果显示，33.15%的转染细胞表达绿色荧光蛋白报告基因。T7内切酶I检测显示插入-缺失率为8.30%。治疗组CBs中P2 X3受体mRNA和蛋白表达水平分别下降56.31%和45.10%。与基线值相比，治疗组的平均收缩压（152.5 ± 3.0 vs 138.0 ± 2.9 mm Hg，P= 0.003）和舒张压（97.8 ± 1.5 vs 87.2 ± 2.3 mm Hg，P= 0.002）在第14天降低，而LIFU治疗或单独注射阳离子微泡未观察到影响。用该策略治疗的犬未表现出局部或全身不良事件。因此，LIFU介导的基因传递到CB成功地调制CB功能和降低血压的犬模型，这表明一个新的可能性，治疗高血压和进一步的临床翻译。
The purinergic P2X3 receptor in the carotid body (CB) is considered a new target for treating hypertension, although approaches for targeted regulating P2X3 receptor expression are lacking. Here, we explored the feasibility of targeted P2X3 receptor down-regulation in CBs by localized low-intensity focused ultrasound (LIFU)-mediated gene delivery to reduce the blood pressure. Thirty-two Kunming canines were randomly assigned to the treatment group (n = 14), negative control group (n = 10), LIFU + cationic microbubbles group (n = 4), and LIFU-only group (n = 4). Plasmid-loaded cationic microbubbles were injected and bilateral CBs were irradiated with a LIFU-based transducer. Flow cytometry showed that 33.15% of transfected cells expressed the green fluorescent protein reporter gene. T7 endonuclease I assays showed an insertion-deletion rate of 8.30%. The P2X3 receptor mRNA- and protein-expression levels in CBs decreased by 56.31% and 45.10%, respectively, in the treatment group. Mean systolic (152.5 ± 3.0 vs 138.0 ± 2.9 mm Hg,P= 0.003) and diastolic (97.8 ± 1.5 vs 87.2 ± 2.3 mm Hg,P= 0.002) blood pressures reduced on day 14 in the treatment group, compared with the baseline values, whereas no effects were observed with LIFU treatment or cationic microbubbles injection alone. Canines treated with this strategy exhibited no local or systemic adverse events. Thus, LIFU-mediated gene delivery to CBs successfully modulated CB function and reduced blood pressure in a canine model, suggesting a new possibility for treating hypertension and further clinical translation.