Immunotherapy for Infarcts: In Vivo Postinfarction Macrophage Modulation Using Intramyocardial Microparticle Delivery of Map4k4 Small Interfering RNA.
Immunotherapy for Infarcts: In Vivo Postinfarction Macrophage Modulation Using Intramyocardial Microparticle Delivery of Map4k4 Small Interfering RNA.
复制标题
DOI：
10.1089/biores.2020.0037
复制
发表时间：
2020
影响因子：
--
通讯作者：
Allen MD
中科院分区：
文献类型：
--
作者：
Luo J;Weaver MS;Fitzgibbons TP;Aouadi M;Czech MP;Allen MD
急性心肌梗死后早期浸润心脏的髓样细胞形成了一个分泌蛋白组，在很大程度上协调了随后的心室壁修复。调节这种先天免疫反应可能是改善梗死愈合的一种手段。为了试验这一概念，我们使用了（β 1，3-d-）葡聚糖封装的含小干扰RNA（siRNA）的颗粒（GeRPs），靶向单核吞噬细胞，在急性梗死后立即一次性心肌内注射给小鼠。研究结果表明，心脏巨噬细胞在体内吞噬GeRPs，几乎没有全身性传播，从而提供了一种手段，提供局部治疗。然后在体内用磷酸盐缓冲盐水（PBS;载体）或装载有针对Map 4k 4的siRNA的GeRP注射急性梗死，并在3天和7天时通过定量聚合酶链反应、流式细胞术和组织学检查切除的心脏。与经PBS处理的梗死心脏相比，梗死内注射siRNA负载的GeRPs的心脏在3天时显示Map 4k 4（促分裂原活化蛋白激酶4）、白细胞介素（IL）-1β和肿瘤坏死因子α的转录物减少69-89%。与基质重塑相关的其他因子-单核细胞趋化蛋白-1（MCP-1）、基质金属蛋白酶、透明质酸酶、基质细胞蛋白和促纤维化因子转化生长因子β（TGF-β）和结缔组织生长因子（CTGF）的表达也降低。大多数作用在第3天达到峰值，但在某些情况下（Map 4k 4、IL-1β、TGF-β、CTGF、多功能蛋白聚糖和骨膜蛋白），抑制持续至第7天。因此，直接心肌内注射GeRP可以作为一种新的和临床上可翻译的平台，在体内RNA递送到心内巨噬细胞局部和选择性免疫调节梗死微环境。
The myeloid cells infiltrating the heart early after acute myocardial infarction elaborate a secretome that largely orchestrates subsequent ventricular wall repair. Regulating this innate immune response could be a means to improve infarct healing. To pilot this concept, we utilized (β1,3-d-) glucan-encapsulated small interfering RNA (siRNA)-containing particles (GeRPs), targeting mononuclear phagocytes, delivered to mice as a one-time intramyocardial injection immediately after acute infarction. Findings demonstrated that cardiac macrophages phagocytosed GeRPs in vivo and had little systemic dissemination, thus providing a means to deliver local therapeutics. Acute infarcts were then injected in vivo with phosphate-buffered saline (PBS; vehicle) or GeRPs loaded with siRNA to Map4k4, and excised hearts were examined at 3 and 7 days by quantitative polymerase chain reaction, flow cytometry, and histology. Compared with infarcted PBS-treated hearts, hearts with intrainfarct injections of siRNA-loaded GeRPs exhibited 69–89% reductions in transcripts for Map4k4 (mitogen-activated protein kinase kinase kinase kinase 4), interleukin (IL)-1β, and tumor necrosis factor α at 3 days. Expression of other factors relevant to matrix remodeling—monocyte chemoattractant protein-1 (MCP-1), matrix metalloproteinases, hyaluronan synthases, matricellular proteins, and profibrotic factors transforming growth factor beta (TGF-β), and connective tissue growth factor (CTGF)—were also decreased. Most effects peaked at 3 days, but, in some instances (Map4k4, IL-1β, TGF-β, CTGF, versican, and periostin), suppression persisted to 7 days. Thus, direct intramyocardial GeRP injection could serve as a novel and clinically translatable platform for in vivo RNA delivery to intracardiac macrophages for local and selective immunomodulation of the infarct microenvironment.