Department of Ophthalmology and Visual Sciences, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, United States.
Department of Biomedical Engineering, Vanderbilt University, Nashville, Tennessee 37235, United States.
ACS Chem Biol. 2020 Nov 20;15(11):3004-3012. doi: 10.1021/acschembio.0c00662. Epub 2020 Oct 20.
Bone marrow-derived progenitor cells and macrophages are known to migrate into the retina in response to inflammation and neovascularization. These migratory cells might play important regulatory roles in the pathogenesis of neovascularization, a common complication observed in diabetic retinopathy, retinopathy of prematurity, and retinal vein occlusion. Hypoxia-inducible factor 1α (HIF-1α) has been shown to contribute to the pathogenesis of retinal inflammation and neovascularization. However, contributions of monocyte-derived macrophages to neovascularization are largely unknown. We hypothesized that selective visualization of these microglia/macrophages could be a powerful method for predicting the onset of neovascularization and its progression at the molecular level. In this report, we describe the synthesis of a new hybrid nanoparticle to visualize HIF-1α mRNA selectively in microglia/macrophages in a mouse model of neovascularization. expression was confirmed in positive monocytes/macrophages as well as in CD4 positive T-cells and CD19 positive B-cells using single-cell RNA sequencing data analysis. The imaging probes (AS- or NS-shRNA-lipid) were synthesized by conjugating diacyl-lipids to short hairpin RNA with an antisense sequence complementary to HIF-1α mRNA and a fluorophore that is quenched by a black hole quencher. We believe that imaging mRNA selectively in tissue specific microglia/macrophages could be a powerful method for predicting the onset of neovascularization, its progression, and its response to therapy.
骨髓源性祖细胞和巨噬细胞已知会在炎症和新生血管形成的情况下迁移到视网膜。这些迁移细胞可能在新生血管形成的发病机制中发挥重要的调节作用，新生血管形成是糖尿病视网膜病变、早产儿视网膜病变和视网膜静脉阻塞等常见并发症。缺氧诱导因子 1α（HIF-1α）已被证明有助于视网膜炎症和新生血管形成的发病机制。然而，单核细胞衍生的巨噬细胞对新生血管形成的贡献在很大程度上是未知的。我们假设，这些小胶质细胞/巨噬细胞的选择性可视化可能是一种强大的方法，可以在分子水平上预测新生血管形成及其进展。在本报告中，我们描述了一种新的杂交纳米颗粒的合成，用于在新生血管形成的小鼠模型中选择性地可视化小胶质细胞/巨噬细胞中的 HIF-1α mRNA。使用单细胞 RNA 测序数据分析，证实了在阳性单核细胞/巨噬细胞以及 CD4 阳性 T 细胞和 CD19 阳性 B 细胞中表达。成像探针（AS 或 NS-shRNA-脂质）是通过将二酰基脂质与具有与 HIF-1α mRNA 互补的反义序列的短发夹 RNA 缀合，并与荧光团缀合而合成的，该荧光团被黑洞猝灭剂猝灭。我们相信，选择性地在组织特异性小胶质细胞/巨噬细胞中成像 mRNA 可能是一种强大的方法，可以预测新生血管形成的发生、其进展及其对治疗的反应。
