Effective silencing of miR-126 after ischemic stroke by means of intravenous α-tocopherol–conjugated heteroduplex oligonucleotide in mice

Abstract

Brain endothelial cells (BECs) are involved in the pathogenesis of ischemic stroke. Recently, several microRNAs (miRNAs) in BECs were reported to regulate the endothelial function in ischemic brain. Therefore, modulation of miRNAs in BECs by a therapeutic oligonucleotide to inhibit miRNA (antimiR) could be a useful strategy for treating ischemic stroke. However, few attempts have been made to achieve this strategy via systemic route due to lack of efficient delivery-method toward BECs. Here, we have developed a new technology for delivering an antimiR into BECs and silencing miRNAs in BECs, using a mouse ischemic stroke model. We designed a heteroduplex oligonucleotide, comprising an antimiR against miRNA-126 (miR-126) known as the endothelial-specific miRNA and its complementary RNA, conjugated to α-tocopherol as a delivery ligand (Toc-HDO targeting miR-126). Intravenous administration of Toc-HDO targeting miR-126 remarkably suppressed miR-126 expression in ischemic brain of the model mice. In addition, we showed that Toc-HDO targeting miR-126 was delivered into BECs more efficiently than the parent antimiR in ischemic brain, and that it was delivered more effectively in ischemic brain than non-ischemic brain of this model mice. Our study highlights the potential of this technology as a new clinical therapeutic option for ischemic stroke.

Therapeutic gene delivery by mesenchymal stem cell for brain ischemia damage: Focus on molecular mechanisms in ischemic stroke

“…regulates the activities of endothelial progenitor cells in hypoxic environments and may prevent inflammation and oxidative stress brought on by ischemia. 198,199 By blocking an increase in MMPs levels and stopping the decline in junctional proteins, such as occludin, claudin-5, and zonula occludens-1 (ZO-1), it lessens the breakdown of the blood-brain barrier. Furthermore, miR-126 promotes neurogenesis and angiogenesis following a stroke.…”

“…MicroRNA (miR)‐126 is a miR unique to endothelial cells that has a role in angiogenesis. MiR‐126 regulates the activities of endothelial progenitor cells in hypoxic environments and may prevent inflammation and oxidative stress brought on by ischemia 198,199 . By blocking an increase in MMPs levels and stopping the decline in junctional proteins, such as occludin, claudin‐5, and zonula occludens‐1 (ZO‐1), it lessens the breakdown of the blood‐brain barrier.…”

DNA/RNA heteroduplex oligonucleotides: An unanticipated twist in the delivery of ASOs

“…The Yokota lab has now published a series of studies investigating HDOs in preclinical models. 5 , 7 , 8 , 9 , 10 , 11 , 12 In this issue of Molecular Therapy: Nucleic Acids , Kaburagi et al. utilized the Yokota lab’s previously published tocopherol-conjugated HDO (Toc-HDO) to investigate the ability of systemically administered Toc-HDOs to efficiently enter dorsal root ganglia (DRG) neurons in the peripheral nervous system.…”