Inducible miR-1224 silences cerebrovascular Serpine1 and restores blood flow to the stroke-affected site of the brain

Ravichand Palakurti; Nirupam Biswas; Sashwati Roy; Surya C Gnyawali; Mithun Sinha; Kanhaiya Singh; Subhadip Ghatak; Chandan K Sen; Savita Khanna

doi:10.1016/j.omtn.2022.12.019

Inducible miR-1224 silences cerebrovascular Serpine1 and restores blood flow to the stroke-affected site of the brain

Mol Ther Nucleic Acids. 2023 Jan 2:31:276-292. doi: 10.1016/j.omtn.2022.12.019. eCollection 2023 Mar 14.

Authors

Ravichand Palakurti¹, Nirupam Biswas¹, Sashwati Roy¹, Surya C Gnyawali¹, Mithun Sinha¹, Kanhaiya Singh¹, Subhadip Ghatak¹, Chandan K Sen^{1

2}, Savita Khanna¹

Affiliations

¹ Department of Surgery, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
² Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, USA.

Abstract

The α-tocotrienol (TCT) form of natural vitamin E is more potent than the better known α-tocopherol against stroke. Angiographic studies of canine stroke have revealed beneficial cerebrovascular effects of TCT. This work seeks to understand the molecular basis of such effect. In mice, TCT supplementation improved perfusion at the stroke-affected site by inducing miR-1224. miRNA profiling of a laser-capture-microdissected stroke-affected brain site identified miR-1224 as the only vascular miR induced. Lentiviral knockdown of miR-1224 significantly blunted the otherwise beneficial effects of TCT on stroke outcomes. Studies on primary brain microvascular endothelial cells revealed direct angiogenic properties of miR-1224. In mice not treated with TCT, advance stereotaxic delivery of an miR-1224 mimic to the stroke site markedly improved stroke outcomes. Mechanistic studies identified Serpine1 as a target of miR-1224. Downregulation of Serpine1 augmented the angiogenic response of the miR-1224 mimic in the brain endothelial cells. The inhibition of Serpine1, by dietary TCT and pharmacologically, increased cerebrovascular blood flow at the stroke-affected site and protected against stroke. This work assigns Serpine1, otherwise known to be of critical significance in stroke, a cerebrovascular function that worsens stroke outcomes. miR-1224-dependent inhibition of Serpine1 can be achieved by dietary TCT as well as by the small-molecule inhibitor TM5441.

Keywords: MCAO; MT: Noncoding RNAs; Serpine1; miR-1224; stroke; tocotrienol.