Although some of the cellular mechanisms of neuronal injury after ischemic stroke are fairly well understood, the neuronal death after ischemic stroke cannot be well controlled because of a lack of efficient treatments. The long-term goal of my research is to develop an effective strategy for ischemic stroke therapy. The proposed research is focused on targeting the post- ischemic inflammation at blood-brain barrier (BBB) in search of new approaches for stroke therapy. We propose a novel RNA nanoparticle, 3WJ-pRNA/2siICAM-FB4, consisting of one RNA aptamer (FB4), two ICAM-1 siRNA and a 3-way junction of packaging RNA (3WJ-pRNA) of bacteriophage phi29. FB4 is a RNA aptamer specific to transferrin receptor (TfR) that can recognize TfRs at the BBB and facilitate RNA nanoparticle delivery into brain endothelial cells. Once the 3WJ-pRNA/2siICAM-FB4 enters the cell, the siRNAs will be released and knockdown ICAM-1 expression leading to inhibition of inflammatory responses at the BBB. The pRNA 3-way junction serves as the core for holding FB4 and siRNAs together without disrupting the folding of either of the attached payload. We will use mouse transient ischemic stroke model in vivo and oxygen-glucose deprivation in primary brain endothelial cells in vitro to demonstrate the feasibility of this vascular targeting strategy for treatment of ischemic stroke. Our central hypothesis is that the 3WJ-pRNA/2siICAM-FB4 has therapeutic effects on ischemic stroke by inhibition of post-ischemic inflammation.
Specific Aims :
Aim 1 : Determine pharmacological activity of 3WJ-pRNA/2siICAM-FB4 in vitro.
Aim 2 : Evaluate protective effect of 3WJ- pRNA/2siICAM-FB4 in transient ischemic stroke model in vivo. This project will develop a novel brain-targeted RNA therapeutics for therapy of ischemic stroke, and this approach could be broadly applied for therapy of diseases affecting the BBB and the central nervous system (CNS).

Public Health Relevance

The proposed studies address an important area of research concerned with the development of better drugs to treat ischemic stroke. The findings can have wide implications for potential RNA-based therapeutics, which are often hampered by their inability to reach the target tissues.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21NS093493-01
Application #
8953140
Study Section
Drug Discovery for the Nervous System Study Section (DDNS)
Program Officer
Bosetti, Francesca
Project Start
2015-06-01
Project End
2017-05-31
Budget Start
2015-06-01
Budget End
2016-05-31
Support Year
1
Fiscal Year
2015
Total Cost
Indirect Cost
Name
University of Cincinnati
Department
Type
Schools of Pharmacy
DUNS #
041064767
City
Cincinnati
State
OH
Country
United States
Zip Code
45221
Sakabe, Masahide; Fan, Jieqing; Odaka, Yoshinobu et al. (2017) YAP/TAZ-CDC42 signaling regulates vascular tip cell migration. Proc Natl Acad Sci U S A 114:10918-10923
Hu, Jing; Al-Waili, Daniah; Hassan, Aishlin et al. (2016) Inhibition of cerebral vascular inflammation by brain endothelium-targeted oligodeoxynucleotide complex. Neuroscience 329:30-42