PROJECT SUMMARY: The objective of this proposal is to test cerebroprotective interventions for the SPAN program by utilizing an intraluminal middle cerebral artery occlusion (MCAO) mouse model or an animal blood clot embolic model. Our lab has been using innovative techniques in producing highly consistent mouse MCAO and clinically relevant animal blood clot embolic stroke models. Our animal surgeons have more than 15 years of experience in various animal ischemic stroke models and have performed surgeries on thousands of animals of various species (e.g., mice, rats, and rabbits). We established an easy-to-use technique to monitor the middle cerebral artery (MCA) blood flow throughout the peri-MCAO periods. Our animal operating and behavioral exam rooms have about 900 square feet of space and were completely renovated in 2019. Our three animal surgical stations with matching monitoring instruments are state-of-the- art. Our Bruker 9.4 T MRI Scanners are advanced and allow for high-resolution quantitative assessment of CNS structures and functions. Our state-of-the-art infrastructure, together with our highly skilled personnel, allows us to run multiple studies in parallel. We have about a decade of experience in performing interactive multi-institutional projects funded by the NIH. We have published more than ten studies about stroke-related comorbidities. Death and disability/dependency are always key primary outcome measures in stroke clinical trials. However, many functional tests in animal stroke studies are not designed to assess animal “natural (i.e., minimal investigator interaction)” disability; rather, animals are required or forced to perform tasks. These tasks are highly useful to test specific deficits but may differ from “natural” disability/dependency that presents in stroke clinical trials. Therefore, we recently established two novel tests to reflect animal long-term “natural” disability (unable to work, eat, and drink by themselves): (i) nest building activity measuring ability to work, and (ii) PhenoTyper monitoring the “total” activity, food and drink intake activities, and time of death. These long-term “natural” behavioral tests are objective, easy-to-use, measurable, and sensitive, and may mimic the clinically relevant disability/dependency benchmarks used in stroke clinical trials. In the first year, we will: (i) set up the required infrastructure and animal models; (ii) sign agreements and participate in all SPAN meetings; (iii) share data with the Coordinating Center (CC); and (iv) execute the animal studies following the assignments and protocols set forth by SPAN. In the second year, we will further: (i) execute the animal studies; (ii) present our results to SPAN for recommendations of go/no-go, and (iii) participate in all SPAN meetings and share resources, infrastructure, and protocols. In the third year, we will continue to execute the animal studies and participate in all scheduled SPAN meetings. We will consult the CC: (i) to get a consensus of the best intervention(s) and, if agreed by the CC, (ii) validate the clinical benefits of the best intervention(s) with a clinically relevant animal blood clot embolic stroke model.

Project Narrative: Ischemic stroke is a devastating neurological disease affecting millions of Americans. There is now an urgent need to develop therapeutics against stroke brain injury. This proposal will use state-of-the-art animal models, animal research infrastructure, and animal facilities to test up to eight cerebroprotective interventions for the SPAN program.