Testing Cerebroprotective Interventions with Rodent Ischemic Stroke Models

项目摘要：这项建议的目标是测试脑保护干预措施 SPAN程序通过利用大脑中动脉阻塞(MCAO)小鼠模型或 动物血栓栓塞术模型。我们的实验室一直在使用创新技术来生产高度一致的 小鼠大脑中动脉闭塞及临床相关动物血栓栓塞性卒中模型。我们的动物外科医生 在各种动物缺血性卒中模型方面有超过15年的经验，并曾在 数以千计的不同物种的动物(如小鼠、大鼠和兔子)。我们建立了一个易于使用的 监测大脑中动脉(MCA)围手术期血流的技术。我们的 动物手术和行为检查室有大约900平方英尺的空间，完全是 于2019年翻新。我们的三个动物外科手术站配有配套的监测仪器，是最先进的- 艺术。我们的Bruker 9.4 T磁共振扫描仪是先进的，可以对 中枢神经系统的结构和功能。我们最先进的基础设施，加上我们的高技能人员， 使我们能够同时进行多项研究。我们在互动表演方面有大约十年的经验 由美国国立卫生研究院资助的多机构项目。我们已经发表了十多项与中风相关的研究 合并症。死亡和残疾/依赖一直是中风临床的主要预后指标 审判。然而，动物中风研究中的许多功能测试并不是为了评估动物的自然(即， 最低限度的调查人员互动)“残疾；相反，动物被要求或被迫执行任务。这些 任务对于测试特定的缺陷非常有用，但可能不同于呈现的“自然”残疾/依赖 在中风临床试验中。因此，我们最近建立了两个新的测试来反映动物长期的“自然” 残疾(不能自己工作、吃和喝)：(I)筑巢活动衡量工作能力， 以及(Ii)PhenoTyper监控“总”活动、食物和饮料摄入活动以及死亡时间。这些 长期的“自然”行为测试是客观的、易于使用的、可测量的和敏感的，并且可以模仿 中风临床试验中使用的与临床相关的残疾/依赖基准。在第一年，我们会：(I) 建立所需的基础设施和动物模型；(Ii)签署协议并参与所有项目 会议；(3)与协调中心(CC)共享数据；以及(4)根据 SPAN制定的任务和协议。在第二年，我们将进一步：(I)处死动物 研究；(Ii)将我们的结果提交给SPAN，以获得关于去/不去的建议，以及(Iii)参与所有SPAN 会议和共享资源、基础设施和协议。在第三年，我们将继续执行 动物研究，并参加所有预定的SPAN会议。我们会征询消委会的意见：(I)达成共识 最佳干预措施(S)，如果经CC同意，(Ii)确认最佳干预措施的临床益处(S) 与临床相关的动物血栓栓塞性中风模型。

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.