Comparative transcriptome of brain blood vessels: age, hypertension and diabetes

描述（申请人提供）：脑血管在脑卒中的病理生理中起关键作用。我们都“知道”，脑血管很可能不同于其他器官的血管。例如，脑血管具有独特的紧密连接，有助于形成血脑屏障。但除了这种屏障表型，我们对其他的知之甚少。令人惊讶的是，对脑血管进行系统分析并将其与其他器官的血管进行比较的研究从未进行过。在本RC2提案中，我们试图填补这一重要的知识空白。我们的总体目标是组装小鼠脑、心脏和肾脏内皮细胞和平滑肌细胞的比较转录组（mRNA表达谱）。我们将提出以下问题：1。小鼠脑灰质与白质血管基因表达是否存在差异？2. 脑血管的基因表达谱与心脏和肾脏的不同吗？3. 年龄、高血压和糖尿病对脑血管的影响与对心脏和肾脏血管的影响不同吗？内皮细胞和平滑肌细胞将通过两种互补的方式收集：激光捕获新鲜冷冻组织的显微解剖，以及从新鲜制备的整个组织匀浆中分离原代细胞。使用Affymetrix小鼠基因组430 2.0芯片分析基因表达。对于年龄，我们将比较“年轻”（5个月）和“年轻”（5个月）。“老”（15个月）C57Bl6小鼠。对于高血压，我们将比较正常BPN/3J小鼠和自发高血压BPH/2J小鼠。对于糖尿病，我们将比较匹配野生型对照与糖尿病db/db小鼠。这个项目应该产生一个新的数据库，可以被所有中风研究人员用来研究新的假设、机制和目标。就血管功能障碍可能在神经退行性变中起作用而言，这些结果也可以影响其他广泛的中枢神经系统疾病。最后，虽然我们主要关注的是中风，但了解两种主要疾病（高血压和糖尿病）是如何系统地影响血管基因表达的，对许多其他生物医学领域也会有广泛的帮助。

DESCRIPTION (Provided by Applicant): Cerebral blood vessels play a key role in the pathophysiology of stroke. We all "know" that brain blood vessels are likely to be different from vessels in other organs. For example, brain blood vessels have unique tight junctions that contribute to the blood-brain barrier. But beyond this barrier phenotype, not much else is known. Surprisingly, a systematic analysis of brain blood vessels comparing them with those from other organs has never been performed. In this RC2 proposal, we seek to fill this important gap in knowledge. Our overall goal is to assemble a comparative transcriptome (mRNA expression profiles) of endothelial and smooth muscle cells in mouse brain, heart and kidney. We will ask the following questions: 1. Are there differences in vascular gene expression in gray matter vs. white matter of mouse brain? 2. Are gene expression profiles of brain blood vessels different from those in heart and kidney? 3. Will age, hypertension and diabetes affect brain blood vessels differently compared to those in heart and kidney? Endothelial and smooth muscle cells will be collected in two complementary ways: laser capture microdissection of fresh-frozen tissue, and primary cell isolation from freshly prepared whole tissue homogenates. Gene expression will be analyzed with the Affymetrix Mouse Genome 430 2.0 chip. For age, we will compare "young" (5 month) vs. "old" (15 month) C57Bl6 mice. For hypertension, we will compare normotensive BPN/3J mice vs. spontaneously hypertensive BPH/2J mice. For diabetes, we will compare matching wildtype controls vs. diabetic db/db mice. This project should yield a novel database that can be used by all stroke researchers to investigate new hypotheses, mechanisms and targets. Insofar as vascular dysfunction may play a role in neurodegeneration, these results can also impact a wide spectrum of other CNS disorders. And finally, although our primary focus here is on stroke, gaining an understanding of how two major diseases (hypertension and diabetes) systematically affect vascular gene expression should be broadly useful for many other biomedical fields as well. PUBLIC HEALTH RELEVANCE: There is no clinically effective neuroprotective therapy for stroke. Recent data suggest that focusing on neurons alone is not enough, and that blood vessels within the brain are very important. Yet, no one knows how these brain blood vessels are different from blood vessels in other organs in our body. Here, we seek to find out the full gene expression profile of brain blood vessels, and compare it with heart and kidney. And we will also find out how aging, hypertension and diabetes alter these gene patterns. This database should be invaluable for stroke research, in terms of finding new therapeutic targets in the future.