Investigating the Role of Dach1 in Artery Specification and Collateral Artery Development

研究 Dach1 在动脉规格和侧支动脉发育中的作用

• 批准号: 9759081
• 负责人: Brian Christopher Raftrey
• 金额: $ 3.38万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9759081
• 关键词: Adult; Affect; Alleles; Arteries; Blood; Blood Vessels; Blood flow; Candidate Disease Gene; Cause of Death; Cell Cycle; Cells; Collaborations; Confocal Microscopy; Coronary; Coronary Arteriosclerosis; Coronary Circulation; Coronary artery; Development; Embryo; Endothelial Cells; Endothelium; Equus caballus; Gene Expression; Genes; Genetic; Goals; Growth; Heart; Heart Diseases; Heart failure; Human; In Vitro; Institution; Knock-out; Laboratories; Lead; Left; Ligation; Measures; Medical; Mentorship; Modeling; Molecular; Mus; Myocardial Infarction; Myocardial Ischemia; Natural regeneration; Operative Surgical Procedures; Outcomes Research; Pathway interactions; Patients; Pharmacology; Population; Protocols documentation; Recovery; Research; Role; Route; Tamoxifen; Testing; Therapeutic; Transcriptional Activation; Transgenic Mice; Universities; Veins; Venous; Wild Type Mouse; Writing; cardiac regeneration; cardiogenesis; career development; experimental study; faculty mentor; graduate student; improved; in vivo; meetings; mortality; mutant; novel; novel therapeutic intervention; overexpression; prevent; targeted treatment; transcription factor; transcriptome sequencing

Coronary artery disease is the leading cause of death worldwide and development of new therapeutic strategies could reduce mortality. Collateral arteries connect arteries to one another and provide multiple routes for coronary circulation. In patients with coronary artery disease, those that have well-developed collateral arteries have a reduced rate of mortality. Unfortunately, due to our poor understanding of the basic mechanisms responsible for artery growth, there are no therapies to increase collateral artery coverage. In order to find novel genes which regulate artery development, we generated a transgenic mouse line that over expressed the transcription factor Dach1 in endothelial cells. Using whole mouse immunostaining and confocal microscopy, we found that Dach1 over expression mice had increased artery and collateral artery coverage during development. Through a combination of in vitro and in vivo experiments we went on to find that Dach1 over expression increased expression increased the expression of arterial genes and decreased venous gene expression supporting a model where Dach1 induces the specification of artery endothelial cells. The goal of this proposal is to 1) understand how the pathway activated by Dach1 increases arterial cell specification and 2) test if increasing collateral artery coverage though Dach1 overexpression can protect against myocardial infarction. We will find the factors that act downstream of Dach1 to initiate artery specification by genetically or pharmacologically inhibiting candidate factors in the Dach1 overexpression background and visualizing artery formation using confocal microscopy. In addition, we will use the mouse coronary artery ligation model to test if adult mice over expressing Dach1 can make more collateral arteries post- ligation and increase survival. Answering these questions will elucidate new molecular regulators of artery growth that could be targeted therapeutically in patients with coronary artery disease. This research will be conducted in the laboratory of Dr. Kristy Red-Horse at Stanford University. Both the faculty mentor and institution are well suited for the scientific and career development of the applicant. Specifically, the applicant will benefit from abundant opportunities for scientific presentations and writing practice, mentorship of junior graduate students, access to state of the art facilities for experimenting with mouse genetics, weekly meetings with Dr. Red-Horse, and close collaboration with Dr. Dan Bernstein who will lead the surgical studies.

冠状动脉疾病是全世界死亡的主要原因，新的治疗方法的开发 治疗策略可以降低死亡率。侧支动脉将动脉相互连接 并为冠脉循环提供多种途径。对于患有冠状动脉疾病的患者来说， 侧支动脉发达的人死亡率较低。 不幸的是，由于我们对动脉的基本机制了解甚少。 生长，没有治疗方法可以增加侧支动脉覆盖范围。 为了找到调节动脉发育的新基因，我们生成了一个 在内皮细胞中过度表达转录因子 Dach1 的转基因小鼠系。 使用全小鼠免疫染色和共聚焦显微镜，我们发现 Dach1 超过 表达小鼠在发育过程中动脉和侧支动脉覆盖率增加。 通过体外和体内实验的结合，我们发现 Dach1 超过 表达增加 表达增加动脉基因的表达并减少 静脉基因表达支持 Dach1 诱导动脉特化的模型 内皮细胞。 该提案的目标是 1) 了解 Dach1 激活的途径 增加动脉细胞规格，2) 测试是否增加侧支动脉覆盖 Dach1 过度表达可以预防心肌梗塞。我们会找到起作用的因素 Dach1 下游通过遗传或药理学启动动脉规范 抑制 Dach1 过表达背景中的候选因子并可视化动脉 使用共焦显微镜形成。另外，我们将使用小鼠冠状动脉结扎术 模型来测试过度表达 Dach1 的成年小鼠是否可以在术后产生更多的侧支动脉 连接并提高存活率。回答这些问题将阐明新的分子 动脉生长调节剂可作为冠状动脉患者治疗的靶点 疾病。 这项研究将在斯坦福大学 Kristy Red-Horse 博士的实验室进行 大学。教师导师和机构都非常适合科学和职业 申请人的发展。具体来说，申请人将受益于丰富的机会 用于科学演示和写作练习、初级研究生的指导、访问 使用最先进的小鼠遗传学实验设施，每周与 Dr. 会面。 Red-Horse 以及与领导外科研究的 Dan Bernstein 博士密切合作。