Bioengineering Strategies for Cardiovascular Disease

项目摘要 血管疾病对数百万美国人来说是常见和致命的。当前医学 血管疾病的治疗方法有限， and mortality.因此，血管疾病需要新的和新颖的疗法。长 范围目标和临床意义的建议是使用我们的新 开发了ETV 2基因敲除猪作为宿主，最终用于生产个性化的 用于临床应用的人体脉管系统。当前修订版应用程序的目标是 是在猪身上建立一个非人类灵长类动物平台， 在基因编辑的猪中工程化人源化脉管系统。我们的实验室发现 Etv 2作为Nkx 2 -5的下游靶标，并确定Etv 2突变小鼠胚胎 不能存活，缺乏内皮/血管和造血谱系。使用 CRISPR/Cas9基因编辑技术，我们进一步确定了ETV 2突变体 猪胚胎缺乏血管和血液谱系。根据我们的调查结果， 假设Etv 2是主分子程序的必要因素， 维管谱系发育过程中。在这些拟议的研究中，我们将利用 许多新兴技术可以改变非人类灵长类动物的模式 转基因动物替代品中的脉管系统。为了验证我们的假设，我们 具体目标#1：确定联合国的能力， 囊胚互补，使用GFP标记的猪卵裂球， 拯救ETV 2无效猪宿主;具体目标#2：定义 用于猪胚泡互补的非人灵长类干细胞群体 具体目标#3：在ETV 2中设计非人灵长类血管系统 变异猪宿主在这些研究中，我们将使用最先进的基因技术 和猕猴GFP标记的干细胞群来改造非人类灵长类动物 在大型动物模型中的血管。这种非人类灵长类大型动物模型将是 再生医学的重要资源，并将作为一个平台， 产生个性化的人源化猪模型。这一战略能够 对慢性血管疾病新兴疗法的发展产生深远影响， 疾病和移植。鉴于巨大的发病率和死亡率， 心血管疾病在我们的社会，这一建议可能会有重要的临床影响。

PROJECT SUMMARY Vascular disease is common and deadly for millions of Americans. Current medical therapies for vascular disease are limited and are associated with significant morbidity and mortality. Therefore, vascular diseases warrant new and novel therapies. The long range goal and the clinical significance of this proposal are to use our newly developed ETV2 knockout pigs as hosts ultimately for the production of personalized human vasculature for clinical applications. The goal of this current revised application is to establish a nonhuman primate platform in a pig that would provide the feasibility for engineering humanized vasculature in a gene edited pig. Our laboratory discovered Etv2 as a downstream target of Nkx2-5 and defined that Etv2 mutant mouse embryos were nonviable and lacked endothelial/vascular and hematopoietic lineages. Using CRISPR/Cas9 gene editing technology, we have further established that ETV2 mutant porcine embryos lack vascular and blood lineages. Based on our results, our overall hypothesis is that Etv2 is an essential factor for the master molecular program for vascular lineages during development. In these proposed studies, we will utilize a number of emerging technologies to engineer a paradigm shifting nonhuman primate vasculature in a genetically modified animal surrogate. To examine our hypotheses, we will address the following specific aims: Specific Aim #1: To define the capacity of blastocyst complementation, using GFP labeled porcine blastomeres, to fully rescue the ETV2 null porcine host; Specific Aim #2: To define the capacity of nonhuman primate stem cell populations for porcine blastocyst complementation and Specific Aim #3: To engineer nonhuman primate vasculature in the ETV2 mutant porcine host. In these studies, we will use state-of-the-art gene technologies and macaque GFP-labeled stem cell populations to engineer a nonhuman primate vasculature in a large animal model. This nonhuman primate large animal model will be an important resource for regenerative medicine and will serve as a platform for generating personalized humanized porcine models. This strategy has the capacity to have a profound impact on the development of emerging therapies for chronic vascular diseases and transplantation. Given the tremendous morbidity and mortality of cardiovascular disease in our society, this proposal could have important clinical impact.