Bioengineering Strategies for Cardiovascular Disease

心血管疾病的生物工程策略

• 批准号: 10227924
• 负责人: Daniel J. Garry
• 金额: $ 76.94万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10227924
• 关键词: Address; Adolescent; Adult; Affect; American; Animal Model; Biomedical Engineering; Blood; Blood Vessels; Bypass; CRISPR/Cas technology; Cardiovascular Diseases; Cells; Chimera organism; Chronic; Chronic Disease; Clinical; Cloning; Complement; Coronary Artery Bypass; Data; Development; Diabetes Mellitus; Disease; Embryo; Emerging Technologies; Endothelium; Engineering; Family suidae; Future Generations; Genes; Genetic Engineering; Genetically Modified Animals; Goals; Hematological Disease; Hematopoietic; Human; In Vitro; Incidence; Inner Cell Mass; Knock-out; Label; Laboratories; Macaca; Medical; Modeling; Molecular; Morbidity - disease rate; Morphology; Mutant Strains Mice; Obesity; Operative Surgical Procedures; Patients; Peripheral arterial disease; Pharmacology; Physiology; Production; Publications; Publishing; Regenerative Medicine; Regenerative research; Research; Resources; Signal Transduction; Societies; Source; Technology; Technology Transfer; Testing; Therapeutic Intervention; Tissues; Transplantation; Vascular Diseases; Vascular Endothelium; base; blastocyst; boar; body system; clinical application; clinically significant; embryo stage 2; immunological status; in vivo; innovation; innovative technologies; limb amputation; mortality; mutant; nonhuman primate; novel; novel therapeutics; porcine model; programs; response; somatic cell nuclear transfer; stem cell population; stem cells; transcription factor; transcriptome

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.

项目摘要 血管疾病对于数百万美国人来说是常见的，而且致命。当前的医疗 血管疾病的疗法受到限制，与明显的发病率有关 和死亡率。因此，血管疾病需要新的新型疗法。长 该提议的范围目标和临床意义是使用我们的新 开发的ETV2基因敲除猪作为宿主最终用于生产个性化 用于临床应用的人体脉管系统。此当前修订的应用程序的目标 是在猪中建立一个非人类灵长类动物平台，以提供可行性 在基因编辑的猪中工程人性化脉管系统。我们的实验室发现 ETV2作为NKX2-5的下游靶标，并定义为ETV2突变小鼠胚胎 是不可行的，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ETV2突变体 猪胚胎缺乏血管和血统。根据我们的结果，总体 假设是ETV2是主分子程序的重要因素 发育过程中的血管谱系。在这些拟议的研究中，我们将利用 设计范式转移非人类灵长类动物的新兴技术数量 在转基因动物替代物中的脉管系统。为了审查我们的假设，我们 将解决以下特定目的：特定目标＃1：定义能力 使用GFP标记为猪囊泡的胚泡互补，以完全 营救ETV2无效猪宿主；特定目的＃2：定义能力 非人类灵长类动物干细胞群，用于猪胚泡互补 和特定目的＃3：ETV2中的非人类灵长类动物脉管系统 突变猪宿主。在这些研究中，我们将使用最先进的基因技术 和猕猴GFP标记的干细胞种群来设计非人类灵长类动物 大型动物模型中的脉管系统。这个非人类灵长类动物模型将是 再生医学的重要资源，并将作为 产生个性化的人源猪模型。该策略有能力 对慢性血管的新兴疗法的发展产生深远影响 疾病和移植。考虑到巨大的发病率和死亡率 在我们社会的心血管疾病中，该提案可能会产生重要的临床影响。