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.

项目总结 血管疾病对数百万美国人来说是常见的，也是致命的。现代医学 血管疾病的治疗是有限的，并且与显著的发病率有关。 和死亡率。因此，血管疾病需要新的和新的治疗方法。《长河》 这项建议的范围、目标和临床意义是使用我们新的 开发了ETV2淘汰型猪作为宿主，最终用于生产个性化 临床应用的人体血管系统。当前修订后的申请的目标是 是在猪身上建立一个非人类灵长类平台，这将为 在基因编辑猪中设计人源化血管系统。我们实验室发现了 ETV2作为Nkx2-5的下游靶点，并定义了ETV2突变小鼠胚胎 不能存活，缺乏内皮/血管和造血系。vbl.使用 CRISPR/Cas9基因编辑技术，我们进一步建立了ETV2突变体 猪胚胎缺乏血管和血液谱系。根据我们的结果，我们的整体 假设ETV2是主分子程序的关键因素 发育过程中的血管谱系。在这些拟议的研究中，我们将利用 设计范式转换的非人类灵长类的新兴技术的数量 转基因动物代孕体内的血管系统。为了检验我们的假设，我们 将针对以下具体目标：具体目标1：确定 利用GFP标记的猪卵裂球进行囊胚补充，以充分 抢救ETV2缺失型猪宿主；具体目标2：确定 非人灵长类干细胞群体在猪囊胚互补中的应用 具体目标#3：在ETV2中设计非人类灵长类血管系统 突变的猪宿主。在这些研究中，我们将使用最先进的基因技术 和猕猴GFP标记的干细胞群体来设计非人类灵长类动物 在大型动物模型中的血管系统。这个非人类的灵长类大型动物模型将是 再生医学的重要资源，并将成为 生成个性化、人性化的猪模型。这一战略有能力 对慢性血管疾病新兴疗法的发展产生了深远的影响 疾病和移植。考虑到巨大的发病率和死亡率， 在我们的社会，这项建议可能会对心血管疾病产生重要的临床影响。