Bioengineering Strategies for Cardiovascular Disease

心血管疾病的生物工程策略

• 批准号: 10468711
• 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 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10468711
• 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.

项目概要 血管疾病对于数百万美国人来说是常见且致命的。目前医疗 血管疾病的治疗方法有限并且与显着的发病率相关 和死亡率。因此，血管疾病需要新的治疗方法。长的 该提案的范围目标和临床意义是使用我们新的 开发了 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。当前修订的应用程序的目标 是在猪身上建立一个非人类灵长类动物平台，这将为 在基因编辑的猪中设计人性化脉管系统。我们的实验室发现 Etv2作为Nkx2-5的下游靶标并定义Etv2突变小鼠胚胎 无法存活并且缺乏内皮/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ETV2突变体 猪胚胎缺乏血管和血统。根据我们的结果，我们的总体 假设 Etv2 是主分子程序的一个重要因素 发育过程中的维管谱系。在这些拟议的研究中，我们将利用 大量新兴技术可改变非人类灵长类动物的范式 转基因动物替代物中的脉管系统。为了检验我们的假设，我们 将实现以下具体目标： 具体目标#1：定义 囊胚互补，使用 GFP 标记的猪卵裂球，以充分 拯救ETV2无效猪宿主；具体目标#2：定义能力 用于猪囊胚互补的非人灵长类干细胞群 具体目标#3：在 ETV2 中设计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 标记的干细胞群来改造非人类灵长类动物 大型动物模型中的脉管系统。这种非人类灵长类大型动物模型将 再生医学的重要资源，并将作为一个平台 生成个性化的人性化猪模型。该策略有能力 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 在我们社会的心血管疾病中，这一提议可能会产生重要的临床影响。

项目成果

Chimeric Humanized Vasculature and Blood: The Intersection of Science and Ethics.

嵌合人源化脉管系统和血液：科学与伦理的交叉点。

• 发表时间: 2020
• 影响因子: 5.9
• 作者: Garry, Daniel J;Caplan, Arthur L;Garry, Mary G
• 通讯作者: Garry, Mary G

Interspecies chimeras as a platform for exogenic organ production and transplantation.

种间嵌合体作为外源器官生产和移植的平台。

• 发表时间: 2021
• 作者: Garry, Daniel J;Garry, Mary G
• 通讯作者: Garry, Mary G

ETV2-null porcine embryos survive to post-implantation following incomplete enucleation.

ETV2 缺失的猪胚胎在不完全摘除后可存活至植入后。

• 发表时间: 2020
• 作者: Maeng, Geunho;Gong, Wuming;Das, Satyabrata;Yannopoulos, Demetris;Garry, Daniel J;Garry, Mary G
• 通讯作者: Garry, Mary G