Cardiac Revascularization with Direct Reprogramming Approaches

通过直接重编程方法进行心脏血运重建

基本信息

批准号：
9903989
负责人：
Young-Sup Yoon
金额：
$ 46.31万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-12 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9903989
关键词：
Adenovirus Vector Adult American Animals Area Biocompatible Materials Biological Biomedical Engineering Blood Vessels Bone Marrow Cardiac Cardiac Myocytes Cardiovascular Diseases Cell Lineage Cell Survival Cell Therapy Cell Transplantation Cells Clinical Coronary Arteriosclerosis Disease Disease model Echocardiography Embryo Encapsulated Endothelial Cells Endothelium Engineering Ethics Family Fibroblasts Functional disorder Gel Generations Genes Genomics Goals Growth Heart Hepatocyte Histologic Human Impairment Injections Insertion Mutation Lentivirus Vector Magnetic Resonance Imaging Medical Modeling Molecular Morbidity - disease rate Mus Myocardial Ischemia Neurons Nitric Oxide Operative Surgical Procedures Patients Peptides Play Population Problem Solving Research Risk Rodent Role Somatic Cell Technology Therapeutic Therapeutic Effect Time Tissue Engineering Tissues Variant Vascularization base blood vessel development cardiac regeneration cell type clinical application clinical development clinical translation cost design heart function improved in vivo induced pluripotent stem cell member microCT mortality neovascularization next generation novel novel strategies overexpression postnatal human regenerative therapy side effect success transcription factor tumor vector

项目摘要

Project Summary Ischemic heart diseases are the leading cause of morbidity and mortality. The underlying problems of these diseases are loss or dysfunction of blood vessels and insufficient new vessel formation. While cell therapy has emerged as a promising option to promote blood vessel growth, no therapy is yet clinically available and there is much room for improvement. The potential of bone marrow (BM)-derived cells turned out to be minimal9, and embryonic or induced pluripotent stem cell (ESC/iPSC)-derived ECs are difficult to maintain, costly to produce, and may cause side effects. To avoid these problems, a new approach, called direct reprogramming or direct conversion has been developed, in which somatic cells are converted into other lineage cells by overexpression of lineage- or cell-type specific transcription factors (TFs). This approach allows simpler and safer target cell generation and has the potential for more convenient clinical translation. We have attempted this direct reprogramming toward ECs using combinations of seven endothelial-related TFs and demonstrated for the first time that ETV2 alone is sufficient to convert human fibroblasts into ECs. However, since we used a lentiviral vector like others, these rECs have restrictions in clinical applicability. The direct EC reprogramming approach for therapy has two options: cell therapy or direct in vivo reprogramming. For clinical application, both require a safer delivery vector to minimize the possibility of genomic integration. Thus, we developed an adenoviral-ETV2 (Ad-ETV2) vector. Another important obstacle for cell therapy is short- term survival of the transplanted cells. To overcome this problem, we have investigated the potential of biomaterial for prolongation of the cell survival and therapeutic effects. We recently showed that peptide amphiphile (PA) nanomatrix gel is very effective, extending survival of human iPSC-derived ECs longer than 10 months and inducing continuous vessel formation in vivo. In this study, first, we will first develop cell-based therapy. We will generate clinically compatible rECs from human fibroblasts and generate an optimal PA construct combining these rECs with various types of PA nanomatrix gel including a newly developed nitric oxide (NO)-releasing PA. We will then determine the vascularization and therapeutic effects of the selected rEC-PA nanomatrix constructs on rodent ischemic heart disease models. Second, we will investigate whether direct delivery of Ad-ETV2 into cardiac ischemia models can directly reprogram fibroblasts into functional endothelial cells and induce vascularization in vivo. We will use genetically modified mice to track the fate of fibroblasts toward ECs in vivo. The goal of this project is to develop clinically applicable cardiac revascularization strategy using a novel direct reprogramming approach combined with tissue engineering technologies. If successful, this study will provide next-generation platforms for broad areas of research and therapy for ischemic heart diseases.

项目概要缺血性心脏病是发病和死亡的主要原因。这些问题的根本问题疾病是血管损失或功能障碍以及新血管形成不足。虽然细胞疗法已成为促进血管生长的一种有前途的选择，但临床上尚无可用的治疗方法有很大的改进空间。骨髓（BM）来源的细胞的潜力被证明是最小的9，并且胚胎或诱导多能干细胞 (ESC/iPSC) 衍生的 EC 难以维持、生产成本昂贵、并可能引起副作用。为了避免这些问题，一种新方法被称为直接重编程或直接已经开发出转化，其中体细胞通过过度表达转化为其他谱系细胞谱系或细胞类型特异性转录因子 (TF)。这种方法允许更简单、更安全的靶细胞生成并具有更方便的临床转化的潜力。我们已经尝试过这种直接使用七种内皮相关转录因子的组合对 EC 进行重编程，并首次得到证实单独使用 ETV2 就足以将人成纤维细胞转化为 EC 的时间。然而，由于我们使用的是慢病毒与其他载体一样，这些 rEC 在临床应用方面也受到限制。直接内皮细胞重编程治疗方法有两种选择：细胞疗法或直接体内重编程。对于临床应用，两者都需要更安全的递送载体，以尽量减少基因组整合的可能性。因此，我们开发了腺病毒-ETV2 (Ad-ETV2) 载体。细胞疗法的另一个重要障碍是短期移植细胞的长期存活。为了解决这个问题，我们研究了用于延长细胞存活和治疗效果的生物材料。我们最近证明肽两亲物 (PA) 纳米基质凝胶非常有效，可将人类 iPSC 衍生的 EC 的存活时间延长 10 年以上几个月并诱导体内连续血管形成。在这项研究中，首先，我们将首先开发基于细胞的疗法。我们将从以下来源生成临床相容的 rEC 人成纤维细胞，并将这些 rEC 与各种类型的 PA 相结合，生成最佳的 PA 构建体纳米基质凝胶包含新开发的释放一氧化氮 (NO) 的 PA。然后我们将确定所选 rEC-PA 纳米基质结构对啮齿动物缺血性心脏的血管化和治疗作用疾病模型。其次，我们将研究Ad-ETV2是否直接递送至心脏缺血模型中可以直接将成纤维细胞重编程为功能性内皮细胞并诱导体内血管化。我们将使用转基因小鼠在体内追踪成纤维细胞向 EC 的命运。该项目的目标是开发使用新型直接重编程方法结合临床适用的心脏血运重建策略与组织工程技术。如果成功，这项研究将为广泛的研究提供下一代平台缺血性心脏病的研究和治疗领域。