Vascular Regeneration with Direct Reprogramming and Engineering Strategies

直接重编程和工程策略的血管再生

基本信息

批准号：
10530784
负责人：
Young-Sup Yoon
金额：
$ 53.85万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10530784
关键词：
Alginates American Animal Model Area Biocompatible Materials Biological Biomedical Engineering Blood Vessels Bone Marrow Bypass Cardiac Myocytes Cardiovascular Diseases Cell Adhesion Cell Differentiation process Cell Lineage Cell Survival Cell Therapy Cell Transplantation Cells Clinical Clinical Trials Dermal Disease Embryo Encapsulated Endothelial Cells Endothelium Engineering Family Fibroblasts Functional disorder Gelatin Genes Genomics Goals Hindlimb Histologic Human Hybrids Hydrogels Imaging technology Impairment Inflammatory Inflammatory Response Injections Insertion Mutation Ischemia Lentivirus Vector Measurement Microspheres Modeling Molecular Morbidity - disease rate Mus Myocardial Ischemia Neurons Patients Peripheral Vascular Diseases Play Polymers Reporting Research Residual state Role Side Somatic Cell Technology Therapeutic Therapeutic Effect Time Tissues Transgenic Mice Transplantation Tumorigenicity Undifferentiated Variant Vascular regeneration Viral Viral Vector adult stem cell blastomere structure blood vessel development cell type clinical application clinical translation copolymer cost delivery vehicle design in vivo induced pluripotent stem cell inflammatory milieu innovative technologies member mortality neovascularization next generation novel novel strategies organ injury particle poly(glycerol-sebacate)postnatal human regenerative therapy stem cells therapy development tissue regeneration tissue repair transcription factor tumorigenic vector

项目摘要

Project Summary Ischemic cardiovascular diseases are the leading cause of morbidity and mortality. Underlying pathophysiology of these diseases is associated with loss or dysfunction of blood vessels and/or impaired new vessel formation (neovascularization). While cell therapy has emerged as a promising option to form blood vessels, the effects of adult stem cells are uncertain and embryonic or induced pluripotent stem cells (ESCs/iPSCs) are potentially tumorigenic. To avoid these problems, a new approach has been developed using lineage- or cell-type specific transcription factors (TFs) for direct conversion or reprogramming of somatic cells into other lineage cells. We have attempted this direct reprogramming toward endothelial cells (ECs) using combinations of seven TFs and found for the first time that ETV2, alone, is sufficient to convert human fibroblasts into ECs. However, since we used a lentiviral vector, these reprogrammed ECs (rECs) have limited clinical applicability. The direct reprogramming approach allows two therapeutic strategies: cell-based therapy or direct in vivo reprogramming. For clinical application, both approaches require a safer delivery vector to minimize the possibility of genomic integration. Thus, we developed an adenoviral-ETV2 (Ad-ETV2) vector and generated rECs (Adeno-rECs). Another important barrier for cell therapy is short-term survival of the transplanted cells. To overcome this problem, we have been investigating bioengineered cell therapy. In this study, first, we will generate an optimal construct combining these Adeno-rECs with novel biomaterials. We have developed a novel biodegradable hybrid copolymer consisting of gelatin and poly glycerol sebacate (PGS), which was further made into a microbead form with alginate. We refer to this co-polymer as AlGPM. This hybrid polymer is biodegradable and elicits minimal inflammatory response. Furthermore, its microbead form promotes wide distribution of encapsulated cells after injection. The composition of AlGPM will be optimized to promote cell survival and maximize function of rECs. We will then determine the neovascularization and therapeutic effects of the selected AlGPM microbeads encapsulating rECs using ischemic animal models. Second, we will determine whether local injection of viral particles of ETV2 into animal models can directly reprogram somatic cells into endothelial cells and promote vascular regeneration and tissue repair in vivo. Moreover, by using various transgenic mice, we will genetically track the fate of somatic cells toward ECs in vivo. Together, the goal of this project is to develop clinically applicable vascular regenerative therapy using direct reprogramming approaches and bioengineering technologies.

项目摘要缺血性心血管疾病是发病率和死亡率的主要原因。潜在的这些疾病的病理生理学与血管的损失或功能障碍有关血管形成（新血管形成）。虽然细胞疗法已成为形成血液的有前途的选择血管，成年干细胞的作用不确定，胚胎或诱导的多能干细胞（ESC/IPSC）可能是肿瘤的。为了避免这些问题，已经开发了一种新方法谱系或细胞类型特异性转录因子（TFS），用于直接转换或重新编程进入其他谱系细胞。我们已经尝试使用这种直接重编程对内皮细胞（EC）使用七个TF的组合，首次发现ETV2足以转换人成纤维细胞进入ECS。但是，由于我们使用了慢病毒载体，因此这些重编程的EC（REC）的临床有限适用性。直接重编程方法允许两种治疗策略：基于细胞的治疗或直接体内重新编程。对于临床应用，这两种方法都需要更安全的输送矢量来最大程度地减少基因组整合的可能性。因此，我们开发了腺病毒-ETV2（AD-ETV2）向量并生成 recs（Adeno-Recs）。细胞治疗的另一个重要障碍是移植细胞的短期存活。到克服这个问题，我们一直在研究生物工程细胞疗法。在这项研究中，首先，我们将生成一种结合这些腺样体与新颖的最佳结构生物材料。我们已经开发了一种新型的可生物降解杂交共聚物，由明胶和聚甘油组成黑甲酸酯（PGS），进一步形成具有藻酸盐的微粒形式。我们将此共聚物称为 algpm。该混合聚合物是可生物降解的，并且引起最小的炎症反应。此外，它的 Microbead形式促进注射后封装的细胞的广泛分布。 ALGPM的组成将优化以促进细胞存活并最大化REC的功能。然后，我们将确定选定的ALGPM微粒的新血管形成和治疗效应，使用REC封装Recs 缺血性动物模型。其次，我们将确定是否将ETV2病毒颗粒局部注射到动物中模型可以将体细胞直接重新编程为内皮细胞，并促进血管再生和组织在体内维修。此外，通过使用各种转基因小鼠，我们将基因跟踪体细胞的命运朝着体内的ECS。该项目的目标是开发临床适用的血管再生使用直接重编程方法和生物工程技术的治疗。