Adaptable tissue-specific endothelial cells for organ regeneration

用于器官再生的适应性组织特异性内皮细胞

• 批准号: 10397474
• 负责人: Shahin Rafii
• 金额: $ 101.78万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10397474
• 关键词: Adult; Allogenic; Award; Blood; Blood Vessels; Cardiac; Cells; Cicatrix; Clinic; Clinical Trials; Coculture Techniques; Code; Endothelial Cells; Engineering; FDA approved; Fibrosis; Functional disorder; Genetic Transcription; Goals; Growth Factor; Heart; Hematopoietic; Hepatic; Heterogeneity; Homeostasis; Human; In Vitro; Infusion procedures; Intention; Lung; Macaca nemestrina; Mission; Molecular; Monkeys; Morphogenesis; Mus; National Heart, Lung, and Blood Institute; Natural regeneration; Organ; Organoids; Outcome; Pathway interactions; Production; RNA; Recovery; Regenerative Medicine; Research; Safety; Signal Transduction; Specific qualifier value; Technology; Testing; Therapeutic; Tissue Engineering; Tissue Transplantation; Tissues; Translating; Translations; Transplantation; cellular transduction; efficacious treatment; experimental study; fetal; follow-up; healing; in vivo; induced pluripotent stem cell; innovation; knock-down; lung repair; nonhuman primate; novel; organ injury; organ regeneration; organ repair; overexpression; programs; reconstitution; regenerative; repaired; replacement tissue; stem cells; tissue repair; tumorigenesis; vascular contributions; Adult; Allogenic; Award; Blood; Blood Vessels; Cardiac; Cells; Cicatrix; Clinic; Clinical Trials; Coculture Techniques; Code; Endothelial Cells; Engineering; FDA approved; Fibrosis; Functional disorder; Genetic Transcription; Goals; Growth Factor; Heart; Hematopoietic; Hepatic; Heterogeneity; Homeostasis; Human; In Vitro; Infusion procedures; Intention; Lung; Macaca nemestrina; Mission; Molecular; Monkeys; Morphogenesis; Mus; National Heart, Lung, and Blood Institute; Natural regeneration; Organ; Organoids; Outcome; Pathway interactions; Production; RNA; Recovery; Regenerative Medicine; Research; Safety; Signal Transduction; Specific qualifier value; Technology; Testing; Therapeutic; Tissue Engineering; Tissue Transplantation; Tissues; Translating; Translations; Transplantation; cellular transduction; efficacious treatment; experimental study; fetal; follow-up; healing; in vivo; induced pluripotent stem cell; innovation; knock-down; lung repair; nonhuman primate; novel; organ injury; organ regeneration; organ repair; overexpression; programs; reconstitution; regenerative; repaired; replacement tissue; stem cells; tissue repair; tumorigenesis; vascular contributions

PROJECT ABSTRACT The overarching goal of our proposed research program is to develop a discovery pipeline that will enable identification of transcriptional codes for engineering tissue-specific endothelial cells (ECs) for therapeutic organ regeneration of heart, lung and blood. Therapies for organ regeneration promises unlimited access to the replacement tissues. However, despite breakthroughs in uncovering the molecular underpinnings of organ morphogenesis and organoid technology, translation of regenerative medicine to the clinic has confronted with hurdles. These bottlenecks are in part due to the lack of understanding as to how niche cells coordinate organ repair. Specifically, contribution of vascular niche cells that supply regenerative signals has not been realized. This R35 application builds upon the novel proposition that poor healing after organ damage is due to the dysfunction and loss of the tissue-specific ECs. This programmatic proposal examines the hypothesis that reconstitution of stem cells in injured organs is dependent on the pro-regenerative angiocrine signals supplied by tissue-specific vascular niche ECs. We have shown that organotypic ECs by deploying defined angiocrine factors support lung, cardiac, hepatic and hematopoietic regeneration. Thus, ECs perform actively as dynamic, tissue-specified niche cells critical for tissue homeostasis and repair. To test this and to set up the stage for therapies, we have engineered adaptable mouse, nonhuman primate and human ECs by transducing the transduction factor (TF) ETV2 into adult mature ECs (R-VECs) and differentiating human induced pluripotent stem cells (iPSCs) into generic fetal-like ECs (iVECs) that could inform on the pathways that induce organotypic TFs. These adaptive iVECs and R-VECs will be cocultured with heart, lung, and blood organoids in vitro or infused in vivo in mice undergoing organ repair to identify the induction of organotypic TFs in these cells. The educated iVECs and R-VECs will be recovered and subjected to RNA profiling and de novo motif discovery to identify induced tissue-specific TF(s). The identified TFs will be overexpressed or knocked down in ECs, to validate their function in sustaining organotypic and angiocrine profile for organ repair. We anticipate that transplantation of organotypic ECs will promote long-lasting tissue repair without provoking tumorigenesis or fibrosis. We have initiated FDA-approved human clinical trials to examine the safety and efficacy of allogeneic generic EC infusion for hematopoietic recovery. As a follow up, we intend to assess the contribution of R-VECs or iVECs-derived from nonhuman primates to regeneration in the pigtail macaque monkeys with the intention of translating the potential of organotypic ECs to clinic. The expected outcomes of the proposed research are identification of molecular signals and transcriptional determinants of tissue-specific vascular and angiocrine heterogeneity. Goals of this proposal fit with the mission of NHLBI R35 award to develop innovative regenerative discovery pipeline to promote safe and efficacious treatments for cardiac, pulmonary and blood maladies.

项目摘要 我们拟议的研究计划的总体目标是开发一种发现管道，以实现 鉴定用于治疗的工程组织特异性内皮细胞（EC）的转录代码 器官的心脏，肺和血液的再生。器官再生的疗法承诺无限访问 替代组织。然而，尽管突破了器官的分子基础 形态发生和器官技术，再生医学向诊所的翻译已面临 障碍。这些瓶颈部分是由于缺乏对小裂细胞如何坐标器官的理解 维修。具体而言，尚未实现供应再生信号的血管生态位细胞的贡献。 此R35应用于新颖的主张，即器官后的愈合不良是由于 功能障碍和组织特异性EC的丧失。该计划提案探讨了以下假设 受伤器官中干细胞的重建取决于提供的促再生血管分泌信号 通过组织特异性的血管生态位EC。我们已经通过部署定义的血管分泌表明了器官型EC 因素支持肺，心脏，肝和造血再生。因此，EC的表现为动态， 组织指定的小众细胞对于组织稳态和修复至关重要。测试这一点并设置舞台 疗法，我们通过传输来设计适应性的老鼠，非人类灵长类动物和人类ECS 转导因子（TF）ETV2进入成年成熟EC（R-VEC），并区分人类诱导的多能 干细胞（IPSC）进入通用的胎儿样EC（IVEC），可以告知诱导的途径 器官TFS。这些自适应IVEC和R-VEC将与心脏，肺和血型共培养 体外或在接受器官维修的小鼠中体内注入体内，以鉴定在这些细胞型TF中的诱导 细胞。受过教育的IVEC和R-VEC将被恢复并进行RNA分析和从头图案 鉴定诱导组织特异性TF的发现。确定的TF将过表达或击倒 在EC中，为了验证其在维持器官修复的器官和血管分泌轮廓方面的功能。我们期待 器官型EC的移植将促进持久的组织修复而不会引起肿瘤发生 或纤维化。我们已经启动了FDA批准的人类临床试验，以检查 同种异体通用EC输注用于造血恢复。作为后续，我们打算评估贡献 从非人类灵长类动物衍生的R-Vecs或Ivecs的衍生作用，到辫子猕猴的再生 将器官EC的潜力转化为诊所的意图。提议的预期结果 研究是组织特异性血管和分子信号和转录决定因素的鉴定 血管分泌异质性。该提案的目标符合NHLBI R35奖的使命，以发展创新 再生发现管道，以促进心脏，肺和血液的安全有效治疗 疾病。